Surfaceome analysis of extracellular vesicles from senescent cells uncovers uptake repressor DPP4.

Senescent cells are beneficial for repairing acute tissue damage, but they are harmful when they accumulate in tissues, as occurs with advancing age. Senescence-associated extracellular vesicles (S-EVs) can mediate cell-to-cell communication and export intracellular content to the microenvironment of aging tissues. Here, we studied the uptake of EVs from senescent cells (S-EVs) and proliferating cells (P-EVs) and found that P-EVs were readily taken up by proliferating cells (fibroblasts and cervical cancer cells) while S-EVs were not. We thus investigated the surface proteome (surfaceome) of P-EVs relative to S-EVs derived from cells that had reached senescence via replicative exhaustion, exposure to ionizing radiation, or treatment with etoposide. We found that relative to P-EVs, S-EVs from all senescence models were enriched in proteins DPP4, ANXA1, ANXA6, S10AB, AT1A1, and EPHB2. Among them, DPP4 was found to selectively prevent uptake by proliferating cells, as ectopic overexpression of DPP4 in HeLa cells rendered DPP4-expressing EVs that were no longer taken up by other proliferating cells. We propose that DPP4 on the surface of S-EVs makes these EVs refractory to internalization by proliferating cells, advancing our knowledge of the impact of senescent cells in aging-associated processes.

ΔNp63 overexpression promotes oral cancer cell migration through hyperactivated Activin A signaling.

Oral cancer is a common malignant tumor of the oral cavity that affects many countries with a prevalent distribution in the Indian subcontinent, with poor prognosis rate on account of locoregional metastases. Gain-of-function mutations in p53 and overexpression of its related transcription factor, p63 are both widely reported events in oral cancers. However, targeting these alterations remains a far-achieved aim due to lack of knowledge on their downstream signaling pathways. In the present study, we characterize the isoforms of p63 and using knockdown strategy, decipher the functions and oncogenic signaling of p63 in oral cancers. Using Microarray and Chromatin Immunoprecipitation experiments, we decipher a novel transcriptional regulatory axis between p63 and Activin A and establish its functional significance in migration of oral cancer cells. Using an orally bioavailable inhibitor of the Activin A pathway to attenuate oral cancer cell migration and invasion, we further demonstrate the targetability of this signaling axis. Our study highlights the oncogenic role of ΔNp63 - Activin A - SMAD2/3 signaling and provides a basis for targeting this oncogenic pathway in oral cancers.

Comparison of clinical and laboratory profile of survivors and non-survivors of SARS-CoV-2-related multisystem inflammatory syndrome of childhood in India: An observational study.

AIM: The SARS-CoV-2 pandemic is characterised by multiple reports of paediatric multisystem inflammatory disease or multisystem inflammatory syndrome in children (MIS-C) with Kawasaki disease-like features often complicated by myocarditis, shock and macrophage activation syndrome. Certain clinical and laboratory markers may be used to identify high risk cases. METHODS: All sequentially admitted patients hospitalised between April 2020 and October 2020, who met the WHO case definition for MIS-C were included. Data included patient demographic information, presenting symptoms, organ dysfunction and laboratory parameters. SARS-CoV-2 infection was diagnosed by nasopharyngeal swab real-time reverse transcription-polymerase chain reaction and/or rapid antibody test for SARS-CoV-2 as recommended. The clinical and laboratory criteria were compared in the survival and non-survival groups. RESULTS: A total of 29 patients with MIS-C were treated during the study period. There were 21 survivors and 8 non-survivors. The non-survivors had more neurocognitive and respiratory symptoms along with increased incidence of myocarditis compared with survivors. The serum levels of CPK-MB, D-dimer, ferritin and triglyceride were significantly raised in non-survivors as compared to survivors. CONCLUSION: The non-survivor group had higher CPK and greater proportion of children with troponin-T elevation indicating higher incidence of myocardial injury and necrosis. The D-dimer, ferritin and triglyceride were also higher in the mortality group, indicating the greater extent of inflammatory damage in this group.

Lipocalin 2 expression promotes tumor progression and therapy resistance by inhibiting ferroptosis in colorectal cancer.

Lipocalin 2 is a siderophore-binding protein that regulates iron homeostasis. Lipocalin 2 expression is elevated in multiple tumor types; however, the mechanisms that drive tumor progression upon Lipocalin 2 expression remain unclear. When Lipocalin 2 is over-expressed, it leads to resistance to 5-fluorouracil in colon cancer cell lines in vitro and in vivo by inhibiting ferroptosis. Lipocalin 2 inhibits ferroptosis by decreasing intracellular iron levels and stimulating the expression of glutathione peroxidase4 and a component of the cysteine glutamate antiporter, xCT. The increase in xCT levels is dependent on increased levels of ETS1 in Lipocalin 2 over-expressing cells. Inhibiting Lipocalin 2 function with a monoclonal antibody leads to a decrease in chemo-resistance and transformation in vitro, and a decrease in tumor progression and chemo-resistance in xenograft mouse models. Lipocalin 2 and xCT levels exhibit a positive correlation in human tumor samples suggesting that the pathway we have identified in cell lines is operative in human tumor samples. These results indicate that Lipocalin 2 is a potential therapeutic target and that the monoclonal antibody described in our study can serve as the basis for a potential therapeutic in patients who do not respond to chemotherapy.

Plakophilin3 loss leads to an increase in lipocalin2 expression, which is required for tumour formation.

An increase in tumour formation and metastasis are observed upon plakophilin3 (PKP3) loss. To identify pathways downstream of PKP3 loss that are required for increased tumour formation, a gene expression analysis was performed, which demonstrated that the expression of lipocalin2 (LCN2) was elevated upon PKP3 loss and this is consistent with expression data from human tumour samples suggesting that PKP3 loss correlates with an increase in LCN2 expression. PKP3 loss leads to an increase in invasion, tumour formation and metastasis and these phenotypes were dependent on the increase in LCN2 expression. The increased LCN2 expression was due to an increase in the activation of p38 MAPK in the HCT116 derived PKP3 knockdown clones as LCN2 expression decreased upon inhibition of p38 MAPK. The phosphorylated active form of p38 MAPK is translocated to the nucleus upon PKP3 loss and is dependent on complex formation between p38 MAPK and PKP3. WT PKP3 inhibits LCN2 reporter activity in PKP3 knockdown cells but a PKP3 mutant that fails to form a complex with p38 MAPK cannot suppress LCN2 promoter activity. Further, LCN2 expression is decreased upon loss of p38ß, but not p38α, in the PKP3 knockdown cells. These results suggest that PKP3 loss leads to an increase in the nuclear translocation of p38 MAPK and p38ß MAPK is required for the increase in LCN2 expression.

Stevens-Johnson syndrome and toxic epidermal necrolysis-like cutaneous presentation of chikungunya fever: A case series.

Chikungunya fever is a benign, self-limiting, acute viral illness. An epidemic occurred in New Delhi, India, in August and September 2016. We observed many cases with atypical cutaneous features mimicking Stevens-Johnson syndrome and toxic epidermal necrolysis during this epidemic, especially in infants and children. Twenty-one children (13 [61.9%] boys, 8 [38%] girls) presenting with vesico-bullous and necrotic lesions were reviewed. Cutaneous presentation included vesicles and bullae with purpuric macules and necrosis, seen in 16 (76%) patients. Skin lesions resolved in 5-7 days, leaving behind hyperpigmentation in seven (33.3%) patients and hypopigmentation in three (14.2%). Minor oral erosions were observed in three (14.2%) patients, and palmoplantar erythema was seen in four (19.04%). It is essential for dermatologists to understand the Stevens-Johnson syndrome and toxic epidermal necrolysis-like presentation of chikungunya and not to misinterpret it as true Stevens-Johnson syndrome and toxic epidermal necrolysis, which will lead to unnecessary intervention and management.

14-3-3γ-Mediated transport of plakoglobin to the cell border is required for the initiation of desmosome assembly in vitro and in vivo.

The regulation of cell-cell adhesion is important for the processes of tissue formation and morphogenesis. Here, we report that loss of 14-3-3γ leads to a decrease in cell-cell adhesion and a defect in the transport of plakoglobin and other desmosomal proteins to the cell border in HCT116 cells and cells of the mouse testis. 14-3-3γ binds to plakoglobin in a PKCµ-dependent fashion, resulting in microtubule-dependent transport of plakoglobin to cell borders. Transport of plakoglobin to the border is dependent on the KIF5B-KLC1 complex. Knockdown of KIF5B in HCT116 cells, or in the mouse testis, results in a phenotype similar to that observed upon 14-3-3γ knockdown. Our results suggest that loss of 14-3-3γ leads to decreased desmosome formation and a decrease in cell-cell adhesion in vitro, and in the mouse testis in vivo, leading to defects in testis organization and spermatogenesis.

High Dependency Units (HDUs) in Pediatrics: Need of the Hour in Resource-Limited Settings.

Background Critically ill children, being vulnerable and having higher mortality as compared to adults, require specialized intensive care. However, the focus of critical care remains on adults, especially in resource-limited countries. Limited beds in the pediatric intensive care unit (PICU) along with the limitation of infrastructure and staff add to the challenge in pediatric critical care. In such scenarios, high-dependency units (HDUs) can help save a few more lives, who could not be provided with the PICU facility. HDU provides a level of care that is intermediate to that of the PICU and the general ward providing close observation, monitoring, and intervention to children who are critically ill. Our study highlighted that critically ill children can be given a chance of survival in resource-limited settings through HDU care. Materials and methods In our single-center prospective observational study, 204 children (less than 18 years) admitted to the HDU over 11 months and fulfilling the inclusion criteria were included. Blood samples were drawn for baseline investigations. The child's clinical course in the HDU along with the total duration of stay were recorded in a proforma. Children were reviewed for the requirement of invasive, non-invasive respiratory support along with inotropic support. Various parameters of the pediatric risk of mortality (PRISM) IV score were recorded within a time period of two hours prior and four hours following admission to HDU. The final outcome of the children was recorded. All data were analyzed and reviewed. Results Among the 204 patients admitted to HDU 136 (66.7%) children were treated successfully, whereas 63 (30.9%) children succumbed to their disease and complications, and five children were transferred to the PICU. Among various factors of age less than one year, the primary indication of admission being respiratory distress, the need of >2 inotropes had higher odds of mortality. Odds of mortality were eight times in patients with shock and altered sensorium, three times in children with respiratory distress, and two times in those having seizures. Those patients with a PRISM IV score of >15 had almost 100 times higher odds of mortality as compared to those with a score of <15. Conclusion In a resource-limited setting like ours, there's a scarcity of PICU beds for the provision of critical care. We envisage that providing intensive care in HDU will help save a few more lives, who could not be provided PICU facility for any reason.

Clinical and Laboratory Profile of Macrophage Activation Syndrome in Kawasaki Disease: A Single Centre Cross-Sectional Study.

OBJECTIVE: To study the prevalence of macrophage activation syndrome (MAS) in children with Kawasaki disease (KD) and to devise a classification tree for predicting MAS in early KD based on easily available clinical and laboratory information using artificial intelligence (AI) technology. METHODS: A cross-sectional observational study was conducted (March 2020 - October 2021) during which hospitalized children aged 1-18 years with KD were consecutively enrolled. Those with a positive RTPCR test or IgM/IgG serology for COVID-19 were excluded. The clinical and laboratory profiles of children with and without MAS were studied. A multivariable logistic regression (LR) model was developed utilizing backward elimination method to determine the relationship between select candidate predictor variables and MAS in patients with KD. A classification tree was created based on these using artificial intelligence algorithms. RESULTS: Sixty-two children were diagnosed with KD during the study period, of these, 42 children with KD were included; 14 (33.3 %) were diagnosed with MAS. The median (IQR) duration of fever (days) was significantly more in MAS than those without MAS [7 (5, 15) vs 5 (5, 9), P < 0.05]. Serum albumin (g/dL) was significantly lower in those with MAS [2.3 (2.2, 2.7) vs 2.8 (2.3, 3.1), P = 0.03]. The classification tree constructed using the AI-based algorithm predicted that in children with KD who had myocardial dysfunction, serum albumin < 2.8 g/dL and fever > 6 days duration at admission had an increased likelihood of developing MAS. In children without myocardial dysfunction, alanine transaminase (ALT) levels > 70 U/L and fever > 5 days were equally predictive of MAS. CONCLUSION: Nearly one-third of the children with KD had MAS. Clinicians should consider screening all children with KD for MAS at admission. A classification tree based on the presence of myocardial dysfunction, duration of fever > 6 days, ALT levels and hypoalbuminemia can identify MAS in the course of KD.

Utility of Subjective Global Nutritional Assessment Tool for the Assessment of Malnutrition in Pediatric Patients with Chronic Liver Disease.

Purpose: Approximately 30% of children with chronic liver disease (CLD) are malnourished. However, proper assessment of their nutritional status is difficult. The subjective global nutritional assessment (SGNA) is a comprehensive approach that uses nutrition-focused history and examination, followed by grading of malnourishment. We aimed to study the prevalence of malnutrition in children with CLD using the SGNA tool. Methods: This cross-sectional observational study included patients aged <18 years with CLD. Nutritional assessments were recorded using SGNA tool. Conventional anthropometric measurements were performed and corroborated with nutritional status using SGNA tool. Results: A total of 85 children with CLD and mean age of 62 months were enrolled in this study. The prevalence of malnourished children according to SGNA was 34%; 22% were moderately malnourished and 12% were severely malnourished. We found statistically significant differences in anthropometric parameters among the three groups. A moderate degree of agreement was found between SGNA and weight-for-age (W/A) (p=0.020), mid-upper arm circumference (MUAC) (p<0.001), and triceps skin-fold thickness (TSF)-for-age (p=0.029). Furthermore, a fair degree of agreement was found between height-for-age (H/A) (p=0.001) and weight-for-height (W/H) (p<0.001). The sensitivity of W/A for detecting malnutrition was 93%, H/A was 90%, MUAC was 86%, and TSF was 88%. The sensitivity was much lower for W/H and body mass index for age (55% for both). Conclusion: In our study, more than one-third of children with CLD were malnourished. Nutritional assessment using SGNA is a reliable method for evaluating nutritional status and is significantly correlated with common anthropometric measurements.

Indian Academy of Pediatrics Revised Guidelines on Evaluation, Prevention and Management of Childhood Obesity.

JUSTIFICATION: The last guidelines for pediatric obesity were released in 2004 by Indian Academy of Pediatrics (IAP). Since then, there has been an alarming increase in prevalence and a significant shift in our understanding in the pathogenesis, risk factors, evaluation, and management of pediatric obesity and its complications. Thus, it was decided to revise and update the previous recommendations. OBJECTIVES: To review the existing literature on the burden of childhood obesity and its underlying etiology and risk factors. To recommend evaluation of childhood obesity and suggest optimum prevention and management strategies of childhood obesity. PROCESS: The following IAP chapters (Pediatric and Adolescent Endocrinology, Infant and Young Child feeding, Nutrition, Non-Communicable Disease and Adolescent Health Academy) were invited to nominate members to become part of the writing committee. The Committee held discussions on various aspects of childhood obesity through online meetings between February and August, 2023. Recommendations were then formulated, which were analyzed, revised and approved by all members of the Committee. RECOMMENDATIONS: Exogenous or primary obesity accounts for the majority of cases of childhood obesity. It is important to differentiate it from endogenous or secondary obesity as evaluation and management changes depending on the cause. In Indian, in children under 5 years of age, weight for length/height using WHO charts, and in children 5-18 years, BMI using IAP 2015 charts is used to diagnose overweight and obesity. Waist circumference should be routinely measured in all overweight and obese children and plotted on India specific charts, as it is a key measure of cardio-metabolic risk. Routine evaluation for endocrine causes is not recommended, except in short and obese children with additional diagnostic clues. All obese children more than ten years old should be evaluated for comorbidities like hypertension, dyslipidemia, hyperglycemia and non-alcoholic fatty liver disease/metabolic dysfunction associated steatotic liver disease (NAFLD/ MASLD). Prevention and management of childhood obesity mainly involves healthy diet practices, daily moderate to vigorous physical activity and reduced screen time. Pharmacotherapy may be offered as an addition to lifestyle interventions only in cases of class 3 obesity or if there are any life-threatening comorbidities. Finally, surgical management may be offered in children older than 12 years of age with class 2 obesity and associated comorbidities or class 3 obesity with/without comorbidities, only after failure of a proper trial of intense lifestyle modifications and pharmacotherapy for at least 6 months.

Myelofibrosis: a cause of increased transfusion requirement in a child with ß-thalassemia intermedia.

A 10-year-old girl presented with gradually increasing pallor and abdominal distension for 1 year and fever for 6 months. She required 3 packed cell transfusions during this interval. Investigations done revealed that the child had thalassemia intermedia and parents were carrier of ß-thalassemia gene. The increased transfusion requirement in this case prompted further investigations, which revealed the presence of myelofibrosis. Case reports of myelofibrosis and myelodysplastic syndromes with acquired hemoglobin disorders exist in literature, especially α-thalassemia. To the best of our knowledge, this is the first reported case of ß-thalassemia intermedia in association with myelofibrosis.

Steroid-resistant nephrotic syndrome associated with steroid sulfatase deficiency-x-linked recessive ichthyosis: a case report and review of literature.

UNLABELLED: Nephrotic syndrome associated with X-linked recessive ichthyosis due to steroid sulfatase deficiency has rarely been reported in English literature. We describe a 4 and a half-year-old boy presenting with steroid-resistant nephrotic syndrome (SRNS) with an underlying ichthyotic skin present since birth. Renal biopsy revealed minimal change disease. As many of the male members of the family also showed similar skin manifestations, genetic analysis was done on the patient, which revealed deletion of the steroid sulfatase (STS) gene spanning both the 3' as well as the 5'ends. The patient was thus diagnosed with SRNS associated with X-linked recessive ichthyosis. He was started on cyclosporine regimen, and remission was achieved in 5 weeks. We speculate that the deficiency of STS resulting in increased cholesterol sulfate accumulation interferes with the integrity of adherens junctions present between glomerular epithelial cells of the slit diaphragm, and this results in proteinuria and nephrotic syndrome. The nephrotic syndrome remitted with a calcineurin inhibitor medication. CONCLUSION: We suggest that the deficiency of STS is another one in an increasing list of genetic causes of podocytopathy and nephrotic syndrome. Remission of proteinuria in such a case may be achieved with immunosuppressive medication.

Refractory pancytopenia and megaloblastic anemia due to falciparum malaria.

Anemia is a common complication in malarial infection. Direct destruction and ineffective erythropoesis does not adequately explain the cause of anemia in malaria. We present a case with refractory megaloblastic anemia with asymptomatic falciparum malaria. We hypothesize that promoter variants in the inducible nitric oxide synthase gene might be the cause of severe refractory megaloblastic anemia and pancytopenia in our patient. Malaria should always be kept in mind as a cause of anemia especially in endemic areas even if the child is asymptomatic or there is no demonstrable parasite on routine smear examination.

Vitamin D-dependent rickets (VDDR) type 1: case series of two siblings with a CYP27B1 mutation and review of the literature.

Two siblings presented with clinical and biochemical features of rickets, initially suspected as hypophosphatemic rickets. There was no improvement initially, hence the siblings were reinvestigated and later diagnosed as having vitamin D-dependent rickets (VDDR) type 1 due to a rare mutation in the CYP27B1 gene encoding the 1α-hydroxylase enzyme. Both siblings improved with calcitriol supplementation. The initial presentation of VDDR is often confusing and algorithmic evaluation helps in diagnosis. We also present a brief review of the literature, including genetics.

Outcomes of Hospitalized Infants Aged One to Six Months in Relation to Different Anthropometric Indices - An Observational Cohort Study.

OBJECTIVES: Malnutrition in infants less than six months is increasingly recognized. However, the WHO criteria for identifying malnutrition have not been fully evaluated against the risk of in-patient mortality. The observational study was conducted to evaluate the predictability of in-patient mortality of different anthropometric criteria and combination of criteria in order to understand which diagnostic criteria or combination of criteria most accurately predict in-patient mortality. METHODS: Data from a cohort of infants aged one to six months, admitted to Kalawati Saran Children's Hospital, New Delhi between February and December 2018 was analyzed. The discriminatory ability of different anthropometric indexes [weight-for-age Z score (WAZ), weight-for-length Z score (WLZ) and mid-upper arm circumference (MUAC)] and their combinations to predict in-patient mortality was assessed using Receiver operating characteristic (ROC) curves. RESULTS: A total of 1813 infants aged one to six months were admitted during the 11 mo period, of which 107 (5.9%) died in the hospital. Of all admissions, 39.9%, 26% and 23.4% were severely underweight, severely wasted and severely stunted, respectively. WAZ < -3 was the most sensitive predictor of mortality [sensitivity: 74.8%; specificity: 62.3%; area under the curve (AUC): 0.69, 95% CI: 0.64-0.74]. CONCLUSIONS: WAZ < -3 was the most sensitive predictor out of all individual and combined parameters/indexes in identifying infants less than six months at high risk of mortality which suggests that, it should be used to identify at-risk infants between one to six months on admission to in-patient care. Children identified as falling into this category should be properly evaluated and treated during their in-patient stay.

Medication errors in neonates admitted in intensive care unit and emergency department.

BACKGROUND: Medication is the most common health-care intervention, and the errors arising out of its usage are potentially an avoidable cause of iatrogenic injuries. There are reports of medication errors from neonatal emergency setups. AIMS: To study the medication errors of ordering, dispensing and administering in neonates admitted for emergency care and to compare the errors occurring in the emergency department (ED) with those occurring in the neonatal intensive care unit (NICU) of a teaching hospital in north India. PRIMARY OBJECTIVE: To study the medication errors in ordering and dispensing for neonates. SECONDARY OBJECTIVE: To compare these errors in 2 different settings--ED and NICU. MATERIALS AND METHODS: We did a retrospective chart review of neonatal prescriptions written in the 4 months from January to April 2004 in the neonatal intensive care unit and the pediatric emergency department. The prescriptions were analyzed from the case records bearing an even registration number, obtained from the hospital 'medical records' section. Medication error was defined as 'any preventable event that occurs in the process of ordering, transcribing, dispensing, administering or monitoring a drug irrespective of whether the injury occurred or potential for injury was present! RESULTS: A total of 821 prescriptions were analyzed and 81 (9.6%) errors were detected. The error rate was found to be 1.5 (54/38) and 0.7 (27/38) per patient in ED and NICU, respectively, being highly significant in ED. Every tenth prescription had medication error in ordering or dispensing; of this, every sixth prescription in ED and nineteenth prescription in NICU had medication error. Dosing errors were the commonest form of detected errors. None of the errors caused any significant harm to the patient but had the potential to cause severe injury, and majority of these errors were preventable. CONCLUSION: Medication errors are common in neonatology; more so, in emergency departments than in the neonatal intensive care units.

Comparison of needle autopsy with conventional autopsy in neonates.

Tru-cut needle biopsy postmortems were compared with the standard conventional autopsy in neonates. Twenty-five fresh deceased neonates between 33 weeks of gestation to below 28 days of life were examined by both the techniques. Tissue collection by needle biopsy varied from 92% for liver to 20% for spleen while lung, brain, and kidney gave intermediate results. The cause of death could be determined in 17 cases (68%) by needle biopsy and in 24 cases (96%) by conventional postmortem. In 14 of the 17 cases (56%) cause of death could be determined by biopsy which was in full concordance with the findings of the full autopsy, whereas 3 (12%) cases showed only partial concordance and the diagnosis were discrepant in 32% cases. Needle autopsy missed diagnosis of various congenital malformations, which can be discerned only after meticulous gross examination. Needle autopsy is of value when permission for a complete autopsy cannot be obtained.