Pakistan Journal of Biological Sciences最新文献

<b>Background and Objective:</b> The rising global challenge of drug resistance and cancer demands the discovery of novel therapeutic agents. Endophytic microorganisms, particularly those from medicinal plants like <i>Ocimum basilicum</i>, represent a promising, yet underexplored, source of natural products. This study aimed to isolate and screen bioactive endophytic actinomycetes from <i>O. basilicum</i> roots. The research focused on characterizing the most active isolate, purifying its secondary metabolites and evaluating their antibacterial and anticancer properties. <b>Materials and Methods:</b> The potent strain SB03 was isolated from basil roots and identified as <i>Streptomyces griseoincarnatus</i> using 16S rDNA sequencing. Two compounds were purified from its crude extract and structurally characterized by spectroscopy. Their bioactivities were assessed by determining MIC/MBC against pathogenic bacteria (including MRSA) and IC₅₀ against human cancer cells (HeLa, HepG2 and MDA-MB-231). Molecular docking investigated their interaction with topoisomerase II, complemented by <i>in silico</i> ADMET analysis. <b>Results:</b> A potent actinomycete strain, <i>S. griseoincarnatus</i> SB03, was successfully isolated from sweet basil roots. Two known anthraquinone compounds, 2-butyryl-1,8-dihydroxy-3-methyl-anthraquinone (Compound <b>1</b>) and 2-butyryl-1,4,8-trihydroxy-3-methyl-anthraquinone (Compound <b>2</b>), were isolated from the culture extract. Both exhibited strong antibacterial activity (MIC 64-128 μg/mL against Gram-positive strains) and significant cytotoxicity (IC₅₀ 35.01-97.68 μg/mL) against all tested cancer lines. Importantly, they showed a higher selectivity index against the HepG2 line than the control, doxorubicin. Docking studies indicated favorable binding to topoisomerase II. <b>Conclusion:</b> <i>Streptomyces griseoincarnatus</i> SB03 from sweet basil is a promising source of therapeutic agents. Its anthraquinone metabolites possess valuable dual antibacterial and selective anticancer properties. These compounds are excellent lead candidates for future drug development against infectious diseases and hepatocellular carcinoma.

<b>Background and Objective:</b> Endophytic actinomycetes associated with medicinal plants constitute a valuable yet underinvestigated source of bioactive secondary metabolites. <i>Tinospora cordifolia</i>, a widely respected ethnomedicinal plant, harbors diverse microbial endophytes with strong therapeutic potential. This study aimed to isolate and characterize endophytic actinomycetes from <i>T. cordifolia</i>, identify their major metabolites and evaluate the antibacterial, anticancer and molecular inhibitory properties of the purified compounds. <b>Materials and Methods:</b> Actinomycetes were isolated from surface-sterilized root, stem and leaf tissues using selective media. The most potent isolate, TTCF1, was identified through morphological characteristics, chemotaxonomic profiling and 16S rRNA gene sequencing. Bioactive metabolites were extracted and purified via column chromatography, followed by structural characterization using advanced spectroscopic techniques. Antibacterial activity was assessed by determining MIC and MBC values against human pathogens, including MRSA. Cytotoxicity was evaluated using the MTT assay on HeLa, HepG2 and MDA-MB-231 cancer cell lines, along with Vero cells as the non-cancerous control. Molecular docking was performed against EGFR, accompanied by ADMET property prediction. Statistical significance was determined using one-way ANOVA with Tukey's <i>post hoc</i> test (p<0.05). <b>Results:</b> Ten actinomycete isolates were obtained, all exclusively from root tissues. The strongest producer, TTCF1, showed 99.54% 16S rRNA similarity to <i>Streptomyces triticiradicis</i>. Chemical analysis yielded two diketopiperazines: Cyclo-(D-Pro-L-Tyr) and Cyclo-(D-Pro-L-Leu). Both compounds demonstrated potent antibacterial activity against Gram-positive pathogens (MIC 32-64 μg/mL) and cytotoxicity toward cancer cell lines (IC₅₀ 58.16-362.71 μg/mL). Compound 1 showed selective toxicity toward HepG2 cells and exhibited stronger predicted EGFR binding affinity (-7.188 kcal/mol) than the reference inhibitor AQ4 (-6.703 kcal/mol). The ADMET profiles indicated good oral absorption. <b>Conclusion:</b> <i>Streptomyces triticiradicis</i> TTCF1 is a promising source of pharmacologically relevant diketopiperazines. Cyclo-(D-Pro-L-Tyr) emerges as a potential lead molecule with notable antimicrobial and selective anticancer activity, supported by strong EGFR-binding predictions.

<b>Background and Objective:</b> Diabetic cardiomyopathy (DCM) is a serious complication of type 2 diabetes, characterized by metabolic imbalance, oxidative stress, inflammation and apoptosis, leading to impaired cardiac function. Despite available treatments, safer and more effective natural alternatives remain underexplored. This study investigated the active composition of <i>Stevia rebaudiana</i> leaf extract (SLE) and evaluated its prophylactic effects against diabetic cardiomyopathy in an experimental rat model. <b>Materials and Methods:</b> Twenty-four male rats were randomly divided into four groups: Control, SLE-treated (400 mg/kg Stevia glycosides for 30 days), STZ-induced diabetic (110 mg/kg Nicotinamide followed by 60 mg/kg Streptozotocin) and SLE+STZ co-treated. Biochemical assays were performed to assess glucose metabolism, insulin sensitivity, serum electrolytes, protein and cardiac enzymes (LDH, CK-MB). Oxidative stress markers, antioxidant levels, inflammatory cytokines, apoptotic markers and cardiac histopathology were evaluated. Data were analyzed using One-way ANOVA followed by <i>post hoc</i> tests, with p<0.05 considered significant. <b>Results:</b> The SLE significantly reduced serum glucose and insulin resistance while improving QUICKI and regulating electrolytes, total protein, LDH and CK-MB compared to diabetic rats. It restored redox balance by lowering PCO, NO and MDA, while enhancing SOD, CAT, GSH and TAC. Inflammatory cytokines (TNF-α, IL-6, IL-1β, VCAM-1 and ICAM-1) and apoptotic markers (Bax, P53, Cytochrome C, Caspase-9 and Caspase-3) were markedly downregulated, whereas anti-apoptotic Bcl-2 was upregulated. Histological analysis confirmed preserved cardiac structure, reduced fibrosis and diminished DNA fragmentation, further supported by decreased PARP levels. <b>Conclusion:</b> <i>Stevia</i> <i>rebaudiana</i> leaf extract exerts potent cardioprotective effects in diabetic rats by improving metabolic indices, attenuating oxidative stress, inflammation and apoptosis and preserving cardiac architecture. These findings highlight its therapeutic potential as a natural prophylactic agent against diabetic cardiomyopathy, warranting further clinical validation.

<b>Background and Objective:</b> Up to now, evidence remains inconclusive about how vitamin D (VD) status relates to the severity of COVID-19 among hospitalized patients. The present study aims to explore the impact of VD status on the severity of COVID-19 among hospitalized patients in Qatar and examine its effects on health factors related to the disease. <b>Materials and Methods:</b> A retrospective, cross-sectional, multicenter study was conducted at Hamad Medical Corporation in Qatar from March, 2020 to May, 2021. Demographics, laboratory values, comorbidities, clinical outcomes and some inflammatory markers were retrieved from the hospital's system. The severity of COVID-19 disease was determined by an elevated neutrophil-lymphocyte ratio (NLR). A patient with VD levels less than 20 ng/mL was considered deficient. Data were analyzed using SPSS v22, with significance set at p<0.05, employing Mann-Whitney, Chi-squared and correlation tests as appropriate. <b>Results:</b> The VD deficiency was common (68.6%), with longer ICU stays observed in deficient patients compared to those with normal VD status (19.7 vs. 11.2; p = 0.04). There was a trend of higher NLR with lower VD levels. No significant difference in interleukin-6 levels was found between the deficient and sufficient VD groups (60.6±64.1 vs. 41.2±45.2 pg/mL; p = 0.472). <b>Conclusion:</b> This study highlights the link between VD status and COVID-19 disease severity. The findings suggest that suboptimal levels of VD in COVID-19 patients may be potentially associated with increased length of stay in the ICU and have a minor impact on the pro-inflammatory state.

A comprehensive review of published information on ethnomedicinal uses, phytochemical and pharmacological properties of <i>Terminalia mollis</i> M.A. Lawson, a medium-sized to large deciduous tree, is presented. This study revealed that <i>T. mollis</i> is used as ethnoveterinary medicine and traditional medicine against sexually transmitted infections, respiratory infections, gastrointestinal problems, bilharzia, malaria, haemorrhoids, abdominal pains, measles, jaundice, cryptococcal meningitis and back pain. Phytochemical research identified flavonoids, polyphenols, ellagitannin, pentacyclic triterpenoids, trihydroxybenzoic acid, tannins, steroids, saponins, anthraquinones, cardiac glycosides and anthocyanins from leaves and root bark of <i>T. mollis</i>. Ethnopharmacological research revealed that the phytochemical compounds isolated from <i>T. mollis</i> and crude extracts of the species showed antibacterial, antimycobacterial, antimycoplasmal, antifungal, antiviral, anticonvulsant, antileishmanial, antioxidant, antiplasmodial, antitrypanosomal and cytotoxicity activities. Since <i>T. mollis</i> extracts are widely used as sources of traditional medicines, there is a need for extensive phytochemical, pharmacological, toxicological evaluations, <i>in vivo</i> and clinical studies.

<b>Background and Objective:</b> Fishmeal in Indonesia still mostly comes from abroad, while the available fish waste has not been widely processed as an alternative feed ingredient to replace it. This study aimed to evaluate the effect of feeding fermented fish waste meals in the ration on the physical quality of quail eggs and reproductive organ characteristics. <b>Materials and Methods:</b> One hundred and sixty female quails, 25 weeks old, were reared until 33 weeks. The diet treatments were P0 = Control ration, P1 = Ration containing 2% fermented fish waste meal, P2 = Ration containing 4% fermented fish waste meal and P3 = Ration containing 6% fermented fish waste meal. A completely randomized design (CRD) was used with four treatments and four replications, with ten female quails per replicate. The data were analyzed using ANOVA and if there were significant differences, the data were further analyzed using Duncan's Multiple Range Test. Significance was considered at a probability level of p<0.05. The variables measured were the egg's physical quality and reproductive organs. <b>Results:</b> The treatments did not significantly (p>0.05) affect egg weight, egg shape index, shell thickness, eggshell, albumen, yolk weight percentage, yolk color index, the Haugh unit, the carcass percentage and reproductive tract of quails. However, fermented fish waste meal increased the yolk color index and carcass percentage. <b>Conclusion:</b> Fermented fish waste meal up to 6% had no negative effect on egg quality or reproductive organs of quails. Fermented fish waste meal from pineapple can be considered an alternative feed ingredient, utilizing bromelain enzymes to reduce the use of fish meal.

<b>Background and Objective:</b> The liver is a crucial organ in the human body that contributes to metabolic processes, immune function and nutrient regulation. The objective of this study is to evaluate the effects of mulberry leaves (<i>Morus nigra</i> L.) on rats with acute liver injury induced by CCl₄. <b>Materials and Methods:</b> Twenty-four male albino Sprague Dawley rats (~195±10 g) were divided into control and CCl₄-induced liver injury groups. The treated group received a monthly basal diet of 2.5 and 5% mulberry leaves. Liver histopathology, body weight, organ-to-body weight ratios and feed intake were assessed. Lipid profiles and serum markers (glucose, urea, AST and ALT) were analyzed using SPSS (Mean±SD) with a significance level of p<0.05. <b>Results:</b> Total serum cholesterol and blood triglyceride levels were significantly lower (p<0.05) in all mulberry-fed groups, with the most notable reduction in the 5% group. The 5% mulberry leaf group also observed the best lipoprotein profile. The 5% mulberry leaf diet improved AST, ALT and serum glucose levels in rats with acute liver disease. <b>Conclusion:</b> Based on the results shown here, it appears that taking 5% ML daily can help improve liver functioning and shield rats from acute damage caused by CCL₄.

<b>Background and Objective:</b> Silverleaf whitefly (<i>Bemisia tabaci</i> ) is one of the most notable pest attacks on tomato plants. A viable eco-friendly option for pest control involves using pesticides specifically derived from the methanol extracts of <i>Tithonia diversifolia</i> and <i>Pachyrhizus erosus</i>. This study aims to determine the concentration of methanol extracts from <i>T. diversifolia</i> leaves and <i>P. erosus</i>. <b>Materials and Methods:</b> The study employed a Randomized Complete Block Design (RCBD) with 9 treatments and 3 replications. The toxicity of various extract concentrations was assessed through probit analysis, while plant growth and pest population density were monitored across multiple stages, with manual spraying of the extracts. This study uses a controlled greenhouse environment and detailed monitoring of environmental conditions, plant height and pest density throughout the two-week treatment period. A one-way ANOVA was conducted to evaluate differences between treatment groups, with significance set at p<0.05. <b>Results:</b> The LC₅₀ and LC₉₅ values for <i>T. diversifolia</i> leaf methanol extract at 3.8 and 7.9 g/L, respectively. The LC₅₀ and LC₉₅ values for <i>P. erosus</i> seed methanol extract were 4.3 and 8.7 g/L, respectively. The toxicity test results of both extracts have relatively high LC₅₀ and LC₉₅ values, indicating moderate toxicity levels. The most significant reduction in nymph population density occurred with the highest concentrations in <i>T. diversifolia</i> leaf (15.9 and 23.9 g/L) and <i>P. erosus</i> seed (17.4 and 26.2 g/L) extracts. In addition, both of the extracts can enhance tomato fruit height and harvest yield. <b>Conclusion:</b> Methanol extracts of <i>T. diversifolia</i> leaves and <i>P. erosus</i> seeds effectively reduce the <i>B. tabaci</i> population on tomato plants, with the highest suppression (100%) observed at concentrations of 15.9 g/L for <i>T. diversifolia</i> leaves and 26.2 g/L for <i>P. erosus</i> seeds, leading to increased fruit production compared to the control.

<b>Background and Objective:</b> Gonorrhoea, caused by <i>Neisseria gonorrhoeae</i>, continues to pose a major global health threat due to the rising incidence of antibiotic resistance. This study aimed to explore the potential of metabolites derived from immunized larvae of <i>Rhynchophorus</i> sp., as alternative therapeutic agents for the treatment of gonorrhea. <b>Materials and Methods:</b> Liquid Chromatography-Mass Spectrometry/Mass Spectrometry (LC-MS/MS) was used to identify metabolites from the hemolymph of larvae immunized with non-pathogenic <i>Escherichia coli</i>. Subsequently, key molecular targets of these metabolites were identified through network pharmacology and molecular docking approaches. <b>Results:</b> Metabolite interaction networks revealed L-glutamic acid as the compound with the highest degree of interaction (25) among the 15 identified amino acid-derived metabolites. Further analysis of gene interactions identified <i>guaA</i>, which encodes guanosine monophosphate synthetase, as having the highest degree of interaction (27). Binding-free energy of identified metabolites, with L-tryptophan showing the highest binding free energy of -5.9 kcal/mol in the guanosine monophosphate receptor. <b>Conclusion:</b> The study identifies guanosine monophosphate synthetase (GMPS) as a promising target for combating gonorrhea, with L-tryptophan showing potential as a lead compound. Targeting GMPS disrupts critical metabolic pathways in <i>N. gonorrhea</i>, offering a novel approach against antibiotic-resistant strains. By integrating metabolite analysis, network pharmacology and molecular docking, the study provides a comprehensive framework for drug discovery. This multidisciplinary strategy advances the development of effective therapies for antibiotic-resistant pathogens.

<b>Background and Objective:</b> Endophytic actinomycetes represent a promising source of novel bioactive compounds. This study aimed to isolate and characterize endophytic actinomycetes from <i>Aloe vera</i> leaves, identify their bioactive metabolites and assess their antimicrobial and anticancer properties. <b>Materials and Methods:</b> Endophytic actinomycetes were isolated from <i>Aloe vera</i> leaf sections and screened for antimicrobial activity. Isolate AVL08, exhibiting potent activity against <i>Staphylococcus aureus</i>, was identified using 16S rRNA gene sequencing and morphological characterization. Bioactive compounds were extracted, purified and identified using spectroscopic methods. Antimicrobial activity was evaluated using microbroth dilution assays and cytotoxicity was assessed against Vero, HeLa, HepG2 and MDA-MB-231 cell lines. Molecular docking studies were performed to investigate interactions with human DNA topoisomerase II. <i>In silico</i>, ADMET predictions were conducted using SwissADME, PreADMET and pkCSM. Statistical analysis was performed using SPSS (version 11.01) with One-way ANOVA and Tukey's <i>post hoc</i> test, considering differences significant at p<0.05. <b>Results:</b> Isolate AVL08 was identified as <i>Streptomyces galbus</i> and produced two anthraquinone compounds, 1,4,6-trihydroxy-8-(2'-oxopentyl)-9,10-anthraquinone (<b>1</b>) and 1,4,6-trihydroxy-8-pentacarboxyl-9,10-anthraquinone (<b>2</b>). These compounds exhibited selective antibacterial activity against Gram-positive bacteria and significant cytotoxicity against MDA-MB-231 and HeLa cancer cell lines, with IC₅₀ values ranging from 94.00 to 154.52 μg/mL. Compound <b>2</b> demonstrated enhanced potency and selectivity against HepG2 cells. Molecular docking revealed favorable interactions of the anthraquinones with topoisomerase II. The ADMET predictions indicated favorable pharmacokinetic profiles but highlighted potential hERG channel inhibition and mutagenicity in specific bacterial strains. <b>Conclusion:</b> <i>Streptomyces galbus</i> AVL08 is a promising source of bioactive anthraquinones with potent antimicrobial and anticancer properties. The identified compounds, particularly Compound <b>2</b>, warrant further investigation for therapeutic applications. This study highlights the potential of endophytic actinomycetes from <i>Aloe vera</i> as a valuable resource for drug discovery.