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BACKGROUND Alagille syndrome is an autosomal dominant disorder associated with variable clinical phenotypic features including cholestasis, congenital heart defects, vertebral defects, and dysmorphic facies. OBJECTIVE Whole exome sequencing (WES) has become technically feasible due to the recent advances in next-generation sequencing technologies, therefore offering new possibilities for mutations or genes identification. METHODS WES was used to identify pathogenic variants, which may have significant prognostic implications for patients’ clinical presentation of the proband. In this paper, we have uncovered a novel JAGGED1 gene (JAG1) mutation associated with Alagille syndrome in a 5-year-old girl presented with conjugated hyperbilirubinemia and infantile cholestasis. RESULTS The exome sequencing analysis revealed the presence of a novel JAG1 heterozygous c.3080delC variant in exon 25. The detected variant introduced a stop codon (p.P1027RfsTer9) in the gene sequence, encoding a truncated protein. Our exome observations were confirmed through Sanger sequencing as well. CONCLUSIONS Here, we report a case of a patient diagnosed with Alagille syndrome, conjugated hyperbilirubinemia, and infantile cholestasis, with emphasis on its association with the detection of the novel JAG1 mutation, thereby establishing the genetic diagnosis of the disease.

Hemorrhagic cystitis (HC) has been reported with increased frequency following post-transplantation cyclophosphamide (PTCy)–based haploidentical hematopoietic cell transplantation (HCT) along with a strong association with BK viruria. We prospectively evaluated the incidence of BK viruria and HC in 115 patients (median age 20 years, 2–65) undergoing PTCy-based haploidentical HCT with (n = 71) or without (n = 44) CTLA4Ig. HC prophylaxis consisted of a continuous infusion of mesna 30 min prior and 48 h post-PTCy. The overall incidence of BK viruria was 65.7%. None with BK viruria

Background Alagille syndrome is an autosomal dominant disorder associated with variable clinical phenotypic features including cholestasis, congenital heart defects, vertebral defects, and dysmorphic facies. Objective Whole exome sequencing (WES) has become technically feasible due to the recent advances in next-generation sequencing technologies, therefore offering new possibilities for mutations or genes identification. Methods WES was used to identify pathogenic variants, which may have significant prognostic implications for patients’ clinical presentation of the proband. In this paper, we have uncovered a novel JAGGED1 gene (JAG1) mutation associated with Alagille syndrome in a 5-year-old girl presented with conjugated hyperbilirubinemia and infantile cholestasis. Results The exome sequencing analysis revealed the presence of a novel JAG1 heterozygous c.3080delC variant in exon 25. The detected variant introduced a stop codon (p.P1027RfsTer9) in the gene sequence, encoding a truncated protein. Our exome observations were confirmed through Sanger sequencing as well. Conclusions Here, we report a case of a patient diagnosed with Alagille syndrome, conjugated hyperbilirubinemia, and infantile cholestasis, with emphasis on its association with the detection of the novel JAG1 mutation, thereby establishing the genetic diagnosis of the disease.

Direct Acting Antivirals (DAA) have changed the landscape of hepatitis C virus (HCV) infection with high cure rates across genotypes. However, the use of these agents in the setting of allogeneic hematopoietic cell transplantation (HCT) has been limited. In this context, we report the outcome of five children (5-12 years) with relapsed and refractory leukemia and active HCV infection (genotype 1b), who underwent urgent haploidentical HCT and were treated with Sofosbuvir and Velpatasvir (Sof-Vel) from initiation of treatment to 24 weeks post-HCT. All achieved complete virologic response (VR) at a median of 2 weeks, with normalization of liver enzymes. There were no adverse events related to the use of Sof-Vel, with no major fluctuations in cyclosporine levels. Two of the patients developed chronic GVHD and one relapsed. Sof-Vel was continued in one of them along with sirolimus without affecting drug levels. With a median follow-up of 15 months, four patients are disease free with sustained VR. Our study shows that combination of Sof-Vel might be effective in inducing rapid complete and sustained VR during HCT without any major untoward drug interaction.

Following a major seasonal outbreak of H1N1 influenza in 2018 September, prophylactic oseltamivir for six months was initiated in children undergoing haploidentical HCT with regular monitoring for influenza and other respiratory virus infections. Influenza was not detected in 22 children undergoing prophylaxis, compared to 8 H1N1 infections in 21 adults without prophylaxis (P = .01). Four children on prophylaxis were detected to have other respiratory viruses, compared to 8 in those without prophylaxis. Invasive pulmonary aspergillosis (IPA) was observed only in association with H1N1 (4/8 with H1N1 vs 0/35 without H1N1, P = .001) and was thus lower in the prophylaxis group (P = .04). The overall incidence of episodes of respiratory illness and hospital stay were also lower in those on prophylaxis (P = .001). There were no untoward side effects associated with prophylactic oseltamivir. Prophylactic oseltamivir was safe and effective in prevention of H1N1 infection and subsequent IPA in children at-risk, early after haploidentical HCT.

Background & Methods Post-transplantation cyclophosphamide (PTCy)-based HCT has been associated with an increased frequency of hemorrhagic cystitis (HC), dominantly associated with BK viruria. We prospectively evaluated the incidence of BK viruria and HC in 115 patients [(median age 20 years, 2-65), malignant(87) and non-malignant(28)], undergoing PTCy-based haploidentical HCT with CTLA4Ig (n=71) or without (n=44). Conditioning was predominantly myeloablative (74%). As prophylaxis for HC, continuous infusion of Mesna was used from 30 minutes prior to PTCy at 50% of the daily dose of Cy every 8 hourly for 72 hours. Development of HC before day+15 was termed as ‘early HC’ and later was referred to as ‘HC’. Results The overall incidence of BK viruria was 65.7%. None with BK viruria Overall BK viruria was significantly lower in those receiving CTLA4Ig (54.1% vs 89.4% in those without CTLA4Ig, p=0.001, Figure 2A). The incidences of both high BK viruria (4.3% with vs 11.9% without CTLA4Ig, p=0.08) and HC (2.9% with vs 9.5% without CTLA4Ig p=0.09, Figure 2B) tended to be lower in those receiving CTLA4Ig-based protocols. A higher NRM was observed in those with BK viruria ≥ 104 copies/ml, related to GVHD and its complications (41.7% vs 12.6%, p=0.04). Conclusions In conclusion, our study demonstrates that extended infusion of Mesna, along with reduction in alloreactivity with the use of CTLA4Ig were probably responsible for a much lower incidence of BK viruria and resultant HC, than previously reported following PTCy-based haploidentical HCT.

Background: Tuberculosis (TB), an infectious disease caused by bacterium Mycobacterium tuberculosis, has crossed the borders of developing nations and has now emerged as disease of global emergency due to an alarming increase in cases of co-infection with human immunodeficiency virus (HIV). Conventional diagnostics used for TB diagnosis like microscopy and culture although reliable are time consuming. Rapid diagnosis of the disease is required for early commencement of treatment of TB patients. The diagnosis of extra pulmonary tuberculosis (EPTB) is more troublesome due to limited accessibility of infection sites and lack of accurate differentiation from other granulomatous diseases. Currently, WHO advises use of Xpert MTB/RIF assay for EPTB diagnosis, but this assay cannot be used in resource constrained settings prevalent in most of parts of India. Aim: We aimed to develop a multiplex real-time PCR assay which not only detects Mycobacterium tuberculosis complex (MTBC) but also differentiates them from nontuberculous mycobacteria (NTM). Methods: For development and validation of TBSURE, we included 3, 709 EPTB samples from adult Indian patients suspected for TB. TBSURE is a real time PCR diagnostic which uses three Taqman probes based on IS6110, MPB64 and 16S rRNA genes. The sensitivity and specificity of TBSURE was compared with the goldstandard culture test and XpertMTB/RIF assay for TB detection. Results and Conclusion: TBSURE gave a sensitivity of 92% and specificity of 91% as compared to Xpert MTB/RIF assay, when samples from TB suspected individuals were subjected to both the tests. As compared to AFB culture, TBSURE gave a sensitivity of 93.1% and specificity of 96.5%. Thus, TBSURE is an in-house efficient and affordable diagnostic test for EPTB diagnosis.

Cyclodextrin glucanotransferase (CGTase) is a commercially important enzyme which catalyses the formation of cyclodextrins (CDs) from starch. A CGTase producing bacterium was isolated from soil which gave a fairly high enzyme activity of 7.5 U mL(-1) after 24 h of growth which was further increased to 22 U mL(-1) by proper media design. The enzyme was purified to homogeneity by a novel single affinity precipitation step which resulted in a very high recovery of more than 90%. The molecular weight, as determined by SDS-PAGE, was found to be 68 kDa. The pH optimum was found to be 6.6 while a temperature optimum of 65 degrees C was observed. The enzyme exhibited a fairly high degree of functional stability with little loss of activity, even after 8 h of incubation at 65 degrees C. Ca++ had little effect on the activity of the enzyme while urea at 10 mM concentration increased the activity of the enzyme by more than 200%, suggesting that it is a unique enzyme.