Biosynthesis of antimicrobial nanoparticle from Swertia spp. (Chirayita) against bacterial pathogens of Poultry- A way forward to Green Nanotechnology and Nano-medicines.

The increasing prevalence of multidrug-resistant microorganisms in poultry has led to a rise in bacterial infections, causing significant economic loss. Green nanotechnology, such as silver nanoparticles (AgNPs), has the potential to address this issue by providing potent antifungal, antiviral, and antibacterial properties. This study explored the combined potential of AgNPs and the local herb Swertia chirayita against established poultry pathogens, employing a non-factorial Central Composite Design (CCD) to evaluate the factors affecting the production of nanoparticles induced by silver nitrate from the selected herb. The optimal values for temperature, wavelength, silver nitrate concentration, incubation duration, and pH were found to produce the highest nanoparticles. The functional groups in Swertia chirayita stimulated nanoparticles were confirmed using FTIR spectroscopy, and the stability of ScNPs was elucidated using zeta potential. The crystalline structure of ScNPs was confirmed using diffraction intensity patterns. Silver nanoparticles demonstrated antibacterial activity against Salmonella spp. and Escherichia coli (E.coli), both known as significant poultry pathogens, using the agar well diffusion method, with inhibition zones of 25.0 mm and 35.0 mm, respectively.This study explored the green manufacturing of silver nanoparticles by using plants and microorganisms, focusing on their antibacterial properties. The exact mechanism of synthesis and action in AgNPs is still poorly understood. Researchers should prioritize the use of accessible, easy-to-extract plants or bacteria, especially non-pathogenic and fast-growing microorganisms for safe handling. Analyzing biomolecules in plant extract, microbial biomass, or culture supernatants, including probiotic bacteria, is crucial for creating and stabilizing AgNPs, which could be effective synthetic agents. It is crucial to optimize conditions for rapid, stable, and large-scale synthesis. Based on this research, Sc-NPs may be proposed as nanomedicine for treating infections in poultry caused by E. coli and Salmonella spp.

Detection, quantification and genotype distribution of HCV patients in Lahore, Pakistan by real-time PCR.

BACKGROUND: Hepatitis C virus (HCV) is considered as "Viral Time Bomb" suggested by the World Health Organization and if it is not treated timely, it will lead towards cirrhosis and hepatocellular carcinoma (HCC). OBJECTIVE: The purpose of the present research is to study possible risk factors, frequent genotypes of HCV and its association with different age groups. METHODS: Suspected blood samples from HCV patients were collected from different hospitals of Lahore, Pakistan. Out of 1000 HCV suspected samples, 920 samples were found HCV positive detected by Anti-HCV ELISA, CobasR. kit. The quantification of HCV load was determined by HCV quantification kit and LINEAR ARRAY KIT (Roche) was used for genotype determination by Real-Time PCR (ABI). Statistical analysis was done by using Microsoft Excel. RESULTS: Out of 920 subjects, 77 subjects (8.4%) were false positive and they were not detected by nested PCR. Three PCR positive samples were untypeable. Genotype 3 was predominant in Lahore which was 83.5%, whereas type 1 and 2 were 5.1% and 0.7% respectively. There were also mixed genotypes detected, 1 and 3 were 0.4%, 2 and 3 were 1.41% and 3 and 4 were 0.2% only. Male were more infected of HCV in the age <40 years and females >40years. CONCLUSION: The major risk factor for HCV transmission is by use of unsterilized razors/blades. It is necessary to spread awareness among the general population of Pakistan about HCV transmission risk factors. Regular physical examination at least once a year is recommended, so that early detection of HCV could be done.

Determination of expression profile of p53 gene in different grades of breast cancer tissues by real time PCR.

BACKGROUND: Pakistan has a high incidence of breast cancer in Asia, where annually 16,232 deaths are reported. There are many exogenous and endogenous risk factors that affect the tumor suppressor genes and oncogenes. The p53 gene is a tumor suppressor gene and it has a role to protect the whole genome from external and internal stresses, which causes damages to the genome. OBJECTIVE: The aim of the current study was to investigate the p53 gene expression by using the real-time PCR technique in different grades of breast cancer as compared to the normal tissue. METHODS: Fresh Modified Radical Mastectomy (MRM) samples (grade1-grade3) were collected from different hospitals of the Lahore. The project was approved by an ethical review committee of Jinnah Hospital, Lahore. And before sampling an informed consent was obtained from patients and clinicians. RNA from fresh biopsies was extracted by Qiagen extraction kit and cDNA was formed. Real time PCR performed by using SYBR green master mix (ABI) and the data was evaluated by using Livak method. Statistical analysis was done by using Microsoft Excel. RESULTS: There was an abnormal gene expression of p53 in all grades of the breast tumors. Non-significant (p>0.05) difference of down and up regulation of p53 in different grades of breast tumor was found. However, as a whole up-regulation was more than down-regulation with significant difference (p<0.0011). CONCLUSION: The abnormal expression of p53 shows that there are some genetic and epigenetic factors which are the primal cause of an abnormal gene expression. It is recommended that perform next generation sequencing (NGS) of the gene to find out the mutations causing the abnormal behavior of p53 gene.