Background Escherichia coli lineage ST131 predominates across various spectra of extra-intestinal infections, including urinary tract infection (UTI). The distinctive resistance profile, diverse armamentarium of virulence factors and rapid global dissemination of ST131 E. coli makes it an intriguing pathogen. However, not much is known about the prevalence and genetic attributes of ST131 lineage in Pakistan. Methods We estimated prevalence and genetic attributes of E. coli ST131 isolates causing UTI among 155 randomly selected samples. Samples were analyzed for phylogenetic grouping, O-typing and fum C/ fim H typing. Isolates were further tested for the ESBL and virulence factors using PCR. Results Overall, 59% of the UPEC isolates belonged to the phylogenetic group B2, followed by D = 28%, B1 = 8% and A = 5%. Among 18 different Sequence-types, ST131 was the dominant lineage ( n = 71; 46%) out of which 72% of the isolates were assigned to the phylogenetic group B2, while 61% adhered to the serogroup O25b. Fum C/ fim H typing confirmed 49% of the ST131 as H30 sub-types. In this study, significant numbers of the identified ST131 isolates were MDR and 42% showed ESBL phenotypes, out of which 37% carried bla - CTX-M-15 . Moreover, different virulence factors were detected in following percentages: fim H,155(100%), iut A 86 (55%), feo B 76 (49%), pap C 75 (48%), papGII 70 (45%), kpsMTII 40 (26%), pap EF 37 (24%), fyu A 37 (24%), usp 22 (14%), pap A 20 (13%), sfa/foc 20 (13%), hly A 18 (12%), afa 15 (10%), cdt B 11 (7%), papGI 6 (4%), papGIII 6 (4%), kpsMTIII 4 (3%) and bma E2 (1%). Conclusion Conclusively, this study provides important insight into the genetic and virulence attributes of pandemic MDR ST131 strains involved in UTIs. It also highlights higher prevalence of ST131-O25b-H30 UPEC isolates in patients, which was previously unreported from this part of globe.
Intellectual disability (ID) is a condition of significant limitation of cognitive functioning and adaptive behavior, with 50% of etiology attributed to genetic predisposition. We recruited two consanguineous Pakistani families manifesting severe ID and developmental delay. The probands were subjected to whole exome sequencing (WES) and variants were further prioritized based on population frequency, predicted pathogenicity and functional relevance. The WES data analysis identified homozygous pathogenic variants in genes MBOAT7 and TRAPPC9. The pathogenicity of the variants was supported by co-segregation analysis and in silico tool. The findings of this study expand mutation spectrum and provide additional evidence to the role of MBOAT7 and TRAPPC9 in causation of ID.
The current research aimed to evaluate in vitro biological activities of green-synthesized silver nanoparticles using the Allium sativum clove extract. The stability of green-synthesized silver nanoparticles was evaluated via storage at 4°C, room temperature (37°C), and calcined at 300°C, 500°C, and 700°C. The antibacterial effect was evaluated using agar well, spread plate, biofilm reduction, and cell proliferation inhibition assays. The cytotoxic and antidiabetic effects were determined via brine shrimp lethality, protein kinase inhibition, and α-amylase inhibition assays. DPPH scavenging, iron-chelating, anticoagulant, and hemolytic effects were evaluated. The highest inhibition of Klebsiella pneumoniae was observed when freshly prepared, calcined (300°C), and stored nanoparticles (4°C and 37°C) were applied (9.66, 9.55, 7.33, and 6.65 mm) against freshly prepared and calcined at 700°C which showed the highest inhibition of Pseudomonas aeruginosa (8.55 and 7.66 mm). Cell viability assay, biofilm reduction assay, and spread plate method showed a significant antibacterial effect of freshly prepared silver nanoparticles. Freshly prepared and calcined nanoparticles at 300°C and 500°C possessed strong antioxidant and iron-chelating activity. Among all the synthesized silver nanoparticles, freshly prepared and calcined nanoparticles (300°C and 500°C) increases the prothrombin time. Silver nanoparticles possessed significant anticoagulant properties and less toxic at least concentration toward human RBCs. In brine shrimp lethality assay, freshly prepared nanoparticles showed a stronger toxic effect and caused high mortality of larvae. Protein kinase inhibition assay revealed that freshly prepared nanoparticles had the highest zone of inhibition (18.0 mm) at 50 µg/disc. Green-synthesized nanoparticles would be used as potential therapeutic agents to overcome both infectious and noninfectious diseases.
Background Escherichia coli lineage ST131 predominates across various spectra of extra-intestinal infections, including urinary tract infection (UTI). The distinctive resistance profile, diverse armamentarium of virulence factors and rapid global dissemination of ST131 E. coli makes it an intriguing pathogen. However, not much is known about the prevalence and genetic attributes of ST131 lineage in Pakistan. Methods We estimated the prevalence and genetic attributes of E. coli ST131 isolates causing UTI among 155 randomly selected samples. Samples were analyzed by phylogenetic grouping, O-typing, fumC/fimH typing. Isolates were further tested for ESBL and virulence factors using PCR. Results Overall, 59% of the UPEC isolates belonged to the phylogenetic group B2, followed by D=28%, B1= 8% and A= 5%. Among 18 different Sequence-types, ST131 was the dominant lineage (n=71; 46%) out of which 72% of the isolates were assigned to phylogenetic group B2 while 61% adhered to serogroup O25b. FumC/fimH typing confirmed 49% of the ST131 as H30 sub-types. In this study, significant numbers of the identified ST131 isolates were MDR and 42% showed ESBL phenotypes, out of which 57% carried blaCTX-M-15. Moreover, different virulence factors were detected in following percentages: fimH,155 (100%), iutA 86 (55%), feoB 76 (49%), papC 75 (48%), papGII 70 (45%), kpsMTII 40 (26%), papEF 37 (24%), fyuA 37 (24%), usp 22 (14%), papA 20 (13%), sfa/foc 20 (13%), hlyA 18 (12%), afa 15 (10%), cdtB 11 (7%), papGI 6 (4%), papGIII 6 (4%), kpsMTIII 4 (3%) and bmaE2 (1%). Conclusion Conclusively, this study provides important insights into the genetic and virulence attributes of pandemic MDR ST131 strains involved in UTIs. It also highlights high prevalence of ST131-O25b-H30 UPEC isolates in local population, which was previously unreported from this part of globe.
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