Uncovering the growing burden of enteric fever: A molecular analysis of Salmonella Typhi antimicrobial resistance.

Enteric fever, a persistent public health challenge in developing regions, is exacerbated by suboptimal socioeconomic conditions, contaminated water and food sources, and insufficient sanitation. This study delves into the antimicrobial susceptibility of Salmonella Typhi, uncovering the genetic underpinnings of its resistance. Analyzing 897 suspected cases, we identified a significant prevalence of typhoid fever, predominantly in males (58.3 %) and younger demographics. Alarmingly, our data reveals an escalation in resistance to both primary and secondary antibiotics, with cases of multi-drug resistant (MDR) and extensively drug-resistant (XDR) S. Typhi reaching 14.7 % and 43.4 %, respectively, in 2021. The Multiple Antibiotic Resistance (MAR) index exceeded 0.2 in over half of the isolates, signaling widespread antibiotic misuse. The study discerned 47 unique antibiotic resistance patterns and pinpointed carbapenem and macrolide antibiotics as the remaining effective treatments against XDR strains, underlining the critical need to preserve these drugs for severe cases. Molecular examinations identified blaTEM, blaSHV, and blaCTX-M genes in ceftriaxone-resistant strains, while qnrS was specific to ciprofloxacin-resistant variants. Notably, all examined strains exhibited a singular mutation in the gyrA gene, maintaining wild-type gyrB and parC genes. The erm(B) gene emerged as the primary determinant of azithromycin resistance. Furthermore, a distressing increase in resistance genes was observed over three years, with erm(B), blaTEM and qnrS showing significant upward trends. These findings are a clarion call for robust antimicrobial stewardship programs to curtail inappropriate antibiotic use and forestall the burgeoning threat of antibiotic resistance in S. Typhi.

Efficacy of cell-free DNA as a diagnostic biomarker in breast cancer patients.

Breast cancer is the most prevalent and leading cause of mortality worldwide among women. Cell-free DNA (cfDNA) analysis is an alternative quantitative approach to conventional methods for cancer diagnosis. The current research project aimed to determine the efficacy of cfDNA as a diagnostic biomarker in breast cancer patients in Pakistan. Eighty-four female breast cancer patients were selected as cases, and 152 healthy females as controls. Immunohistochemistry was performed to identify tumor biomarkers along with clinical profiling. cfDNA was extracted from serum using the phenol-chloroform method. The cfDNA level in the serum was estimated using Agarose Gel Electrophoresis and Nanodrop. SPPS version 25.0 was used to perform statistical analyses. The results showed that the cancer biomarkers were significantly associated with breast cancer. The changes in hematological parameters were insignificant, whereas the biochemical parameter variations between the cases and controls were statistically significant. A significant association of cfDNA level with breast cancer was observed. Further cfDNA levels and cancer biomarkers were not statistically significant. A significant correlation was observed between cfDNA and biochemical parameters, except for creatinine, whereas hematological parameters showed no significant correlation.ROC analysis declared cfDNA as an authentic diagnostic marker for breast cancer. It was concluded that the level of cfDNA is significantly increased in breast cancer patients and can be utilized as a diagnostic biomarker.

Dyslipidemia and impaired liver function biomarkers in patients with hepatitis B liver cirrhosis.

Objective: This study was conducted to determine changes in lipid metabolism and liver enzyme status among HBV-positive patients with liver cirrhosis. Methods: A total of 300 HBV-positive patients with liver cirrhosis and 200 healthy controls were included in this case-control study. The patients were recruited from several tertiary care hospitals in Lahore from March to October 2021. Their blood samples were collected and analyzed for HBsAg, HBeAg, liver function biomarkers, and serum lipids. Liver cirrhosis was confirmed by ultrasonography and liver biopsy. The data were analyzed with chi-square test, Student's t-test, logistic regression, and ROC curve analysis. Results: Serum liver function biomarkers were significantly higher, and serum lipid levels were substantially lower, in HBV-infected patients with liver cirrhosis than in controls. No significant associations of sex and age with dyslipidemia were observed in patients with cirrhosis. Grading and staging scores for liver cirrhosis were negatively associated with total cholesterol levels. Moreover, sex and high levels of liver enzymes were significant risk factors associated with dyslipidemia in HBV-positive patients with liver cirrhosis. The optimum cut-off values of liver enzymes and serum lipids for the prognosis of liver cirrhosis exceeded normal ranges. Conclusion: Serum lipid concentrations may serve as a clinical index to assess liver damage in HBV-positive patients.

Genetic Diversity and Virulence Profiling of Multi-Drug Resistant Escherichia coli of Human, Animal, and Environmental Origins.

Rapid urbanization has increased human-animal interaction and consequently enhanced the chances to acquire zoonotic diseases. The current investigation is focused to uncover the genetic diversity of multidrug-resistant E. coli strains between different ecologies (i.e., humans, livestock, and environment) at the molecular level by employing antimicrobial resistance profiling, virulence genes profiling, and microbial typing approach using ERIC PCR. Based on multiple antibiotic resistance, overall, 19 antibiotic resistance patterns (R1-R19) were observed. Most of the strains (49/60) were detected to have the combinations of stx, eaeA, and hlyA genes and considered STEC/EPEC/EHEC. A total of 18 unique genetic profiles were identified based on ERIC-PCR fingerprints and most of the strains (13) belong to P1 whereas the least number of strains were showing profiles P7 and P8-P11 (one member each profile). The calculated values for Shannon index (H) for human, animal, and environment are 1.70, 1.82, and 1.78, respectively revealing the highest genetic diversity among the E. coli strains of animal origin. The study revealed that drug-resistant pathogenic E. coli strains could be transmitted bidirectionally among the environment, humans, and animals.

Assessment of serum biochemical derangements and associated risk factors of chronic kidney disease.

Objective: Chronic kidney disease and/or disturbance in renal excretory function may lead to nitrogenous waste collection beyond the term as well as derangements of several serum biochemicals. There is no previous study from Pakistan that reveals serum electrolyte derangements in confirmed chronic kidney disease (CKD) patients and other biochemicals associated with CKD. This study aims to examine the derangements of serum biochemicals and the association of several risk factors with CKD. Methods: The study enrolled 612 confirmed CKD patients with a glomerular filtration rate (GFR) < 15 ml/min that were treated as a part of the integrated care programme at Mayo Hospital Lahore (one of the largest hospitals in Pakistan). Serum biochemicals were estimated on AU 680 (Beckman Coulter) using the spectrophotometric technique. Results: All the CKD patients had elevated creatinine and urea levels, but only 63.4% were suffering from hyperuricemia. The incidence of diabetes and malnutrition assessed by serum albumin (hypoalbuminemia) was 27.4% and 72%, respectively. Among electrolyte disorders, hyperphosphatemia (71.8%) and hypocalcaemia (61.9%) were found to be more prevalent. Furthermore, gender, malnutrition, diabetes, hyperuricemia, and phosphorus and magnesium derangements were found to be statistically significant risk factors for CKD, whereas malnutrition and magnesium derangement were associated with hyperuricemia. Conclusion: It is imperative to improve dietary protein and monitor serum electrolyte concentration in renal dysfunction patients to slow the progression of CKD to end-stage renal disease (ESRD) and other serious complications.

Antibiotic Resistance Pattern Of Acinetobacter Baumannii Isolated From Bacteremia Patients In Pakistan.

BACKGROUND: Acinetobacter baumannii causes a number of life-threatening infections in Hospitalized patients attributed to its ability to develop resistance against multiple antibiotics. The current scrutinisation is aimed to observe the prevalence and antibiotic resistance profile of A. baumannii strains isolated from blood of tertiary care Hospitalized patients in Lahore, Pakistan. METHODS: This research is a retrospective study conducted over a period of one year where 1864 blood samples were collected from both male and female patients with septicaemia. Total 156 A. baumannii species were identified by conventional method and their antimicrobial resistance pattern against 22 antimicrobials (representing all known classes of antibiotics) was evaluated by Kirby Bauer disc diffusion method. MICs of colistin, polymyxin B and vancomycin against A. baumannii were calculated by E test and broth dilution method. RESULTS: More males (n=97, 62%) were found infected than females (n=59, 38%). The spreading rate of A. baumannii was highest (n=101, 65%) in patients of age ≤20 years, and lowest (n=12, 7%) in the patients with the age of 41-60 years. Most of the strains of A. baumannii (n=118, 75.6%) were found to be MDR (multi drug resistant), 37 (23.7%) strains were XDR (Extensively drug-resistant) and only 1 (0.05%) strain was PDR (pandrug resistant). All the strains were sensitive to minocycline and tigecycline whereas highest non-susceptibility (n=144, 92%) was seen against Ampicillin-Sulbactam. Most of the strains demonstrated resistance against carbapenem and cephalosporin beckoning that A. baumannii can no longer be considered for salvage therapy by carbapenem. MICs of colistin, polymyxin B and vancomycin against A. baumannii divulged polymixin B as the most effective drug. CONCLUSIONS: Use of wide range of drugs has made A. baumannii multidrug resistant. Colistin, polymyxin B and vancomycin are the preferable drugs for the treatment of A. baumannii infections.

Cadmium induces GAPDH- and- MDH mediated delayed cell aging and dysfunction in Candida tropicalis 3Aer.

Eukaryotes employ various mechanisms to survive environmental stress conditions. Multicellular organisms eliminate permanently damaged cells by apoptosis, while unicellular eukaryotes like yeast react by decelerating cell aging. In the present study, transcriptomic and proteomic approaches were employed to elucidate the underlying mechanism of delayed apoptosis. Our findings suggest that Candida tropicalis 3Aer has a set of tightly controlled genes that are activated under Cd+2 exposition. Acute exposure to Cd+2 halts the cell cycle at the G2/M phase checkpoint and activates multiple cytoplasmic proteins that overcome effects of Cd+2-induced reactive oxygen species. Prolonged Cd+2 stress damages DNA and initiates GAPDH amyloid formation. This is the first report that Cd+2 challenge initiates dynamic redistribution of GAPDH and MDH and alters various metabolic pathways including the pentose phosphate pathway. In conclusion, the intracellular redistribution of GAPDH and MDH induced by prolonged cadmium stress modulates various cellular reactions, which facilitate delayed aging in the yeast cell.

Degradation of azo dyes by Alcaligenes aquatilis 3c and its potential use in the wastewater treatment.

In the present study, Alcaligenes aquatilis was found to decolorize 82% Synazol red 6HBN after incubation of 4 days at 37 °C and pH 7. Maximum decolorization was found under static conditions by using saw dust and yeast extract as carbon and nitrogen source. It also showed promising potential to decolorize mixture of multiple dyes at a rate of more than 86% in 5 days. Decolorization of dye had positive influence on the growth of bacterium as growth rate was increased along with decolorization. The cleavage of azo bond was confirmed through TLC, HPLC and GC-MS analysis. The dye metabolites produced during bacterial treatment are linked to various pathways including ATP synthesis process. The absence of peaks of wavelength 1612/cm and 1532/cm in bacterially treated FTIR sample demonstrated the cleavage of azo bond. Microbial growth in decolorized dye wastewater shows that bacterially decolorized wastewater is unharmful for the growth of micro-flora. The high decolorization ability of A. aquatilis 3c to convert toxic azo dyes into useful end products may find potential applications in the environmental biotechnology.

Molecular basis of Cd+2 stress response in Candida tropicalis.

This study examines the bioremediation potential and cadmium-induced cellular response on a molecular level in Candida tropicalis 3Aer. Spectroscopic analysis clearly illustrated the involvement of yeast cell wall components in biosorption. Cadmium bioaccumulation was confirmed by TEM, SEM, and EDX examination. TEM images revealed extracellular as well as cytoplasmic and vacuolar cadmium nanoparticle formation, further validated by presence of ycf1 gene and increased biosynthesis of GSH under cadmium stress. Fourteen proteins exhibited differential expression and during cellular redox homeostasis are found to involve in nitrogen metabolism, nucleotide biosynthesis, and carbohydrate catabolism. Interestingly, C. tropicalis 3Aer is equipped with nitrile hydratase enzyme, rarely been reported in yeast. It has the potential to remove nitriles from the environment. The Cd+2 toxicity not only caused growth stasis but also upregulated the cysteine biosynthesis, protein folding and cytoplasmic detoxification response elements. The present study suggests that C. tropicalis 3Aer is a potential candidate for bioremediating environmental pollution by Cd+2.

Biosorption behavior and proteomic analysis of Escherichia coli P4 under cadmium stress.

Bacteria develop a variety of adaptations at transcriptomic, metabolomic and proteomic levels in order to survive potentially damaging environmental perturbations. Present study is exploring the fluctuations in proteome of E. coli P4 to knob Cd+2-induced cytotoxicity. An attempt was also made to integrate all these approaches to gain comprehensive insight of Cd+2 stress response in E. coli P4. This study is exposing the altered behavior of various proteins and their underlying metabolic pathways which have previously not been reported with reference to Cd+2 stress such as sulfoquinovose biosynthesis and degradation pathway. Some of the responses studied on all integrated levels followed same dynamics and strategies to conserve energy by down regulating carbohydrate metabolism (depicted by the repression of succinyl-CoA ligase) and growth stasis (down regulation of ftsZ). Moreover, proteomic analysis clearly revealed the affection of Cd+2 stress on various proteins expression including Rrf, MdaB, DapA, GpmA,Cdd, FabI, DsbA, ZnuA and YihW found modulating key cellular metabolic pathways enabling E. coli P4 to withstand Cd+2-induced toxic effects. Furthermore, over-expression of Mn-SOD provided evidence that Cd+2exposure induces superoxide free radicals mediated oxidative stress rather than hydrogen peroxide (H2O2). EnvZ/OmpR -a two component cell envelope regulatory system was observed operating to homeostat the cell's internal environment. Cd+2 bioremediation potential of E. coli P4 and its kinetic and thermodynamic basis were studied by applying different isotherm models which nominated E. coli P4 a good bioresource for green chemistry to eradicate environmental Cd+2.

Cadmium resistance and uptake by bacterium, Salmonella enterica 43C, isolated from industrial effluent.

Cadmium resistant bacterium, isolated from industrial wastewater, was characterized as Salmonella enterica 43C on the basis of biochemical and 16S rRNA ribotyping. It is first ever reported S. enterica 43C bared extreme resistance against heavy metal consortia in order of Pb(2+)>Cd(2+)>As(3+)>Zn(2+)>Cr(6+)>Cu(2+)>Hg(2+). Cd(2+) stress altered growth pattern of the bacterium in time dependent manner. It could remove nearly 57 % Cd(2+) from the medium over a period of 8 days. Kinetic and thermodynamic studies based on various adsorption isotherm models (Langmuir and Freundlich) depicted the Cd(2+) biosorption as spontaneous, feasible and endothermic in nature. Interestingly, the bacterium followed pseudo first order kinetics, making it a good biosorbent for heavy metal ions. The S. enterica 43C Cd(2+) processivity was significantly influenced by temperature, pH, initial Cd(2+) concentration, biomass dosage and co-metal ions. FTIR analysis of the bacterium revealed the active participation of amide and carbonyl moieties in Cd(2+) adsorption confirmed by EDX analysis. Electron micrographs beckoned further surface adsorption and increased bacterial size due to intracellular Cd(2+) accumulation. An overwhelming increase in glutathione and other non-protein thiols levels played a significant role in thriving oxidative stress generated by metal cations. Presence of metallothionein clearly depicted the role of such proteins in bacterial metal resistance mechanism. The present study results clearly declare S. enterica 43C a suitable candidate for green chemistry to bioremediate environmental Cd(2+).

Resistance and uptake of cadmium by yeast, Pichia hampshirensis 4Aer, isolated from industrial effluent and its potential use in decontamination of wastewater.

Pichia hampshirensis 4Aer is first ever used yeast for the bioremediation of environmental cadmium (Cd(+2)) which could maximally remove 22 mM/g and 28 mM/g Cd(+2) from aqueous medium at lab and large scales, respectively. The biosorption was found to be the function of temperature, pH of solution, initial Cd(+2) concentration and biomass dosage. Competitive biosorption was investigated in binary and multi-metal system which indicated the decrease in Cd(+2) biosorption with increasing the competitive metal ions attributed to their higher electronegativity and larger radius. FTIR analysis revealed the active participation of amide and carbonyl moieties in Cd(+2) adsorption confirmed by EDX analysis. Electron micrographs summoned further surface adsorption and increased cell size due to intracellular Cd(+2) accumulation. Cd(+2) was the causative agent of some metal binding proteins as well as prodigious increase in glutathione and other non-protein thiols levels which is the crucial for the yeast to thrive oxidative stress generated by Cd(+2). Our experimental data were consistent with Langmuir as well as Freundlich isotherm models. The yeast obeyed pseudo second order kinetic model which makes it an effective biosorbent for Cd(+2). High bioremediation potential and spontaneity and feasibility of the process make P. hampshirensis 4Aer an impending foundation for green chemistry to exterminate environmental Cd(+2).

Cadmium resistance mechanism in Escherichia coli P4 and its potential use to bioremediate environmental cadmium.

A cadmium-resistant bacterium was isolated from industrial wastewater and identified as Escherichia coli (dubbed as P4) on the basis of morphological, biochemical tests and 16S rRNA ribotyping. It showed optimum growth at 30 °C and pH 7. E. coli P4 found to resist Cd(+2) (10.6 mM) as well as Zn(+2) (4.4 mM), Pb(+2) (17 mM), Cu(+2) (3.5 mM), Cr(+6) (4.4 mM), As(+2) (10.6 mM), and Hg(+2) (0.53 mM). It could remove 18.8, 37, and 56 % Cd(+2) from aqueous medium after 48, 96, and 144 h, respectively. Fourier transform infrared spectroscopy (FTIR), scanning electron microscope (SEM), and Energy-dispersive X-ray (EDX) analysis also confirmed the biosorption of Cd(+2) by E. coli P4. However, temperature and pH were found to be the most critical factors in biosorption of Cd(+2) by E. coli P4. Cd(+2) stress altered E. coli P4 cell physiology analyzed by measuring glutathione (GSH) and non-protein thiol (cysteine) levels which were increased up to 130 and 48 %, respectively. Quantitative real-time polymerase chain reaction (qRT-PCR) showed alteration in the expression levels of ftsZ, mutS, clpB, ef-tu, and dnaK genes in the presence of Cd(+2). Total protein profiles of E. coli P4 in the absence and presence of Cd(+2) were compared by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), which showed remarkable difference in the banding pattern. czcB gene, a component of czcCBA operon, was amplified from genomic DNA which suggested the chromosomal-borne Cd(+2) resistance in E. coli P4. Furthermore, it harbors smtAB gene which plays a significant role in Cd(+2) resistance.

Multiple carpometacarpal volar dislocation.

A case of multiple carpometacarpal joints volar dislocation in a young man of 25 years of age is reported. He presented with pain and deformity of his left hand immediately after road traffic accident. He was unable to move his left hand. X-rays of the affected hand showed volar dislocation of all four medial carpometacarpal joints. Dislocated joints were reduced, fixed with K-wires and immobilized in slab. K-wires were removed after 8 weeks. Hand function is satisfactory on the follow-up.