A common presentation of uncommon disease - Biliary Fasciola Hepatica.

Hepatobiliary fascioliasis is one of the rare but important parasitic infection in endemic areas such as Pakistan. Clinical manifestations overlap with common clinical conditions such as hepatitis and biliary stones causing common bile duct obstruction. Fascioliasis is a zoonotic infection caused by a leaf-shaped organism known as Fasciola Hepatica. Humans are accidentally infected by consuming contaminated water or food, especially undercooked cattle and sheep liver or raw green vegetables. We report three cases with similar clinical manifestations presenting in two different cities in Pakistan. After detailed clinical evaluation and laboratory investigations, the diagnosis was established by abdominal ultrasound and diagnostic and therapeutic cholangiography. Patients were followed till complete resolution of their symptoms and normalisation of liver function tests.

Predictive Potential Of Il28b Gene In Hcv Patients, Resistant To Daclatasvir And Sofosbuvir In KPK Population.

Background: Recently various combinations of direct acting antivirals (DAAs) have been tried successfully. The Sofosbuvir + Daclatasvir combination has been used with promising results. Recently, resistance has been noticed against DAAs. Therefore, polymorphism at particular sites in the interleukin 28B gene are under study to find possible association with resistance. This study was aimed at finding out any association of SNPs rs8099917 and rs12979860 (IL28B gene) with response and resistance to treatment in HCV genotype 3 patients in Khyber Pakhtunkhwa. Methods: This cross sectional, Analytical study was conducted at Gastroenterology/hepatology OPD of Prime Teaching Hospital, Peshawar Medical College. Collected Samples were stored at -20o C in PCR Lab of the College. DNA extraction and genotyping was carried out at BJ Molecular Biology Lab in Rawalpindi. Data was analyzed by using SPSS version 21. Chi-Square Test was used to see the statistically significant differences between rs8099917 T/G and rs12979860 T/C model. Results: In the IL28-B gene, single nucleotide polymorphism at rs12979860 T/C model, we observed that there are 37.5% CC homozygous, 12.5% TT homozygous and 50% CT heterozygous genotypes in resistant patients and 42.85% CC homozygous, 28.57% TT and 28.57% CT genotype in responder group. In rs12979860 T/C model, genotype of IL28-B in the responder and resistant group significantly varies at p-value =0.00572. Conclusion: We conclude that in SNP at rs12979860, CC genotype is associated with clearance of HCV, while CT genotype was more prevalent in the resistant group and associated with chronicity.

Interaction of LysM BON family protein domain with carbapenems: A putative mechanism of carbapenem resistance.

Carbapenem resistance in Gram-negative pathogens has become a global concern for health workers worldwide. In one of our earlier studies, a Klebsiella pneumoniae-carbapenemase-2 producing strain was induced with meropenem to explore differentially expressed proteins under induced and uninduced conditions. There is, LysM domain BON family protein, was found over 12-fold expressed under the induced state. A hypothesis was proposed that LysM domain protein might have an affinity towards carbapenem antibiotics making them unavailable to bind with their target. Hence, we initiated a study to understand the binding mode of carbapenem with LysM domain protein. MICs of imipenem and meropenem against LysM clone were increased by several folds as compared to NP-6 clinical strain as well as DH5 α (PET-28a KPC-2) clone. This study further revealed a strong binding of both antibiotics to LysM domain protein. Molecular simulation studies of LysM domain protein with meropenem and imipenem for 80 ns has also showed stable structure. We concluded that overexpressed LysM domain under induced condition interacted with carbapenems, leading to enhanced resistance as proved by high MIC values. Hence, the study proved the proposed hypothesis that the LysM domain plays a significant role in the putative mechanism of antibiotics resistance.

Proteomic analysis of a carbapenem-resistant Klebsiella pneumoniae strain in response to meropenem stress.

OBJECTIVES: Antibiotic resistance has become a major problem in treating bacterial infections. The aim of this study was to elucidate the effects of meropenem on a blaKPC-2-harbouring multidrug-resistant clinical strain of Klebsiella pneumoniae through a proteomics approach in order to attain a deeper understanding of bacterial resistance strategies. METHODS: Analysis was performed by two-dimensional gel electrophoresis of whole-cell extracts of bacteria exposed to a sublethal concentration of meropenem compared with the untreated control. Differentially expressed proteins were identified by matrix-assisted laser desorption/ionisation time-of-flight (MALDI-TOF). RESULTS: Based on Quantity One® software and MALDI-TOF analysis, 16 overexpressed proteins were identified in meropenem-treated bacteria. These proteins were primarily enzymes involved in defence against oxidative stress as well as glycolytic enzymes. LysM domain/BON superfamily protein was found overexpressed by >12-fold. STRING-10 was used to determine protein-protein interaction among the overexpressed proteins and to predict their functional associations. This study demonstrated that treatment with meropenem resulted in upregulation of various proteins involved in defence and repair mechanisms along with enzymes of energy metabolism. CONCLUSIONS: These overexpressed proteins may play an important role in bacterial resistance mechanisms against carbapenems, however their role in resistance needs to be further validated. High expression of lysine M domain/BON superfamily protein may indicate its possible involvement in modulating the bacterial response to antibiotic stress, but its actual role requires more investigation. These findings may also help in the development of newer therapeutic agents or diagnostic markers against carbapenem resistance.

Evaluation of inhibitory action of novel non ß-lactam inhibitor against Klebsiella pneumoniae carbapenemase (KPC-2).

The use of three classical ß-lactamase inhibitors (Clavulanic acid, tazobactam and sulbactam) in combination with ß-lactam antibiotics is presently the mainstay of antibiotic therapy against Gram-negative bacterial infections. However these inhibitors are unable to inhibit carbapenemase KPC-2 effectively. They being ß-lactam derivatives behave as substrates for this enzyme instead of inactivating it. We have initiated our study to check the in vitro inhibition activity of the two novel screened inhibitors (ZINC01807204 and ZINC02318494) in combination with carbapenems against KPC-2 expressing bacterial strain and their effect on purified enzyme KPC-2. The MIC values of meropenem and ertapenem showed maximum reduction (8 folds) in combination with screened compounds (ZINC01807204 and ZINC02318494). CLSM images also depicted their strong antibacterial activity in comparison to conventional ß-lactamase inhibitors. Moreover no toxic effect has been shown on HeLa cell line. Though the IC50 value of ZINC01807204 was high (200 µM), it exhibited fairly good affinity for KPC-2 (Ki = 43.82 µM). With promising results this study identifies ZINC01807204 as a lead molecule for further optimization and development of more potent non ß-lactam inhibitors against KPC-2.

Structure-based screening of inhibitors against KPC-2: designing potential drug candidates against multidrug-resistant bacteria.

KPC-2 ß-lactamase demonstrates a wide substrate spectrum that includes carbapenamases, oxyimino-cephalosporins, and cephamycins. In addition, strains harboring KPC-type ß-lactamases are often identified as resistant to standard ß-lactamase inhibitors. Thus, KPC-2 carbapenems present a significant clinical challenge, as the mechanistic bases for KPC-2-associated phenotypes remain mysterious. Inhibiting the function of these resistance enzymes could control the hydrolysis of antibiotics. In the present study, we have reported two novel (non-ß-lactatam) compounds that inhibit the activity of the KPC-2 enzyme. These compounds were identified by structure-based virtual screening using computational docking programs and molecular dynamics simulations with the solved crystal structure. Two compounds (ZINC01807204 and ZINC02318494) were selected on the basis of fitness scores from docking program and 5 ns molecular dynamics simulations. These commercially available compounds have been procured and their biological activity was experimentally evaluated on the E. coli strain carrying recombinant KPC-2. These new compounds in combination with ceftazidime and cefoxitin exhibited the Minimum Inhibitory Concentration (MIC) values of 2 and 8 µg/ml respectively, which were found to be lower as compared to known ß-lactamase inhibitors. Moreover, these compounds were also found to have comparable MICs values being 64 µg/ml in combination with ceftriaxone. This study explored novel inhibitors against KPC-2, a class A ß-lactamase, which may be putative drug candidates against KPC-2 producing bacterial infection.

Development, characterization and efficacy of niosomal diallyl disulfide in treatment of disseminated murine candidiasis.

In the current study, a novel niosome based formulation of diallyl disulfide (DADS) was evaluated for its potential to treat disseminated candidiasis in mouse model. Among various non-ionic surfactants tested, niosome formulation prepared using Span 80 was found to be most efficient in the entrapment of DADS. The DADS loaded niosomes had size dimensions in the range of 140 ± 30 nm with zeta potential of -30.67 ± 4.5. Liver/kidney function tests as well as histopathologic studies suggested that noisome-based DADS formulations are safe at the dose investigated. When administered to Candida albicans infected animals, the DADS bearing niosomal formulation cleared the fungal burden and increased their survival much efficiently than its free form. FROM THE CLINICAL EDITOR: In this study, a novel niosomal formulation of the antifungal DADS was utilized in a murine candidiasis model, resulting in more efficient fungal clearance compared to the free formulation.

Histiocytic Sarcoma of the Terminal Ileum Presenting As a Large Ulcerating Lesion: CT Diagnosis.

We report the case of an 84-year-old woman with histiocytic sarcoma arising in the terminal ileum. The lesion was identified on CT of the abdomen as a 10 cm segment of terminal ileum with mild circumferential wall thickening, isoattenuation, and diffuse homogeneous enhancement. The patient was treated with surgery and adjuvant chemotherapy. This case provides an example of when imaging may be useful for the detection of this rare malignancy and its complications.

Quality evaluation of different honey samples produced in Peshawar valley.

The present study was designed to evaluate different honey samples obtained from local market for their quality parameters for assessment of their feasibility for foreign export by comparing it with international standards. The study was conducted at PCSIR laboratories complex, Peshawar, during 2006. The tested samples were evaluated for moisture content, Ash percentage, acid content, HMF and reducing sugars percentage. The moisture content of locally produced honey was in the range of 14.5 to 18.23%. The ash content of locally produced honey samples ranged between 0.047-0.35 which is within the standard limits. The acid content of the honey samples ranged between 19.5 and 38.0 meq kg(-1). The HMF contents of locally produced honeys ranged from 5.3 to 23.20 mg kg(-1). The content of reducing sugar of the tested samples ranged between 43.14 and 81.40% for the tested samples of locally produced honey. All of the samples were found to be in acceptable range of international standards for all of the tested parameters except for only one sample with lower reducing sugars. These samples were marked to be according to the international standards and are healthy for human consumption.