Intraperitoneal transplant of Hepatocytes co-Encapsulated with mesenchymal stromal cells in modified alginate microbeads for the treatment of acute Liver failure in Pediatric patients (HELP)-An open-label, single-arm Simon's two stage phase 1 study protocol.

BACKGROUND: Pediatric acute liver failure (PALF) carries a high mortality without liver transplantation (LT) in children. Liver transplantation, though lifesaving, is limited by timely donor organ availability, the risks of major surgery and complications of life-long immunosuppression. Hepatocyte transplantation (HT) improves synthetic and detoxification functions in small animal models. The encapsulation of hepatocytes in alginate protects it from the recipient immune system while the intraperitoneal route of administration allows large volumes to be infused. The safety and possibly short-term efficacy of encapsulated hepatocytes has been observed in a named patient use. A novel type of microbeads (HMB002) has been developed, using a modified alginate and mesenchymal stromal cells (MSCs). Its safety and medium-term efficacy need to be studied in the context of clinical study while optimizing the hepatocyte function and viability using modifications of the alginate and MSCs co-encapsulation. METHODS: A single centre, non-randomised, open-label, single-arm Simon's two stage study will be conducted to evaluate the safety, biological activity and tolerability of transplantation of a single intraperitoneal dose of microbeads made from an optimum combination of a modified alginate, MSCs and hepatocytes in 17 patients less than 16 years of age with acute liver failure (Stage 1: 9 patients and Stage 2: 8 patient). Safety will be assessed by documenting moderate to severe (including life threatening and death) adverse events due to HMB002 in the first 52 weeks post-procedure. Tolerability will be assessed by observing the proportion of initiated infusions where >80% of infusion is received by the patient. Biological activity will be reflected in patient survival with native liver at 24 weeks post treatment. DISCUSSION: HMB002, if safe and efficacious in acute liver failure, could be a bridge until the liver regenerates or a suitable organ becomes available. There are multiple advantages to using HT. HT, when delivered by the intraperitoneal route, is less invasive than LT. Hepatocytes from a single donor liver can be used to treat multiple patients. Cryopreserved cells provide an off-the-shelf emergency treatment in PALF. When encapsulated, alginate encapsulation of hepatocytes precludes the need for immunosuppression unlike in LT.

Outbreak of indeterminate acute liver failure in children with adenoviraemia - Not a new disease.

BACKGROUND & AIMS: In the year 2022, an outbreak of indeterminate acute hepatitis and indeterminate paediatric acute liver failure (ID-PALF) in association with adenoviraemia in immunocompetent children was reported in the UK. We postulate that this association is not a new disease in immunocompetent children. METHODS: Children with acute hepatitis during the outbreak who were referred to King's College Hospital, London for advice and management were included in the study. Data on the frequency of ID-PALF in 2022, as well as transplantation rates and the association with adenovirus infection, were obtained from electronic health records. The clinical presentation, histology and outcomes of children with ID-PALF and adenoviraemia in 2017-2021 were compared with those in 2022. RESULTS: From January to June 2022, 65 patients with acute hepatitis were referred. Ten children were admitted with ID-PALF. ID-PALF constituted 26% of all PALF cases in 2017-2021, in contrast to 58.8% during the 2022 outbreak. During the outbreak, adenoviraemia was present in 52% of children with acute hepatitis without liver failure (in whom adenoviraemia test results were available) and in 100% of ID-PALF cases. Adenoviraemia was seen in immunocompetent children in 6/13 (46%) of all ID-PALF cases between 2017-2019, with a clear absence of adenoviraemia in the 6 ID-PALF cases during 2020-2021. Compared to ID-PALF with adenoviraemia in 2017-2019 (n = 6), ID-PALF with adenoviraemia during the outbreak (n = 10) was associated with more frequent hepatic encephalopathy, hypotension requiring vasoactive medications and higher plasma ammonia levels (admission and peak), with similar native liver survival. CONCLUSIONS: The recent outbreak of ID-PALF with adenoviraemia in immunocompetent children does not appear to be a new disease, contrary to perception and other reports. The frequency of such cases over the years could be linked to background rates of adenovirus infections. IMPACT AND IMPLICATIONS: Indeterminate paediatric acute liver failure (ID-PALF) associated with adenoviraemia in immunocompetent children is not a new disease specific to 2022. The exclusive role of human adenovirus infection in the causation of this outbreak of acute hepatitis seems unlikely. Indeed, we provide histological data from explants in transplanted patients that do not support direct viral cytotoxicity. The disease is probably mediated by immunological injury directed towards adenovirus infection and/or adeno-associated virus-2.

Emergencies in paediatric hepatology.

The aetiology of several liver diseases in children is age specific and many of these conditions have significant and potentially long-term clinical repercussions if not diagnosed early and managed in a timely fashion. We address 5 clinical scenarios that cover most of the diagnostic and therapeutic emergencies in children: infants with liver disease; acute liver failure; management of bleeding varices; liver-based metabolic disorders; and liver tumours and trauma. A wide spectrum of conditions that cause liver disease in infants may present as conjugated jaundice, which could be the only symptom of time-sensitive disorders - such as biliary atresia, metabolic disorders, infections, and haematological/alloimmune disorders - wherein algorithmic multistage testing is required for accurate diagnosis. In infantile cholestasis, algorithmic multistage tests are necessary for an accurate early diagnosis, while vitamin K, specific milk formulae and disease-specific medications are essential to avoid mortality and long-term morbidity. Management of paediatric acute liver failure requires co-ordination with a liver transplant centre, safe transport and detailed age-specific aetiological work-up - clinical stabilisation with appropriate supportive care is central to survival if transplantation is indicated. Gastrointestinal bleeding may present as the initial manifestation or during follow-up in patients with portal vein thrombosis or chronic liver disease and can be managed pharmacologically, or with endoscopic/radiological interventions. Liver-based inborn errors of metabolism may present as encephalopathy that needs to be recognised and treated early to avoid further neurological sequelae and death. Liver tumours and liver trauma are both rare occurrences in children and are best managed by a multidisciplinary team in a specialist centre.

Biliary Atresia Screening in India-Strategies and Challenges in Implementation.

OBJECTIVE: To assess the utilization of SCC implemented in southern India and the effect on SCC utilization of face-to-face verbal education versus video-based content delivery. METHODS: The study included newborns with postnatal age of less than 2 wk at discharge. Mothers were administered SCCs and provided standardized verbal or video health education based on the time-period of enrollment. Home based monitoring of stool color and return of SCC on postnatal day 21 was advised. Telephone surveys were conducted to identify SCC use among families that did not return the SCC by post. RESULTS: Of the 2254 newborns enrolled, 1130 were in the verbal-counseling group and 1124 in the video-counseling group. No newborns with pale stools and biliary atresia were identified. SCC return rates were 3.8% and 2.8%. Comparing the verbal and video-counseling groups, there were no differences in the conservative (81.8% vs. 81.5%) and optimistic estimates (97.1% vs. 97.3%) of SCC utilization rates. Mothers with better educational status had higher optimistic estimates of SCC utilization. CONCLUSIONS: The use of a validated SCC in Tamil with standardized information delivery leads to good utilization rates in southern India, with video content delivery being as effective as face-to-face verbal content delivery. SCC return by post is not a feasible mode of identification of card use. TRIAL REGISTRATION: The study is registered under Clinical Trials Registry - India (CTRI/2018/01/011285).

Evaluation of LAMP for detection of Shigella from stool samples in children.

BACKGROUND: To assess the diagnostic accuracy of loop-mediated isothermal amplification (LAMP) for the detection of Shigella from stool samples from children. METHODS: Consecutive stool samples from children aged <13 years old who presented with acute watery diarrhoea or dysentery to the Department of Paediatrics were collected and processed in the Department of Microbiology. All the stool samples were subjected to culture, conventional PCR and LAMP. Genomic sequencing was performed for samples that were positive by LAMP but negative by both culture and conventional PCR. The LAMP results were compared to those from culture and to a composite reference standard based on culture and conventional PCR. RESULTS: Amongst the 374 stool samples tested, 291 samples were positive by LAMP and 213 were positive by the composite reference standard. The sensitivity of LAMP was 100 % (98.3-100 %) and its specificity was 51.6 % (43.6-59.5 %) with a disease prevalence of 57 %. The sensitivity and specificity of LAMP improved to 99.3 % (94.2-100) and 98.2 % (94.5-99.9), respectively, using latent class analysis, while assuming that genomic sequencing has perfect specificity. DISCUSSION: The authors have standardized the LAMP procedure for direct application to clinical stool samples. LAMP is a sensitive and specific method for the diagnosis of Shigella from stool samples of children as compared to both culture and conventional PCR.

Splenic stiffness and platelet count to predict varices needing treatment in pediatric extrahepatic portal vein obstruction.

BACKGROUND: Surveillance endoscopy to detect varices needing treatment (VNT) is important to prevent bleeding and morbidity in portal hypertension. In adult and pediatric cirrhosis, platelet count and liver stiffness measurement (LSM) are useful in selecting patients for endoscopy. Such recommendations do not exist for extrahepatic portal vein obstruction (EHPVO). Splenic stiffness measurement (SSM) has been studied in adult and pediatric EHPVO with conflicting results and methodological errors. This study evaluates the role of platelet counts and SSM to predict VNT and bleeding in pediatric EHPVO while comparing LSM and SSM between pediatric EHPVO and controls. METHODS: One hundred and seven children (55 with EHPVO and 52 controls) were recruited. Clinical, biochemical, hematological, and radiographic parameters of all children were noted. All children with EHPVO underwent endoscopy. RESULTS: Of the 55 children with EHPVO, 48 (87.3%) had VNT. There was no difference in the platelet counts (85,000/mm3 vs. 120,000/mm3, p = 0.58) and SSM (3.62 vs. 3.19, p = 0.05) between EHPVO children with VNT and those without. They had poor sensitivity and specificity to predict VNT. EHPVO children with bleeding had higher SSM that those without. LSM was higher among EHPVO than among controls (1.19 vs. 1.10, p = 0.003). Those with LSM higher than controls had normal liver histology. CONCLUSION: SSM is higher in EHPVO bleeders but SSM and platelet counts are unreliable to predict VNT in pediatric EHPVO. Surveillance endoscopies may be needed in all pediatric EHPVO until better screening strategies are available. TRIAL REGISTRATION: Not applicable.

Paediatric inflammatory bowel disease in India: a prospective multicentre study.

BACKGROUND: Paediatric inflammatory bowel disease (PIBD) is increasing across the world. However, information from India is sparse. This multicentre study evaluated the demographics, clinical phenotype and outcome of PIBD from India. METHODS: Data of children (≤18 years) with PIBD were collected using a proforma containing details of demographics, clinical profile, extraintestinal manifestations (EIM), investigations, disease extent and treatment. RESULTS: Three hundred twenty-five children [Crohn's disease: 65.2%, ulcerative colitis: 28.0%, IBD unclassified (IBDU): 6.7%, median age at diagnosis: 11 (interquartile range 6.3) years] were enrolled. 6.9% children had family history of IBD. Pancolitis (E4) was predominant in ulcerative colitis (57.8%) and ileocolonic (L3, 55.7%) in Crohn's disease. Perianal disease was present in 10.9% and growth failure in 20.9% of Crohn's disease cases. Steroids were the initial therapy in 84.2%, 5-amino salicylic acid in 67.3% and exclusive enteral nutrition (EEN) in 1.3% cases. Overall, immunomodulators and biologics were given to 84.3 and 17.9% cases, respectively, and 2.9% cases underwent surgery. Very early onset IBD (VEOIBD) was seen in 60 (19.2%) children. IBDU was commoner in the VEOIBD than the older-PIBD (18/60 vs 4/253; P < 0.001). VEOIBD-Crohn's disease patients more often had isolated colonic disease than the older Crohn's disease (45.4% vs 11.8%; P < 0.001). Prevalence of perianal disease, EIM, therapeutic requirements and outcome were not different between VEOIBD and older-PIBD. CONCLUSION: Disease location and phenotype of PIBD in Indian children is similar to the children from the west. However, the therapeutic options of EEN, biologics and surgery are underutilized. VEOIBD accounted for 19.2% of PIBD.

Hepatitis B Seroprotection in Pediatric Nephrotic Syndrome.

OBJECTIVE: To study the prevalence of Hepatitis B seroprotection in children (>1 y) with nephrotic syndrome vaccinated against Hepatitis B vaccine as per the Universal Immunization Program schedule (0,6,10,14 wk); to compare the Hepatitis B seroprotection rates and anti-HBs titers among different phenotypes of nephrotic syndrome; to evaluate the association between Hepatitis B seroprotection status and the immunosuppressive agents; and to study the correlation between anti-HBs titres and proteinuria. METHODS: Hepatitis B serology and anti-HBs titers were analyzed in 100 children (age-1-18 y) with different clinical phenotypes of nephrotic syndrome (cases) and 100 healthy controls. RESULTS: The proportion of seroprotected children among the cases and controls was 37% (n=37) and 61% (n=61), respectively (P<0.04). The median (IQR) anti- HBs antibodies titers among the cases was 75 (62.5, 81) mIU/mL and 112 (56, 367) mIU/mL among the controls (P=0.001). The proportion of seroprotected children among the steroid sensitive nephrotic syndrome, steroid-resistant nephrotic syndrome and controls was 40% (n=28), 30% (n=9) and 61% (n=61), respectively (P<0.01). No differences in the anti-HBs titers between children receiving steroids versus steroids along with other immunosuppressants were found. Weak negative correlation was noted between proteinuria and protective titers (r = -0.155; P=0.039). CONCLUSIONS: Children with nephrotic syndrome, in general, and steroid-resistant nephrotic syndrome in particular, show poor seroprotection with Hepatitis B vaccination.

Wilson disease and lupus nephritis: is it coincidence or a true association?

A 12-year-old girl born to third-degree consanguineous parents presented with recurrent episodes of haematuria for 8 months in association with peri-orbital and lower limb oedema for 20 days. There was no jaundice, hepatomegaly or neurological abnormality at presentation. An older brother had died following jaundice at 10 years of age. Urinalysis showed multiple dysmorphic erythrocytes without proteinuria and there was leucopenia, thrombocytopenia and hypo-albuminaemia (23 g/L). C3 component of complementaemia was low and anti-nuclear antibodies and anti-double-stranded DNA antibodies were strongly positive by immunofluorescence. Systemic lupus erythematosus (SLE) was considered but the severe hypo-albuminaemia was unexplained. During the pre-renal biopsy work-up, a deranged coagulation profile with raised transaminases prompted evaluation for chronic liver disease which culminated in the diagnosis of Wilson disease. Treatment with penicillamine and immunosuppressants was initiated, but there was neurological deterioration on Day 30 of admission and she died owing to worsening liver failure on the Day 41. Post-mortem liver biopsy demonstrated cirrhosis and post-mortem renal biopsy showed features of class-II lupus nephritis. Auto-immune antibodies and autoimmune disorders have been reported in Wilson disease and there are anecdotal reports of an association of SLE with Wilson disease. However, this case is unique in that lupus nephritis was the presenting manifestation before Wilson disease was diagnosed. The underlying pathophysiological mechanisms of this association requires further research.

Acute Motor Axonal Polyneuropathy Following Mumps Infection in a 9-Year-Old Girl.

A 9-year-old girl presented with lower motor neuron type of paralysis involving limbs, trunk and multiple cranial nerves (7, 9 and 10) with preceding history of mumps 1 week before the onset of weakness. There were no features to suggest either a meningitis or encephalitis in the child. Cerebrospinal fluid showed hypoglycorrhachia and mild protein elevation; magnetic resonance imaging of the brain was normal. Nerve conduction study showed motor axonal neuropathy. Serology for mumps IgM was positive, consistent with a diagnosis of post-mumps acute motor axonal polyneuropathy. The girl made a complete recovery within 3 weeks.

Digital rectal examination training among Indian medical graduates.

BACKGROUND: Digital rectal examination (DRE) is an important component of physical examination and an essential skill for medical graduates. DRE is often underutilised in clinical practice. The lack of confidence and expertise and also underutilization of DRE have been associated with inadequate training of medical students during their undergraduate studies. The training of Indian undergraduates in DRE has not been studied. METHODS: A questionnaire on undergraduate training in DRE was administered to students from various medical colleges joining specialty postgraduate courses in Jawaharlal Institute of Postgraduate Medical Education and Research. RESULTS: A total of 101 out of 131 students participated in the survey. Ninety-one percent of students were taught DRE as undergraduates but only three-quarters had performed DRE on patients. Among the respondents who had performed DRE, two-thirds had performed fewer than five DREs before the completion of their medical education. Respondents who had performed fewer DREs were less confident about performing DRE (p < 0.05). Only 8% had performed DRE with manikins. Patients declining DRE and the need to minimise DRE influenced the decisions to perform DRE during training. DRE was never taught in paediatrics. DREs were most often performed only in the final year of the Bachelor of Medicine and Bachelor of Surgery (MBBS) degree, and mostly without supervision (49.3%); 61.4% were unsatisfied with their training in DRE and would like to be trained better. A lack of confidence, expertise and use of DRE are associated with inadequate training of medical students CONCLUSION: The survey indicates a lack of importance given to DRE training of undergraduate students and huge gaps in imparting this clinical skill. Training may be improved by introducing manikins, changing attitudes to DRE by incorporating it in clinical problem solving, and with more frequent opportunities to practise under supervision.