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Screening of Rhamnus Purpurea (Edgew.) Leaves for Antimicrobial, Antioxidant, and Cytotoxic Potential
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  • Fazli Khuda*
    Fazli Khuda
    Department of Pharmacy, University of Peshawar, Peshawar 25120, Pakistan
    *Email: [email protected]. Tel: +92 91 9216750. Fax: +92 91 9218131.
    More by Fazli Khuda
    Orcidhttps://orcid.org/0000-0002-6847-1744
  • Nida Alam
    Nida Alam
    Department of Pharmacy, University of Peshawar, Peshawar 25120, Pakistan
    More by Nida Alam
  • Atif Ali Khan Khalil
    Atif Ali Khan Khalil
    Department of Biological Sciences, National University of Medical Sciences, Rawalpindi 46000, Pakistan
    More by Atif Ali Khan Khalil
  • Asif Jan
    Asif Jan
    Department of Pharmacy, University of Peshawar, Peshawar 25120, Pakistan
    More by Asif Jan
  • Faiza Naureen
    Faiza Naureen
    Department of Pharmacy, Abdul Wali Khan University, Mardan 23200, Pakistan
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  • Zaki Ullah
    Zaki Ullah
    Department of Pharmacy, University of Peshawar, Peshawar 25120, Pakistan
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  • Amal Alotaibi
    Amal Alotaibi
    Department of Basic Science, College of Medicine, Princess Nourah Bint Abdulrahman University, P.O. Box 84428, Riyadh 11671, Saudi Arabia
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  • Riaz Ullah
    Riaz Ullah
    Department of Pharmacognosy (Medicinal Aromatic and Poisonous Plants Research Center) College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia
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    Orcidhttps://orcid.org/0000-0002-2860-467X
  • Sami Ullah
    Sami Ullah
    Department of Pharmacy, University of Peshawar, Peshawar 25120, Pakistan
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  • Yasar Shah
    Yasar Shah
    Department of Pharmacy, Abdul Wali Khan University, Mardan 23200, Pakistan
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  • Sayyed Ibrahim Shah
    Sayyed Ibrahim Shah
    Department of Pharmacy, Abdul Wali Khan University, Mardan 23200, Pakistan
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  • Sultan Mehtap Büyüker
    Sultan Mehtap Büyüker
    Department of Pharmacy Services, Üsküdar University, İstanbul 34664, Turkey
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ACS Omega

Cite this: ACS Omega 2022, 7, 26, 22977–22985
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https://doi.org/10.1021/acsomega.2c03094
Published June 16, 2022

Copyright © 2022 The Authors. Published by American Chemical Society. This publication is licensed under

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Abstract

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Exploring new antimicrobial and cytotoxic drugs has been one of the most active areas of research. Rhamnus purpurea (Edgew.) buckthorn (Rhamnaceae) is a wild shrub traditionally used in Pakistan for the treatment of various ailments including cancer and infectious diseases. The aim of this study is to find novel antimicrobial and cytotoxic agents of plant origin. The crude methanol extract and full range of fractions of R. purpurea leaves were screened for the said activities using in vitro antimicrobial, antioxidant, and cytotoxic models following standard protocols. The antimicrobial activity was evaluated using the agar well diffusion method, while the antioxidant activity was assessed with 1,1-diphenyl-2-picryl hydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assays. The cytotoxic effect was investigated against the human cancer cell lines i.e. Caco-2 (gut), A549 (lung), HepG2 (liver), and MDA-MB-231 (breast) by MTS assay. In addition, toxicity studies were conducted on renal and alveolar primary epithelial cells (HRPTEpiC and HPAEpiC, respectively). Phytochemical investigation showed the presence of secondary metabolites such as alkaloids, saponins, tannins, glycosides, phenols, carbohydrates, proteins, and flavonoids. The n-hexane and chloroform fractions showed significant activity against Staphylococcus aureus (MIC 0.60 and 0.68 mg/mL, respectively), Salmonella typhi (MIC 0.48 and 0.45 mg/mL, respectively), and Bacillus subtilis (MIC 0.54 and 0.76 mg/mL, respectively). Among fungal strains, crude methanol and chloroform fractions exhibited significant activity against Fusarium solani (MIC 0.53 and 0.44 mg/mL, respectively) and Aspergillus niger (MIC 0.47 and 0.42 mg/mL, respectively). The crude methanol, n-hexane and chloroform fractions revealed the highest antioxidant activity at 1000 μg/mL, compared to that of ascorbic acid. The n-hexane fraction showed a significant cytotoxic effect against Caco-2, A549, and HepG2 cell lines with IC50 values of 5.65 ± 0.88, 5.50 ± 0.90, and 4.95 ± 1.0 μg/mL, respectively. Similarly, the chloroform fraction depicted significant activity against Caco-2, A549, and HepG2 cell lines with IC50 values of 4.55 ± 1.25, 4.65 ± 1.55, and 2.85 ± 0.98 μg/mL, respectively. The crude methanol extract and almost all fractions exhibited the highest selectivity index (>2.0) for Caco-2, A549, and HepG2 cancer cell lines, providing safety data for this study. The results showed that R. purpurea leaves have excellent antimicrobial, antioxidant, and cytotoxic potential and warrant further studies to search for novel compounds for the said activities.

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1. Introduction

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Antimicrobial resistance (AMR) is a serious issue across the globe, particularly in developing countries such as those of South Asia. (1) The major reasons include its availability over the counter that has caused approximately 5.3 million deaths worldwide annually. In Pakistan, both extensively drug-resistant (XDR) and multidrug-resistant (MDR) bacteria have been identified in recent years. (2,3) It has been reported that Enterobacteriaceae has developed a significant resistance against quinolones. Similarly, Salmonella species has shown almost 100% resistance against fluoroquinolones. (4) Furthermore, in the US (1999–2012), 68.4% Staphylococcus epidermidis, 47.9% Acinetobacter baumannii, and 13.7% Escherichia coli were ciprofloxacin-, carbapenem-, and β-lactam-resistant, respectively. (5,6) A similar resistance has emerged in Canada (2007–2011), where 27% E. coli, 19.3% Staphylococcus aureus, and 16.8% Streptococcus pneumoniae were ciprofloxacin-, methicillin-, and penicillin-resistant, respectively. (7)
Likewise, cancer is one of the leading causes of mortality around the globe. According to the World Health Organization (WHO), approximately 13 million deaths are expected from this fatal disease by the end of 2030. (8,9) Among these, breast cancer is the most common malignancy worldwide. It is noteworthy that Pakistan has the highest rate of breast cancer in Asia with more than 40,000 deaths and 90,000 cases reported annually. (10) It has 25% more incidence in women; however, men are also susceptible, but fortunately, the mortality rate is very low in the latter (1:1000). (9) Hepatocellular carcinoma (HCC) is the sixth most common cancer worldwide, with approximately 5.7% new cases annually. Globally, it remains the third leading cause of cancer-related deaths with an overall survival rate of only 3–5%. It is more prevalent in South Asia, with 55% cases in China only. Hepatitis B and C are the well-known factors for HCC. (11,12) Lung cancer accounts for 470,000 cases in Europe each year. In Pakistan, its incidence, prevalence, and mortality are not known due to the absence of population-based cancer registry. According to regional cancer registries, this is the second most common cancer in men and third most common cancer in both sexes. (13) Colorectal cancer is the third most common type of cancer worldwide. This type of cancer is closely associated to the age, as it is more common in people over the age of 65 years and accounts for approximately 70% of all cancer cases. (14,15) It is more prevalent in developed countries; however, its prevalence is on rise in developing countries such as Pakistan and India. Various chemotherapeutic drugs are used for the treatment of these cancers. However, they are associated with resistance and severe side effects such as nausea, vomiting, hair loss, and fatigue. Therefore, there is a need to search for safe and more selective/specific cytotoxic agents.
Production of free radicals is among the several factors responsible for causing a variety of cancers. According to the literature, oxidation is one the important processes in the production of free radicals, i.e., hydroxyl and perhydroxyl. (16,17) Synthetic antioxidants are frequently used to prevent the generation of free radicals and subsequently stop the oxidation process. However, they are usually associated with several side effects. Butylated hydroxy toluene and hydroxy anisole are frequently linked to liver cirrhosis and cancer. (18) Therefore, there is a need to search for more safe and effective antioxidants especially of natural origin.
Rhamnus is a genus of approximately 110 species, commonly known as buckthorns. Some of these species are Rhamnus alaternus, Rhamnus caroliniana, Rhamnus cathartica, Rhamnus diffusus, Rhamnus globosa, Rhamnus japonica, and Rhamnus saxatilis. Rhamnus purpurea (Figure 1), commonly known as buckthorns, is a wild shrub, widely distributed in East Asia and North America with species ranging in height from 1 to 10 meter. Several phytochemicals have been isolated from the Rhamnus genus including flavonoids, alkaloids, glycosides, phenols, and anthraquinones. To date, no pharmacological activity of this plant has been reported. However, in folk medicine, it has been widely used in the treatment of cancer, wound and throat infections, inflammation, and other health problems. (19−21) Due to its widespread traditional use, the current study scientifically investigates different fractions of the said plant for antimicrobial, antioxidant, and cytotoxic potential.

Figure 1

Figure 1. Rhamnus purpurea Edgew. (photograph courtesy of Dr. Atif Ali Khan Khalil; the photo is from a free domain).

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2. Materials and Methods

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2.1. Chemicals

Imipenem and Miconazole were donated by Ferozsons Laboratories Limited, Nowshera, Pakistan, and 1,1-diphenyl-2-picryl hydrazyl (DPPH), ferric reducing antioxidant power (FRAP) reagents, acetate buffer, HCl, methanol, chloroform, n-hexane, and ethyl acetate were purchased from Sigma-Aldrich, Germany.

2.2. Plant Materials

The plant was collected (July 2018) from Dir and Swat valleys (latitude 35° 3′ 11″ N; longitude 72° 33′ 42″ E), Khyber Pakhtunkhwa (KPK), Pakistan, and was identified by Dr. Fazal Hadi, Department of Botany, University of Peshawar. (Voucher no. F.Khuda-32522).

2.2.1. Preparation of the Extract

The leaves were thoroughly washed, dried under shade, and subsequently powdered. The powdered material (1000 g) was macerated for at least 3 days in methanol and stirred occasionally. The macerate was filtered and concentrated to dryness under vacuum (40 °C) using a rotary evaporator.

2.2.2. Fractionation

The dark-green extract was dissolved in distilled water and subsequently partitioned with different solvents to obtain chloroform, n-hexane, ethyl acetate, and aqueous fractions. The crude methanol extract and its fractions were then screened for the following biological activities.

2.3. Phytochemical Screening

The extract was screened for the presence of the following phytochemicals: alkaloids, saponins, tannins, glycosides, phenols, carbohydrates, proteins, and flavonoids. The method of Farzana et al. was used for conducting the following assays. (22)

2.3.1. Tests for Alkaloids

2.3.1.1. Mayer’s Reagent Test
In this test, 2 mL each of methanol crude extract and conc. hydrochloric acid was mixed, followed by the dropwise addition of Mayer’s reagent. The appearance of the green color or white precipitate confirmed the presence of alkaloids.
2.3.1.2. Hager’s Test
In this test, 2–3 drops of Hager’s reagent (picric acid) were added to the plant extract (2 mL). The appearance of a yellow precipitate revealed the presence of alkaloids.

2.3.2. Test for Saponins

In a test tube, 2 mL each of the plant extract and distilled water was mixed and shaked vigorously for 5 min. The formation of a thick foam at the top of the tube indicted the presence of saponins.

2.3.3. Test for Flavonoids

This test was performed by the addition of a few drops of ferric chloride to about 2 mL of the plant extract. The appearance of blackish-red precipitates confirmed the presence of flavonoids.

2.3.4. Test for Tannins

2.3.4.1. Alkaline Reagent Test
In this test, 2 mL each of the extract and sodium hydroxide (1N) was added and mixed thoroughly. The formation of red or yellow precipitates showed the presence of tannins.

2.3.5. Test for Glycosides

2.3.5.1. Keller Killiani Test
This test was performed by the addition of 1 mL each of the extract and glacial acetic acid, followed by cooling and adding 2–3 drops of ferric chloride. Finally, sulfuric acid (0.5 mL) was added along the sides of the test tube. The formation of a reddish-brown ring at the junction of the two layers confirmed the presence of glycosides.

2.3.6. Test for Phenols

2.3.6.1. Ellagic Acid Test
To 1 mL of the plant extract, few drops of glacial acetic acid (5%) were added and mixed, followed by the addition of few drops of sodium nitrite (5%). The appearance of muddy brown color indicated the presence of phenols.

2.3.7. Test for Carbohydrates

2.3.7.1. Benedict’s Test
A few drops of Benedict’s reagent were mixed with the plant extract, followed by boiling. The formation of reddish-brown precipitates showed the presence of carbohydrates.

2.3.8. Test for Proteins

2.3.8.1. Xanthoproteic Test
This test was conducted by the addition of few drops of conc. nitric acid to about 1 mL of the extract. The appearance of yellow color revealed the presence of proteins in the extract.

2.4. Biological Activities

2.4.1. Antimicrobial Assay

2.4.1.1. Microorganisms
The following reference strains of bacteria were tested: Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853), S. aureus (ATCC 25923), Salmonella typhi (ATCC 19430), and Bacillus subtilis (ATCC 6633). Fungal strains include the following: Fusarium solani (ATCC 11712), Aspergillus terreus (ATCC 20542), Aspergillus flavus (ATCC 32611), and Aspergillus niger (ATCC10549).
2.4.1.2. Antibacterial Activity
The agar well diffusion method was used for conducting the assay. (23) The stated bacterial strains were cultured on nutrient broth media at 37 °C for 24 h. After incubation, the bacterial suspension (100 μL) was spread on sterilized medium plates. Wells were made using a sterilized cork borer for both the test samples and control. Test samples were prepared by dissolving 5 mg of the crude methanol extract and its fractions in dimethyl sulfoxide (DMSO). These samples (100 μL) were then added to the respective wells along with negative (DMSO) and positive (Imipenem) controls. The plates were incubated for 48 h at 37 °C. The percent inhibition growth was calculated by measuring the diameter around cleared zones created by the sample and positive control. The experiment was conducted in triplicate, and the mean values were recorded.
2.4.1.3. Antifungal Activity
This activity was conducted using the agar slanting method. (24) The cultured medium was poured into sterilized test tubes. Test samples were prepared by dissolving 25 mg of the crude methanol extract and its fractions in DMSO (3%). About 100 μl of each sample was added into each test tube containing Sabouraud dextrose agar media (SDA) and allowed to solidify in the tilted position. Subsequently, it was incubated at 30 °C for at least 1 week. DMSO and miconazole were used as negative and positive controls, respectively. Finally, the test tubes were measured for linear growth inhibition, and % inhibition was determined using eq 1.
(1)
2.4.1.4. Determination of the Minimum Inhibitory Concentration (MIC)
The MIC of different fractions of the methanolic extract was determined by the microdilution method, using 96-well microtiter plates. (25) The extract and fractions were dissolved in DMSO and serially diluted with the respective broth to obtain different dilutions (0.1–30 mg/mL). Susceptible strains of bacterial and fungal cultures were then added to each well and incubated at 37 °C for 24 and 72 h, respectively. Tetrazolium salt was added to each well as an indicator; the appearance of violet color in culture media indicates the growth of microorganisms. Imipenem and miconazole serve as positive controls, while DMSO serves as a negative control.

2.5. Antioxidant Assay

2.5.1. 1,1-Diphenyl-2-Picryl Hydrazyl (DPPH) Assay

This assay was performed with DPPH following a well-established procedure with slight modifications. (26) The reagent solution was prepared (0.01 mM) by dissolving it in methyl alcohol (95 mL). Similarly, stock solutions of the crude extract and its fractions were prepared in methyl alcohol (1 mg/mL) and subsequently diluted to get working solutions of different concentrations (62.5, 125, 250, 500, and 100 μg/mL). From each working solution, 100 μl was mixed with 3 mL of DPPH solution and allowed to incubate for 15 min at 23 °C. Finally, the maximum absorbance was measured using a spectrophotometer (517 nm), and % radical scavenging activity of the extract and its fractions was calculated using eq 2.
(2)
where CAbs and SAbs are the absorbance of the control and test samples/standard, respectively.

2.5.2. Ferric Reducing Antioxidant Power (FRAP) Assay

The method of Iris and Strain was adopted for the conduction of this assay. (27) The assay is used to determine the reducing power of the tested compound. Compounds with antioxidant activity reduces the ferric ion (Fe3+) to the ferrous ion (Fe2+); the latter form a blue complex (Fe2+/TPTZ) that significantly increases the total absorption at 593 nm. The assay mixture comprised TPTZ solution (0.25 mL; 10 mM) in HCl (40 mM), acetate buffer (2.5 mL; 300 mM, pH 3.6), FeCl3 (0.25 mL; 20 mM), and crude methanol extract along with its fractions at various concentrations (62.5, 125, 250, 500, and 100 μg/mL). The FRAP reagent (4 μL) and sample solutions (100 μL) were added to each well and mixed thoroughly. Following incubation for 30 min, the absorbance was measured at 593 nm, and % radical scavenging activity of the extract and its fractions was calculated. The assay was performed in triplicate.

2.6. Cytotoxicity Assay

The cytotoxic potential against four human cancer cell lines, i.e., hepatocellular carcinoma (HepG2), lung adenocarcinoma (A549), intestinal epithelial (Caco-2), and breast adenocarcinoma cells (MDA-MB-321) (Korea cell line bank, Seoul, Korea), and two normal cell lines, i.e., primary epithelial cells (HPAEpiC and HRPTEpiC), was investigated by the MTS assay with slight modifications. (28) The cells were cultured in RPMI-1640 media containing streptomycin/penicillin solutions (1% v/v) and fetal bovine serum (10%) (FBS) and subsequently incubated at 37 °C with 5% CO2. Before seeding, cell lines with 80–90% confluency were expanded up to three passages. Prior to the addition of fresh media, Dulbecco’s phosphate buffer saline (DPBS) was used to wash the cells to remove cell debris or any metabolites. Cells showing 90% confluency were trypsinized (0.05% trypsin-EDTA solution) and suspended in RPMI-1640 media containing FBS. About 24 h prior to the treatment, cell lines were seeded in 96-well microplates having 2000 cells/well. Following an overnight incubation, the cells were treated with various concentrations of the crude methanol extract and its fractions. Wells containing media, cells, and tested drugs served as positive controls, while those containing only cells and media served as the negative control. After incubation for 48 h, the MTS reagent (25 μL) was added to each well and allowed to incubate for 30 min. Using a multimode microplate reader (Varioskan Flash, Thermo Scientific, Waltham, MA), the absorbance was measured at 490 nm. Finally, the percent cell viability for the extract and fractions was determined using eq 3. The concentration required to kill 50% of the cell population or IC50 was computed using linear regression of the graph of absorbance against concentration. To find out the specificity of the extract and fractions to cancer cells, the data were analyzed to calculate the selectivity index (SI). It was calculated by dividing the IC50 values for the noncancer (HPAEpiC and HRPTEpiC) cell lines by the values of IC50 for cancer (Caco-2, A549, HepG2 and MDA-MB-231) cell lines. A value of 2 or more indicated high specificity.
(3)

2.7. Statistical Analysis

One-way ANOVA (GraphPad Software) was used for data analysis, and it was presented as the mean ± standard error of the mean (SEM) (n = 5).

3. Results

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3.1. Phytochemical Screening

The results of phytochemical screening are summarized in Table 1. Based on phytochemical analysis, the R. purpurea crude methanol extract contains alkaloids, glycosides, saponins, phenols, and flavonoids as the major secondary metabolites. Ellagic acid and Xanthoproteic tests were unable to confirm the presence of carbohydrates and proteins, respectively.
Table 1. Phytochemical Screening of the Crude Extract of R. Purpureaa
phytochemicalschemical testsresults
alkaloidsMayer’s reagent test+
 Hager’s test+
saponinsFoam test+
tanninsalkaline reagent test+
glycosidesKeller Killiani test+
phenolsEllagic acid test+
carbohydratesBenedict’s testN
proteinsXanthoproteic testN
flavonoidsgeneral test+
a

Abbreviations: + sign, present; N, not indicated.

3.2. Antimicrobial Assays

The antibacterial and antifungal activities were characterized using agar well diffusion and agar slanting methods, respectively.

3.2.1. Antibacterial Activity

The antibacterial activity of the R. purpurea leaf extract and fractions was investigated according to their inhibition zones and MIC values against the selected bacterial strains (Table 2). The methanol extract, chloroform, and n-hexane fractions showed significant inhibitory zones against P. aeruginosa, S. aureus, S. typhi, and B. subtilis. Among the tested strains, S. typhi remained the most sensitive organism against chloroform and n-hexane fractions (% IZ: 71 and 76%, respectively) with MIC values of 0.48 and 0.45 mg/mL, respectively, as compared to the standard. Likewise, the said fractions showed significant activity against B. subtilis (% IZ: 64 and 56%, respectively) with MIC values of 0.54 and 0.76 mg/mL, respectively. The crude methanol extract and n-hexane and chloroform fractions also depicted significant activity against S. aureus (% IZ: 52, 63, and 55%, respectively) with MIC values of 0.77, 0.60, and 0.68 mg/mL, respectively. Results of antibacterial activity suggest that E. coli remained the most resistant strain to the crude methanol extract and all of its fractions. On the other hand, ethyl acetate and aqueous fractions did not show considerable activity against any of the tested bacteria.
Table 2. Antibacterial Activity of the Crude Extract and Its Fractions against Selected Bacterial Strainsa
 crude extractn-hexaneChloroformethyl acetateaqueousstd. 
test microorganismsIZ% IMICIZ%IMICIZ%IMICIZ% IMICIZ% IMICIZMIC
E. coli13380.9211320.6810290.728231.8813301.87340.0020
P. aeruginosa18510.7722620.5520570.6911311.76   350.0019
S. aureus14520.7717630.6015550.68   5181.90270.0014
S. typhi19500.7927710.4829760.4514361.08211.92380.0018
B. subtilis10400.9016640.5414560.767281.2514560.68250.0022
a

IZ, inhibition zone (mm); % I, percent inhibition; MIC, minimum inhibitory concentration (mg/mL); Std, imipenem.

3.2.2. Antifungal Activity

The zones of inhibitions exhibited by the crude methanol extract, its fractions, and standard against different fungal strains are shown in Table 3. The crude methanol extract and chloroform fractions exhibited significant activity against F. solani and A. niger (% I: 48, 58 and 55, 62, respectively) with MIC values of 0.53 and 0.47 mg/mL for the former and 0.44 and 0.42 mg/mL for the latter. Among the fungal strains, A. terreus remained the most resistant strain and did not show considerable sensitivity against the crude methanol extract and any of its fractions. The n-hexane, ethyl acetate, and aqueous fractions did not show any considerable activity against the tested fungal strains.
Table 3. Antifungal Activity of the Crude Extract and Its Fractions against the Selected Fungal Strainsa
 crude extractn-hexanechloroformethyl acetateaqueousstd.
Test microorganisms% IMIC% IMIC% IMIC% IMIC% IMIC% IMIC
F. solani480.53171.55580.44321.15181.50720.0004
A. terreus231.78181.88161.70101.88  580.0018
A. flavus380.84440.64400.68281.0  660.0012
A. niger550.47211.45620.42330.89111.98750.0005
a

% I, percent inhibition; MIC, minimum inhibitory concentration (mg/mL); Std, miconazole.

3.3. Antioxidant Activity

The antioxidant activity values of the extract, its fractions, and standard are shown in Tables 4 and 5. The DPPH radical scavenging assay demonstrated dose-dependent activity for the extract and fractions, i.e., n-hexane, chloroform, ethyl acetate, and aqueous with IC50 values of 22.35, 25.20, 19.52, 25.78, and 28.70 μg/mL, respectively, compared to that of the standard (14.55 μg/mL). Among various fractions, the chloroform fraction showed significant antioxidant activity particularly at higher concentrations (65% at 1000 μg/mL with an IC50 of 19.52) followed by the crude methanol extract, which depicted 47% inhibition (IC50 22.35) at the same concentration. In the present study, the aqueous fraction demonstrated the lowest free radical scavenging activity at all concentrations (Table 4). Similarly, the FRAP assay demonstrated a dose-dependent activity for the crude methanol extract and all of its fractions (Table 5). The n-hexane and chloroform fractions showed significant activity (68 and 72% inhibition at 1000 μg/mL, respectively) with IC50 values of 17.68 and 15.34 μg/mL, respectively. The ethyl acetate (IC50; 24.48 μg/mL) and aqueous (IC50; 27.34 μg/mL) fractions did not show any considerable activity compared to that of the standard (IC50; 9.25 μg/mL). Likewise, the aqueous fraction remained the most inactive at all tested concentrations. Results of both assays depicted that the crude methanol extract and n-hexane and chloroform fractions had comparable free radical scavenging activities, as evident from the IC50 values. On the other hand, ethyl acetate and aqueous fractions did not show any considerable antioxidant activities.
Table 4. DPPH Free Radical Scavenging Activity
 DPPH scavenging activity (%)
concentration (μg/mL)extractn-hexanechloroformethyl acetateaqueousascorbic acid
62.57.20 ± 2.03.12 ± 0.8011.56 ± 1.554.25 ± 0.56 22.32 ± 4.67
12511.65 ± 2.08.45 ± 1.6523.65 ± 3.1011.34 ± 1.204.45 ± 0.6548.34 ± 5.20
25022.55 ± 2.5318.50 ± 2.2041.80 ± 5.5517.87 ± 1.8811.23 ± 1.2163.30 ± 2.52
50039.70 ± 4.031.78 ± 3.8956.70 ± 7.2830.38 ± 3.4321.50 ± 2.3973.80 ± 4.55
100047.0 ± 2.5639.90 ± 5.3265.98 ± 7.1037.72 ± 4.4530.88 ± 3.1085 ± 5.87
IC5022.3525.2019.5225.7828.7014.55
Table 5. FRAP Scavenging Activity
 FRAP scavenging activity (%)
concentration (μg/mL)extractn-hexanechloroformethyl acetateaqueousascorbic acid
62.59.12 ± 1.1011.55 ± 1.1019.45 ± 0.88 2.11 ± 0.4529.45 ± 3.23
12517.23 ± 2.3419.15 ± 1.7033.18 ± 1.09.18 ± 0.889.88 ± 0.8947.52 ± 4.20
25029.80 ±1.5538.22 ± 3.5247.90 ± 1.8014.54 ± 1.9015.20 ± 1.069.90 ± 7.89
50044.65 ± 3.4056.60 ± 5.1066.38 ± 4.2423.52 ± 1.8821.52 ± 1.8981.35 ± 7.38
100052.0 ± 6.7568.48 ± 7.7572.70 ± 5.7631.60 ± 4.1229.98 ± 2.2092.78 ± 8.80
IC5020.4517.6815.3424.4827.349.25

3.4. Cytotoxicity Assay

In the present study, the crude methanol extract and different fractions of R. purpurea were investigated against various cancer cell lines including Caco-2, A549, HepG2, and MDA-MB-321 and two normal cell lines, i.e., primary epithelial cells (HPAEpiC and HRPTEpiC) using the MTS assay. The IC50 values are depicted in Figure 2.

Figure 2

Figure 2. IC50 values of the plant extract and fractions against different cell lines using cyclophosphamide and doxorubicin as positive controls and DMSO as a negative control.

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The chloroform fractions revealed significant activity against Caco-2, A549, and HepG2 cell lines with IC50 values of 4.55, 4.65, and 2.85 μg/mL, respectively, as compared to standard drugs, i.e, cyclophosphamide and doxorubicin (IC50 values of 2.15 and 1.98; 0.98 and 1.86; and 1.45 and 2.10 μg/mL against the mentioned cell lines, respectively). Similarly, the n-hexane fraction showed significant activity against the same cell lines with IC50 values of 5.65, 5.50, and 4.95 μg/mL, respectively. The corresponding percent cell viability values are depicted in Figures S1–S4. A lower percent cell viability value shows a greater potential for cytotoxic activity. In terms of percent cell viability, the n-hexane fraction showed significant cytotoxic activity (at 10 mg/mL dose) against Caco-2 (11%), A549 (17%), and HepG2 (13%) cell lines. Similarly, the chloroform fraction depicted significant activity against Caco-2 (9%), A549 (18%), and HepG2 (22%) cell lines at the same concentration. The ethyl acetate and aqueous fractions did not show any considerable cytotoxic activity against the studied cell lines. It is pertinent to mention that the crude methanol extract and fractions showed a dose-dependent activity. Compounds that exhibit potent cytotoxicity should be highly selective against cancer cells and remain safe or noncytotoxic against normal human cell lines. To investigate the safety profile or selectivity of the extract and fractions, they were tested against the primary epithelial cells HPAEpiC and HRPTEpiC (Figures S5 and S6). The percentage cell viability of all fractions and the crude methanol extract remained high compared to that of the standard drugs, i.e., cyclophosphamide and doxorubicin, when tested against HPAEpiC and HRPTEpiC cells. To assess and elaborate on the selectivity of the tested samples for cancer cell lines, the selectivity index was calculated (Figures 3 and 4). In the present study, the crude methanol extract and all fractions except chloroform exhibited high selectivity index values for Caco-2, A549, and HepG2 cancer cell lines, compared to those of standard drugs.

Figure 3

Figure 3. Selectivity indices of the standards, extract, and fractions against HPAEpiC.

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Figure 4

Figure 4. Selectivity indices of the standards, extract, and fractions against HRPTEpiC.

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4. Discussion

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The literature regarding R. purpurea is scarce; therefore, this study may validate the traditional use of this plant as antimicrobial and cytotoxic agents. In the present study, the crude methanol extract and n-hexane and chloroform fractions revealed significant activity against P. aeruginosa, S. aureus, S. typhi, and B. subtilis. It has been documented that typhoid fever, throat, skin, and other frequent infections are caused by these pathogens. S. aureus is a well-known bacteria that causes infections in wounds and throat. (29) Overall, the results suggest that the R. purpurea extract may be used to mitigate infections caused by the mentioned pathogens. In addition, the traditional use to treat several infections, particularly throat and skin, supports the antibacterial findings of this study. Phytochemical analysis revealed the presence of phenolic compounds in the extract, which may be possibly involved in the antibacterial activity observed in the present study. The literature review showed that phenolics are the major plant constituents exhibiting antibacterial activity. (30) The bactericidal effect depicted by these substances may involve several mechanisms such as pore formation, inhibition of DNA gyrase and nucleic acid synthesis, and alteration in the physicochemical properties of bacterial membrane. (31) However, further studies are needed to screen n-hexane and chloroform fractions and to isolate the specific compounds responsible for the said activity and to elucidate the specific mechanism for the same.
Among the fungal strains, the crude methanol extract and chloroform fractions exhibited significant activity against F. solani and A. niger as compared to the standard. The same fractions showed considerable activity against A. flavus. The literature review showed that about half of the human diseases involving Fusarium are caused by F. solani. (32) These infections include, but not limited to, keratitis and skin infections. Similarly, the Aspergillus species are the common opportunistic filamentous fungi and are highly resistant to most of the available antifungal drugs. (33) In this study, the R. purpurea extract showed significant activity against the mentioned fungal strains. In addition, this plant is famous for the treatment of skin-related disorders; therefore, the extract and/or fractions derived from the said plant may be considered as an alternative effective therapy for the mitigation of the above-mentioned disorders. Possible mechanisms for antifungal drugs may include, but not limited to, the inhibition of cell wall synthesis and pore formation; however, further studies are required to elucidate the exact mechanism for the same.
According to the WHO, most of the world population (approx. 80%) rely on herbal medicines for their primary health care needs. Under stress conditions, the human body produces more reactive oxygen species such as hydroxyl and superoxide anion radicals, compared to enzymatic antioxidants. At high concentrations, these radicals are very much hazardous to the body and damage most of the cell components including DNA, cell proteins, and membranes, which eventually leads to the development of cancer and other health conditions. (34) Synthetic antioxidants are widely used in the food industry; however, they are associated with several side effects including carcinogenesis and liver cirrhosis. Therefore, in recent decades, the use of natural antioxidants has got much attention. The DPPH and FRAP assays showed that R. purpurea contains a considerable amount of antioxidants that can be useful in the treatment of several diseases including cancer. Furthermore, as chloroform and n-hexane fractions depicted significant antioxidant activity, the same fractions may be further investigated for the isolation of specific antioxidant compounds.
Despite advances in cancer research, it remains one of the most devastating diseases, being a major public health problem and one of the foremost causes of death worldwide. (35) To date, a number of anticancer drugs have been developed; however, the emergence of resistance and serious side effects associated with the existing drugs demands for the search of safe and effective cytotoxic compounds. Since ancient times, the plant kingdom has provided a number of diverse secondary metabolites for the treatment of several human aliments including different cancers. According to Cao et al., flavonoids isolated from different plants showed significant cytotoxic activity against human cancer cell lines such as breast (MCF-7) and liver (HepG2) cancers. (36) Alkaloids isolated from Catharanthus roseus (vinblastine, vincristine), Solanum mauritianum (solasodine), Camptotheca acuminata (Campothecin), etc. were effectively used in the treatment of ovarian and colorectal cancers. (37) Several saponins obtained from P. japonocus, Panax ginseng, and P. quiquefolius showed excellent cytotoxic properties against oral squamous carcinoma and other related cancers. (38) Similarly, other secondary metabolites such as polyphenols, tannins, brassinosteroids, anthocyanin, podophyllotoxin derivatives, terpenoids, and essential oils exhibited significant cytotoxic activity against various cancer cell lines. (39)
Fortunately, Pakistan has the world’s richest natural flora. There are almost 5700 species of different plants, of which around 2000 species have medicinal values. (40) Exploring the medicinal values of these plants may not only contribute to drug discovery for various aliments but also play a vital role in the revenue generation. The present study was therefore designed to screen the R. purpurea extract, in search for new lead compounds with potential cytotoxic properties. Different fractions of the said plant showed significant activities against various malignancies. The crude methanol extract and chloroform and n-hexane fractions showed almost similar significant activities against gut, liver, and lung cancers. The results suggest further studies to explore new chemical entities that will help discover new therapeutic agents to treat a variety of cancers and infectious diseases.

5. Conclusions

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To the best of our knowledge, this is the first detailed study regarding the antimicrobial, antioxidant, and cytotoxic potential of R. purpurea. The crude methanol extract and n-hexane and chloroform fractions revealed significant activity against the well-known pathogens of bacterial and fungal origin including P. aeruginosa, S. aureus, S. typhi, F. solani, A. flavus, and A. niger. The same fractions showed considerable antioxidant activity particularly at high concentrations (1000 μg/mL). In addition, the n-hexane and chloroform fractions demonstrated significant cytotoxic activity against Caco-2, A549, and HepG2 cell lines. Furthermore, the same fractions exhibited high selectivity against the mentioned cell lines, which confirms their safety for normal human cells. Based on the results, R. purpurea warrants further studies to isolate new leads for the effective treatment of a variety of cancers and infectious diseases. This study also suggests evaluating the in vivo cytotoxic potential of R. purpurea when used alone or in combination with cyclophosphamide or doxorubicin to lessen the side effects of the latter.

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Author Information

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  • Corresponding Author
  • Authors
    • Nida Alam - Department of Pharmacy, University of Peshawar, Peshawar 25120, Pakistan
    • Atif Ali Khan Khalil - Department of Biological Sciences, National University of Medical Sciences, Rawalpindi 46000, Pakistan
    • Asif Jan - Department of Pharmacy, University of Peshawar, Peshawar 25120, Pakistan
    • Faiza Naureen - Department of Pharmacy, Abdul Wali Khan University, Mardan 23200, Pakistan
    • Zaki Ullah - Department of Pharmacy, University of Peshawar, Peshawar 25120, Pakistan
    • Amal Alotaibi - Department of Basic Science, College of Medicine, Princess Nourah Bint Abdulrahman University, P.O. Box 84428, Riyadh 11671, Saudi Arabia
    • Riaz Ullah - Department of Pharmacognosy (Medicinal Aromatic and Poisonous Plants Research Center) College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia;  Orcidhttps://orcid.org/0000-0002-2860-467X
    • Sami Ullah - Department of Pharmacy, University of Peshawar, Peshawar 25120, Pakistan
    • Yasar Shah - Department of Pharmacy, Abdul Wali Khan University, Mardan 23200, Pakistan
    • Sayyed Ibrahim Shah - Department of Pharmacy, Abdul Wali Khan University, Mardan 23200, Pakistan
    • Sultan Mehtap Büyüker - Department of Pharmacy Services, Üsküdar University, İstanbul 34664, Turkey
  • Notes
    The authors declare no competing financial interest.

Acknowledgments

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This research was funded by Princess Nourah Bint Abdulrahman University, Researchers Supporting Project number (PNURSP2022R33), Riyadh, Saudi Arabia.

References

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This article references 40 other publications.

  1. 1
    Richard, J. F.; Yitzhak, T. Antibiotics and bacterial resistance in the 21st century. Perspect. Med. Chem. 2014, 6, 25– 64,  DOI: 10.4137/PMC.S13209
    Google Scholar
    There is no corresponding record for this reference.
  2. 2
    Javaid, N.; Sultana, Q.; Rasool, K.; Gandra, S.; Ahmad, F.; Chaudhary, S. U.; Mirza, S. Trends in antimicrobial resistance amongst pathogens isolated from blood and cerebrospinal fluid cultures in Pakistan (2011-2015): A retrospective cross-sectional study. PLoS One 2021, 16, e0250226  DOI: 10.1371/journal.pone.0250226
    Google Scholar
    2
    Trends in antimicrobial resistance amongst pathogens isolated from blood and cerebrospinal fluid cultures in Pakistan (2011-2015): A retrospective cross-sectional study
    Javaid, Nida; Sultana, Qamar; Rasool, Karam; Gandra, Sumanth; Ahmad, Fayyaz; Chaudhary, Safee Ullah; Mirza, Shaper
    PLoS One (2021), 16 (4), e0250226CODEN: POLNCL; ISSN:1932-6203. (Public Library of Science)
    While antimicrobial resistance (AMR) continues to be a major public health problem in Pakistan, data regarding trends of resistance among pathogenic bacteria remains scarce, with few studies presenting long-term trends in AMR. This study was therefore designed to analyze long-term AMR trends at a national level in Pakistan. We report here results of a comprehensive anal. of resistance, among pathogens isolated from blood and cerebrospinal fluid (CSF), between 2011 and 2015. Susceptibility data was obtained from a local lab. with collection points all across Pakistan (Chughtai Lab.). Resistance proportions to most commonly used antimicrobials were calcd. for each pathogen over a period of five years. While Acinetobacter species demonstrated highest resistance rates to all tested antimicrobials, a sharp increase in carbapenem resistance was the most noticeable (50%-95%) between 2011-2015. Our results also highlight the presence of third and fourth generation cephalosporins resistance in Salmonella enterica serovar Typhi in Pakistan. Interestingly, where rise in AMR was being obsd. in some major invasive pathogens, decreasing resistance trends were obsd. in Staphylococcus aureus, against commonly used antimicrobials. Overall pathogens isolated from blood and CSF between 2011-2015, showed an increase in resistance towards commonly used antimicrobials.
  3. 3
    Cox, H. S.; Sibilia, C.; Feuerriegel, S.; Kalon, S.; Polonsky, J.; Khamraev, A. K.; Rüsch-Gerdes, S.; Mills, C.; Niemann, S. Emergence of extensive drug resistance during treatment for multidrug-resistant tuberculosis. N. Engl. J. Med. 2008, 359, 2398– 2400,  DOI: 10.1056/NEJMc0805644
    Google Scholar
    3
    Emergence of extensive drug resistance during treatment for multidrug-resistant tuberculosis
    Cox, Helen S.; Sibilia, Katia; Feuerriegel, Silke; Kalon, Stobdan; Polonsky, Jonny; Khamraev, Atadjan K.; Rusch-Gerdes, Sabine; Mills, Clair; Niemann, Stefan
    New England Journal of Medicine (2008), 359 (22), 2398-2400CODEN: NEJMAG; ISSN:0028-4793. (Massachusetts Medical Society)
    There is no expanded citation for this reference.
  4. 4
    Qamar, F. N.; Yousafzai, M. T.; Khalid, M.; Kazi, A. M.; Lohana, H.; Karim, S.; Khan, A.; Hotwani, A.; Qureshi, S.; Kabir, F. Outbreak investigation of ceftriaxone-resistant Salmonella enterica serotype Typhi and its risk factors among the general population in S Hyderabad, Pakistan: a matched case-control study. Lancet Infect. Dis. 2018, 18, 1368– 1376,  DOI: 10.1016/S1473-3099(18)30483-3
    Google Scholar
    4
    Outbreak investigation of ceftriaxone-resistant Salmonella enterica serotype Typhi and its risk factors among the general population in Hyderabad, Pakistan: a matched case-control study
    Qamar Farah Naz; Yousafzai Mohammad Tahir; Khalid Muhammad; Kazi Abdul Momin; Karim Sultan; Khan Ayub; Hotwani Aneeta; Qureshi Shahida; Kabir Furqan; Aziz Fatima; Lohana Heeramani; Memon Naveed Masood; Domki Mudassar Hussain; Hasan Rumina
    The Lancet. Infectious diseases (2018), 18 (12), 1368-1376 ISSN:.
    BACKGROUND: Pakistan is currently facing the largest outbreak of ceftriaxone-resistant Salmonella enterica serotype Typhi described to date. Here we aimed to report the outbreak investigation done in Hyderabad, Pakistan, and identify disease risk factors. METHODS: We did an age-matched case-control (1:4) study, in which cases of ceftriaxone-resistant S Typhi were identified from active sentinal sites (three hospitals in Hyderabad, Pakistan), community, and laboratory-based surveillance. Ceftriaxone-resistant S Typhi infection (ie, resistance to ampicillin, chloramphenicol, co-trimoxazole, fluoroquinole, and ceftriaxone) was confirmed using blood culture. Healthy participants (controls) were enrolled for the first 200 people (cases) with ceftriaxone-resistant S Typhi. A structured questionnaire was administered to identify exposures 4 weeks before the illness (cases) or enrolment (controls). Cases were included if written informed consent was provided. Four controls were selected from the same community as the corresponding case, matched on age, being healthy at the time of enrolment, and with no febrile illness in the 4 weeks before enrolment. Samples of drinking water from households and community water sources (ie, hand pumps or taps in common areas outside households) were collected for testing. Conditional logistic regression analysis was used to assess the risk factors for ceftriaxone-resistant S Typhi outbreak in Hyderabad. FINDINGS: Between Nov 30, 2016, and Dec 30, 2017, 486 people with ceftriaxone-resistant S Typhi were identified from Hyderabad. Of the 486 cases, 296 (61%) were male and 447 (92%) were aged 15 years or younger. Several factors were significantly associated with acquisition of ceftriaxone-resistant S Typhi, including male sex (adjusted odds ratio [aOR] 1·53, 95% CI 1·06-2·21), eating outside of the house (aOR 1·48, 1·01-2·19), exposure to a patient with S Typhi infection (aOR 3·81, 2·21-6·83), and a history of antimicrobial use (aOR 4·25, 2·53-7·13). Nine (69%) of 13 water samples taken from the households of people with ceftriaxone-resistant S Typhi infection were positive for Escherichia coli, indicating faecal contamination. S Typhi DNA was detected in 12 (22%) of 55 water samples from community water sources. Geospatial mapping showed clustering of cases around sewerage lines. INTERPRETATION: Hyderabad faces the largest reported outbreak of ceftriaxone-resistant S Typhi. The outbreak is suspected to be attributed to the contaminated drinking water, especially the mixing of sewage with drinking water. The risk of ceftriaxone-resistant S Typhi infection is increased among children aged 15 years and younger, male individuals, and those eating outside the house. Vaccination and chlorination of water are recommended for the containment of the outbreak. FUNDING: None.
  5. 5
    Zilberberg, M. D.; Kollef, M. H.; Shorr, A. F. Secular trends in Acinetobacter baumannii resistance in respiratory and blood stream specimens in the United States, 2003 to 2012: A survey study. J. Hosp. Med. 2016, 11, 21– 26,  DOI: 10.1002/jhm.2477
    Google Scholar
    5
    Secular trends in Acinetobacter baumannii resistance in respiratory and blood stream specimens in the United States, 2003 to 2012: A survey study
    Zilberberg Marya D; Zilberberg Marya D; Kollef Marin H; Shorr Andrew F
    Journal of hospital medicine (2016), 11 (1), 21-6 ISSN:.
    BACKGROUND: Acinetobacter baumannii (AB) has evolved a variety of resistance mechanisms and exhibits unpredictable susceptibility patterns, making it difficult to select empiric therapy. OBJECTIVE: To examine US secular trends in the resistance of AB in respiratory infections and blood stream infections (BSI) to antimicrobial agents whose effectiveness is supported in the literature DESIGN: Survey. METHODS: We analyzed 3 time periods (2003-2005, 2006-2008, 2009-2012) in Eurofins' The Surveillance Network for resistance of AB to the following antimicrobials: carbapenems (imipenem, meropenem, doripenem), aminoglycosides (tobramycin, amikacin), tetracyclines (minocycline, doxycycline), polymyxins (colistin, polymyxin B), ampicillin-sulbactam, and trimethoprim-sulfamethoxazole. Resistance to ≥3 drug classes defined multidrug resistance (MDR). RESULTS: We identified 39,320 AB specimens (81.1% respiratory, 18.9% BSI). The highest prevalence of resistance was to doripenem (90.3%) followed by trimethoprim-sulfamethoxazole (55.3%), and the lowest to colistin (5.3%). Resistance to carbapenems (21.0% in 2003-2005 and 47.9% in 2009-2012) and colistin (2.8% in 2006-2008 to 6.9% in 2009-2012) more than doubled. Prevalence of MDR AB rose from 21.4% in 2003 to 2005 to 33.7% in 2006 to 2008, and remained stable at 35.2% in 2009 to 2012. In contrast, resistance to minocycline diminished from 56.5% (2003-2005) to 30.5% (2009-2012). MDR organisms were most frequent in nursing homes (46.5%), followed by general ward (29.2%), intensive care unit (28.7%), and outpatient setting (26.2%). CONCLUSIONS: Resistance rates among AB to such last-resort antimicrobials as carbapenems and colistin are on the rise, whereas that to minocycline has declined. Nursing homes are a reservoir of resistant AB. These trends should inform not only empiric treatment of serious infections, but also approaches to infection control.
  6. 6
    Nida, J.; Qamar, S.; Karam, R.; Sumanth, G.; Fayyaz, A.; Safee, U. C.; Shaper, M. Trends in antimicrobial resistance amongst pathogens isolated from blood and cerebrospinal fluid cultures in Pakistan (2011–2015): A retrospective cross-sectional study. PLoS One 2021, 1– 26,  DOI: 10.1371/journal.pone.0250226
    Google Scholar
    There is no corresponding record for this reference.
  7. 7
    Lagacé-Wiens, P. R. S.; Adam, H. J.; Low, D. E.; Blondeau, J. M.; Baxter, M. R.; Denisuik, A. J. Trends in antibiotic resistance over time among pathogens from Canadian hospitals: results of the CANWARD study 2007–11. J. Antimicrob. Chemother. 2013, 68, i23– i29,  DOI: 10.1093/jac/dkt023
    Google Scholar
    7
    Trends in antibiotic resistance over time among pathogens from Canadian hospitals: results of the CANWARD study 2007-11
    Lagace-Wiens, Philippe R. S.; Adam, Heather J.; Low, Donald E.; Blondeau, Joseph M.; Baxter, Melanie R.; Denisuik, Andrew J.; Nichol, Kimberly A.; Walkty, Andrew; Karlowsky, James A.; Mulvey, Michael R.; Hoban, Daryl J.; Zhanel, George G.
    Journal of Antimicrobial Chemotherapy (2013), 68 (Suppl. 1), i23-i29CODEN: JACHDX; ISSN:0305-7453. (Oxford University Press)
    Antimicrobial resistance patterns change over time and longitudinal surveillance studies provide insight into these trends. We sought to describe the important trends in antimicrobial resistance in key pathogens across Canada to provide useful information to clinicians, policy makers and industry, to assist in optimizing antimicrobial therapy, formulary choices and drug development. We analyzed longitudinal data from the CANWARD study using a multivariate regression model to control for possible effects of patient demographics on resistance, in order to assess the impact of time on antimicrobial resistance independent of other measured variables. We identified several key trends in common pathogens. In particular, we obsd. a statistically significant increase in the proportion of Escherichia coli isolates that were resistant to extended-spectrum cephalosporins and fluoroquinolones, an increase in the proportion of Klebsiella pneumoniae isolates that were resistant to extended-spectrum cephalosporins, a redn. in the proportion of Staphylococcus aureus that were methicillin, clindamycin and trimethoprim/sulfamethoxazole resistant, and a redn. in the proportion of Pseudomonas aeruginosa that were fluoroquinolone and gentamicin resistant. Conclusions Although some of these trends, such as the dramatic increase in fluoroquinolone and cephalosporin resistance in E. coli, can be attributed to the emergence and global spread of resistant clones (e.g. ST131 E. coli), others remain unexplained. However, recognizing these trends remains important to guide changes in empirical antimicrobial therapy and drug development.
  8. 8
    Sharma, S. V.; Haber, D. A.; Settleman, J. Cell line-based platforms to evaluate the therapeutic efficacy of candidate anticancer agents. Nat. Rev. Cancer 2010, 10, 241– 253,  DOI: 10.1038/nrc2820
    Google Scholar
    8
    Cell line-based platforms to evaluate the therapeutic efficacy of candidate anticancer agents
    Sharma, Sreenath V.; Haber, Daniel A.; Settleman, Jeff
    Nature Reviews Cancer (2010), 10 (4), 241-253CODEN: NRCAC4; ISSN:1474-175X. (Nature Publishing Group)
    A review. Efforts to discover new cancer drugs and predict their clin. activity are limited by the fact that lab. models to test drug efficacy do not faithfully recapitulate this complex disease. One important model system for evaluating candidate anticancer agents is human tumor-derived cell lines. Although cultured cancer cells can exhibit distinct properties compared with their naturally growing counterparts, recent technologies that facilitate the parallel anal. of large panels of such lines, together with genomic technologies that define their genetic constitution, have revitalized efforts to use cancer cell lines to assess the clin. utility of new investigational cancer drugs and to discover predictive biomarkers.
  9. 9
    McPherson, K.; Steel, C.; Dixon, J. Breast cancer─epidemiology, risk factors, and genetics. Br. Med. J. 2000, 321, 624– 628,  DOI: 10.1136/bmj.321.7261.624
    Google Scholar
    9
    ABC of breast diseases. Breast cancer-epidemiology, risk factors, and genetics
    McPherson K; Steel C M; Dixon J M
    BMJ (Clinical research ed.) (2000), 321 (7261), 624-8 ISSN:0959-8138.
    There is no expanded citation for this reference.
  10. 10
    Menhas, R.; Umer, S. Breast cancer among Pakistani women. Iran. J. Public Health 2015, 44, 586– 587
    Google Scholar
    10
    Breast Cancer among Pakistani Women
    Menhas Rashid; Umer Shumaila
    Iranian journal of public health (2015), 44 (4), 586-7 ISSN:2251-6085.
    There is no expanded citation for this reference.
  11. 11
    Amna, S. B.; Zaigham, A.; Wasim, J. Hepatocellular Carcinoma in Pakistan: Where do We Stand?. Hepatitis Mon. 2012, 12, e6023  DOI: 10.5812/hepatmon.6023
    Google Scholar
    There is no corresponding record for this reference.
  12. 12
    Bosch, F. X.; Ribes, J.; Díaz, M.; Cléries, R. Primary liver cancer, worldwide incidence and trends. Gastroenterology 2004, 127, S5– S16,  DOI: 10.1053/j.gastro.2004.09.011
    Google Scholar
    12
    Primary liver cancer: worldwide incidence and trends
    Bosch F Xavier; Ribes Josepa; Diaz Mireia; Cleries Ramon
    Gastroenterology (2004), 127 (5 Suppl 1), S5-S16 ISSN:0016-5085.
    Estimates from the year 2000 indicate that liver cancer remains the fifth most common malignancy in men and the eighth in women worldwide. The number of new cases is estimated to be 564,000 per year, including 398,000 in men and 166,000 in women. In high-risk countries, liver cancer can arise before the age of 20 years, whereas, in countries at low risk, liver cancer is rare before the age of 50 years. Rates of liver cancer in men are typically 2 to 4 times higher than in women. The incidence of primary liver cancer is increasing in several developed countries, including the United States, and the increase will likely continue for some decades. The trend is a result of a cohort effect related to infection with hepatitis B and C viruses, the incidence of which peaked in the 1950s to 1980s. In selected areas of some developing countries, the incidence of primary liver cancer has decreased, possibly as a result of the introduction of hepatitis B virus vaccine. The geographic variability in incidence of primary liver cancer is largely explained by the distribution and the natural history of the hepatitis B and C viruses. The attributable risk estimates for the combined effects of these infections account for well over 80% of liver cancer cases worldwide. Primary liver cancer is the first human cancer largely amenable to prevention using hepatitis B virus vaccines and screening of blood and blood products for hepatitis B and C viruses.
  13. 13
    Sheikh, H. S.; Kiran, M.; Fareeha, S.; Muhammad, F. U. Q. Lung Cancer in Pakistan. J. Thorac. Oncol. 2022, 17, 602– 607,  DOI: 10.1016/j.jtho.2022.01.009
    Google Scholar
    13
    Lung Cancer in Pakistan
    Sheikh Hassan Shahryar; Munawar Kiran; Sheikh Fareeha; Qamar Muhammad Fawad Ul
    Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer (2022), 17 (5), 602-607 ISSN:.
    There is no expanded citation for this reference.
  14. 14
    Prashanth, R.; Tagore, S.; Adam, B. Epidemiology of colorectal cancer: incidence, mortality, survival, and risk factors. Przegl. Gastroenterol. 2019, 14, 89– 103,  DOI: 10.5114/pg.2018.81072
    Google Scholar
    There is no corresponding record for this reference.
  15. 15
    Malik, A. I.; Farhana, B.; Aamir, A. S.; Muhammed, A. Y. Surgically treated rectal cancer patients─Outcomes at a tertiary care cancer hospital in Pakistan. Asian J. Surg. 2015, 38, 13– 20,  DOI: 10.1016/j.asjsur.2014.05.004
    Google Scholar
    15
    Surgically treated rectal cancer patients--outcomes at a tertiary care cancer hospital in Pakistan
    Malik Anum Imran; Badar Farhana; Syed Aamir Ali; Yusuf Muhammed Aasim
    Asian journal of surgery (2015), 38 (1), 13-20 ISSN:.
    AIM: The aim of this study was to analyze our experience with rectal cancer patients who underwent surgical excision at our institution. METHODS: Data on 112 rectal cancer patients who underwent surgical resection with total mesorectal excision, from January 2005 to December 2008, were evaluated retrospectively. RESULTS: We achieved an initial complete remission rate of 74.1%. Overall, 92.8% of patients had a complete total mesorectal excision. The overall survival analysis for all patients showed a 1-year survival rate of 98%, a 3-year survival rate of 82%, and a 5-year survival rate of 70%. We report a 41.9% rate of postoperative complications. The 1-, 3-, and 5-year survival rates for females were 100%, 90%, and 72%, respectively and for males, they were 90%, 80%, and 68%, respectively. Differences in overall survival by sex were not statistically significant (p > 0.05). Those patients who were treated with only surgery had the best outcomes with survival being worse in those treated with surgery and adjuvant therapy. Neoadjuvant treatment followed by surgery led to better results. CONCLUSION: We conclude that we have been successful in achieving high rates of curative resection, complete remission, and overall survival. Neoadjuvant and adjuvant chemotherapy significantly impact rates of remission.
  16. 16
    Engwa, G. A. Free radicals and the role of plant phytochemicals as antioxidants against oxidative stress-related diseases. Phytochemicals 2018, 7, 49– 74,  DOI: 10.5772/intechopen.76719
    Google Scholar
    There is no corresponding record for this reference.
  17. 17
    Alkadi, H. A review on free radicals and antioxidants. Infect. Disord.: Drug Targets 2020, 20, 16– 26,  DOI: 10.2174/1871526518666180628124323
    Google Scholar
    17
    A Review on Free Radicals and Antioxidants
    Alkadi, Hourieh
    Infectious Disorders: Drug Targets (2020), 20 (1), 16-26CODEN: IDDTAD; ISSN:1871-5265. (Bentham Science Publishers Ltd.)
    Free radicals are generated in our body by several systems. A balance among free radicals and antioxidants is an important matter for appropriate physiol. function. If free radicals become greater than the ability of the body to control them, a case known as oxidative stress appears, as a result of that, a no. of human diseases spread in the body. Antioxidants can contribute to facingthis oxidative stress. The present review provides a brief overview of free radicals, oxidative stress, some natural antioxidants and the relationship between them.
  18. 18
    Fazli, K.; Madiha, A.; Suleman, K.; Hamayun, K.; Muhammad, U. K. S.; Ameer, K.; Asif, J.; Naveed, U.; Yasar, S.; Zaki, U.; Muhammad, A.; Tayyaba, I.; Abubakr, M. I.; Mayeen, U. K.; Talha, B. E. Antimicrobial, anti-inflammatory and antioxidant activities of natural organic matter extracted from cretaceous shales in district Nowshera-Pakistan. Arab. J. Chem. 2022, 15, 1036033  DOI: 10.1016/j.arabjc.2021.103633
    Google Scholar
    There is no corresponding record for this reference.
  19. 19
    Stocker, P.; Yousfi, M.; Djerridane, O.; Perrier, J.; Amziani, R.; El Boustani, S.; Moulin, A. Effect of flavonoids from various Mediterranean plants on enzymatic activity of intestinal carboxylesterase. Biochimie 2004, 86, 919– 925,  DOI: 10.1016/j.biochi.2004.09.005
    Google Scholar
    19
    Effect of flavonoids from various Mediterranean plants on enzymatic activity of intestinal carboxylesterase
    Stocker, P.; Yousfi, M.; Djerridane, O.; Perrier, J.; Amziani, R.; El Boustani, S.; Moulin, A.
    Biochimie (2004), 86 (12), 919-925CODEN: BICMBE; ISSN:0300-9084. (Elsevier B.V.)
    Flavonol compds. of three Mediterranean plants from the Algerian Atlas used traditionally in Arab folk medicine, Arenaria serpyllifolia, Rhamnus alaternus and Thapsia garganica, were found to inhibit the enzymic activities of both rat intestine and purified porcine liver carboxylesterase in a concn.-dependent manner. Results indicate that the flavonol compds. from the aerial part of these plants lead to the inactivation of the CE pI = 5.1 with Ki of micromolar range. These results encourage us to perform further biol. investigation.
  20. 20
    Kosalec, I.; Kremer, D.; Locatelli, M.; Epifano, F.; Genovese, S.; Carlucci, G.; Končić, M. Z. Anthraquinone profile, antioxidant and antimicrobial activity of bark extracts of Rhamnus alaternus, Rhamnus fallax, Rhamnus intermedia and Rhamnus pumila. Food chem. 2013, 136, 335– 341,  DOI: 10.1016/j.foodchem.2012.08.026
    Google Scholar
    20
    Anthraquinone profile, antioxidant and antimicrobial activity of bark extracts of Rhamnus alaternus, R. fallax, R. intermedia and R. pumila
    Kosalec, I.; Kremer, D.; Locatelli, M.; Epifano, F.; Genovese, S.; Carlucci, G.; Randic, M.; Zovko Koncic, M.
    Food Chemistry (2013), 136 (2), 335-341CODEN: FOCHDJ; ISSN:0308-8146. (Elsevier Ltd.)
    The quantity of phenols, as well as antioxidant and antimicrobial activities, were investigated in bark of Rhamnus alaternus L., R. fallax Boiss., R. intermedia Steud. et Hochst., and R. pumila Turra from natural stands in Croatia. The most abundant anthraquinones in the investigated exts. were chrysophanol in R. alaternus (3.14 mg/g), emodin in R. pumila (0.339 mg/g), and physcion in R. fallax (2.70 mg/g) and R. intermedia (0.285 mg/g). The species exhibiting the highest antioxidant activity were R. fallax and R. pumila. A pos. correlation was obsd. between total phenolic and flavonoid levels of the exts. and antioxidant activity in some of the assays. All species showed antimicrobial activity against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Candida albicans, Aspergillus niger and Microsporum gypseum with minimal inhibitory concns. equal to or below 2.500 mg/mL. The results indicate that the investigated Rhamnus species are a source of anthraquinones and other phenols, which act as multifunctional antioxidants with antimicrobial activity.
  21. 21
    Marzouk, M. S.; El-Toumy, S. A.; Merfort, I.; Nawwar, M. A. Polyphenolic metabolites of Rhamnus disperma. Phytochemistry 1999, 52, 943– 946,  DOI: 10.1016/S0031-9422(99)00262-9
    Google Scholar
    21
    Polyphenolic metabolites of Rhamnus disperma
    Marzouk, Mohamed S.; El-Toumy, Sayed A. A.; Merfort, Irmgard; Nawwar, Mahmoud A. M.
    Phytochemistry (1999), 52 (5), 943-946CODEN: PYTCAS; ISSN:0031-9422. (Elsevier Science Ltd.)
    A new quercetin dirhamnoside has been isolated and identified as quercetin 3,4'-di-O-α-L-rhamnopyranoside (I) together with 24 structurally known phenolic metabolites from the fruits and aerial parts of Rhamnus disperma for the first time. The known compds. have been characterized as kaempferol 3-O-robinoside, 3-O-rhamninoside, 4'-O-rhamninoside, rhamnocitrin 3-O-rhamninoside, 4'-O-rhamninoside, quercetin 3-O-rhamnoside, 3-O-galactoside, 7-O-galactoside, 3-O-Me 7-O-galactoside, 3-O-robinoside, 3-O-rhamninoside, rhamnetin 3-O-rhamninoside, rhamnazin 3-O-robinoside, 3-O-rhamninoside, two phenolic acids 2,5-dihydroxybenzoic and protocatechuic acids, a coumarin isofraxetin, three aglycons identified as quercetin 3-Me ether, eriodictyol and taxifolin, together with five flavonols kaempferol, quercetin, rhamnocitrin, rhamnetin and rhamnazin. The structures of the isolated compds. have been established on the basis of chem. and NMR spectroscopic evidence as well as neg. ESI-MS in somes cases.
  22. 22
    Farzana, K.; Kyung-Hwan, K.; Hafiz, M. U. F.; Muhammad, A. F.; Muhammad, K.; Rooma, W.; Atif, A. K. H.; Fazli, K.; Chethikkattuveli, S. A. R.; Kinam, H.; Kyung-Hyun, C.; Abdul, S. M. Evaluation of antimicrobial and anticancer activities of selected medicinal plants of himalayas, Pakistan. Plants 2022, 11, 48  DOI: 10.3390/plants11010048
    Google Scholar
    There is no corresponding record for this reference.
  23. 23
    Boyanova, L.; Gergova, G.; Nikolov, R.; Derejian, S.; Lazarova, E.; Katsarov, N.; Mitov, I.; Krastev, Z. Activity of Bulgarian propolis against 94 Helicobacter pylori strains in vitro by agar-well diffusion, agar dilution and disc diffusion methods. J. Med. Microbiol. 2005, 54, 481– 483,  DOI: 10.1099/jmm.0.45880-0
    Google Scholar
    23
    Activity of Bulgarian propolis against 94 Helicobacter pylori strains in vitro by agar-well diffusion, agar dilution and disc diffusion methods
    Boyanova Lyudmila; Gergova Galina; Nikolov Rossen; Derejian Sirigan; Lazarova Elena; Katsarov Nikolai; Mitov Ivan; Krastev Zacharii
    Journal of medical microbiology (2005), 54 (Pt 5), 481-483 ISSN:0022-2615.
    Propolis exhibits antimicrobial, anti-inflammatory and other biological effects. The aim of this study was to evaluate the activity of 30 % ethanolic extract of Bulgarian propolis against 94 Helicobacter pylori strains by three methods. By the agar-well diffusion method, only 13.8 % of the strains exhibited no inhibition by 30 microl propolis extract (containing 9 mg propolis) and all isolates were inhibited to some extent by 90 microl of the extract (27 mg propolis) per well. The mean diameters of growth inhibition by 30, 60 or 90 microl propolis extract or 30 microl 96 % ethanol per well were 16.8, 19.2, 27.5 and 8.3 mm, respectively. The propolis extract was more active than the ethanol (P < 0.001). With 90 microl propolis extract per well, 69.4 % of the strains exhibited large diameters of growth inhibition (> or =20 mm) versus 26.6 % with 30 mul per well (P < 0.001). With moist propolis discs, inhibition was detected in more strains (92.1 %) than with dried discs (78.2 %, P < 0.05), with mean inhibitory diameters of 18.7 and 13.8 mm, respectively. By the agar dilution method, 100 and 300 microg propolis ml(-1) inhibited the growth of 57.1 % and 76.2 %, respectively, of the 21 strains tested. In conclusion, Bulgarian propolis had a strong and dose-dependent activity against most of the H. pylori strains tested. Although the effect of propolis on H. pylori in vitro is promising, further microbiological, pharmacological and clinical trials are required.
  24. 24
    Fazli, K.; Zafar, I.; Ayub, K.; Zakiullah; Fazli, N.; Muhammad, S. K. Validation of some of the ethnopharmacological uses of Xanthium strumarium and Duchesnea indica. Pak. J. Bot. 2012, 44, 1199– 1201
    Google Scholar
    There is no corresponding record for this reference.
  25. 25
    CLSI. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically, 9th ed.; Clinical and Laboratory Standards Institute, CLSI Document M07A9, 2012; Vol. 32.
    Google Scholar
    There is no corresponding record for this reference.
  26. 26
    Kedare, S. B.; Sing, R. P. Genesis and development of DPPH method of antioxidant assay. J. Food Sci. Technol. 2011, 48, 412– 422,  DOI: 10.1007/s13197-011-0251-1
    Google Scholar
    26
    Genesis and development of DPPH method of antioxidant assay
    Kedare, Sagar B.; Singh, R. P.
    Journal of Food Science and Technology (2011), 48 (4), 412-422CODEN: JFSTAB; ISSN:0022-1155. (Springer (India) Private Ltd.)
    A review. α, α-Diphenyl-β-picrylhydrazyl (DPPH) free radical scavenging method offers the first approach for evaluating the antioxidant potential of a compd., an ext. or other biol. sources. This is the simplest method, wherein the prospective compd. or ext. is mixed with DPPH soln. and absorbance is recorded after a defined period. However, with the advancement and sophistication in instrumental techniques, the method has undergone various modifications to suit the requirements, even though the basic approach remains same in all of them. This article presents a crit. review on various developments to the DPPH method.
  27. 27
    Iris, F. F. B.; Strain, J. J. Simultaneous measurement of total antioxidant power and ascorbic acid concentration. Methods Enzymol. 1999, 299, 15– 27,  DOI: 10.1016/S0076-6879(99)99005-5
    Google Scholar
    There is no corresponding record for this reference.
  28. 28
    Janice, M.; Joel, J.; Holly, H.; Kerry, W.; Paul, N. In vitri cytotoxic properties of crude polar extracts of plants sourced from Australia. Clin. Complementary Med. Pharmacol. 2022, 2, 100022
    Google Scholar
    There is no corresponding record for this reference.
  29. 29
    Tong, S. Y. C.; Joshua, S. D.; Emily, E.; Thomas, L. H.; Vance, G. F. J. Staphylococcus aureus Infections: Epidemiology, pathophysiology, clinical manifestations, and management. Clin. Microbiol. Rev. 2015, 28, 603– 661,  DOI: 10.1128/CMR.00134-14
    Google Scholar
    29
    Staphylococcus aureus infections: epidemiology, pathophysiology, clinical manifestations, and management
    Tong, Steven Y. C.; Davis, Joshua S.; Eichenberger, Emily; Holland, Thomas L.; Fowler, Vance G., Jr.
    Clinical Microbiology Reviews (2015), 28 (3), 603-661CODEN: CMIREX; ISSN:1098-6618. (American Society for Microbiology)
    Staphylococcus aureus is a major human pathogen that causes a wide range of clin. infections. It is a leading cause of bacteremia and infective endocarditis as well as osteoarticular, skin and soft tissue, pleuropulmonary, and device-related infections. This review comprehensively covers the epidemiol., pathophysiol., clin. manifestations, and management of each of these clin. entities. The past 2 decades have witnessed two clear shifts in the epidemiol. of S. aureus infections: first, a growing no. of health care-assocd. infections, particularly seen in infective endocarditis and prosthetic device infections, and second, an epidemic of community-assocd. skin and soft tissue infections driven by strains with certain virulence factors and resistance to β-lactam antibiotics. In reviewing the literature to support management strategies for these clin. manifestations, we also highlight the paucity of high-quality evidence for many key clin. questions.
  30. 30
    de Araújo, A. A.; Luiz, A. L. S.; Magda, R. A. F.; Manoel, A. S. N.; Giselle, R. S.; Raimundo, F. A. J.; Gerlane, C. B. G.; Maria, C. N. M. Quantification of polyphenols and evaluation of antimicrobial, analgesic and anti-inflammatory activities of aqueous and acetone–water extracts of Libidibia ferrea, Parapiptadenia rigida and Psidium guajava. J. Ethnopharmacol. 2014, 156, 88– 96,  DOI: 10.1016/j.jep.2014.07.031
    Google Scholar
    30
    Quantification of polyphenols and evaluation of antimicrobial, analgesic and anti-inflammatory activities of aqueous and acetone-water extracts of Libidibia ferrea, Parapiptadenia rigida and Psidium guajava
    de Araujo, Aurigena Antunes; Soares, Luiz Alberto Lira; Assuncao Ferreira, Magda Rhayanny; de Souza Neto, Manoel Andre; da Silva, Giselle Ribeiro; de Araujo, Raimundo Fernandes; Guerra, Gerlane Coelho Bernardo; de Melo, Maria Celeste Nunes
    Journal of Ethnopharmacology (2014), 156 (), 88-96CODEN: JOETD7; ISSN:0378-8741. (Elsevier Ireland Ltd.)
    Vast nos. of plant species from northeastern Brazil have not yet been phytochem. or biol. evaluated.The goal of this work was to obtain, characterize and show the antimicrobial, analgesic and anti-inflammatory activities of aq. and acetone-water exts. of Libidibia ferrea, Parapiptadenia rigida and Psidium guajava.The plant material (100 g) was dried, and the crude exts. were obtained by using turbo-extn. (10%; w/v) with water or acetone:water (7:3, vol./vol.) as the extn. solvent. High-performance liq. chromatog. (HPLC) methods were used to screen the crude exts. for hydrolysable tannins (gallic acid) and condensed tannins (catechins). The antibacterial activity was evaluated by agar-diffusion and microdilution methods against Gram-pos. strains (Staphylococcus aureus ATCC 25923, Staphylococcus epidermidis INCQS 00016, Enterococcus faecalis ATCC 29212 and a clin. isolate of methicillin-resistant Staphylococcus aureus) as well as Gram-neg. strains (Escherichia coli ATCC 25922, Salmonella enteritidis INCQS 00258, Shigella flexneri and Klebsiella pneumoniae). To evaluate the anti-inflammatory activity, a leukocyte migration model was used. Analgesic activity was detd. by the hot plate test and the acetic acid-induced abdominal writhing test. Data were analyzed by anal. of variance (ANOVA) at a significance level of 5%.Parapiptadenia rigida presented the highest amt. of total polyphenols (35.82±0.20%), while the greatest catechin content was found in the acetone-water ext. of Psidium guajava (EAWPg; 1.04 μg/g). The largest amts. of catechins were found in the aq. ext. of Libidibia ferrea (EALf; 1.07 μg/g) and the acetone-water ext. of Parapiptadenia rigida (EAWPr; 1.0 μg/g). All exts. showed activity against Gram-pos. bacteria. The aq. and acetone-water exts. of Psidium guajava showed the greatest inhibition zones in the agar diffusion tests. In the evaluation of the min. inhibitory concn. (MIC), the most susceptible Gram-pos. bacterium was Staphylococcus epidermidis and the most susceptible Gram-neg. bacterium was Shigella flexneri. EAPg and EAWPg showed the greatest MIC values. All exts. were significant inhibitors of leukocyte migration (p<0.05). Using the writhing test, significant analgesic activity was found for EAPr (50 mg/kg), EAWPr (100 mg/kg and 200 mg/kg) and EAWPg (50 mg/kg) (p<0.05).Thus, the appropriate extn. procedure preserves the chem. components such as gallic acid and catechin, and showed antimicrobial, anti-inflammatory and analgesic properties.
  31. 31
    Gradišar, H.; Pristovsek, P.; Plaper, A.; Jerala, R. Green tea catechins inhibit bacterial DNA gyrase by interaction with its ATP binding site. J. Med. Chem. 2007, 50, 264– 271,  DOI: 10.1021/jm060817o
    Google Scholar
    31
    Green Tea Catechins Inhibit Bacterial DNA Gyrase by Interaction with Its ATP Binding Site
    Gradisar, Helena; Pristovsek, Primoz; Plaper, Andreja; Jerala, Roman
    Journal of Medicinal Chemistry (2007), 50 (2), 264-271CODEN: JMCMAR; ISSN:0022-2623. (American Chemical Society)
    Catechins are the main ingredients of green tea exts. and have been shown to possess versatile biol. activities, including antimicrobial. We detd. that the catechins inhibit bacterial DNA gyrase by binding to the ATP binding site of the gyrase B subunit. In the group of four tested catechins, epigallocatechin gallate (EGCG) had the highest activity, followed by epicatechin gallate (ECG) and epigallocatechin (EGC). Specific binding to the N-terminal 24 kDa fragment of gyrase B was detd. by fluorescence spectroscopy and confirmed using heteronuclear two-dimensional NMR spectroscopy of the EGCG-15N-labeled gyrase B fragment complex. Protein residues affected by binding to EGCG were identified through chem. shift perturbation. Mol. docking calcns. suggest that the benzopyran ring of EGCG penetrates deeply into the active site while the galloyl moiety anchors it to the cleft through interactions with its hydroxyl groups, which explains the higher activity of EGCG and ECG.
  32. 32
    Laura, B. D.; Carla, W.; Lara, B. I.; Paula, C. S.; Janio, M. S. Activity of MSI-78, h-Lf1- 11 and cecropin B antimicrobial peptides alone and in combination with voriconazole and amphotericin B against clinical isolates of Fusarium solani. J. Med. Mycol. 2021, 31, 101119  DOI: 10.1016/j.mycmed.2021.101119
    Google Scholar
    There is no corresponding record for this reference.
  33. 33
    Shishodia, S. K.; Shraddha, T.; Jata, S. Resistance mechanism and proteins in Aspergillus species against antifungal agents. Mycology 2019, 10, 151– 165,  DOI: 10.1080/21501203.2019.1574927
    Google Scholar
    33
    Resistance mechanism and proteins in Aspergillus species against antifungal agents
    Shishodia, Sonia Kumari; Tiwari, Shraddha; Shankar, Jata
    Mycology (2019), 10 (3), 151-165CODEN: MYCOCG; ISSN:2150-1203. (Taylor & Francis Ltd.)
    Aspergillus species contain pathogenic and opportunistic fungal pathogens which have the potential to cause mycosis (invasive aspergillosis) in humans. The existing antifungal drugs have limitation largely due to the development of drug-resistant isolates. To gain insight into the mechanism of action and antifungal drug resistance in Aspergillus species including biofilm formation, we have reviewed protein data of Aspergillus species during interaction with antifungals drugs (polynes, azoles and echinocandin) and phytochems. (artemisinin, coumarin and quercetin). Our analyzes provided a list of Aspergillus proteins (72 proteins) that were abundant during interaction with different antifungal agents. On the other hand, there are 26 proteins, expression level of which is affected by more than two antifungal agents, suggesting the more general response to the stress induced by the antifungal agents. Our anal. showed enzymes from cell wall remodelling, oxidative stress response and energy metab. are the responsible factors for providing resistance against antifungal drugs in Aspergillus species and could be explored further in clin. isolates. Also, these findings have clin. importance since the effect of drug targeting different proteins can be potentiated by combination therapy. We have also discussed the opportunities ahead to study the functional role of proteins from environmental and clin. isolates of Aspergillus during its interaction with the antifungal drugs.
  34. 34
    Duduku, K.; Rosalam, S.; Rajesh, N. A review of the antioxidant potential of medicinal plant species. Food Bioprod. Process. 2011, 8, 217– 233,  DOI: 10.1016/j.fbp.2010.04.008
    Google Scholar
    There is no corresponding record for this reference.
  35. 35
    Nagai, H.; Young, H. K. Cancer prevention from the perspective of global cancer burden patterns. J. Thorac. Dis. 2017, 9, 448– 451,  DOI: 10.21037/jtd.2017.02.75
    Google Scholar
    35
    Cancer prevention from the perspective of global cancer burden patterns
    Nagai Hiroki; Nagai Hiroki; Kim Young Hak
    Journal of thoracic disease (2017), 9 (3), 448-451 ISSN:2072-1439.
    There is no expanded citation for this reference.
  36. 36
    Cao, J.; Xia, X.; Chen, X.; Xiao, J.; Wang, Q. Characterization of flavonoids from Dryopteris erythrosora and evaluation of their antioxidant, anticancer and acetylcholine esterase inhibition activities. Food Chem. Toxicol. 2013, 51, 242– 250,  DOI: 10.1016/j.fct.2012.09.039
    Google Scholar
    36
    Characterization of flavonoids from Dryopteris erythrosora and evaluation of their antioxidant, anticancer and acetylcholinesterase inhibition activities
    Cao, Jianguo; Xia, Xian; Chen, Xuefei; Xiao, Jianbo; Wang, Quanxi
    Food and Chemical Toxicology (2013), 51 (), 242-250CODEN: FCTOD7; ISSN:0278-6915. (Elsevier Ltd.)
    The profiles and bioactivities of flavonoids extd. from Dryopteris erythrosora were investigated. The total flavonoid content in full plant of D.Erythrosora is about 14.33%. The main flavonoids in D.Erythrosora were identified as gliricidin 7-O-hexoside, apigenin7-O-glucoside, quercetin 7-O-rutinoside, quercetin 7-O-galactoside, keampferol 7-O-gentiobioside, keampferol-3-O-rutinoside, myricetin 3-O-rhamnoside and quercitrin by means of HPLC-DAD-ESI-MS. Flavonoids (0.36 mg/mL) ext. from D. erythrosora showed similar 2,2-diphenyl-1-picrylhydrazyl free radical (DPPH), 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical (ABTS), superoxide anion scavenging potential and ferric reducing antioxidant potential (FRAP) with that of rutin (0.80 mg/mL). However, the antioxidant power by FRAP assay of 0.36 mg/mL flavonoids ext. from D. erythrosora was much weaker than that of 0.80 mg/mL rutin. Moreover, the flavonoids ext. from D. erythrosora showed obvious cytotoxic effects on A549 cells. The antioxidant activities of flavonoids exts. from 69 ferns showed a significant reciprocal proportion to the total flavonoids contents. The flavonoids ext. from D. erythrosora exhibited a dose-dependent inhibition against acetylcholinesterase. Moreover, the anticancer activity slightly increased with improving antioxidant potential of fern flavonoids. Fern flavonoids are excellent function foods.
  37. 37
    Ascar, I. F.; Al-A’Araji, S. B.; Alshanon, A. F. Cytotoxicity and antioxidant effect of ginger gold nanoparticles on thyroid carcinoma cells. J. Pharm. Sci. 2019, 11, 1044– 1051
    Google Scholar
    37
    Cytotoxicity and antioxidant effect of ginger gold nanoparticles on thyroid carcinoma cells
    Ascar, Israa F.; Al-A'Araji, Sanad B.; Alshanon, Ahemd F.
    Journal of Pharmaceutical Sciences and Research (2019), 11 (3), 1044-1051CODEN: JPSRC9; ISSN:0975-1459. (Pharmainfo Publisher)
    The purpose of this study was to evaluate the antioxidant and cytotoxic effects of the ginger ext.-AuNP on the FTC-133 cell line. The biosynthesis of gold nanoparticles was achieved by redn. of gold (III) chloride hydrate (HAuCl4) using Zingiber officinale ext. Characterization of biosynthetic gold (AuNPs) nanoparticles by UV-visible spectroscopy, at. force microscopy (AFM), EDX and SEM. The cytotoxicity of FTC-133 cell viability showed that the nanoethanol ext. caused a decrease in cell viability in a dose-dependent manner for 48 h, which resulted in a significant (p≤0.05)-dependent decrease in the activity of FTC-133 cells at 400μg mL-1. A cell death of 75% was achieved with an IC50 of 85.2, which was significantly different from the ethanol ext. of 242.1μg mL-1. The antioxidant properties of the synthesized nanoparticles were effective, and the IC 50 for scavenging DPPH radicals was detd. to be 40.55μg/mL. Mechanism of Affected Living Cells to Apoptosis High-content screening (HCS) assay showed a significant increase in cell membrane permeability, cytochrome c and nuclear concn. ((200μg/mL) concn. (dose-dependent) (p <0.0001) Nano ethanol ext. when compared with doxorubicin as std.). The effect of nano-ethanol ext. on the mechanism of apoptosis showed that when the concn. was increased (dose-dependent) cell viability and mitochondrial membrane permeability (200μg/mL) compared with doxorubicin as a pos. control (p < 0.0001) reduce.
  38. 38
    Ali, A. Q.; Farah, M. A.; Abou-Tarboush, F. M.; Al-Anazi, K. M.; Ali, M. A.; Lee, J.; Hailan, W. A.; Mahmoud, A. H. Cytogenotoxic effects of Adenium obesum seeds extracts on breast cancer cells. Saudi J. Biol. Sci. 2019, 26, 547– 553,  DOI: 10.1016/j.sjbs.2018.12.014
    Google Scholar
    38
    Cytogenotoxic effects of Adenium obesum seeds extracts on breast cancer cells
    Ali, Ahmed Qasem; Farah, Mohammad Abul; Abou-Tarboush, Faisal M.; Al-Anazi, Khalid M.; Ali, M. Ajmal; Lee, Joongku; Hailan, Waleed A. Q.; Mahmoud, Ahmed Hossam
    Saudi Journal of Biological Sciences (2019), 26 (3), 547-553CODEN: SJBSAG; ISSN:1319-562X. (Elsevier B.V.)
    The exts. prepd. from various areal parts of the Adenium obesum (Forssk.) Roem. & Schult. (Family: Apocynaceae) including leaves, fruit and seeds ethanolic exts. and seed aq. ext. were evaluated against MCF-7 cells in order to investigate its potential of cytogenotoxicity and induction of apoptosis. The ethanolic seeds ext. had comparatively higher cytotoxicity (IC50 ∼ 337 μg/mL). Further, apoptosis and DNA damaging potential of seeds ethanolic ext. was analyzed by applying multiple sub-lethal concns. and durations. Flow cytometry results revealed that max. percentage of early apoptosis (37%) and late apoptosis (35%) were obsd. after 12 h exposure in concns. 200 μg/mL and 300 μg/mL, resp. Similarly, the higher effect of ext. in terms of DNA damage by alk. comet assay was registered after 12 h treatment at concns. 200 and 300 μg/mL. The calcd. total damage score (TDS) for these concns. were 614 and 617, resp. The above findings indicate that A. obesum ethanolic seeds exts. has cytogenotoxic properties that could be further explored for the potential source of chemotherapeutic lead against cancer.
  39. 39
    Seca, A. M. L.; Pinto, D. C. G. A. Plant secondary metabolites as anticancer agents: Successes in clinical trials and therapeutic application. Int. J. Mol. Sci. 2018, 19, 263  DOI: 10.3390/ijms19010263
    Google Scholar
    39
    Plant secondary metabolites as anticancer agents: successes in clinical trials and therapeutic application
    Seca, Ana M. L.; Pinto, Diana C. G. A.
    International Journal of Molecular Sciences (2018), 19 (1), 263/1-263/22CODEN: IJMCFK; ISSN:1422-0067. (MDPI AG)
    A review. Cancer is a multistage process resulting in an uncontrolled and abrupt division of cells and is one of the leading causes of mortality. The cases reported and the predictions for the near future are unthinkable. Food and Drug Administration data showed that 40% of the approved mols. are natural compds. or inspired by them, from which, 74% are used in anticancer therapy. In fact, natural products are viewed as more biol. friendly, that is less toxic to normal cells. In this review, the most recent and successful cases of secondary metabolites, including alkaloid, diterpene, triterpene and polyphenolic type compds., with great anticancer potential are discussed. Focusing on the ones that are in clin. trial development or already used in anticancer therapy, therefore successful cases such as paclitaxel and homoharringtonine (in clin. use), curcumin and ingenol mebutate (in clin. trials) will be addressed. Each compd.'s natural source, the most important steps in their discovery, their therapeutic targets, as well as the main structural modifications that can improve anticancer properties will be discussed in order to show the role of plants as a source of effective and safe anticancer drugs.
  40. 40
    Nazif, U. Medicinal plants of Pakistan: Challenges and opportunities. Int. J. Complement. Altern. Med. 2017, 6, 00193
    Google Scholar
    There is no corresponding record for this reference.

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  3. Ikrimah A. Alhanbali, Mazen K. Nazal, Amjad B. Khalil. In-Vitro Antibacterial Screening of Punica granatum and Rhamnus sp. Extracts from Saudi Arabia. Arabian Journal for Science and Engineering 2024, 49 (7) , 9149-9163. https://doi.org/10.1007/s13369-024-08936-4
  4. , Beste BALBAL, Baris BITMEZ, , Irem Gulfem ALBAYRAK, , Seda Kusoglu GULTEKIN, , Emine AKALIN, , Kevser SALIHLER, , Murat KARTAL, , Belkis Atasever ARSLAN, . Effects of Vaccinium myrtillus L. extract on TNF-α-induced inflammation human paraoxonases. Revue Roumaine de Chimie 2024, 68 (10-12) , 583-587. https://doi.org/10.33224/rrch.2023.68.10-12.11
  5. Baris Bitmez, Seda K. Gultekin, Irem G. Albayrak, Yigit Deveci, Yusuf Sicak, Emine Akalin, Adami F. Pirhan, Ulas Gurer, Belkis A. Arslan. Effects of Hypericum perforatum extract on 6-hydroxydopamine neurotoxicity in differentiated SH-SY5Y cells. Egyptian Pharmaceutical Journal 2023, 22 (2) , 188-191. https://doi.org/10.4103/epj.epj_180_22
  6. Meliha Merve HIZ, Arzu DALYANCI. In silico Interaction of Rhamnus’ Flavonoids With Fat Mass And Obesity Associated Protein. European Journal of Science and Technology 2023, https://doi.org/10.31590/ejosat.1260495
  7. , Berfin Tugba TURAK. PHYTOCHEMICAL CONSTITUENTS AND ANTILEUKEMIC EFFECTS OF JUNIPERUS OXYCEDRUS EXTRACT. Biotechnologia Acta 2022, 15 (5) , 64-70. https://doi.org/10.15407/biotech15.05.064
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  • Abstract

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    Figure 1

    Figure 1. Rhamnus purpurea Edgew. (photograph courtesy of Dr. Atif Ali Khan Khalil; the photo is from a free domain).

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    Figure 2

    Figure 2. IC50 values of the plant extract and fractions against different cell lines using cyclophosphamide and doxorubicin as positive controls and DMSO as a negative control.

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    Figure 3

    Figure 3. Selectivity indices of the standards, extract, and fractions against HPAEpiC.

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    Figure 4

    Figure 4. Selectivity indices of the standards, extract, and fractions against HRPTEpiC.

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  • References

    This article references 40 other publications.

    1. 1
      Richard, J. F.; Yitzhak, T. Antibiotics and bacterial resistance in the 21st century. Perspect. Med. Chem. 2014, 6, 25– 64,  DOI: 10.4137/PMC.S13209
      There is no corresponding record for this reference.
    2. 2
      Javaid, N.; Sultana, Q.; Rasool, K.; Gandra, S.; Ahmad, F.; Chaudhary, S. U.; Mirza, S. Trends in antimicrobial resistance amongst pathogens isolated from blood and cerebrospinal fluid cultures in Pakistan (2011-2015): A retrospective cross-sectional study. PLoS One 2021, 16, e0250226  DOI: 10.1371/journal.pone.0250226
      2
      Trends in antimicrobial resistance amongst pathogens isolated from blood and cerebrospinal fluid cultures in Pakistan (2011-2015): A retrospective cross-sectional study
      Javaid, Nida; Sultana, Qamar; Rasool, Karam; Gandra, Sumanth; Ahmad, Fayyaz; Chaudhary, Safee Ullah; Mirza, Shaper
      PLoS One (2021), 16 (4), e0250226CODEN: POLNCL; ISSN:1932-6203. (Public Library of Science)
      While antimicrobial resistance (AMR) continues to be a major public health problem in Pakistan, data regarding trends of resistance among pathogenic bacteria remains scarce, with few studies presenting long-term trends in AMR. This study was therefore designed to analyze long-term AMR trends at a national level in Pakistan. We report here results of a comprehensive anal. of resistance, among pathogens isolated from blood and cerebrospinal fluid (CSF), between 2011 and 2015. Susceptibility data was obtained from a local lab. with collection points all across Pakistan (Chughtai Lab.). Resistance proportions to most commonly used antimicrobials were calcd. for each pathogen over a period of five years. While Acinetobacter species demonstrated highest resistance rates to all tested antimicrobials, a sharp increase in carbapenem resistance was the most noticeable (50%-95%) between 2011-2015. Our results also highlight the presence of third and fourth generation cephalosporins resistance in Salmonella enterica serovar Typhi in Pakistan. Interestingly, where rise in AMR was being obsd. in some major invasive pathogens, decreasing resistance trends were obsd. in Staphylococcus aureus, against commonly used antimicrobials. Overall pathogens isolated from blood and CSF between 2011-2015, showed an increase in resistance towards commonly used antimicrobials.
    3. 3
      Cox, H. S.; Sibilia, C.; Feuerriegel, S.; Kalon, S.; Polonsky, J.; Khamraev, A. K.; Rüsch-Gerdes, S.; Mills, C.; Niemann, S. Emergence of extensive drug resistance during treatment for multidrug-resistant tuberculosis. N. Engl. J. Med. 2008, 359, 2398– 2400,  DOI: 10.1056/NEJMc0805644
      3
      Emergence of extensive drug resistance during treatment for multidrug-resistant tuberculosis
      Cox, Helen S.; Sibilia, Katia; Feuerriegel, Silke; Kalon, Stobdan; Polonsky, Jonny; Khamraev, Atadjan K.; Rusch-Gerdes, Sabine; Mills, Clair; Niemann, Stefan
      New England Journal of Medicine (2008), 359 (22), 2398-2400CODEN: NEJMAG; ISSN:0028-4793. (Massachusetts Medical Society)
      There is no expanded citation for this reference.
    4. 4
      Qamar, F. N.; Yousafzai, M. T.; Khalid, M.; Kazi, A. M.; Lohana, H.; Karim, S.; Khan, A.; Hotwani, A.; Qureshi, S.; Kabir, F. Outbreak investigation of ceftriaxone-resistant Salmonella enterica serotype Typhi and its risk factors among the general population in S Hyderabad, Pakistan: a matched case-control study. Lancet Infect. Dis. 2018, 18, 1368– 1376,  DOI: 10.1016/S1473-3099(18)30483-3
      4
      Outbreak investigation of ceftriaxone-resistant Salmonella enterica serotype Typhi and its risk factors among the general population in Hyderabad, Pakistan: a matched case-control study
      Qamar Farah Naz; Yousafzai Mohammad Tahir; Khalid Muhammad; Kazi Abdul Momin; Karim Sultan; Khan Ayub; Hotwani Aneeta; Qureshi Shahida; Kabir Furqan; Aziz Fatima; Lohana Heeramani; Memon Naveed Masood; Domki Mudassar Hussain; Hasan Rumina
      The Lancet. Infectious diseases (2018), 18 (12), 1368-1376 ISSN:.
      BACKGROUND: Pakistan is currently facing the largest outbreak of ceftriaxone-resistant Salmonella enterica serotype Typhi described to date. Here we aimed to report the outbreak investigation done in Hyderabad, Pakistan, and identify disease risk factors. METHODS: We did an age-matched case-control (1:4) study, in which cases of ceftriaxone-resistant S Typhi were identified from active sentinal sites (three hospitals in Hyderabad, Pakistan), community, and laboratory-based surveillance. Ceftriaxone-resistant S Typhi infection (ie, resistance to ampicillin, chloramphenicol, co-trimoxazole, fluoroquinole, and ceftriaxone) was confirmed using blood culture. Healthy participants (controls) were enrolled for the first 200 people (cases) with ceftriaxone-resistant S Typhi. A structured questionnaire was administered to identify exposures 4 weeks before the illness (cases) or enrolment (controls). Cases were included if written informed consent was provided. Four controls were selected from the same community as the corresponding case, matched on age, being healthy at the time of enrolment, and with no febrile illness in the 4 weeks before enrolment. Samples of drinking water from households and community water sources (ie, hand pumps or taps in common areas outside households) were collected for testing. Conditional logistic regression analysis was used to assess the risk factors for ceftriaxone-resistant S Typhi outbreak in Hyderabad. FINDINGS: Between Nov 30, 2016, and Dec 30, 2017, 486 people with ceftriaxone-resistant S Typhi were identified from Hyderabad. Of the 486 cases, 296 (61%) were male and 447 (92%) were aged 15 years or younger. Several factors were significantly associated with acquisition of ceftriaxone-resistant S Typhi, including male sex (adjusted odds ratio [aOR] 1·53, 95% CI 1·06-2·21), eating outside of the house (aOR 1·48, 1·01-2·19), exposure to a patient with S Typhi infection (aOR 3·81, 2·21-6·83), and a history of antimicrobial use (aOR 4·25, 2·53-7·13). Nine (69%) of 13 water samples taken from the households of people with ceftriaxone-resistant S Typhi infection were positive for Escherichia coli, indicating faecal contamination. S Typhi DNA was detected in 12 (22%) of 55 water samples from community water sources. Geospatial mapping showed clustering of cases around sewerage lines. INTERPRETATION: Hyderabad faces the largest reported outbreak of ceftriaxone-resistant S Typhi. The outbreak is suspected to be attributed to the contaminated drinking water, especially the mixing of sewage with drinking water. The risk of ceftriaxone-resistant S Typhi infection is increased among children aged 15 years and younger, male individuals, and those eating outside the house. Vaccination and chlorination of water are recommended for the containment of the outbreak. FUNDING: None.
    5. 5
      Zilberberg, M. D.; Kollef, M. H.; Shorr, A. F. Secular trends in Acinetobacter baumannii resistance in respiratory and blood stream specimens in the United States, 2003 to 2012: A survey study. J. Hosp. Med. 2016, 11, 21– 26,  DOI: 10.1002/jhm.2477
      5
      Secular trends in Acinetobacter baumannii resistance in respiratory and blood stream specimens in the United States, 2003 to 2012: A survey study
      Zilberberg Marya D; Zilberberg Marya D; Kollef Marin H; Shorr Andrew F
      Journal of hospital medicine (2016), 11 (1), 21-6 ISSN:.
      BACKGROUND: Acinetobacter baumannii (AB) has evolved a variety of resistance mechanisms and exhibits unpredictable susceptibility patterns, making it difficult to select empiric therapy. OBJECTIVE: To examine US secular trends in the resistance of AB in respiratory infections and blood stream infections (BSI) to antimicrobial agents whose effectiveness is supported in the literature DESIGN: Survey. METHODS: We analyzed 3 time periods (2003-2005, 2006-2008, 2009-2012) in Eurofins' The Surveillance Network for resistance of AB to the following antimicrobials: carbapenems (imipenem, meropenem, doripenem), aminoglycosides (tobramycin, amikacin), tetracyclines (minocycline, doxycycline), polymyxins (colistin, polymyxin B), ampicillin-sulbactam, and trimethoprim-sulfamethoxazole. Resistance to ≥3 drug classes defined multidrug resistance (MDR). RESULTS: We identified 39,320 AB specimens (81.1% respiratory, 18.9% BSI). The highest prevalence of resistance was to doripenem (90.3%) followed by trimethoprim-sulfamethoxazole (55.3%), and the lowest to colistin (5.3%). Resistance to carbapenems (21.0% in 2003-2005 and 47.9% in 2009-2012) and colistin (2.8% in 2006-2008 to 6.9% in 2009-2012) more than doubled. Prevalence of MDR AB rose from 21.4% in 2003 to 2005 to 33.7% in 2006 to 2008, and remained stable at 35.2% in 2009 to 2012. In contrast, resistance to minocycline diminished from 56.5% (2003-2005) to 30.5% (2009-2012). MDR organisms were most frequent in nursing homes (46.5%), followed by general ward (29.2%), intensive care unit (28.7%), and outpatient setting (26.2%). CONCLUSIONS: Resistance rates among AB to such last-resort antimicrobials as carbapenems and colistin are on the rise, whereas that to minocycline has declined. Nursing homes are a reservoir of resistant AB. These trends should inform not only empiric treatment of serious infections, but also approaches to infection control.
    6. 6
      Nida, J.; Qamar, S.; Karam, R.; Sumanth, G.; Fayyaz, A.; Safee, U. C.; Shaper, M. Trends in antimicrobial resistance amongst pathogens isolated from blood and cerebrospinal fluid cultures in Pakistan (2011–2015): A retrospective cross-sectional study. PLoS One 2021, 1– 26,  DOI: 10.1371/journal.pone.0250226
      There is no corresponding record for this reference.
    7. 7
      Lagacé-Wiens, P. R. S.; Adam, H. J.; Low, D. E.; Blondeau, J. M.; Baxter, M. R.; Denisuik, A. J. Trends in antibiotic resistance over time among pathogens from Canadian hospitals: results of the CANWARD study 2007–11. J. Antimicrob. Chemother. 2013, 68, i23– i29,  DOI: 10.1093/jac/dkt023
      7
      Trends in antibiotic resistance over time among pathogens from Canadian hospitals: results of the CANWARD study 2007-11
      Lagace-Wiens, Philippe R. S.; Adam, Heather J.; Low, Donald E.; Blondeau, Joseph M.; Baxter, Melanie R.; Denisuik, Andrew J.; Nichol, Kimberly A.; Walkty, Andrew; Karlowsky, James A.; Mulvey, Michael R.; Hoban, Daryl J.; Zhanel, George G.
      Journal of Antimicrobial Chemotherapy (2013), 68 (Suppl. 1), i23-i29CODEN: JACHDX; ISSN:0305-7453. (Oxford University Press)
      Antimicrobial resistance patterns change over time and longitudinal surveillance studies provide insight into these trends. We sought to describe the important trends in antimicrobial resistance in key pathogens across Canada to provide useful information to clinicians, policy makers and industry, to assist in optimizing antimicrobial therapy, formulary choices and drug development. We analyzed longitudinal data from the CANWARD study using a multivariate regression model to control for possible effects of patient demographics on resistance, in order to assess the impact of time on antimicrobial resistance independent of other measured variables. We identified several key trends in common pathogens. In particular, we obsd. a statistically significant increase in the proportion of Escherichia coli isolates that were resistant to extended-spectrum cephalosporins and fluoroquinolones, an increase in the proportion of Klebsiella pneumoniae isolates that were resistant to extended-spectrum cephalosporins, a redn. in the proportion of Staphylococcus aureus that were methicillin, clindamycin and trimethoprim/sulfamethoxazole resistant, and a redn. in the proportion of Pseudomonas aeruginosa that were fluoroquinolone and gentamicin resistant. Conclusions Although some of these trends, such as the dramatic increase in fluoroquinolone and cephalosporin resistance in E. coli, can be attributed to the emergence and global spread of resistant clones (e.g. ST131 E. coli), others remain unexplained. However, recognizing these trends remains important to guide changes in empirical antimicrobial therapy and drug development.
    8. 8
      Sharma, S. V.; Haber, D. A.; Settleman, J. Cell line-based platforms to evaluate the therapeutic efficacy of candidate anticancer agents. Nat. Rev. Cancer 2010, 10, 241– 253,  DOI: 10.1038/nrc2820
      8
      Cell line-based platforms to evaluate the therapeutic efficacy of candidate anticancer agents
      Sharma, Sreenath V.; Haber, Daniel A.; Settleman, Jeff
      Nature Reviews Cancer (2010), 10 (4), 241-253CODEN: NRCAC4; ISSN:1474-175X. (Nature Publishing Group)
      A review. Efforts to discover new cancer drugs and predict their clin. activity are limited by the fact that lab. models to test drug efficacy do not faithfully recapitulate this complex disease. One important model system for evaluating candidate anticancer agents is human tumor-derived cell lines. Although cultured cancer cells can exhibit distinct properties compared with their naturally growing counterparts, recent technologies that facilitate the parallel anal. of large panels of such lines, together with genomic technologies that define their genetic constitution, have revitalized efforts to use cancer cell lines to assess the clin. utility of new investigational cancer drugs and to discover predictive biomarkers.
    9. 9
      McPherson, K.; Steel, C.; Dixon, J. Breast cancer─epidemiology, risk factors, and genetics. Br. Med. J. 2000, 321, 624– 628,  DOI: 10.1136/bmj.321.7261.624
      9
      ABC of breast diseases. Breast cancer-epidemiology, risk factors, and genetics
      McPherson K; Steel C M; Dixon J M
      BMJ (Clinical research ed.) (2000), 321 (7261), 624-8 ISSN:0959-8138.
      There is no expanded citation for this reference.
    10. 10
      Menhas, R.; Umer, S. Breast cancer among Pakistani women. Iran. J. Public Health 2015, 44, 586– 587
      10
      Breast Cancer among Pakistani Women
      Menhas Rashid; Umer Shumaila
      Iranian journal of public health (2015), 44 (4), 586-7 ISSN:2251-6085.
      There is no expanded citation for this reference.
    11. 11
      Amna, S. B.; Zaigham, A.; Wasim, J. Hepatocellular Carcinoma in Pakistan: Where do We Stand?. Hepatitis Mon. 2012, 12, e6023  DOI: 10.5812/hepatmon.6023
      There is no corresponding record for this reference.
    12. 12
      Bosch, F. X.; Ribes, J.; Díaz, M.; Cléries, R. Primary liver cancer, worldwide incidence and trends. Gastroenterology 2004, 127, S5– S16,  DOI: 10.1053/j.gastro.2004.09.011
      12
      Primary liver cancer: worldwide incidence and trends
      Bosch F Xavier; Ribes Josepa; Diaz Mireia; Cleries Ramon
      Gastroenterology (2004), 127 (5 Suppl 1), S5-S16 ISSN:0016-5085.
      Estimates from the year 2000 indicate that liver cancer remains the fifth most common malignancy in men and the eighth in women worldwide. The number of new cases is estimated to be 564,000 per year, including 398,000 in men and 166,000 in women. In high-risk countries, liver cancer can arise before the age of 20 years, whereas, in countries at low risk, liver cancer is rare before the age of 50 years. Rates of liver cancer in men are typically 2 to 4 times higher than in women. The incidence of primary liver cancer is increasing in several developed countries, including the United States, and the increase will likely continue for some decades. The trend is a result of a cohort effect related to infection with hepatitis B and C viruses, the incidence of which peaked in the 1950s to 1980s. In selected areas of some developing countries, the incidence of primary liver cancer has decreased, possibly as a result of the introduction of hepatitis B virus vaccine. The geographic variability in incidence of primary liver cancer is largely explained by the distribution and the natural history of the hepatitis B and C viruses. The attributable risk estimates for the combined effects of these infections account for well over 80% of liver cancer cases worldwide. Primary liver cancer is the first human cancer largely amenable to prevention using hepatitis B virus vaccines and screening of blood and blood products for hepatitis B and C viruses.
    13. 13
      Sheikh, H. S.; Kiran, M.; Fareeha, S.; Muhammad, F. U. Q. Lung Cancer in Pakistan. J. Thorac. Oncol. 2022, 17, 602– 607,  DOI: 10.1016/j.jtho.2022.01.009
      13
      Lung Cancer in Pakistan
      Sheikh Hassan Shahryar; Munawar Kiran; Sheikh Fareeha; Qamar Muhammad Fawad Ul
      Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer (2022), 17 (5), 602-607 ISSN:.
      There is no expanded citation for this reference.
    14. 14
      Prashanth, R.; Tagore, S.; Adam, B. Epidemiology of colorectal cancer: incidence, mortality, survival, and risk factors. Przegl. Gastroenterol. 2019, 14, 89– 103,  DOI: 10.5114/pg.2018.81072
      There is no corresponding record for this reference.
    15. 15
      Malik, A. I.; Farhana, B.; Aamir, A. S.; Muhammed, A. Y. Surgically treated rectal cancer patients─Outcomes at a tertiary care cancer hospital in Pakistan. Asian J. Surg. 2015, 38, 13– 20,  DOI: 10.1016/j.asjsur.2014.05.004
      15
      Surgically treated rectal cancer patients--outcomes at a tertiary care cancer hospital in Pakistan
      Malik Anum Imran; Badar Farhana; Syed Aamir Ali; Yusuf Muhammed Aasim
      Asian journal of surgery (2015), 38 (1), 13-20 ISSN:.
      AIM: The aim of this study was to analyze our experience with rectal cancer patients who underwent surgical excision at our institution. METHODS: Data on 112 rectal cancer patients who underwent surgical resection with total mesorectal excision, from January 2005 to December 2008, were evaluated retrospectively. RESULTS: We achieved an initial complete remission rate of 74.1%. Overall, 92.8% of patients had a complete total mesorectal excision. The overall survival analysis for all patients showed a 1-year survival rate of 98%, a 3-year survival rate of 82%, and a 5-year survival rate of 70%. We report a 41.9% rate of postoperative complications. The 1-, 3-, and 5-year survival rates for females were 100%, 90%, and 72%, respectively and for males, they were 90%, 80%, and 68%, respectively. Differences in overall survival by sex were not statistically significant (p > 0.05). Those patients who were treated with only surgery had the best outcomes with survival being worse in those treated with surgery and adjuvant therapy. Neoadjuvant treatment followed by surgery led to better results. CONCLUSION: We conclude that we have been successful in achieving high rates of curative resection, complete remission, and overall survival. Neoadjuvant and adjuvant chemotherapy significantly impact rates of remission.
    16. 16
      Engwa, G. A. Free radicals and the role of plant phytochemicals as antioxidants against oxidative stress-related diseases. Phytochemicals 2018, 7, 49– 74,  DOI: 10.5772/intechopen.76719
      There is no corresponding record for this reference.
    17. 17
      Alkadi, H. A review on free radicals and antioxidants. Infect. Disord.: Drug Targets 2020, 20, 16– 26,  DOI: 10.2174/1871526518666180628124323
      17
      A Review on Free Radicals and Antioxidants
      Alkadi, Hourieh
      Infectious Disorders: Drug Targets (2020), 20 (1), 16-26CODEN: IDDTAD; ISSN:1871-5265. (Bentham Science Publishers Ltd.)
      Free radicals are generated in our body by several systems. A balance among free radicals and antioxidants is an important matter for appropriate physiol. function. If free radicals become greater than the ability of the body to control them, a case known as oxidative stress appears, as a result of that, a no. of human diseases spread in the body. Antioxidants can contribute to facingthis oxidative stress. The present review provides a brief overview of free radicals, oxidative stress, some natural antioxidants and the relationship between them.
    18. 18
      Fazli, K.; Madiha, A.; Suleman, K.; Hamayun, K.; Muhammad, U. K. S.; Ameer, K.; Asif, J.; Naveed, U.; Yasar, S.; Zaki, U.; Muhammad, A.; Tayyaba, I.; Abubakr, M. I.; Mayeen, U. K.; Talha, B. E. Antimicrobial, anti-inflammatory and antioxidant activities of natural organic matter extracted from cretaceous shales in district Nowshera-Pakistan. Arab. J. Chem. 2022, 15, 1036033  DOI: 10.1016/j.arabjc.2021.103633
      There is no corresponding record for this reference.
    19. 19
      Stocker, P.; Yousfi, M.; Djerridane, O.; Perrier, J.; Amziani, R.; El Boustani, S.; Moulin, A. Effect of flavonoids from various Mediterranean plants on enzymatic activity of intestinal carboxylesterase. Biochimie 2004, 86, 919– 925,  DOI: 10.1016/j.biochi.2004.09.005
      19
      Effect of flavonoids from various Mediterranean plants on enzymatic activity of intestinal carboxylesterase
      Stocker, P.; Yousfi, M.; Djerridane, O.; Perrier, J.; Amziani, R.; El Boustani, S.; Moulin, A.
      Biochimie (2004), 86 (12), 919-925CODEN: BICMBE; ISSN:0300-9084. (Elsevier B.V.)
      Flavonol compds. of three Mediterranean plants from the Algerian Atlas used traditionally in Arab folk medicine, Arenaria serpyllifolia, Rhamnus alaternus and Thapsia garganica, were found to inhibit the enzymic activities of both rat intestine and purified porcine liver carboxylesterase in a concn.-dependent manner. Results indicate that the flavonol compds. from the aerial part of these plants lead to the inactivation of the CE pI = 5.1 with Ki of micromolar range. These results encourage us to perform further biol. investigation.
    20. 20
      Kosalec, I.; Kremer, D.; Locatelli, M.; Epifano, F.; Genovese, S.; Carlucci, G.; Končić, M. Z. Anthraquinone profile, antioxidant and antimicrobial activity of bark extracts of Rhamnus alaternus, Rhamnus fallax, Rhamnus intermedia and Rhamnus pumila. Food chem. 2013, 136, 335– 341,  DOI: 10.1016/j.foodchem.2012.08.026
      20
      Anthraquinone profile, antioxidant and antimicrobial activity of bark extracts of Rhamnus alaternus, R. fallax, R. intermedia and R. pumila
      Kosalec, I.; Kremer, D.; Locatelli, M.; Epifano, F.; Genovese, S.; Carlucci, G.; Randic, M.; Zovko Koncic, M.
      Food Chemistry (2013), 136 (2), 335-341CODEN: FOCHDJ; ISSN:0308-8146. (Elsevier Ltd.)
      The quantity of phenols, as well as antioxidant and antimicrobial activities, were investigated in bark of Rhamnus alaternus L., R. fallax Boiss., R. intermedia Steud. et Hochst., and R. pumila Turra from natural stands in Croatia. The most abundant anthraquinones in the investigated exts. were chrysophanol in R. alaternus (3.14 mg/g), emodin in R. pumila (0.339 mg/g), and physcion in R. fallax (2.70 mg/g) and R. intermedia (0.285 mg/g). The species exhibiting the highest antioxidant activity were R. fallax and R. pumila. A pos. correlation was obsd. between total phenolic and flavonoid levels of the exts. and antioxidant activity in some of the assays. All species showed antimicrobial activity against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Candida albicans, Aspergillus niger and Microsporum gypseum with minimal inhibitory concns. equal to or below 2.500 mg/mL. The results indicate that the investigated Rhamnus species are a source of anthraquinones and other phenols, which act as multifunctional antioxidants with antimicrobial activity.
    21. 21
      Marzouk, M. S.; El-Toumy, S. A.; Merfort, I.; Nawwar, M. A. Polyphenolic metabolites of Rhamnus disperma. Phytochemistry 1999, 52, 943– 946,  DOI: 10.1016/S0031-9422(99)00262-9
      21
      Polyphenolic metabolites of Rhamnus disperma
      Marzouk, Mohamed S.; El-Toumy, Sayed A. A.; Merfort, Irmgard; Nawwar, Mahmoud A. M.
      Phytochemistry (1999), 52 (5), 943-946CODEN: PYTCAS; ISSN:0031-9422. (Elsevier Science Ltd.)
      A new quercetin dirhamnoside has been isolated and identified as quercetin 3,4'-di-O-α-L-rhamnopyranoside (I) together with 24 structurally known phenolic metabolites from the fruits and aerial parts of Rhamnus disperma for the first time. The known compds. have been characterized as kaempferol 3-O-robinoside, 3-O-rhamninoside, 4'-O-rhamninoside, rhamnocitrin 3-O-rhamninoside, 4'-O-rhamninoside, quercetin 3-O-rhamnoside, 3-O-galactoside, 7-O-galactoside, 3-O-Me 7-O-galactoside, 3-O-robinoside, 3-O-rhamninoside, rhamnetin 3-O-rhamninoside, rhamnazin 3-O-robinoside, 3-O-rhamninoside, two phenolic acids 2,5-dihydroxybenzoic and protocatechuic acids, a coumarin isofraxetin, three aglycons identified as quercetin 3-Me ether, eriodictyol and taxifolin, together with five flavonols kaempferol, quercetin, rhamnocitrin, rhamnetin and rhamnazin. The structures of the isolated compds. have been established on the basis of chem. and NMR spectroscopic evidence as well as neg. ESI-MS in somes cases.
    22. 22
      Farzana, K.; Kyung-Hwan, K.; Hafiz, M. U. F.; Muhammad, A. F.; Muhammad, K.; Rooma, W.; Atif, A. K. H.; Fazli, K.; Chethikkattuveli, S. A. R.; Kinam, H.; Kyung-Hyun, C.; Abdul, S. M. Evaluation of antimicrobial and anticancer activities of selected medicinal plants of himalayas, Pakistan. Plants 2022, 11, 48  DOI: 10.3390/plants11010048
      There is no corresponding record for this reference.
    23. 23
      Boyanova, L.; Gergova, G.; Nikolov, R.; Derejian, S.; Lazarova, E.; Katsarov, N.; Mitov, I.; Krastev, Z. Activity of Bulgarian propolis against 94 Helicobacter pylori strains in vitro by agar-well diffusion, agar dilution and disc diffusion methods. J. Med. Microbiol. 2005, 54, 481– 483,  DOI: 10.1099/jmm.0.45880-0
      23
      Activity of Bulgarian propolis against 94 Helicobacter pylori strains in vitro by agar-well diffusion, agar dilution and disc diffusion methods
      Boyanova Lyudmila; Gergova Galina; Nikolov Rossen; Derejian Sirigan; Lazarova Elena; Katsarov Nikolai; Mitov Ivan; Krastev Zacharii
      Journal of medical microbiology (2005), 54 (Pt 5), 481-483 ISSN:0022-2615.
      Propolis exhibits antimicrobial, anti-inflammatory and other biological effects. The aim of this study was to evaluate the activity of 30 % ethanolic extract of Bulgarian propolis against 94 Helicobacter pylori strains by three methods. By the agar-well diffusion method, only 13.8 % of the strains exhibited no inhibition by 30 microl propolis extract (containing 9 mg propolis) and all isolates were inhibited to some extent by 90 microl of the extract (27 mg propolis) per well. The mean diameters of growth inhibition by 30, 60 or 90 microl propolis extract or 30 microl 96 % ethanol per well were 16.8, 19.2, 27.5 and 8.3 mm, respectively. The propolis extract was more active than the ethanol (P < 0.001). With 90 microl propolis extract per well, 69.4 % of the strains exhibited large diameters of growth inhibition (> or =20 mm) versus 26.6 % with 30 mul per well (P < 0.001). With moist propolis discs, inhibition was detected in more strains (92.1 %) than with dried discs (78.2 %, P < 0.05), with mean inhibitory diameters of 18.7 and 13.8 mm, respectively. By the agar dilution method, 100 and 300 microg propolis ml(-1) inhibited the growth of 57.1 % and 76.2 %, respectively, of the 21 strains tested. In conclusion, Bulgarian propolis had a strong and dose-dependent activity against most of the H. pylori strains tested. Although the effect of propolis on H. pylori in vitro is promising, further microbiological, pharmacological and clinical trials are required.
    24. 24
      Fazli, K.; Zafar, I.; Ayub, K.; Zakiullah; Fazli, N.; Muhammad, S. K. Validation of some of the ethnopharmacological uses of Xanthium strumarium and Duchesnea indica. Pak. J. Bot. 2012, 44, 1199– 1201
      There is no corresponding record for this reference.
    25. 25
      CLSI. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically, 9th ed.; Clinical and Laboratory Standards Institute, CLSI Document M07A9, 2012; Vol. 32.
      There is no corresponding record for this reference.
    26. 26
      Kedare, S. B.; Sing, R. P. Genesis and development of DPPH method of antioxidant assay. J. Food Sci. Technol. 2011, 48, 412– 422,  DOI: 10.1007/s13197-011-0251-1
      26
      Genesis and development of DPPH method of antioxidant assay
      Kedare, Sagar B.; Singh, R. P.
      Journal of Food Science and Technology (2011), 48 (4), 412-422CODEN: JFSTAB; ISSN:0022-1155. (Springer (India) Private Ltd.)
      A review. α, α-Diphenyl-β-picrylhydrazyl (DPPH) free radical scavenging method offers the first approach for evaluating the antioxidant potential of a compd., an ext. or other biol. sources. This is the simplest method, wherein the prospective compd. or ext. is mixed with DPPH soln. and absorbance is recorded after a defined period. However, with the advancement and sophistication in instrumental techniques, the method has undergone various modifications to suit the requirements, even though the basic approach remains same in all of them. This article presents a crit. review on various developments to the DPPH method.
    27. 27
      Iris, F. F. B.; Strain, J. J. Simultaneous measurement of total antioxidant power and ascorbic acid concentration. Methods Enzymol. 1999, 299, 15– 27,  DOI: 10.1016/S0076-6879(99)99005-5
      There is no corresponding record for this reference.
    28. 28
      Janice, M.; Joel, J.; Holly, H.; Kerry, W.; Paul, N. In vitri cytotoxic properties of crude polar extracts of plants sourced from Australia. Clin. Complementary Med. Pharmacol. 2022, 2, 100022
      There is no corresponding record for this reference.
    29. 29
      Tong, S. Y. C.; Joshua, S. D.; Emily, E.; Thomas, L. H.; Vance, G. F. J. Staphylococcus aureus Infections: Epidemiology, pathophysiology, clinical manifestations, and management. Clin. Microbiol. Rev. 2015, 28, 603– 661,  DOI: 10.1128/CMR.00134-14
      29
      Staphylococcus aureus infections: epidemiology, pathophysiology, clinical manifestations, and management
      Tong, Steven Y. C.; Davis, Joshua S.; Eichenberger, Emily; Holland, Thomas L.; Fowler, Vance G., Jr.
      Clinical Microbiology Reviews (2015), 28 (3), 603-661CODEN: CMIREX; ISSN:1098-6618. (American Society for Microbiology)
      Staphylococcus aureus is a major human pathogen that causes a wide range of clin. infections. It is a leading cause of bacteremia and infective endocarditis as well as osteoarticular, skin and soft tissue, pleuropulmonary, and device-related infections. This review comprehensively covers the epidemiol., pathophysiol., clin. manifestations, and management of each of these clin. entities. The past 2 decades have witnessed two clear shifts in the epidemiol. of S. aureus infections: first, a growing no. of health care-assocd. infections, particularly seen in infective endocarditis and prosthetic device infections, and second, an epidemic of community-assocd. skin and soft tissue infections driven by strains with certain virulence factors and resistance to β-lactam antibiotics. In reviewing the literature to support management strategies for these clin. manifestations, we also highlight the paucity of high-quality evidence for many key clin. questions.
    30. 30
      de Araújo, A. A.; Luiz, A. L. S.; Magda, R. A. F.; Manoel, A. S. N.; Giselle, R. S.; Raimundo, F. A. J.; Gerlane, C. B. G.; Maria, C. N. M. Quantification of polyphenols and evaluation of antimicrobial, analgesic and anti-inflammatory activities of aqueous and acetone–water extracts of Libidibia ferrea, Parapiptadenia rigida and Psidium guajava. J. Ethnopharmacol. 2014, 156, 88– 96,  DOI: 10.1016/j.jep.2014.07.031
      30
      Quantification of polyphenols and evaluation of antimicrobial, analgesic and anti-inflammatory activities of aqueous and acetone-water extracts of Libidibia ferrea, Parapiptadenia rigida and Psidium guajava
      de Araujo, Aurigena Antunes; Soares, Luiz Alberto Lira; Assuncao Ferreira, Magda Rhayanny; de Souza Neto, Manoel Andre; da Silva, Giselle Ribeiro; de Araujo, Raimundo Fernandes; Guerra, Gerlane Coelho Bernardo; de Melo, Maria Celeste Nunes
      Journal of Ethnopharmacology (2014), 156 (), 88-96CODEN: JOETD7; ISSN:0378-8741. (Elsevier Ireland Ltd.)
      Vast nos. of plant species from northeastern Brazil have not yet been phytochem. or biol. evaluated.The goal of this work was to obtain, characterize and show the antimicrobial, analgesic and anti-inflammatory activities of aq. and acetone-water exts. of Libidibia ferrea, Parapiptadenia rigida and Psidium guajava.The plant material (100 g) was dried, and the crude exts. were obtained by using turbo-extn. (10%; w/v) with water or acetone:water (7:3, vol./vol.) as the extn. solvent. High-performance liq. chromatog. (HPLC) methods were used to screen the crude exts. for hydrolysable tannins (gallic acid) and condensed tannins (catechins). The antibacterial activity was evaluated by agar-diffusion and microdilution methods against Gram-pos. strains (Staphylococcus aureus ATCC 25923, Staphylococcus epidermidis INCQS 00016, Enterococcus faecalis ATCC 29212 and a clin. isolate of methicillin-resistant Staphylococcus aureus) as well as Gram-neg. strains (Escherichia coli ATCC 25922, Salmonella enteritidis INCQS 00258, Shigella flexneri and Klebsiella pneumoniae). To evaluate the anti-inflammatory activity, a leukocyte migration model was used. Analgesic activity was detd. by the hot plate test and the acetic acid-induced abdominal writhing test. Data were analyzed by anal. of variance (ANOVA) at a significance level of 5%.Parapiptadenia rigida presented the highest amt. of total polyphenols (35.82±0.20%), while the greatest catechin content was found in the acetone-water ext. of Psidium guajava (EAWPg; 1.04 μg/g). The largest amts. of catechins were found in the aq. ext. of Libidibia ferrea (EALf; 1.07 μg/g) and the acetone-water ext. of Parapiptadenia rigida (EAWPr; 1.0 μg/g). All exts. showed activity against Gram-pos. bacteria. The aq. and acetone-water exts. of Psidium guajava showed the greatest inhibition zones in the agar diffusion tests. In the evaluation of the min. inhibitory concn. (MIC), the most susceptible Gram-pos. bacterium was Staphylococcus epidermidis and the most susceptible Gram-neg. bacterium was Shigella flexneri. EAPg and EAWPg showed the greatest MIC values. All exts. were significant inhibitors of leukocyte migration (p<0.05). Using the writhing test, significant analgesic activity was found for EAPr (50 mg/kg), EAWPr (100 mg/kg and 200 mg/kg) and EAWPg (50 mg/kg) (p<0.05).Thus, the appropriate extn. procedure preserves the chem. components such as gallic acid and catechin, and showed antimicrobial, anti-inflammatory and analgesic properties.
    31. 31
      Gradišar, H.; Pristovsek, P.; Plaper, A.; Jerala, R. Green tea catechins inhibit bacterial DNA gyrase by interaction with its ATP binding site. J. Med. Chem. 2007, 50, 264– 271,  DOI: 10.1021/jm060817o
      31
      Green Tea Catechins Inhibit Bacterial DNA Gyrase by Interaction with Its ATP Binding Site
      Gradisar, Helena; Pristovsek, Primoz; Plaper, Andreja; Jerala, Roman
      Journal of Medicinal Chemistry (2007), 50 (2), 264-271CODEN: JMCMAR; ISSN:0022-2623. (American Chemical Society)
      Catechins are the main ingredients of green tea exts. and have been shown to possess versatile biol. activities, including antimicrobial. We detd. that the catechins inhibit bacterial DNA gyrase by binding to the ATP binding site of the gyrase B subunit. In the group of four tested catechins, epigallocatechin gallate (EGCG) had the highest activity, followed by epicatechin gallate (ECG) and epigallocatechin (EGC). Specific binding to the N-terminal 24 kDa fragment of gyrase B was detd. by fluorescence spectroscopy and confirmed using heteronuclear two-dimensional NMR spectroscopy of the EGCG-15N-labeled gyrase B fragment complex. Protein residues affected by binding to EGCG were identified through chem. shift perturbation. Mol. docking calcns. suggest that the benzopyran ring of EGCG penetrates deeply into the active site while the galloyl moiety anchors it to the cleft through interactions with its hydroxyl groups, which explains the higher activity of EGCG and ECG.
    32. 32
      Laura, B. D.; Carla, W.; Lara, B. I.; Paula, C. S.; Janio, M. S. Activity of MSI-78, h-Lf1- 11 and cecropin B antimicrobial peptides alone and in combination with voriconazole and amphotericin B against clinical isolates of Fusarium solani. J. Med. Mycol. 2021, 31, 101119  DOI: 10.1016/j.mycmed.2021.101119
      There is no corresponding record for this reference.
    33. 33
      Shishodia, S. K.; Shraddha, T.; Jata, S. Resistance mechanism and proteins in Aspergillus species against antifungal agents. Mycology 2019, 10, 151– 165,  DOI: 10.1080/21501203.2019.1574927
      33
      Resistance mechanism and proteins in Aspergillus species against antifungal agents
      Shishodia, Sonia Kumari; Tiwari, Shraddha; Shankar, Jata
      Mycology (2019), 10 (3), 151-165CODEN: MYCOCG; ISSN:2150-1203. (Taylor & Francis Ltd.)
      Aspergillus species contain pathogenic and opportunistic fungal pathogens which have the potential to cause mycosis (invasive aspergillosis) in humans. The existing antifungal drugs have limitation largely due to the development of drug-resistant isolates. To gain insight into the mechanism of action and antifungal drug resistance in Aspergillus species including biofilm formation, we have reviewed protein data of Aspergillus species during interaction with antifungals drugs (polynes, azoles and echinocandin) and phytochems. (artemisinin, coumarin and quercetin). Our analyzes provided a list of Aspergillus proteins (72 proteins) that were abundant during interaction with different antifungal agents. On the other hand, there are 26 proteins, expression level of which is affected by more than two antifungal agents, suggesting the more general response to the stress induced by the antifungal agents. Our anal. showed enzymes from cell wall remodelling, oxidative stress response and energy metab. are the responsible factors for providing resistance against antifungal drugs in Aspergillus species and could be explored further in clin. isolates. Also, these findings have clin. importance since the effect of drug targeting different proteins can be potentiated by combination therapy. We have also discussed the opportunities ahead to study the functional role of proteins from environmental and clin. isolates of Aspergillus during its interaction with the antifungal drugs.
    34. 34
      Duduku, K.; Rosalam, S.; Rajesh, N. A review of the antioxidant potential of medicinal plant species. Food Bioprod. Process. 2011, 8, 217– 233,  DOI: 10.1016/j.fbp.2010.04.008
      There is no corresponding record for this reference.
    35. 35
      Nagai, H.; Young, H. K. Cancer prevention from the perspective of global cancer burden patterns. J. Thorac. Dis. 2017, 9, 448– 451,  DOI: 10.21037/jtd.2017.02.75
      35
      Cancer prevention from the perspective of global cancer burden patterns
      Nagai Hiroki; Nagai Hiroki; Kim Young Hak
      Journal of thoracic disease (2017), 9 (3), 448-451 ISSN:2072-1439.
      There is no expanded citation for this reference.
    36. 36
      Cao, J.; Xia, X.; Chen, X.; Xiao, J.; Wang, Q. Characterization of flavonoids from Dryopteris erythrosora and evaluation of their antioxidant, anticancer and acetylcholine esterase inhibition activities. Food Chem. Toxicol. 2013, 51, 242– 250,  DOI: 10.1016/j.fct.2012.09.039
      36
      Characterization of flavonoids from Dryopteris erythrosora and evaluation of their antioxidant, anticancer and acetylcholinesterase inhibition activities
      Cao, Jianguo; Xia, Xian; Chen, Xuefei; Xiao, Jianbo; Wang, Quanxi
      Food and Chemical Toxicology (2013), 51 (), 242-250CODEN: FCTOD7; ISSN:0278-6915. (Elsevier Ltd.)
      The profiles and bioactivities of flavonoids extd. from Dryopteris erythrosora were investigated. The total flavonoid content in full plant of D.Erythrosora is about 14.33%. The main flavonoids in D.Erythrosora were identified as gliricidin 7-O-hexoside, apigenin7-O-glucoside, quercetin 7-O-rutinoside, quercetin 7-O-galactoside, keampferol 7-O-gentiobioside, keampferol-3-O-rutinoside, myricetin 3-O-rhamnoside and quercitrin by means of HPLC-DAD-ESI-MS. Flavonoids (0.36 mg/mL) ext. from D. erythrosora showed similar 2,2-diphenyl-1-picrylhydrazyl free radical (DPPH), 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical (ABTS), superoxide anion scavenging potential and ferric reducing antioxidant potential (FRAP) with that of rutin (0.80 mg/mL). However, the antioxidant power by FRAP assay of 0.36 mg/mL flavonoids ext. from D. erythrosora was much weaker than that of 0.80 mg/mL rutin. Moreover, the flavonoids ext. from D. erythrosora showed obvious cytotoxic effects on A549 cells. The antioxidant activities of flavonoids exts. from 69 ferns showed a significant reciprocal proportion to the total flavonoids contents. The flavonoids ext. from D. erythrosora exhibited a dose-dependent inhibition against acetylcholinesterase. Moreover, the anticancer activity slightly increased with improving antioxidant potential of fern flavonoids. Fern flavonoids are excellent function foods.
    37. 37
      Ascar, I. F.; Al-A’Araji, S. B.; Alshanon, A. F. Cytotoxicity and antioxidant effect of ginger gold nanoparticles on thyroid carcinoma cells. J. Pharm. Sci. 2019, 11, 1044– 1051
      37
      Cytotoxicity and antioxidant effect of ginger gold nanoparticles on thyroid carcinoma cells
      Ascar, Israa F.; Al-A'Araji, Sanad B.; Alshanon, Ahemd F.
      Journal of Pharmaceutical Sciences and Research (2019), 11 (3), 1044-1051CODEN: JPSRC9; ISSN:0975-1459. (Pharmainfo Publisher)
      The purpose of this study was to evaluate the antioxidant and cytotoxic effects of the ginger ext.-AuNP on the FTC-133 cell line. The biosynthesis of gold nanoparticles was achieved by redn. of gold (III) chloride hydrate (HAuCl4) using Zingiber officinale ext. Characterization of biosynthetic gold (AuNPs) nanoparticles by UV-visible spectroscopy, at. force microscopy (AFM), EDX and SEM. The cytotoxicity of FTC-133 cell viability showed that the nanoethanol ext. caused a decrease in cell viability in a dose-dependent manner for 48 h, which resulted in a significant (p≤0.05)-dependent decrease in the activity of FTC-133 cells at 400μg mL-1. A cell death of 75% was achieved with an IC50 of 85.2, which was significantly different from the ethanol ext. of 242.1μg mL-1. The antioxidant properties of the synthesized nanoparticles were effective, and the IC 50 for scavenging DPPH radicals was detd. to be 40.55μg/mL. Mechanism of Affected Living Cells to Apoptosis High-content screening (HCS) assay showed a significant increase in cell membrane permeability, cytochrome c and nuclear concn. ((200μg/mL) concn. (dose-dependent) (p <0.0001) Nano ethanol ext. when compared with doxorubicin as std.). The effect of nano-ethanol ext. on the mechanism of apoptosis showed that when the concn. was increased (dose-dependent) cell viability and mitochondrial membrane permeability (200μg/mL) compared with doxorubicin as a pos. control (p < 0.0001) reduce.
    38. 38
      Ali, A. Q.; Farah, M. A.; Abou-Tarboush, F. M.; Al-Anazi, K. M.; Ali, M. A.; Lee, J.; Hailan, W. A.; Mahmoud, A. H. Cytogenotoxic effects of Adenium obesum seeds extracts on breast cancer cells. Saudi J. Biol. Sci. 2019, 26, 547– 553,  DOI: 10.1016/j.sjbs.2018.12.014
      38
      Cytogenotoxic effects of Adenium obesum seeds extracts on breast cancer cells
      Ali, Ahmed Qasem; Farah, Mohammad Abul; Abou-Tarboush, Faisal M.; Al-Anazi, Khalid M.; Ali, M. Ajmal; Lee, Joongku; Hailan, Waleed A. Q.; Mahmoud, Ahmed Hossam
      Saudi Journal of Biological Sciences (2019), 26 (3), 547-553CODEN: SJBSAG; ISSN:1319-562X. (Elsevier B.V.)
      The exts. prepd. from various areal parts of the Adenium obesum (Forssk.) Roem. & Schult. (Family: Apocynaceae) including leaves, fruit and seeds ethanolic exts. and seed aq. ext. were evaluated against MCF-7 cells in order to investigate its potential of cytogenotoxicity and induction of apoptosis. The ethanolic seeds ext. had comparatively higher cytotoxicity (IC50 ∼ 337 μg/mL). Further, apoptosis and DNA damaging potential of seeds ethanolic ext. was analyzed by applying multiple sub-lethal concns. and durations. Flow cytometry results revealed that max. percentage of early apoptosis (37%) and late apoptosis (35%) were obsd. after 12 h exposure in concns. 200 μg/mL and 300 μg/mL, resp. Similarly, the higher effect of ext. in terms of DNA damage by alk. comet assay was registered after 12 h treatment at concns. 200 and 300 μg/mL. The calcd. total damage score (TDS) for these concns. were 614 and 617, resp. The above findings indicate that A. obesum ethanolic seeds exts. has cytogenotoxic properties that could be further explored for the potential source of chemotherapeutic lead against cancer.
    39. 39
      Seca, A. M. L.; Pinto, D. C. G. A. Plant secondary metabolites as anticancer agents: Successes in clinical trials and therapeutic application. Int. J. Mol. Sci. 2018, 19, 263  DOI: 10.3390/ijms19010263
      39
      Plant secondary metabolites as anticancer agents: successes in clinical trials and therapeutic application
      Seca, Ana M. L.; Pinto, Diana C. G. A.
      International Journal of Molecular Sciences (2018), 19 (1), 263/1-263/22CODEN: IJMCFK; ISSN:1422-0067. (MDPI AG)
      A review. Cancer is a multistage process resulting in an uncontrolled and abrupt division of cells and is one of the leading causes of mortality. The cases reported and the predictions for the near future are unthinkable. Food and Drug Administration data showed that 40% of the approved mols. are natural compds. or inspired by them, from which, 74% are used in anticancer therapy. In fact, natural products are viewed as more biol. friendly, that is less toxic to normal cells. In this review, the most recent and successful cases of secondary metabolites, including alkaloid, diterpene, triterpene and polyphenolic type compds., with great anticancer potential are discussed. Focusing on the ones that are in clin. trial development or already used in anticancer therapy, therefore successful cases such as paclitaxel and homoharringtonine (in clin. use), curcumin and ingenol mebutate (in clin. trials) will be addressed. Each compd.'s natural source, the most important steps in their discovery, their therapeutic targets, as well as the main structural modifications that can improve anticancer properties will be discussed in order to show the role of plants as a source of effective and safe anticancer drugs.
    40. 40
      Nazif, U. Medicinal plants of Pakistan: Challenges and opportunities. Int. J. Complement. Altern. Med. 2017, 6, 00193
      There is no corresponding record for this reference.

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