Muhammad Rizwan

Objectives: The purpose of the study was to determine the prevalence of biofilm production and resistance to several antibiotics in P. aeruginosa isolated from post-surgical wound infections at a tertiary hospital. It also investigated the relationship between biofilm formation and drug resistance.Methods: The research was a cross-sectional study carried out between November 2024 and October 2025 on 116 non-duplicate P. aeruginosa isolates in patients with clinically diagnosed PSWIs. Standard microbiological techniques were used to identify isolates. The Kirby-Bauer disk diffusion technique was used to test antimicrobial susceptibility according to CLSI 2024 standards. The Tissue Culture Plate (microtiter plate) method was used to quantitatively measure biofilm production. Thestatistical analysis was conducted with SPSS 26.0, and the p-value was considered significant below 0.05.Results: Polymyxins (colistin and polymyxin B) exhibited the highest susceptibility rates (96.6% and 94.8%, respectively), while the lowest susceptibility was observed against ciprofloxacin (44.0%). Overall, 52 (44.8%; 95% CI: 35.9–53.9%) isolates were MDR, and 18 (15.5%; 95% CI: 9.5–23.5%) were XDR. Biofilm production was detected in 93 (80.2%) isolates, with 24 (20.7%) classified as strong producers. Using the Chi-square test for trend, we found a statistically significant relationship between biofilm formation and MDR (p < 0.001), with strong biofilm producers exhibiting markedly higher resistance to multiple antibiotics, including ciprofloxacin (87.5%) and meropenem (62.5%).Conclusion: The study shows high biofilm production and resistance to antimicrobials (MDR and XDR) of P. aeruginosa in post-surgical wound infections (PSWIs). This observation reveals a therapeutic dilemma, which suggests screening for biofilm and judicious use of antibiotics, along with adjunct therapies, such as biofilm-disrupting agents, to treat PSWIs.

The human gastrointestinal microbiome is a highly intricate network of trillions of biological microorganisms that include complex microbial ecosystems that are useful to the human health status by governing digestion, immunity, metabolism, and neurological health. Microbial imbalance, also known as dysbiosis, has been associated with diverse pathologies, such as gastrointestinal diseases (e.g. IBD, IBS), metabolic disorders (e.g. obesity, diabetes) and neuropsychiatric disorders. Future microbiota-based interventions including probiotics, prebiotics, synbiotics, postbiotics, and fecal microbiota transplantation (FMT) provide hopeful avenues that solve this problem by restoring microbe-biotic balance and alleviating these disorders. FMT proved to be amazing in frequent infections of Clostridioides difficile, whereas probiotics and prebiotics have revealed promise in treating IBS and metabolic disorders. There are, however, some difficulties such as the absence of clear mechanisms of actions, strain-specific variability, and the necessity of personalized methods. As well, the functions of non-bacterial microbiota (e.g., fungi, viruses) are understudied. This article will summarize the recent literature, show the current achievements in treatment, and outline research gaps in light of necessity to conduct more clinical trials and interdisciplinary research to realize the full potential of microbiome-based medicine. Filling these gaps may transform the treatment approaches to the disease of the gut and bodywide diseases.

Aims: The research aims to determine quantitative values of HBeAg and HBeAb concentrations in chronic hepatitis B patients while assessing their relationship with serum hepatitis B DNA levels. Place and Duration of Study: This study was conducted at Saidu Teaching Hospital, Swat. This study lasts for 6 months, from December 2023 to April 2024. Methodology: This study comprised 100 patients with confirmed HBV DNA positive and a chronic hepatitis B. Exclusion criteria included those using antiviral medication, those with autoimmune or metabolic liver disorders, and those with co-infections (HCV, HIV, or HDV). HBeAg and HBeAb levels were assessed using CLIA, and HBV DNA was processed using qRT-PCR. Accuracy was ensured using internal quality control measures. To evaluate the correlations between HBV DNA, HBeAg, and HBeAb, data analysis was done using SPSS Version 17.0 and Pearson correlation. P-values less than 0.05 were regarded as statistically significant. Results: The mean viral load of the 100 patients was 60,800 IU/mL, and all of them tested positive for HBV DNA. HBeAb was 8.75 IU/mL on average, while HBeAg was 12.82 IU/mL on average. HBV DNA and HBeAg were found to be 59% associated, HBV DNA and HBeAb to be 46%, and HBeAg and HBeAb to be 77%, according to the correlation study. Epidemiological insights into the distribution and correlation of HBV markers in chronic hepatitis B patients are provided by these findings. Conclusion: This study concluded that not all chronic HBV patients have HBeAg and HBeAb levels, which are lower than those of HBV DNA. These findings offer valuable information on the serological trends of chronic hepatitis B, which might aid in comprehending how the illness develops.

The current study aimed to evaluate the effect of extraction solvents on the anti-bacterial and anti-fungal efficacy of the stem and roots of Ajuga integrifolia Buch. Ham. ex D. Extracts were prepared in different solvents and tested against fungi and bacteria species including, Agrobacterium tumefaciens, Xanthomonos oryzae, Citrobacter freundi, Alternaria alternata, A. solani and Aspergillus niger. Antibacterial efficacy of the Ajuga integrifolia was carried out by disc diffusion susceptibility method and antifungal efficacy by well diffusion susceptibility method. Methanol stem extract revealed efficacy against C. freundi by producing a 63% zone of inhibition at 3000µg/disc while methanol roots extract produced 77% ZI against A. tumifaciens. Methanol root and stem extracts produced an equal zone of inhibition (97%) at 1000 µg/ml concentration against A. alternata and A. solani. The results of the study clearly stated that the polarity of the solvents used in the extraction procedure affects the bioactivities of the extracts.

Nanotechnology is a research hotspot that has gained considerable interest due to its potential inferences in the bioscience, medical, and engineering disciplines. The present study uses biomass from the Enterobacter hormaechei EAF63 strain to create bio-inspired metallic tin oxide nanoparticles (SnO₂ NPs). The biosynthesized NPs were extensively analyzed using UV spectroscopy, X-ray diffraction (XRD), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), and Fourier transform infrared (FTIR) techniques. The identification of the crystalline phase was confirmed by XRD. The SEM technique elucidated the morphological characteristics and size of SnO₂ NPs. SEM investigation revealed that the SnO₂ NPs have a size of 10 nm with spherical morphology. The capping of NPs was confirmed by FTIR analysis that revealed the presence of different compounds found in the biomass of the E. hormaechei EAF63 strain. Later, EDX confirmed the elemental composition of NPs. Moreover, the synthesized SnO₂ NPs were employed for important applications including anti-aging, anti-Alzheimer's, anti-inflammatory, anti-larvicidal, and antibacterial action against sinusitis pathogens. The highest value was observed for Streptococcus pyogenes (19.75 ± 0.46), followed by Moraxella catarrhalis (17.49 ± 0.82) and Haemophilus influenzae (15.31 ± 0.73), respectively. Among the used concentrations, the highest inhibition of 76.8 ± 0.93 for 15-lipoxygenase (15-LOX) was observed at 400 μg/mL, followed by 67.4 ± 0.91 for cyclooxygenase-1 (COX-1). So, as an outcome, E. hormaechei-mediated SnO₂ NPs might be considered as the safe and effective nanoplatforms for multifunctional biological applications in the field of nanomedicine.

The current study aims to utilize the bacteria Paraclostridium benzoelyticum strain 5610 to synthesize bio‐genic silver nanoparticles (AgNPs). Biogenic AgNPs were thoroughly examined using various characterization techniques such as UV‐spectroscopy, XRD, FTIR, SEM, and EDX. Synthesis of AgNPs was confirmed by UV–vis analysis resulting in absorption peak at 448.31 nm wavelength. The SEM analysis indicated the morphological characteristics and size of AgNPs which was 25.29 nm. The face centered cubic (FCC) crystallographic structure was confirmed by XRD. Furthermore, FTIR study affirmed the capping of AgNPs by different compounds found in biomass of the Paraclostridium benzoelyticum strain 5610. Later, EDX was used to determine the elemental composition with respective concentration and distribution. Additionally, in the current study the antibacterial, anti‐inflammatory, antioxidant, anti‐aging, and anti‐cancer ability of AgNPs was assessed. The antibacterial activity of AgNPs was tested against four distinct sinusitis pathogens: Haemophilus in‐fluenza, Streptococcus pyogenes, Moraxella catarrhalis and Streptococcus pneumonia. AgNPs shows significant inhibition zone against Streptococcus pyogenes 16.64 ± 0.35 followed by 14.32 ± 071 for Moraxella catarrhalis. Similarly, the antioxidant potential was found maximum (68.37 ± 0.55%) at 400 μg/mL and decrease (5.48 ± 0.65%) at 25 μg/mL, hence the significant antioxidant ability was observed. Furthermore, anti‐inflammatory activity of AgNPs shows the strongest inhibitory action (42.68 ± 0.62%) for 15‐LOX with lowest inhibition activity for COX‐2 (13.16 ± 0.46%). AgNPs have been shown to exhibit significant inhibitory actions against the enzyme elastases AGEs (66.25 ± 0.49%), which are followed by AGEs of visperlysine (63.27 ± 0.69%). Furthermore, the AgNPs show high toxicity against HepG2 cell line which shows 53.543% reduction in the cell viability after 24 h of treatment. The anti‐inflammatory activity demonstrated a potent inhibitory effect of the bio‐inspired AgNPs. Overall, the biogenic AgNPs have the ability to be served for the treatments of anti‐aging and also due to their anti‐cancer, antioxidant abilities NPs may be a useful therapy choice for a variety of disorders including cancer, bacterial infections and other inflammatory diseases. Moreover, further studies are required in the future to evaluate their in vivo biomedical applications.Highlights: Biogenic synthesis of AgNPs using Paraclostridium benzoelyticum Strain for the first time. FTIR analysis confirmed capping of potent biomolecules which are of great use in applied field especially Nanomedicines. Notable antimicrobial activity against sinusitis bacteria and cytotoxic potential of synthesized AgNPs on in vitro basis produce a new idea shifting us to treat cancerous cell lines.

Syphilis, a sexually transmitted infection, is a deadly disease caused by Treponema pallidum. It is a Gram-negative spirochete that can infect nearly every organ of the human body. It can be transmitted both sexually and perinatally. Since syphilis is the second most fatal sexually transmitted disease after AIDS, an efficient vaccine candidate is needed to establish long-term protection against infections by T. pallidum. This study used reverse-vaccinology-based immunoinformatic pathway subtractive proteomics to find the best antigenic proteins for multi-epitope vaccine production. Six essential virulent and antigenic proteins were identified, including the membrane lipoprotein TpN32 (UniProt ID: O07950), DNA translocase FtsK (UniProt ID: O83964), Protein Soj homolog (UniProt ID: O83296), site-determining protein (UniProt ID: F7IVD2), ABC transporter, ATP-binding protein (UniProt ID: O83930), and Sugar ABC superfamily ATP-binding cassette transporter, ABC protein (UniProt ID: O83782). We found that the multiepitope subunit vaccine consisting of 4 CTL, 4 HTL, and 11 B-cell epitopes mixed with the adjuvant TLR-2 agonist ESAT6 has potent antigenic characteristics and does not induce an allergic response. Before being docked at Toll-like receptors 2 and 4, the developed vaccine was modeled, improved, and validated. Docking studies revealed significant binding interactions, whereas molecular dynamics simulations demonstrated its stability. Furthermore, the immune system simulation indicated significant and long-lasting immunological responses. The vaccine was then reverse-transcribed into a DNA sequence and cloned into the pET28a (+) vector to validate translational activity as well as the microbial production process. The vaccine developed in this study requires further scientific consensus before it can be used against T. pallidum to confirm its safety and efficacy.

Nanotechnology is a new field that has gained considerable importance due to its potential uses in the field of biosciences, medicine, engineering, etc. In the present study, bio-inspired metallic iron nanoparticles (FeNPs) were prepared using biomass of Enterobacter train G52. The prepared particles were characterized by UV-spectroscopy, TGA, XRD, SEM, EDX, and FTIR techniques. The crystalline nature of the prepared FeNPs was confirmed by XRD. The SEM techniques revealed the particles size to be 23 nm, whereas in FTIR spectra the peaks in the functional group region indicated the involvement of bioactive compounds of selected bacterial strains in the capping of FeNPs. The EDX confirmed the presence of iron in the engineered FeNPs. The FeNPs were then evaluated for its antibacterial, antifungal, antioxidant, anti-inflammatory, anti-Alzheimer’s, anti-larvicidal, protein kinase inhibition, anti-diabetic, and biocompatibility potentials using standard protocols. Substantial activities were observed in almost all biological assays used. The antioxidant, anti-cholinesterase, and anti-diabetic potential of the prepared nanoparticles were high in comparison to other areas of biological potential, indicating that the FeNPs are capable of targeting meditators of oxidative stress leading to diabetes and Alzheimer’s disease. However, the claim made needs some further experimentation to confirm the observed potential in in vivo animal models.

The apple is the most widely used fruit globally. Apples are more prone to fungal spoilage, which leads to browning and subsequent changes in their flavor and texture. Browning is also caused by the tyrosinase enzyme. By inhibiting tyrosinase initiation and fungal spoilage in fruits, the natural flavor and texture of fruits can be maintained. Biogenic NPs can act as antioxidants to inhibit tyrosinase and due to oxidative stress, it also catalyzes the deformation of fungal hyphae and spores. Nanotechnology is a research hotspot that has gained considerable interest due to its potential inferences in biosciences and food preservation technology. The present study aims to use biomass from the Fagonia cretica to create bio-inspired manganese oxide MnO₂ NPs and to evaluate its bio-catalytic potential for antifungal anti-browning through the inhibition of tyrosinase and its antioxidant potential for preserving apple flavor and texture. The green synthesized nanoparticles were extensively analyzed using UV spectroscopy, XRD, SEM, EDX, and FTIR techniques. Moreover, the synthesized manganese oxide nanoparticles (MnO₂ NPs) were evaluated for their bio-catalytic potential as anti-fungal and anti-spoiling agents. The values of antifungal activity among all the samples were 14.2 ± 86 mm, 8.9 ± 6.0 mm, 17.7 ± 1.26, and 20.7 ± 4.38 mm for Penicillium expansum, Monilinia fructigena, Penicillium chrysogenum, and Aspergillus oryzae at 200 µg/well, respectively. Moreover, the biogenic NPs were evaluated for their anti-browning potential through the inhibition of tyrosinase. MnO₂ NPs have been shown to have considerable inhibitory effects on tyrosinase up to 64.8 ± 0.16 at 200 µg/mL and (27.2 ± 0.58) at 25 µg/mL. Biogenic MnO₂ NPs can also act as antioxidants to inhibit tyrosinase and fungal growth by the formation of free radicals that damage the fungal hyphae and, as a result, slow down browning. The maximum DPPH free radical scavenging activity was 74.5 ± 0.39% at 200 µg/mL, and the minimum was 12.4 ± 0.27 at 25 µg/mL. The biogenic MnO₂ NPs are biocompatible and play a potent role in maintaining the flavor and texture of apples.