Taj Ali Khan

Impact of geographical locations on Mentha spicata antibacterial activities

Department of Pharmacy, Faculty of Life Sciences, Sarhad University of Science and Information Technology, Peshawar, Pakistan. Department of Microbiology, Kohat University of Science and Technology, Kohat, Pakistan. Department of Pharmacology, Kohat Institute of Medical Sciences, Kohat, Pakistan. Department of Medicine, University of Toronto, Canada. Department of Oncology, Hayatabad Medical Complex, Peshawar, Pakistan.

Effects of Luteolin and Quercetin in Combination with Some Conventional Antibiotics against Methicillin-Resistant Staphylococcus aureus

The present study was designed to evaluate the effects of flavonoids luteolin (L) and quercetin + luteolin (Q + L) in combination with commonly used antibacterial agents against methicillin-resistant Staphylococcus aureus (MRSA) clinical isolates and S. aureus (ATCC 43300). Minimum inhibitory concentrations (MICs) of L and Q + L, as well as the MICs of flavonoids in combination with antibiotics were determined and results showed an increased activity of flavonoids with antibiotics. The synergistic, additive, or antagonistic relationships between flavonoids (L and Q + L) and antibiotics were also evaluated, and additive and synergistic effects were observed for some antibiotic + flavonoid combinations. In addition, some combinations were also found to damage the bacterial cytoplasmic membrane, as assessed through potassium leakage assay. The effects of flavonoids and flavonoids + antibiotics on mecA gene mutations were also tested, and no functional variation was detected in the coding region.

Interferon-Gamma Improves Macrophages Function against M. tuberculosis in Multidrug-Resistant Tuberculosis Patients

Background. Mycobacterium tuberculosis (M. tuberculosis) that causes tuberculosis (TB) kills millions of infected people annually especially multidrug-resistant tuberculosis (MDR-TB). On infection, macrophages recognize the mycobacteria by toll-like receptor (TLR) followed by phagocytosis and control of mycobacteria. In addition, macrophages also secrete IL-12 to induce IFN-γ production by T, which, in turn, increases the phagocytosis and oxidative burst. Individuals with defects in innate or adaptive immunity exhibit increased susceptibility to M. tuberculosis. Understanding these immunologic mechanisms will help in TB control. We aimed to investigate the immunopathologic mechanisms in MDR-TB and role of recombinant human interferon-gamma (rhIFN-γ). Study Design and Methods. Monocyte-derived macrophages (MDMs) were generated from peripheral blood mononuclear cells of MDR-TB patients and healthy subjects and were investigated for immunologic response by ELISA and flow cytometry. Results. Different functional and molecular anomalies were observed in macrophages. In addition, a defective immune response to M. tuberculosis from the patients MDMs was characterized, which in turn improved by pretreatment with rhIFN-γ. Conclusion. This work highlights the fact that rhIFN-γ improves macrophages function against M. tuberculosis and treatment of patients with poor responsiveness to TB therapy may be needed in future to include IFN-γ as adjuvant therapy after the full characterization of pathological and molecular mechanisms in these and in other more multidrug-resistant TB patients.

Expanding the clinical and genetic spectrum of G6PD deficiency: The occurrence of BCGitis and novel missense mutation

Glucose-6-phosphate dehydrogenase (G6PD) is a key enzyme in the pentose phosphate pathway that ensures sufficient production of coenzyme nicotinamide adenine dinucleotide phosphate (NADPH) by catalyzing the reduction of NADP+ to NADPH. Noteworthy, the latter mediates the production of reactive oxygen species (ROS) by phagocytic cells such as neutrophils and monocytes. Therefore, patients with severe forms of G6PD deficiency may present impaired NADPH oxidase activity and become susceptible to recurrent infections. This fact, highlights the importance to characterize the immunopathologic mechanisms underlying the susceptibility to infections in patients with G6PD deficiency. Here we report the first two cases of G6PD deficiency with Bacille Calmette-Guérin (BCG) adverse effect, besides jaundice, hemolytic anemia and recurrent infections caused by Staphylococcus aureus. The qualitative G6PD screening was performed and followed by oxidative burst analysis using flow cytometry. Genetic and in silico analyses were carried out by Sanger sequencing and mutation pathogenicity predicted using bioinformatics tools, respectively. Activated neutrophils and monocytes from patients displayed impaired oxidative burst. The genetic analysis revealed the novel missense mutation c.1157T>A/p.L386Q in G6PD. In addition, in silico analysis indicated that this mutation is pathogenic, thereby hampering the oxidative burst of neutrophils and monocytes from patients. Our data expand the clinical and genetic spectrum of G6PD deficiency, and suggest that impaired oxidative burst in this severe primary immune deficiency is an underlying immunopathologic mechanism that predisposes to mycobacterial infections.

First report on molecular characterization of Leishmania species from cutaneous leishmaniasis patients in southern Khyber Pakhtunkhwa province of Pakistan

OBJECTIVE To report presence of Leishmania major in Khyber Pakhtunkhwa of Pakistan, where cutaneous leishmaniasis (CL) is endemic and was thought to be caused by Leishmania tropica only. METHODS Biopsy samples from 432 CL suspected patients were collected from 3 southern districts of Khyber Pakhtunkhwa during years 2011-2016. Microscopy on Giemsa stained slides were done followed by amplification of the ribosomal internal transcribed spacer 1 gene. RESULTS Leishmania amastigotes were detected by microscopy in 308 of 432 samples (71.3%) while 374 out of 432 samples (86.6%) were positive by ribosomal internal transcribed spacer 1 PCR. Subsequent restriction fragment length polymorphism confirmed L. tropica in 351 and L. major in 6 biopsy samples. CONCLUSIONS This study is the first molecular characterization of Leishmania species in southern Khyber Pakhtunkhwa. It confirmed the previous assumptions that anthroponotic CL is the major CL form present in Khyber Pakhtunkhwa province. Furthermore, this is the first report of L. major from a classical anthroponotic CL endemic focus identified in rural areas of Kohat district in southern Khyber Pakhtunkhwa.

Antimicrobial resistance among aerobic biofilm producing bacteria isolated from chronic wounds in the tertiary care hospitals of Peshawar, Pakistan

OBJECTIVE Chronic wound infections impose major medical and economic costs on health-care systems, cause significant morbidity, mortality and prolonged hospitalisation. The presence of biofilm producing bacteria in these wounds is considered as an important virulence factor that leads to chronic implications including ulceration. The undertaken study aimed to isolate and identify the biofilm aerobic bacterial pathogens from patients with chronic wound infections, and determine their antibiotics resistance profiles Method: During this study, swab specimens were collected from patients with chronic wounds at teaching hospitals of Peshawar, Pakistan between May 2013 and June 2014. The isolated aerobic bacterial pathogens were identified on the basis of standard cultural characteristics and biochemical tests. Antibiotics resistance profiles of biofilm producing bacteria against selected antibiotics were then determined. RESULTS Among the chronic wound infections, diabetic foot ulcers were most common 37 (37%), followed by surgical ulcers 27 (27%). Chronic wounds were common in male patients older than 40 years. Among the total 163 isolated bacterial pathogens the most prevalent bacterial species were Pseudomonas aeruginosa 44 (27%), Klebsiella pneumoniae 26 (16%), Staphylococcus species 22 (14%) and Streptococcus spp. 21 (13%). The isolation rate of bacterial pathogens was high among patients with diabetic foot ulcers 83 (50.9%). Among bacterial isolates, 108 (66.2%) were observed as biofilm producers while 55 (33.8%) did not form biofilm in our model. The investigated biofilm producing bacterial isolates showed comparatively high resistance against tested antibiotics compared to non-biofilm producing bacterial isolates. The most effective antibiotics were amikacine and cefepime against all isolates. CONCLUSION Increased multidrug resistance in biofilm producing bacteria associated with chronic wounds was observed in this study. Judicious use of antibiotics is needed to control the wound associated biofilm associated pathogens.

Impact of water losses and maintenance of canal irrigation system on agriculture (case study: Urmar Minor of Warsak Gravity Canal Pakistan).

A study was conducted in May-August 2005 to assess the conveyance losses, seepage losses, maintenance status and impact of water losses and maintenance on the agricultural yield in case of irrigation channels. In this regard, a case study was conducted on three watercourses, at Urmar Minor of Warsak Gravity Canal irrigation system in Pakistan. The Inflow-outflow and pondingmethods were used to determine conveyance and seepage losses respectively. Local farmers were interviewed to inquire about the maintenance condition of the selected water courses. A considerable decrease i.e. more than 50% in the yield of wheat and maize was observedwhile moving along the channels from head to tail. The irrigation system of Urmar Minor is evaluated as a guiding study and remedial measures are suggested to handle widely spread reduction in agricultural production due to these losses.

Epidemiology of Cutaneous Leishmaniasis Outbreak, Waziristan, Pakistan

During 2013–2015, prevalence of cutaneous leishmaniasis in war-affected Waziristan areas was 3.61% by PCR. Youths (1–15 years of age) were more susceptible. Internal transcribed spacer 1 PCR followed by restriction fragment length polymorphism analysis identified Leishmania tropica in 215 samples and Leishmania major in 6 samples.

Molecular detection and antimicrobial resistance profile of zoonotic Salmonella enteritidis isolated from broiler chickens in Kohat, Pakistan

Background Salmonella enteritidis infection is a frequently encountered zoonotic problem, occurring with concerning regularity in recent years on a worldwide basis. The study that we undertook for the first time detected S. enteritidis and associated antimicrobial resistance pattern in broiler chickens. Methods A total of 150 different poultry samples were first enriched and grown on selective media, and then processed for molecular detection of S. enteritidis by amplification of the spvb gene. Results The overall detection rate of S. enteritidis was 23.3% (n = 35), while an increased detection rate of S. enteritidis was found in the chicken breast tissue (n = 9; 30%). When antibiogram was tested for S. enteritidis against common antibiotics, increased resistance to ampicillin (n = 29; 82.2%), tetracycline (n = 28; 80%), augmentin (n = 27; 77.14%), and chloramphenicol (n = 19; 54.2%) was observed. Multidrug resistance was reported in 54.8% (n = 19) of the S. enteritidis isolates, while 20% (n = 07) of isolates were extensively drug resistant. Conclusion The present study for the first time reports S. enteritidis on the basis of spvb gene detection. The increased drug resistance in S. enteritidis is an emerging problem that could negatively impact efforts to prevent and treat broiler‐transmitted S. enteritidis.

Induction of Hypergammaglobulinemia and Autoantibodies by Salmonella Infection in MyD88-Deficient Mice

Growing evidence indicates a link between persistent infections and the development of autoimmune diseases. For instance, the inability to control Salmonella infection due to defective toll-like receptor (TLR)/myeloid differentiation primary response 88 (MyD88) signaling has linked the development of persistent infections to a breakdown in B cell tolerance. However, the extent of immune dysregulation in the absence of TLR-MyD88 signaling remains poorly characterized. Here, we show that MyD88−/− mice are unable to eliminate attenuated Salmonella enterica serovar Typhimurium, even when challenged with a low-dose inoculum (200 CFUs/mouse), developing a persistent and progressive infection when compared to wild-type (MyD88+/+) animals. The splenic niche of MyD88−/− mice revealed increased counts of activated, Sca-1-positive, myeloid subpopulations highly expressing BAFF during persistent Salmonella infection. Likewise, the T cell compartment of Salmonella-infected MyD88−/− mice showed increased levels of CD4+ and CD8+ T cells expressing Sca-1 and CD25 and producing elevated amounts of IL-4, IL-10, and IL-21 in response to CD3/CD28 stimulation. This was associated with increased Tfh cell differentiation and the presence of CD4+ T cells co-expressing IFN-γ/IL-4 and IFN-γ/IL-10. Noteworthy, infected MyD88−/− mice had enhanced serum titers of both anti-Salmonella antibodies as well as autoantibodies directed against double-stranded DNA, thyroglobulin, and IgG rheumatoid factor, positive nuclear staining with HEp-2 cells, and immune complex deposition in the kidneys of MyD88−/− mice infected with live but not heat-killed Salmonella. Infection with other microorganisms (Acinetobacter baumanii, Streptococcus agalactiae, or Escherichia coli) was unable to trigger the autoimmune phenomenon. Our findings suggest that dysregulation of the immune response in the absence of MyD88 is pathogen-dependent and highlight potentially important genotype–environmental factor correlations.