Microbes-mediated synthesis strategies of metal nanoparticles and their potential role in cancer therapeutics.

Past few years have seen a paradigm shift towards ecofriendly, green and biological fabrication of metal nanoparticles (MNPs) for diverse nanomedicinal applications especially in cancer nanotheranostics. Besides, the well-known green synthesis methods of plant materials, the potential of the microbial world (bacteria, fungi, alga, etc.) in biofabrication is equally realized. Biomolecules and enzymes in the microbial cells are capable of catalyzing the biosynthesis process. These microbial derived inorganic nanoparticles have been frequently evaluated as potential agents in cancer therapies revealing exciting results. Through, cellular and molecular pathways, these microbial derived nanoparticles are capable of killing the cancer cells. Considering the recent developments in the anticancer applications of microbial derived inorganic MNPs, a dire need was felt to bring the available information to a single document. This manuscript reviews not only the mechanistic aspects of the microbial derived MNPs but also include the diverse mechanisms that governs their anticancer potential. Besides, an updated literature review is presented that includes studies of 2019-onwards.

Silver nanoparticles biosynthesized from secondary metabolite producing marine actinobacteria and evaluation of their biomedical potential.

Biosynthesis of silver nanoparticles (AgNPs) from marine actinobacteria offers a promising avenue for exploring bacterial extracts as reducing and stabilizing agents. We report extracellular extracts of Rhodococcus rhodochrous (MOSEL-ME29) and Streptomyces sp. (MOSEL-ME28), identified by 16S rRNA gene sequencing for synthesis of AgNPs. Ultrafine silver nanoparticles were biosynthesized using the extracts of R. rhodochrous and Streptomyces sp. and their possible therapeutic applications were studied. The physicochemical properties of nanoparticles were established by HR-SEM/TEM, SAED, UV-Vis, EDS, XRD, and FTIR. UV-Vis spectra displayed characteristic absorption at 430 nm and 412 nm for AgNPs from Streptomyces sp. (S-AgNPs) and Rhodococcus sp. (R-AgNPs), respectively. HR-SEM/TEM, XRD, EDS analysis confirmed the spherical shape, crystalline nature, and elemental formation of silver. Crystallite or grain size was deduced as 5.52 nm for R-AgNPs and 35 nm for S-AgNPs. Zeta-potential indicated electrostatic negative charge for AgNPs, while FTIR revealed the presence of diverse functional groups. Disc diffusion assay indicated the broad-spectrum antibacterial potential of S-AgNPs with the maximum inhibition of B. subtilis while R-AgNPs revealed potency against P. aeruginosa at 10 µg/mL concentration. Biogenic AgNPs revealed antileishmanial activity and the IC50 was calculated as 164 µg/mL and 184 µg/mL for R-AgNPs and S-AgNPs respectively. Similarly, the R-AgNPs and S-AgNPs revealed anti-cancer potential against HepG2 and the IC50 was calculated as 49 µg/mL and 69 µg/mL for R-AgNPs and S-AgNPs, respectively. Moreover, the antioxidant activity showed significant results. MTT assay on RD cells, L20B cells, and Hep-2C indicated intensification in viability by reducing the concentration of R-AgNPs and S-AgNPs. The R-AgNPs and S-AgNPs inhibited sabin-like poliovirus (1TCID50 infection in RD cells). Furthermore, hemocompatibility at low concentrations has been confirmed. Hence, it is concluded that biogenic-AgNPs has the potential to be used in diverse biological applications and that the marine actinobacteria are an excellent resource for fabrication of AgNPs.

Ethics, pandemic and environment; looking at the future of low middle income countries.

COVID-19 which started in Wuhan, China and swiftly expanded geographically worldwide, including to Low to Middle Income Countries (LMICs). This in turn raised numerous ethical concerns in preparedness, knowledge sharing, intellectual property rights, environmental health together with the serious constraints regarding readiness of health care systems in LMICs to respond to this enormous public health crisis. From the restrictions on public freedom and burgeoning socio-economic impacts to the rationing of scarce medical resources, the spread of COVID-19 is an extraordinary ethical dilemma for resource constrained nations with less developed health and research systems. In the current crisis, scientific knowledge and technology has an important role to play in effective response. Emergency preparedness is a shared responsibility of all countries with a moral obligation to support each other. This review discusses the ethical concerns regarding the national capacities and response strategies in LMICs to deal with the COVID-19 pandemic as well as the deep link between the environment and the increasing risk of pandemics.

Endophyte-mediated synthesis of silver nanoparticles and their biological applications.

Biosynthesis has emerged as a frontier technology for fabrication of functionally diverse nanoparticles that possess tremendous therapeutic implications. Various biological resources have already demonstrated their potential to produce nanoparticles with interesting features. Endophytic microbes live in a symbiotic relationship with plants possessing a unique and versatile reservoir of potentially therapeutic secondary metabolites having the tendency to reduce metallic ions into nanoparticles. Successful biosynthesis of AgNPs using endophytic organisms has already been reported; however, the overall picture about its synthesis and applications is still not clear. In the current article, a comprehensive review of literature was performed for comparing different physical and biological properties of endophytic microbe-derived AgNPs. In addition, the present paper mechanistically explains the synthesis of AgNPs and their diverse pharmacognostic properties. Further studies are encouraged to understand the mechanism of biopharmaceutical effects of these endophyte-mediated NPs.

Bio-guided profiling and HPLC-DAD finger printing of Atriplex lasiantha Boiss.

BACKGROUND: Plants represent an intricate and innovative source for the discovery of novel therapeutic remedies for the management of various ailments. The current study has been aimed to validate the therapeutic potential of ethnomedicinally significant plant Atriplex lasiantha Boiss. METHODS: The polarity based extraction process was carried out using fourteen solvents to figure out best extraction solvent and bioactive fractions. Total phenolic-flavonoids contents were quantified colorimetrically and polyphenolics were measured using HPLC-DAD analysis. Moreover, the test samples were tested against several diseases targets following various assays including free radicals scavenging, antibacterial, antifungal, cytotoxic and antileishmanial assay. RESULTS: Among the solvent fractions, maximum yield was obtained with methanol-water extract i.e., 11 ± 0.49%. Maximum quantity of gallic acid equivalent phenolic content and quercetin equivalent flavonoid content were quantified in methanol-ethyl acetate extract of A. lasiantha. Significant quantity of rutin i.e., 0.3 µg/mg was quantified by HPLC analysis. The methanol-ethyl acetate extract of A. lasiantha exhibited maximum total antioxidant and total reducing power with 64.8 ± 1.16 AAE/mg extract respectively, while showing 59.8 ± 1.07% free radical scavenging potential. A significant antibacterial potential was exhibited by acetone-distilled water extract of A. lasiantha with 11 ± 0.65 mm zone of inhibition against B. subtilis. Considerable antifungal activity was exhibited by ethyl acetate-n-hexane extract of aerial part of A. lasiantha with 14 ± 1.94 mm zone of inhibition against A. fumigatus. Highest percentage of α-amylase inhibition (41.8 ± 1.09%) was observed in ethyl acetate-n-hexane extract. Methanol-acetone extract of A. lasiantha demonstrated significant inhibition of hyphae formation with 11 ± 0.49 mm bald zone of inhibition. Significant in-vitro cytotoxicity against Hep G2 cell line has been exhibited by methanol-chloroforms extract of A. lasiantha. CONCLUSION: The current study reveals the prospective potential of Atriplex lasiantha Boiss. for the discovery of biologically active compounds through bioassay guided isolation against various diseases.

Phytochemical Analysis, Ephedra Procera C. A. Mey. Mediated Green Synthesis of Silver Nanoparticles, Their Cytotoxic and Antimicrobial Potentials.

Background and Objectives: The current study focuses on an eco-friendly and cost-effective method of Ephedra procera C. A. Mey. mediated green synthesis of silver nanoparticles as potential cytotoxic, antimicrobial and anti-oxidant agents. Materials and Methods: Plant aqueous extracts were screened for Total Phenolic (TPC), Total Flavonoid contents (TFC), Total Antioxidant Capacity (TAC) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging potentials. Total reducing power estimated by potassium ferricyanide colorimetric assay. The biosynthesized E. procera nanoparticles (EpNPs) were characterized by UV-spectroscopy, Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction and Scanning electron microscopy. EpNPs were evaluated for their antimicrobial, bio-compatibility and cytotoxic potentials. Results: Initial phytocheimcal analysis of plant aqueous extract revealed TFC of 20.7 ± 0.21 µg/mg extract and TPC of 117.01 ± 0.78 µg/mg extract. TAC, DPPH free radical scavenging and reducing power were 73.8 ± 0.32 µg/mg extract, 71.8 ± 0.73% and 105.4 ± 0.65 µg/mg extract respectively. The synthesized EpNPs were observed to possess high cytotoxicity against HepG2 cancer cell lines with IC50 (61.3 µg/mL) as compared aqueous extract with IC50 of (247 µg/mL). EpNPs were found to be biocompatible and have less effect on human erythrocytes. EpNPs exhibited significant antioxidant potentials and exhibited considerable activity against Escherichia coli and Bacillus subtilis with Minimum Inhibitory Concentration (MICs) of 11.12 µg/mL and 11.33 µg/mL respectively. Fungal species Aspergillus niger and Aspergillus flavus were found susceptible to EpNPs. Conclusions: Results of the current study revealed that EpNPs exhibited considerable antibacterial, antifungal and cytotoxic potentials. Aqueous extract possesses significant anti-radical properties and thus can be useful in free radicals induced degenerative disorders.

Ethical Issues Regarding CRISPR Mediated Genome Editing.

CRISPR-Cas9 has emerged as a simple, precise and most rapid genome editing technology. With a number of promising applications ranging from agriculture and environment to clinical therapeutics, it is greatly transforming the field of molecular biology. However, there are certain ethical, moral and safety concerns related to the attractive applications of this technique. The most contentious issues concerning human germline modifications are the challenges to human safety and morality such as risk of unforeseen, undesirable effects in clinical applications particularly to correct or prevent genetic diseases, matter of informed consent and the risk of exploitation for eugenics. Stringent regulations and guidelines as well as worldwide debate and awareness are required to ensure responsible and wise use of CRISPR mediated genome editing technology. There is a need for an extensive dialogue among scientists, ethicists, industrialists and policy makers on its societal implications. The opinion of different elements of the society including the general public as well as religious scholars is also critical. In countries with existing legislative framework, it might be appropriate to allow CRISPR based research to proceed with proper justification. However, much anticipated future clinical applications must be strictly regulated with newly established regulations.

Wound healing applications of biogenic colloidal silver and gold nanoparticles: recent trends and future prospects.

Nanotechnology has emerged as a prominent scientific discipline in the technological revolution of this millennium. The scientific community has focused on the green synthesis of metal nanoparticles as compared to physical and chemical methods due to its eco-friendly nature and high efficacy. Medicinal plants have been proven as the paramount source of various phytochemicals that can be used for the biogenic synthesis of colloidal silver and gold nanoparticles as compared to other living organisms, e.g., microbes and fungi. According to various scientific reports, the biogenic nanoparticles have shown promising potential as wound healing agents. However, not a single broad review article was present that demonstrates the wound healing application of biogenic silver and gold nanoparticles. Foreseeing the overall literature published, we for the first time intended to discuss the current trends in wound healing via biogenic silver and gold nanoparticles. Furthermore, light has been shed on the mechanistic aspects of wound healing along with futuristic discussion on the faith of biogenic silver and gold nanoparticles as potential wound healing agents.

Role of plant phytochemicals and microbial enzymes in biosynthesis of metallic nanoparticles.

Metal-based nanoparticles have gained tremendous popularity because of their interesting physical, biological, optical, and magnetic properties. These nanoparticles can be synthesized using a variety of different physical, chemical, and biological techniques. The biological means are largely preferred as it provides an environmentally benign, green, and cost-effective route for the biosynthesis of nanoparticles. These bioresources can act as a scaffold, thereby playing the role of reducing as well as capping agents in the biosynthesis of nanoparticles. Medicinal plants tend to have a complex phytochemical constituent such as alcohols, phenols, terpenes, alkaloids, saponins, and proteins, while microbes have key enzymes which can act as reducing as well as stabilizing agent for NP synthesis. However, the mechanism of biosynthesis is still highly debatable. Herein, the present review is directed to give an updated comprehensive overview towards the mechanistic aspects in the biosynthesis of nanoparticles via plants and microbes. Various biosynthetic pathways of secondary metabolites in plants and key enzyme production in microbes have been discussed in detail, along with the underlying mechanisms for biogenic NP synthesis.

Nanomedicines for developing cancer nanotherapeutics: from benchtop to bedside and beyond.

Cancer is a devastating disease and remains a significant cause of mortality and morbidity in both developed and developing countries. Although there are large number of drugs that can be used for the treatment of cancer, the problem is selective and specific killing of cancerous cells without harming the normal cells. There are some biological barriers to potential drug delivery in cancer cells like hepatic, renal, abnormal vasculature, dense extracellular matrix, and high interstitial fluid pressure. The physicochemical characteristics of nanoparticles (NPs) such as size, shape, and surface charge may also have significant effects on tumor penetration. NPs coated with drug can be used to overcome these biological barriers to enhance targeted delivery. This literature survey encompasses the biological barriers to potential drug delivery in cancer cells, elaborate on designing strategies to enhance NPs penetration and distribution inside the tumor interstitium. Scientists are now doing great efforts to design next-generation of nanomedicines (NMs) that need to be better targeted with high specificity and efficacy to kill cancer cells. These challenges need to be overcome through collaborations among academia, pharmaceutical industries, and regulatory agencies to eradicate this global menace. Furthermore, this review article has critically discussed the recent developments, controversies, challenges, emerging concepts, and future perspectives in cancer NMs.

Multifunctional theranostic applications of biocompatible green-synthesized colloidal nanoparticles.

Phytochemicals offer immense promise for sustainable development and production of nanotechnology-enabled products. In the present study, Olax nana Wall. ex Benth. (family: Olacaceae) aqueous extract was used as an effective stabilizing agent to produce biogenic silver (ON-AgNPs) and gold nanoparticles (ON-AuNPs), which were investigated for biocompatibility and prospective biomedical applications (antibacterial, anticancer, antileishmanial, enzyme inhibition, antinociceptive, and anti-inflammatory activities). Various characterization techniques (XRD, FTIR, SEM, TEM, DLS, EDX, and SAED) revealed efficient biosynthesis of ON-AgNPs (26 nm) and ON-AuNPs (47 nm). In the toxicological assessment, ON-AgNPs and ON-AuNPs were found biocompatible towards human RBCs and macrophages (IC50 > 200 µg/mL). In a concentration range of 62.5-2000 µg/mL, a strong antibacterial effect was produced by ON-AgNPs against Staphylococcus epidermidis (MIC = 7.14 µg/mL) and Escherichia coli (8.25 µg/mL), while ON-AuNPs was only active against Staphylococcus aureus (9.14 µg/mL). At a concentration of 3.9-500 µg/mL, a dose-dependant inhibition of HepG2 cancer cells was produced by ON-AgNPs (IC50 = 14.93 µg/mL) and ON-AuNPs (2.97 µg/mL). Both ON-AgNPs and ON-AuNPs were found active against Leishmania tropica (KMH23) promastigotes (IC50 = 12.56 and 21.52 µg/mL) and amastigotes (17.44 and 42.20 µg/mL), respectively, after exposure to a concentration range of 1-200 µg/mL for 72 h. Preferential enzyme inhibition against urease and carbonic anhydrase II were noted for ON-AgNPs (39.23 and 8.89%) and ON-AuNPs (31.34 and 6.34%), respectively; however, these were found inactive against xanthine oxidase at 0.2 mg/mL. In the in vivo antinociceptive (acetic acid-induced abdominal constrictions) and anti-inflammatory (carrageenan-induced paw edema) activities, ON-AgNPs and ON-AuNPs at doses of 40 and 80 mg/kg, significantly attenuated the tonic nociception (P < 0.001) and ameliorated the carrageenan-induced inflammation (P < 0.01, P < 0.001). The results of in vitro and in vivo activities indicated that the biogenic nanoparticles can be used as valuable theranostic agents for further exploration of diverse biomedical applications.

HPLC-DAD finger printing, antioxidant, cholinesterase, and α-glucosidase inhibitory potentials of a novel plant Olax nana.

BACKGROUND: The medicinal importance of a novel plant Olax nana Wall. ex Benth. (family: Olacaceae) was revealed for the first time via HPLC-DAD finger printing, qualitative phytochemical analysis, antioxidant, cholinesterase, and α-glucosidase inhibitory assays. METHODS: The crude methanolic extract of O. nana (ON-Cr) was subjected to qualitative phytochemical analysis and HPLC-DAD finger printing. The antioxidant potential of ON-Cr was assessed via 1,1-diphenyl,2-picrylhydrazyl (DPPH), 2,2-azinobis[3-ethylbenzthiazoline]-6-sulfonic acid (ABTS) and hydrogen peroxide (H2O2) free radical scavenging assays. Furthermore, acetylcholinesterase (AChE) & butyrylcholinesterase (BChE) inhibitory activities were performed using Ellman's assay, while α- glucosidase inhibitory assay was carried out using a standard protocol. RESULTS: The qualitative phytochemical analysis of ON-Cr revealed the presence of secondary metabolites like alkaloids, flavonoids, tannins, sterols, saponins and terpenoids. The HPLC-DAD finger printing revealed the presence of 40 potential compounds in ON-Cr. Considerable anti-radical activities was revealed by ON-Cr in the DPPH, ABTS and H2O2 free radical scavenging assays with IC50 values of 71.46, 72.55 and 92.33 µg/mL, respectively. Furthermore, ON-Cr showed potent AChE and BChE inhibitory potentials as indicated by their IC50 values of 33.2 and 55.36 µg/mL, respectively. In the α-glucosidase inhibition assay, ON-Cr exhibited moderate inhibitory propensity with an IC50 value of 639.89 µg/mL. CONCLUSIONS: This study investigated Olax nana for the first time for detailed qualitative phytochemical tests, HPLC-DAD finger printing analysis, antioxidant, anticholinesterase and α-glucosidase inhibition assays. The antioxidant and cholinesterase inhibitory results were considerable and can provide scientific basis for further studies on the neuroprotective and anti-Alzheimer's potentials of this plant. ON-Cr may further be subjected to fractionation and polarity guided fractionation to narrow down the search for isolation of bioactive compounds.

Selected hepatoprotective herbal medicines: Evidence from ethnomedicinal applications, animal models, and possible mechanism of actions.

Insight into the hepatoprotective effects of medicinally important plants is important, both for physicians and researchers. Main reasons for the use of herbal medicine include their lesser cost compared with conventional drugs, lesser undesirable drug reactions and thus high safety, and reduced side effects. The present review focuses on the composition, pharmacology, and results of experimental trials of selected medicinal plants: Silybum marianum (L.) Gaertn., Glycyrrhiza glabra, Phyllanthus amarus Schumach. & Thonn., Salvia miltiorrhiza Bunge., Astragalus membranaceus (Fisch.) Bunge, Capparis spinosa (L.), Cichorium intybus (L.), Solanum nigrum (L.), Sapindus mukorossi Gaertn., Ginkgo biloba (L.), Woodfordia fruticosa (L.) Kurz, Vitex trifolia (L.), Schisandra chinensis (Turcz.) Baill., Cuscuta chinensis (Lam.), Lycium barbarum, Angelica sinensis (Oliv.) Diels, and Litsea coreana (H. Lev.). The probable modes of action of these plants include immunomodulation, stimulation of hepatic DNA synthesis, simulation of superoxide dismutase and glutathione reductase to inhibit oxidation in hepatocytes, reduction of intracellular reactive oxygen species by enhancing levels of antioxidants, suppression of ethanol-induced lipid accumulation, inhibition of nucleic acid polymerases to downregulate viral mRNA transcription and translation, free radical scavenging and reduction of hepatic fibrosis by decreasing the levels of transforming growth factor beta-1, and collagen synthesis in hepatic cells. However, further research is needed to identify, characterize, and standardize the active ingredients, useful compounds, and their preparations for the treatment of liver diseases.

Biosynthesis of Metal Nanoparticles via Microbial Enzymes: A Mechanistic Approach.

During the last decade, metal nanoparticles (MtNPs) have gained immense popularity due to their characteristic physicochemical properties, as well as containing antimicrobial, anti-cancer, catalyzing, optical, electronic and magnetic properties. Primarily, these MtNPs have been synthesized through different physical and chemical methods. However, these conventional methods have various drawbacks, such as high energy consumption, high cost and the involvement of toxic chemical substances. Microbial flora has provided an alternative platform for the biological synthesis of MtNPs in an eco-friendly and cost effective way. In this article we have focused on various microorganisms used for the synthesis of different MtNPs. We also have elaborated on the intracellular and extracellular mechanisms of MtNP synthesis in microorganisms, and have highlighted their advantages along with their challenges. Moreover, due to several advantages over chemically synthesized nanoparticles, the microbial MtNPs, with their exclusive and dynamic characteristics, can be used in different sectors like the agriculture, medicine, cosmetics and biotechnology industries in the near future.

Current state and prospects of the phytosynthesized colloidal gold nanoparticles and their applications in cancer theranostics.

The design, development, and biomedical applications of phytochemical-based green synthesis of biocompatible colloidal gold nanoparticles (AuNPs) are becoming an emerging field due to several advantages (safer, eco-friendly, simple, fast, energy efficient, low-cost, and less toxic) over conventional chemical synthetic procedures. Biosynthesized colloidal gold nanoparticles are remarkably attractive in several biomedical applications including cancer theranostics due to small size, unusual physico-chemical properties, facile surface modification, high biocompatibility, and numerous other advantages. Of late, several researchers have investigated the biosynthesis and prospective applications (diagnostics, imaging, drug delivery, and cancer therapeutics) of AuNPs in health care and medicine. However, not a single review article is available in the literature that demonstrates the anti-cancer potential of biosynthesized colloidal AuNPs with detailed mechanistic study. In the present review article, we for the first time discuss the biointerface of colloidal AuNPs, plants, and cancer mainly (i) comprehensive mechanistic aspects of phytochemical-based synthesis of AuNPs; (ii) proposed anti-cancer mechanisms along with biomedical applications in diagnostics, imaging, and drug delivery; and (iii) key challenges for biogenic AuNPs as future cancer nanomedicine.

Antifungal, phytotoxic and hemagglutination activity of methanolic extracts of Ocimum basilicum.

OBJECTIVE: To report some of the pharmacological features of the medicinal plant Ocimum basilicum. METHODS: In the current studies, Antifungal activity of crude methanolic fraction of Ocimum basilicum was determined against eight pathogenic fungal strains using tube dilution assay. The methanolic fraction was also investigated for phytotoxic and hemagglutination activity. RESULTS: Of the eight strains investigated only Candida albicans and Curvilaria lunata were found to be least affected by plant extract while the rest were significantly inhibited. Moderate phytotoxic activity was observed against lemna minor. Hemagglutination activity showed absence of phytolectins and hence no agglutination of erythrocytes. CONCLUSION: The crude extract of Ocimum basilicum has significant properties against fungi and phytotoxic substances.

Evaluation of antileishmanial, antibacterial and brine shrimp cytotoxic potential of crude methanolic extract of Herb Ocimum basilicum (Lamiacea).

OBJECTIVE: To collect and screen for ethnopharmacological properties (antileishmanial, antibacterial and brine lethality assays) of medicinal plan Ocimum basilicum from Peshawar region (34.008 latitude and 71.57 altitudes). METHODS: In the present study a general antileishmanial activity against Leishmania tropica strair was carried out. The antibacterial potential of the plant was performed against 06 gram positiv and 06 gram negative bacteria. Brine shrimp cyto- toxicity assay at different concentrations were investigated. RESULTS: The anti-promastigotes profile of the plant showed good antileishmanial activity exhibited LC50 value 21.67 µg/mL. The result for gram positive antibacterial activity revealed that the O. basilicum leaves extract possesses significant inhibitory activity at highest two concentrations ranging from 20.66 ± 0.31 to 31.86 ± 0.80 for Clostridium perfringens type C and Bacillus subtitilis, respectively, as compared to the gentamycin (27.36 ± 0.55 and 21.80 ± 0.72, respectively). For gram negative bacteria good activity was observed. A highest zone of inhibition was recorded for Pseudomonas aeroginosa (28.83 ± 0.28) at the highest concentration (10 mg/ mL). The LC50 value obtained for brine shrimp lethality assay was 91.56 µg/mL. CONCLUSION: The herb basil possesses effective cidal activities which make this plant a good candidate for the isolation of antiprotozoal and antibacterial compounds which may lead to the development of novel drug.

Antibacterial activity of honey in north-west Pakistan against select human pathogens.

OBJECTIVE: To investigate the antimicrobial activity of commercially available honey and raw honey samples in Khyber Pakhtunkhwa, Pakistan, against pathogenic bacterial strains. METHODS: Well diffusion assays were performed to screen pure and diluted honey samples for antibacterial activity against six Gram-negative and six Gram-positive bacterial strains. Zones of inhibition were measured and compared with 10 mg Gentamycin. RESULTS: When honey samples were diluted to 20%-70%, the honey samples showed no activity to mild antibacterial activity. The highest antibacterial activity was recorded when 90% and pure undiluted honey samples were tested and compared with a control Gentamycin disc (10 mg). CONCLUSION: Commercially processed honey and raw honey samples from north-west of Pakistan possess good antimicrobial potential.

Insight into the molecular mechanism, cytotoxic, and anticancer activities of phyto-reduced silver nanoparticles in MCF-7 breast cancer cell lines.

This contribution insight on the cytotoxic and anticancer activities and molecular mechanism of phyto-reduced silver nanoparticles (AgNPs) in MCF-7 breast cancer cell lines. A simple, entirely green synthesis process was optimized for the phyto-reduction of AgNP (~12.7 nm) using aqueous leaf extracts of Indigofera heterantha. The structural and vibrational properties of biosynthesized AgNPs were extensively characterized using UV-Vis spectrophotometer, x-ray diffraction (XRD), dynamic light scattering (DLS), and Fourier transform Infrared spectroscopy (FTIR), while their shape and morphology was studied through scanning electron microscopy (SEM). The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell viability assay indicates concentration dependent inhibition with IC50, 27.93 ± 2.10 µg/mL against MCF-7 cells and 294.38 ± 3.87 µg/mL against L929 cells. The manifested anticancer mechanism in MCF-7 cells was extensively studied using Acridine orange/ethidium bromide (AO/EB), 4',6-diamidino-2-phenylindole (DAPI) and Annexin-V/propedium iodide fluorescence microscopic assays. The level of reactive oxygen species (ROS) was measured using DCFH-DA fluorescent spectroscopy. Overall, the results show that AgNPs exhibit cytotoxic and apoptotic effect on breast cancer MCF-7 cells by damaging membrane integrity and nuclear fragmentation due to oxidative stress generated by elevated level of ROS. RESEARCH HIGHLIGHTS: Biomimetic synthesis of nano dimension size silver nanoparticles (AgNPs). Characterization of AgNPs through UV-Vis, DLS, XRD, FTIR, and SEM. Cytotoxic and anticancer effects of the biosynthesized AgNPs in L929 fibroblast cells and MCF7 breast cancer respectively. Determination of morphological, and nuclear changes triggered by AgNPs in MCF 7 breast cancer cells using fluorescent microscopy and flow cytometry. Apoptosis induction by AgNPs in cancer cells through oxidative stress generated by reactive oxygen species (ROS).

Preliminary insights on the mutational spectrum of BRCA1 and BRCA2 genes in Pakhtun ethnicity breast cancer patients from Khyber Pakhtunkhwa (KP), Pakistan.

Gene mutations are a source of genetic instability which fuels the progression of cancer. Mutations in BRCA1 and BRCA2 are considered as major drivers in the progression of breast cancer and their detection indispensable for devising therapeutic and management approaches. The current study aims to identify novel pathogenic and recurrent mutations in BRCA1 and BRCA2 in Pakhtun population from the Khyber Pakhtunkhwa. To determine the BRCA1 and BRCA2 pathogenic mutation prevalence in Pakhtun population from KP, whole exome sequencing of 19 patients along with 6 normal FFPE embedded blocks were performed. The pathogenicity of the mutations were determined and they were further correlated with different hormonal, sociogenetic and clinicopathological features. We obtained a total of 10 mutations (5 somatic and 5 germline) in BRCA1 while 27 mutations (24 somatic and 3 germline) for BRCA2. Five and seventeen pathogenic or deleterious mutations were identified in BRCA1 and BRCA2 respectively by examining the mutational spectrum through SIFT, PolyPhen-2 and Mutation Taster. Among the SNVs, BRCA1 p.P824L, BRCA2 p. P153Q, p.I180F, p.D559Y, p.G1529R, p.L1576F, p.E2229K were identified as mutations of the interaction sites as predicted by the deep algorithm based ISPRED-SEQ prediction tool. SAAFEQ-SEQ web-based algorithm was used to calculate the changes in free energy and effect of SNVs on protein stability. All SNVs were found to have a destabilizing effect on the protein. ConSurf database was used to determine the evolutionary conservation scores and nature of the mutated residues. Gromacs 4.5 was used for the molecular simulations. Ramachandran plots were generated using procheck server. STRING and GeneMania was used for prediction of the gene interactions. The highest number of mutations (BRCA1 7/10, 70 %) were on exon 9 and (BRCA2, 11/27; 40 %) were on exon 11. 40 % and 60 % of the BRCA2 mutations were associated Grade 2 and Grade 3 tumors respectively. The present study reveals unique BRCA1 and BRCA2 mutations in Pakhtun population. We further suggest sequencing of the large cohorts for further characterizing the pathogenic mutations.