Detection of blaKPC gene among carbapenemase producing Klebsiella pneumoniae isolated from different clinical specimens at tertiary care hospital of Nepal.

BACKGROUND: Klebsiella pneumoniae infections have become a major cause of hospital acquired infection worldwide with the increased rate of acquisition of resistance to antibiotics. Carbapenem resistance mainly among Gram negative is an ongoing problem which causes serious outbreaks dramatically limiting treatment options. This prospective cross-sectional study was designed to detect blaKPC gene from carbapenem resistant K. pneumoniae. MATERIALS AND METHODS: A totally of 1118 different clinical specimens were screened and confirmed for KPC producing K. pneumoniae phenotypically using Meropenem (10 µg) disc. The blaKPC gene was amplified from the isolates of K. pneumoniae to detect the presence of this gene. RESULT: Of the total samples processed, 18.6% (n = 36) were K. pneumoniae and among 36 K. pneumoniae, 61.1% (n = 22/36) were meropenem resistant. This study demonstrated the higher level of MDR 91.7% (n = 33) and KPC production 47.2% (n = 17) among K. pneumoniae isolates. The blaKPC gene was detected in 8.3% (n = 3) of meropenem resistant isolates. CONCLUSION: Since the study demonstrates the higher level of MDR and KPC producing K. pneumoniae isolates that has challenged the use of antimicrobial agents, continuous microbiology, and molecular surveillance to assist early detection and minimize the further dissemination of blaKPC should be initiated. We anticipate that the findings of this study will be useful in understanding the prevalence of KPC-producing K. pneumoniae in Nepal.

TMQuery: a database of precomputed template modeling scores for assessment of protein structural similarity.

SUMMARY: Comparisons of protein structures are critical for developing novel protein designs, annotating protein functions and predicting protein structure. The template modeling score (TM-score) is a widely used but computationally expensive measure of protein similarity that is applicable to a wide variety of structural biology problems. We introduce TMQuery-a continuously updated database containing over eight billion pre-computed TM-score values for every pair of proteins in the Protein Data Bank, allowing researchers to quickly query and download TM-scores via a web interface. AVAILABILITY AND IMPLEMENTATION: Publicly available at https://tmquery.gsk.com/.

Cryptococcal meningitis in people living with human immunodeficiency virus in Nepal: Perspectives from resource limited setting.

Early diagnosis of cryptococcal meningitis among people living with HIV (PLHIV) is crucial for its therapeutic success. The objective of this study was to diagnose cryptococcal meningitis in PLHIV cases using the available laboratory techniques for its confirmation in resource limited setting. This cross-sectional prospective study was conducted among 72 PLHIV with clinical suspicion of meningitis. Each cerebrospinal fluid (CSF) sample received at the National Public Health Laboratory, Kathmandu was processed for India ink staining, cryptococcal antigen lateral flow assay, and fungal culture following standard protocols. The laboratory-confirmed cryptococcal meningitis cases were between 24 and 69 years of age (median age 39 years) with 87.5% (12/14) of cases being male. Cryptococcus was detected in 22.22% (16/72) by any of the three tests, 19.44% (14/72) by cryptococcal antigen lateral flow assay, 16.66% (12/72) by India ink staining, and 8.33% (6/72) by culture. High percentage of cryptococcal meningitis among PLHIV warrants early microbiological diagnosis for better case management. Cryptococcal antigen detection immunoassay should be the priority test for laboratory diagnosis of cryptococcal meningitis in PLHIV. Alternatively, very simple and economic India ink staining of CSF specimens could be used in resource limited settings.

High level of persister frequency in clinical staphylococcal isolates.

BACKGROUND: Staphylococcus aureus is a notorious human pathogen that causes often lethal systemic conditions that are mostly medical device associated biofilm infections. Similarly, coagulase negative staphylococci are emerging as leading pathogen for nosocomial infections owing to their ability to form biofilm on implanted medical equipment. Chronic in nature, these infections are difficult to treat. Such recalcitrance of these infections is caused mainly due to the presence of persister cells, which exhibit transient yet extreme tolerance to antibiotics. Despite tremendous clinical significance, there is lack of studies on persister cells formation among clinical bacterial isolates. Considering the importance of factors influencing persister formation, in this study, we evaluate the association of antibiotic tolerance with biofilm production, antibiotic stress, growth phase, specimen type, and dependency on staphylococcal species. Biofilm formation was detected among 375 clinical staphylococcal isolates by quantitative tissue culture plate method (TCP) and icaAD genes by genotypic method. The antibiotic susceptibility was determined by Kirby Bauer disc diffusion method while minimum inhibitory concentration values were obtained by agar dilution method. Persister cells were measured in the susceptible staphylococcal isolates in the presence of clinically relevant antibiotics. RESULTS: In the study, 161 (43%) S. aureus and 214 (57%) coagulase negative staphylococci (CNS) were isolated from different clinical samples. TCP method detected biofilm production in 84 (52.2%) S. aureus and 90 (42.1%) CNS isolates. The genotypic method detected icaAD genes in 86 (22.9%) isolates. Majority (> 90%) of both the biofilm producers and non-producers were sensitive to chloramphenicol and tetracycline but were resistant to penicillin. Interestingly, all isolates were sensitive to vancomycin irrespective of biofilm production. While high persister frequency was observed among all staphylococci isolates in the stationary growth phase, the persister frequency in exponential growth phase was statistically high among isolates possessing icaAD genes compared to icaAD negative isolates. CONCLUSION: The research findings provide strong evidence that the clinical staphylococcal isolates exhibit extreme antibiotic tolerance suggesting their causal link with treatment failures. Understanding the factors influencing the formation and maintenance of persister cells are of utmost important aspect to design therapeutics and control recalcitrant bacterial infections.

Phenotypic and genotypic characterization of biofilm producing clinical coagulase negative staphylococci from Nepal and their antibiotic susceptibility pattern.

BACKGROUND: Coagulase-negative staphylococci (CNS) survive as commensals of skin, anterior nares and external canals of human and were regarded as non-infectious pathogens. However, they are emerging as a major cause of nosocomial infectious due to their ability to form biofilms and high resistance to several classes of antibiotics. This study examines the biofilm forming abilities of 214 clinical CNS isolates using phenotypic and genotypic methods, and determines their antibiotic susceptibility patterns. METHODS: A total of 214 clinical isolates collected from different clinical samples were identified as CNS and their antibiotic susceptibility determined by CLSI guidelines. The biofilm forming ability of all isolates was determined by three phenotypic methods; Congo red agar (CRA) method, tube adherence method (TM) and tissue culture plate (TCP) method and by genotypic method for the detection of icaAD genes. RESULTS: Among all the isolates, S. epidermidis (57.5%) was found the most frequently, followed by S. saprophyticus (18.7%), S. haemolyticus (11.2%), S. hominis (7%), and S. capitis (5.6%). Antibiotic susceptibility pattern demonstrated 91.6% isolates were resistant to penicillin and 66.8% to cefoxitin while 91.1% isolates were susceptible to chloramphenicol. Constitutive and inducible clindamycin resistant phenotype as measured by D-test was seen among 28% and 14.5% of isolates respectively. Tissue culture plate method detected biofilm production in 42.1% isolate followed by 31.8% through tube method while 20.1% isolates were found to produce slime in Congo red agar method. The genotypic assay revealed presence of icaA and icaD genes in 19.2% isolates. CONCLUSION: The study shows a high prevalence of biofilm formation and inducible clindamycin resistance in CNS isolates, indicating the importance of in-vitro biofilm production test and D-test in routine laboratory diagnostics. Implementation of efficient diagnostic techniques for detection of biofilm production in clinical samples can help manage staphylococcal infections and minimize risks of treatment failures in hospitals.

Hydroxyapatite-collagen augments osteogenic differentiation of dental pulp stem cells.

The objectives of this study were to isolate and culture dental pulp stem cells (DPSCs) and to investigate their proliferation and osteogenic differentiation on hydroxyapatite-collagen (HA-Col) scaffold. DPSCs were characterized by fluorescence-activated cell sorting (FACS). Cultured cells were CD73+, CD90+, CD105+ and CD31-, CD45-. A commercially available HA-Col scaffold was used for culture of DPSCs. Cell attachment and viability of DPSCs cultured on scaffold was studied by sulforhodamine assay. Osteoblast differentiation capacity was studied by alkaline phosphatase assay and the effects of growth factors such as PDGF, IGF1 and FGF2 were further studied. Scanning electron microscopy (SEM) of cell seeded scaffolds was also performed. We found that DPSCs cultured exhibited the characteristic mesenchymal stem cells (MSCs) morphology and differentiation properties. Scaffold was found to be non-cytotoxic and had good biocompatibility in vitro. Osteoblast differentiation ability was found to increase at higher concentration of scaffold and additive effects were observed with the use of growth factors. In SEM, cells appeared to cover the entire surface of the scaffold forming continuous cell layer and extending filopodial extensions. HA-Col scaffold is apt for MSCs attachment and proliferation in vitro. Their unique self-renewal and multilineage differential potential make them ideal for use in regenerative medicine. The limitations of currently available bone graft materials have led to the emergence of tissue engineering using mesenchymal stem cells (MSCs). Since, HA-Col scaffold potentiated the proliferation and osteogenic differentiation of DPSCs, this biomimetic material may be an ideal one for maxillofacial and alveolar bone regeneration.

Urodynamic Changes after Valve Fulguration Alone and Valve Fulguration with Bladder Neck Incision.

AIMS: The aim of this study is to compare urodynamic changes after valve fulguration alone and valve fulguration with bladder neck incision (BNI). SETTINGS AND DESIGN: A total of 81 patients with posterior urethral valve were treated at our center from July 2010 to July 2016. Patients were randomized into two groups using simple randomization. Forty patients underwent BNI in addition to valve fulguration (Group I), and the remaining 41 patients underwent conventional transurethral valve fulguration (Group II). SUBJECTS AND METHODS: The exclusion criteria for both the groups were the presence of simultaneous urogenital anomalies, any neurological condition, history of any urethral manipulation, and urinary diversion. Urodynamic changes were compared in both groups postoperatively. All patients were evaluated throughout their follow-up, according to the following protocol: (a) Voiding cystourethrography at 6 weeks after surgery; (b) Renal function test and urine culture at 6 weeks and then 3 monthly; (c) Ultrasound kidney, ureter, and bladder region and urodynamics at 3 and 6 months after surgery and then yearly. Median follow-up period for Group I was 27.5 months (13-72 months) and 14 months (14.5-72 months) for Group II. STATISTICAL ANALYSIS USED: Statistical analysis was done using the Student's t-test for parametric data and Chi-square test for categorical variable. P ≤ 0.05 was considered as statistically significant. RESULTS: The mean age was 7.26 years in Group I and 7.66 years in Group II at the end of follow-up. There was no statistically significant difference found regarding detrusor overactivity (P = 0.68), compliance (P = 0.052), end-filling pressure (P = 0.08), and max Pdet at Qmax (P = 0.08) in the both groups. However, there was a statistically significant difference regarding improvement of peak flow (P = 0.038) and postvoid residue (PVR) (P = 0.045) in Group I in comparison to Group II. CONCLUSIONS: Valve ablation with BNI gives statistically significant better urodynamics in voiding phase regarding flow and lesser PVR in comparison to valve ablation.

Efficacy of arsenic filtration by Kanchan arsenic filter in Nepal.

Groundwater arsenic contamination has caused a significant public health burden in lowland regions of Nepal. For arsenic mitigation purposes, the Kanchan Arsenic Filter (KAF) was developed and validated for use in 2003 after pilot studies showed its effectiveness in removing arsenic. However, its efficacy in field conditions operating for a long period has been scarcely observed. In this study, we observe the efficacy of KAFs running over 6 months in highly arsenic-affected households in Nawalparasi district. We assessed pair-wise arsenic concentrations of 62 randomly selected household tubewells before filtration and after filtration via KAFs. Of 62 tubewells, 41 had influent arsenic concentration exceeding the Nepal drinking water quality standard value (50 µg/L). Of the 41 tubewells having unsafe arsenic levels, KAFs reduced arsenic concentration to the safe level for only 22 tubewells, an efficacy of 54%. In conclusion, we did not find significantly high efficacy of KAFs in reducing unsafe influent arsenic level to the safe level under the in situ field conditions.

CCL18 aggravates atherosclerosis by inducing CCR6-dependent T-cell influx and polarization.

Introduction: The CC chemokine ligand 18 (CCL18) is a chemokine highly expressed in chronic inflammation in humans. Recent observations of elevated CCL18 plasma levels in patients with acute cardiovascular syndromes prompted an investigation into the role of CCL18 in the pathogenesis of human and mouse atherosclerosis. Methods and results: CCL18 was profoundly upregulated in ruptured human atherosclerotic plaque, particularly within macrophages. Repeated administration of CCL18 in Western-type diet-fed ApoE -/- mice or PCSK9mut-overexpressing wild type (WT) mice led to increased plaque burden, enriched in CD3+ T cells. In subsequent experimental and molecular modeling studies, we identified CCR6 as a functional receptor mediating CCL18 chemotaxis, intracellular Ca2+ flux, and downstream signaling in human Jurkat and mouse T cells. CCL18 failed to induce these effects in vitro in murine spleen T cells with CCR6 deficiency. The ability of CCR6 to act as CCL18 receptor was confirmed in vivo in an inflammation model, where subcutaneous CCL18 injection induced profound focal skin inflammation in WT but not in CCR6-/- mice. This inflammation featured edema and marked infiltration of various leukocyte subsets, including T cells with a Th17 signature, supporting CCR6's role as a Th17 chemotactic receptor. Notably, focal overexpression of CCL18 in plaques was associated with an increased presence of CCR6+ (T) cells. Discussion: Our studies are the first to identify the CCL18/CCR6 axis as a regulator of immune responses in advanced murine and human atherosclerosis.

Macrophage-derived, macrophage migration inhibitory factor (MIF) is necessary to induce disease in the K/BxN serum-induced model of arthritis.

Rheumatoid arthritis (RA) is characterized by the interaction of multiple mediators, among the most important of which are cytokines. In recent years, extensive studies demonstrate a pivotal role for one cytokine, macrophage migration inhibitory factor (MIF), in fundamental events in innate and adaptive immunity. MIF has now been demonstrated to be involved in the pathogenesis of many diseases, but in the case of RA the evidence for a role of MIF is very strong. MIF is abundantly expressed in the sera of RA patients and in RA synovial tissue correlating with disease activity. MIF-deficient mice were used to induce arthritis by serum transfer from K/BxN mice. K/BxN serum transfer arthritis was markedly attenuated in MIF(-) mice, with reduction in clinical index and histological severity as well as decrease in synovial cytokines. Macrophage transfers were done to investigate the specific role of macrophage-derived MIF. We show that adoptive transfer of wild-type macrophages into MIF(-) mice restores the sensitivity of MIF(-) mice to arthritis development, and this affect was associated with a restoration in serum IL-1ß and IL-6 production. These results indicate that MIF plays a critical role in inflammation and joint destruction in K/BxN serum-induced arthritis and that the systemic expression of MIF by a subpopulation of macrophages is necessary and sufficient for the full development of arthritis.

Detection of blaoxa-23 Gene from Carbapenem-resistant Acinetobacter Baumannii.

BACKGROUND: Antibiotic resistance is a great concern for public health and Acinetobacter baumannii-associated infections are increasing in many parts of the world, including Nepal. However, limited data is available on the prevalence of A. baumannii harboring blaOXA-23 from Nepal. METHODS: A hospital-based cross-sectional study was designed to detect the blaOXA-23 gene from carbapenem-resistant A. baumannii isolates in Nepal. A total of 380 clinical specimens were collected and processed following standard microbiological procedures. Antibiotic susceptibility test was performed as per the protocol of the Kirby-Bauer disk diffusion technique and the CLSI guidelines, while screening of carbapenemase production was assessed by the Modified Hodge Test using meropenem (10µg) disc. The presence of the blaOXA-23 gene in carbapenemase-positive A. baumannii was confirmed by PCR. RESULTS: Among 380 specimens analyzed, 210 (55.3%) samples were positive for bacterial growth, where 33(15.7% of total growth) of the isolates were A. baumannii, and most of them were isolated from the ICU patients (20/33, 60.6%) and sputum (16/33, 48.5%). Thirty-two isolates (97%) were colistin sensitive, while only four (12.1%) isolates were sensitive to meropenem and imipenem. Twenty-three (69.7%) of A. baumannii were carbapenemase positive as revealed by the Modified Hodge Test test, and 19 of them (57.6% of total A. baumannii) harbored the blaOXA-23 gene. CONCLUSIONS: A high prevalence of the blaOXA-23 gene among carbapenem-resistant A. baumannii isolates were found. Systematic network surveillance should be established to check the spread of such isolates, especially in the intensive care units of tertiary care hospitals in Nepal.

Extended-spectrum ß-lactamases producing Enterobacteriaceae (ESBL-PE) prevalence in Nepal: A systematic review and meta-analysis.

An alarming increase in the occurrence of extended-spectrum ß-lactamase-producing Enterobacteriaceae (ESBL-PE) has threatened the treatment and management of bacterial infections. This systematic review and meta-analysis aimed to provide a quantitative estimate of the prevalence of ESBL among the members of the Enterobacteriaceae family by analyzing the community-based and clinical studies published between 2011 and 2021 from Nepal and determine if ESBL-PE correlates with multidrug resistance (MDR). The Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines were followed for systematic review and meta-analysis and the articles' quality was assessed using the Newcastle-Ottawa scale. Of the 2529 articles screened, 65 articles were systematically reviewed, data extracted, and included in in-depth meta-analysis. The overall pooled prevalence of ESBL-producers in Enterobacteriaceae was 29 % (95 % CI: 26-32 %) with high heterogeneity (I2 = 96 %, p < 0.001). Escherichia coli was the predominant ESBL-producing member of the Enterobacteriaceae family, followed by Citrobacter spp. and Klebsiella spp. The prevalence of ESBL-PE increased from 18.7 % in 2011 to 29.5 % in 2021. A strong positive correlation (r = 0.98) was observed between ESBL production and MDR in Enterobacteriaceae. ESBL-PE isolates showed high resistance to ampicillin, cephalosporins, and amoxicillin-clavulanic acid, and blaCTX-M type was the most reported gene variant among ESBL-PE. In conclusion, this study demonstrated an increased prevalence of ESBL-PE in Nepal over the last decade, and such isolates showed a high level of MDR against the ß-lactams and non-ß-lactam antibiotics. Tackling the rising antibiotic resistance (AR) and MDR in ESBL-PE would require concerted efforts from all stakeholders to institute effective infection control programs in the community and clinical settings.

Characterization of Thermostable Cellulase from Bacillus licheniformis PANG L Isolated from the Himalayan Soil.

This study aimed to isolate, purify, and characterize a potential thermophilic cellulase-producing bacterium from the Himalayan soil. Eleven thermophilic bacteria were isolated, and the strain PANG L was found to be the most potent cellulolytic producer. Morphological, physiological, biochemical, and molecular characterization identified PANG L as Bacillus licheniformis. This is the first study on the isolation of thermostable cellulase-producing Bacillus licheniformis from the Himalayan soil. This bacterium was processed for the production of cellulase enzyme. The optimum conditions for cellulase production were achieved at 45°C after 48 h of incubation at pH 6.5 in media-containing carboxymethyl cellulose (CMC) and yeast extract as carbon and nitrogen sources, respectively, in a thermo-shaker at 100 rpm. The enzyme was partially purified by 80% ammonium sulphate precipitation followed by dialysis, resulting in a 1.52-fold purification. The optimal activity of partially purified cellulase was observed at a temperature of 60°C and pH 5. The cellulase enzyme was stable within the pH ranges of 3-5 and retained 67% of activity even at 55°C. Cellulase activity was found to be enhanced in the presence of metal ions such as Cd2+, Pb2+, and Ba2+. The enzyme showed the highest activity when CMC was used as a substrate, followed by cellobiose. The Km and Vmax values of the enzyme were 1.8 mg/ml and 10.92 µg/ml/min, respectively. The cellulase enzyme obtained from Bacillus licheniformis PANG L had suitable catalytic properties for use in industrial applications.

Decreased levels of sCD21 and sCD23 in blood of patients with systemic-juvenile arthritis, polyarticular-juvenile arthritis, and pauciarticular-juvenile arthritis.

A soluble form of CD21 (sCD21) and CD23 (sCD23) is released from the surface of human white blood cells upon shedding of the extracellular domain. sCD21 circulates in a complex with cleavage fragments of C3 and sCD23, which were previously identified as ligands of membrane and soluble CD21. sCD21 seems to be a marker of chronic inflammatory disease. To assess the sCD21 and sCD23 status in patients with subsets of juvenile arthritis (JA), we determined plasma levels sCD21 and sCD23. Plasma sCD21 levels were significantly decreased in all JA subtypes (O-JA P < 0.0068; P- and S-JA P < 0.0001) compared to healthy controls. Plasma sCD23 levels were significantly decreased in P-JA and S-JA (both P < 0.0001), but not in O-JA (P < 0.3843) in comparison with healthy controls, and data statistically analyzed. Our results suggest that pathological mechanisms relevant to autoimmune disorders interfere with the regulation of both CD21 and CD23 shedding.

Investigation of Visceral LeishmaniasisTransmission in Selected Districts of Nepal.

BACKGROUND: Visceral leishmaniasis is transmitted to humans by Leishmania donovani infected Phlebotomus argentipes sandflies. Nepal has successfully met the elimination target of less than 1 case per 10,000, although recently this threshold has been surpassed demonstrating ongoing transmission. The main objective of the present study was to investigate transmission of visceral leishmaniasis in 4 visceral leishmaniasis endemic districts of Nepal including Palpa, Morang, Saptari and Sarlahi. METHODS: Human blood samples (331), domestic animals blood samples [goats (n =67), dogs (n =1), cows (n = 6), buffaloes (n = 16), and ox (n = 10)] and sandflies samples (3976 from 142 households) were collected from the villages of these 4 districts. Human blood samples were tested for VL antibodies using the rK39 rapid diagnostic test (InBios International, Seattle, WA). kDNA of L.donovani was amplified by PCR from DNA extracted from human blood, animal blood and sandfly samples. RESULTS: Out of 331 screened across 4 districts,32 were positive on rK39 serology and 16 were positive by PCR amplification of kDNA from L. donovani. The majority of the positive serology and PCR tests were from the Ishworpur village in the Sarlahi district where there was an outbreak of 18 cases of VL. This study also revealed the presence of L. donovani DNA in female P. argentipes sandflies collected from the Ishworpur village of Sarlahi, 6 villages in the Saptari,10 villages in the Palpa, and from 9 villages in the Morang. Blood samples from domestic animals in the same villages were negative for kDNA detection by PCR. CONCLUSIONS: The results of human and sandfly findings strongly point towards local transmission of visceral leishmaniasis in these 4 districts of Nepal. Notably, there is a significant level of transmission in the Ishworpur village in the Sarlahi district. The observations from this study suggest that domestic animals are not a reservoir host for L. donovani in these districts in Nepal. Ongoing surveillance is needed to identify new outbreaks such as in the Sarlahi district.

Serum Metabolic Disturbances in Lung Cancer Investigated through an Elaborative NMR-Based Serum Metabolomics Approach.

Detection of metabolic disturbances in lung cancer (LC) has the potential to aid early diagnosis/prognosis and hence improve disease management strategies through reliable grading, staging, and determination of neoadjuvant status in LC. However, a majority of previous metabolomics studies compare the normalized spectral features which not only provide ambiguous information but further limit the clinical translation of this information. Various such issues can be resolved by performing the concentration profiling of various metabolites with respect to formate as an internal reference using commercial software Chenomx. Continuing our efforts in this direction, the serum metabolic profiles were measured on 39 LC patients and 42 normal controls (NCs, comparable in age/sex) using high-field 800 MHz NMR spectroscopy and compared using multivariate statistical analysis tools to identify metabolic disturbances and metabolites of diagnostic potential. Partial least-squares discriminant analysis (PLS-DA) model revealed a distinct separation between LC and NC groups and resulted in excellent discriminatory ability with the area under the receiver-operating characteristic (AUROC) = 0.97 [95% CI = 0.89-1.00]. The metabolic features contributing to the differentiation of LC from NC samples were identified first using variable importance in projection (VIP) score analysis and then checked for their statistical significance (with p-value < 0.05) and diagnostic potential using the ROC curve analysis. The analysis revealed relevant metabolic disturbances associated with LC. Among various circulatory metabolites, six metabolites, including histidine, glutamine, glycine, threonine, alanine, and valine, were found to be of apposite diagnostic potential for clinical implications. These metabolic alterations indicated altered glucose metabolism, aberrant fatty acid synthesis, and augmented utilization of various amino acids including active glutaminolysis in LC.

Feasibility of one-shot dilation access in the pediatric age group.

Objective: To compare sequential fascial dilation (SFD) versus one-shot dilation (OSD) in the pediatric patients undergoing percutaneous nephrolithotomy. Methods: The present study is an observational study. The study subjects were divided into two groups. In group 1, renal dilation was done using the SFD and in group 2, renal dilation was done using the OSD. The amount of time exposed to radiation during access to pelvicalyceal system was estimated. Complications, stone free rates, ancillary procedures for residual stones and hospital stay were compared. Modified Clavien-Dindo classification was used for grading the complications. Results: Radiation exposure and operative time were less in OSD group (95% confidence interval (CI) 3.068 to 14.072, and 2.565 to 12.435, p<0.005). The mean drop of hematocrit was statistically less significant in OSD group (p=0.032). In both groups, complications, stone free rate and hospital stay were statistically insignificant. Conclusions: OSD is feasible in the children with reduced radiation exposure and shorter operative time. The outcome was similar to SFD.

Detection of differential expression of miRNAs in computerized tomography-guided lung biopsy.

Aims: Nonsmall-cell lung carcinoma comprises 85% of lung malignancies and is usually associated with a poor prognosis due to diagnosis at advanced stages. Molecular diagnosis of computerized tomography (CT)-guided biopsy has the potential to identify subtypes of lung carcinoma like adenocarcinoma (AC) and squamous cell carcinoma (SCC) along with its molecular stratification. This approach will help predict the genetic signature of lung cancer in individual patients. Subjects and Methods: Histopathologically proved a CT-guided biopsy sample of lung cancer cases was used to screen for the expression of microRNA (miRNA) earlier quantitated in blood plasma. Primers against hsa-miR2114, hsa-miR2115, hsa-miR2116, hsa-miR2117, hsa-miR449c, and hsa-miR548q with control RNU6 were used to screen 30 AC, 30 SCC, 5 nonspecific granulomatous inflammation, and 8 control samples. Reverse transcription polymerase chain reaction (RT-PCR) data revealed expression of hsa-miR2114 and hsa-miR548q in AC as well as SCC. Results: RT-PCR data revealed that the expression of hsa-miR2116 and hsa-miR449c was found upregulated in AC while hsa-miR2117 was expressed in SCC cases. Bioinformatic analysis revealed that genes, where these miRNAs are located, were also upregulated while targets of these miRNAs were downregulated. Conclusions: miRNAs expression pattern in the CT-guided biopsy samples can be used as a potential tool to differentially diagnose lung cancer subtypes. The expression pattern of miRNAs matches very well in blood plasma and tissue samples, albeit levels were very low in the earlier case than later. This approach can also be used for screening mutations and other molecular markers in a personalized manner for the management of lung cancer patients.

Crossing Vessel in Pelvi Ureteric Junction Obstruction: A Histopathological Analysis.

OBJECTIVE: The aim of the study is to identify whether crossing vessel is a cause or an associated finding in Pelvi Ureteric Junction Obstruction. MATERIAL AND METHODS: This is a prospective study of a total of 128 patients who underwent laparoscopic pyeloplasty from January 2016 to June 2020. All patients who underwent laparoscopic pyeloplasty and pelvi ureteric junction segments were sent for histopathological examination. The presence of crossing vessels is documented intraoperative and patients were divided into two groups, group 1 having pelvi ureteric junction obstruction with crossing vessel, and group 2, pelvi ureteric junction obstruction without crossing vessels. Histopathological examination findings of pelvi ureteric junction segment including inflammation, fibrosis, muscle hypertrophy, muscle disarray, and synaptophysin were recorded. Unpaired Student t-test was used for comparing differences between continuous normally distributed data from 2 samples and non-parametric tests were applied for continuous data. RESULTS: Of the total 128 patients, crossing vessels were identified in 42 (32.8%), and 86 (67.2%) were without crossing vessels. The demographic profile of patients between the 2 groups was comparable. On histopathological examination, moderate-to-severe chronic inflammation was seen in 23.8% and 44.2% (P > .05) in group 1 and group 2, respectively; fibrosis and muscular hypertrophy were higher in group 2 but statistically insignificant (P > .05), and muscle disarray was higher in group 1 but statistically insignificant (P > .05). Synaptophysin was positive in 4.8% and 4.7% in group 1 and group 2, respectively. CONCLUSION: The differences in histopathological examination between the 2 groups were not statistically significant. However, in patients with crossing vessels, there was a higher degree of inflammation, which may lead to early pelvi ureteric junction obstruction.

Factors associated with arsenicosis and arsenic exposure status in Nepal: implications from community based study.

A significant public health problem due to exposure to arsenic via groundwater in communities of lowland Terai region of Nepal has issued forth need to assess the exposure status and factors associated with arsenicosis. We observed arsenical dermal manifestations and collected and assessed total arsenic content in tubewell water, urine, and hair samples of study subjects at arsenic affected communities in Nawalparasi district of Nepal. The explanatory variables associated with arsenicosis were elevated arsenic in tubewell, male gender and increased age (P < 0.05). 67% (73/109) and 66% (77/117) of subjects exceeded the normal urinary and hair arsenic levels respectively. Among them 52% (57/109) and 47% (55/117) exceeded normal urinary and hair arsenic levels having no arsenical dermal manifestations. Males and symptomatic cases had significantly higher hair arsenic levels (P < 0.05). We also observed significant positive correlation of both urine and hair arsenic levels to tubewell arsenic levels (r = 0.27, 0.37, P < 0.01) and negative correlation of urine arsenic levels with the age of the subjects (r = -0.18, P = 0.06). We conclude that elucidating factors associated with arsenicosis could be of prime importance in intervention and preventive measures. In arsenic affected communities of Nepal exposure to arsenic is still a major problem despite mitigation efforts and the potential for sub-clinical effects in exposed population is high.