Comparative proteome analysis identifies species-specific signature proteins in Aspergillus pathogens.

Aspergillus flavus and Aspergillus fumigatus are important human pathogens that can infect the lung and cornea. During infection, Aspergillus dormant conidia are the primary morphotype that comes in contact with the host. As the conidial surface-associated proteins (CSPs) and the extracellular proteins during the early stages of growth play a crucial role in establishing infection, we profiled and compared these proteins between a clinical strain of A. flavus and a clinical strain of A. fumigatus. We identified nearly 100 CSPs in both Aspergillus, and these non-covalently associated surface proteins were able to stimulate the neutrophils to secrete interleukin IL-8. Mass spectrometry analysis identified more than 200 proteins in the extracellular space during the early stages of conidial growth and germination (early exoproteome). The conidial surface proteins and the early exoproteome of A. fumigatus were enriched with immunoreactive proteins and those with pathogenicity-related functions while that of the A. flavus were primarily enzymes involved in cell wall reorganization and binding. Comparative proteome analysis of the CSPs and the early exoproteome between A. flavus and A. fumigatus enabled the identification of a common core proteome and potential species-specific signature proteins. Transcript analysis of selected proteins indicate that the transcript-protein level correlation does not exist for all proteins and might depend on factors such as membrane-anchor signals and protein half-life. The probable signature proteins of A. flavus and A. fumigatus identified in this study can serve as potential candidates for developing species-specific diagnostic tests. KEY POINTS: • CSPs and exoproteins could differentiate A. flavus and A. fumigatus. • A. fumigatus conidial surface harbored more antigenic proteins than A. flavus. • Identified species-specific signature proteins of A. flavus and A. fumigatus.

Audit of management of children and adolescents with congenital adrenal hyperplasia as per recent Endocrine Society guidelines.

INTRODUCTION: In view of the modifications in the endocrine society guidelines on evaluation and management of children with congenital adrenal hyperplasia (CAH), we performed a review of children and adolescents with CAH. MATERIAL AND METHODS: An audit of 35 children with CAH presenting to the pediatric endocrinology clinic between January 2014 to November 2021 was conducted by formulating ten audit questions. The areas of focus included: genital reconstructive surgery, neonatal screening for CAH, stress dosing, need for adrenocorticotrophic hormone (ACTH) stimulation test, growth promoting therapy, bone age assessment, adrenal imaging, bone mineral density assessment, adequacy of hormone replacement and appropriate management of non-classical CAH. RESULTS: Conservative approach to genitoplasty in female children increased from 42.9% to 88.9%. Newborn screening identified 4 babies including two asymptomatic males averting saltwasting crisis. Stress dosing of steroids were advised in all and emergency usage of injectable glucocorticoids was warranted in two children. Gonadotropin-releasing hormone (GnRH) analogue therapy improved the final median predicted height by 7 cm in 5 children. Twenty-three (65.7%) had bone age assessment with 14 (40%) having advanced bone age. ACTH stimulation test, Adrenal imaging, dual energy X-ray absorptiometry (DEXA) scan were done in accordance with the guideline. One child with nonclassical CAH was initiated on hydrocortisone replacement for advanced bone age. CONCLUSIONS: A shift to conservative surgical management of females, utility of neonatal screening for CAH, judicious use of growth promoting therapy is highlighted. Need for bone age testing, emergency hydrocortisone provision is warranted in our series.

Congenital hypothyroidism in Indian preterm babies - screening, prevalence, and aetiology.

INTRODUCTION: Paucity of data on hypothyroidism in Indian preterms. Aim of the study: To describe the prevalence, aetiology, and experience with screening for primary hypothyroidism in preterm babies. MATERIAL AND METHODS: A prospective observational study conducted for 3 years in a tertiary care unit, where all babies born < 37 weeks screened by heel prick for Thyroid Stimulating Hormone (TSH) were included. All screen positive cases (TSH ≥ 6 µIU/ml) underwent venous testing immediately; venous TSH ≥ 20 or Free T4 < 0.9 ng/dl was considered as confirmed positive. All babies underwent venous testing at term. Etiological testing was performed where feasible. Confirmed cases were initiated on thyroxine therapy and followed up. RESULTS: 1167 preterm babies presented during the study period. 1147 (98%) underwent TSH screening and 17 (1.4%) were screen positive; 15 babies underwent confirmatory venous test. Of these 15 babies, 2 were confirmed and started on therapy. The remaining 13 babies underwent retest venous sample at term, and 8 of these were confirmed cases. Of the screen-negative babies, 94% underwent repeat venous testing at term/ prior to discharge. Five were confirmed to have congenital hypothyroidism. Thus, the prevalence of congenital hypothyroidism was 1 in 77 preterm babies. No correlation was observed between screening TSH and venous TSH (p > 0.05). Aetiological evaluation in 8 babies revealed secondary to maternal antibody in 4 cases (50%) and permanent thyroid defects in 4 cases (50%). CONCLUSIONS: We observed a high prevalence (1 in 77), need for repeat venous testing, irrespective of initial screening, and significant permanent congenital hypothyroidism (50%) in our series.

Species-Specific Immunological Reactivities Depend on the Cell-Wall Organization of the Two Aspergillus, Aspergillus fumigatus and A. flavus.

Although belong to the same genus, Aspergillus fumigatus is primarily involved in invasive pulmonary infection, whereas Aspergillus flavus is a common cause of superficial infection. In this study, we compared conidia (the infective propagules) of these two Aspergillus species. In immunocompetent mice, intranasal inoculation with conidia of A. flavus resulted in significantly higher inflammatory responses in the lungs compared to mice inoculated with A. fumigatus conidia. In vitro assays revealed that the dormant conidia of A. flavus, unlike A. fumigatus dormant conidia, are immunostimulatory. The conidial surface of A. fumigatus was covered by a rodlet-layer, while that of A. flavus were presented with exposed polysaccharides. A. flavus harbored significantly higher number of proteins in its conidial cell wall compared to A. fumigatus conidia. Notably, ß-1,3-glucan in the A. flavus conidial cell-wall showed significantly higher percentage of branching compared to that of A. fumigatus. The polysaccharides ensemble of A. flavus conidial cell wall stimulated the secretion of proinflammatory cytokines, and conidial cell wall associated proteins specifically stimulated IL-8 secretion from the host immune cells. Furthermore, the two species exhibited different sensitivities to antifungal drugs targeting cell wall polysaccharides, proposing the efficacy of species-specific treatment strategies. Overall, the species-specific organization of the conidial cell wall could be important in establishing infection by the two Aspergillus species.

Utility of the New Indian Society of Paediatric and Adolescent Endocrinology (ISPAE) Guidelines for Congenital Hypothyroidism Screening in a High Risk Unit.

OBJECTIVE: To study the new ISPAE guidelines in neonates with congenital hypothyroidism and present authors' experience in managing these neonates. METHODS: A retrospective review of all babies who were screened for congenital hypothyroidism in the institution over a period of 5 y was conducted. Details pertaining to maternal risk factors, neonatal risk factors, screening TSH, venous TSH and details of etiological evaluation including: ultrasound thyroid, technetium 99m scintigraphy and anti thyroid peroxidase antibody estimation were retrieved. The cases were assessed using the new ISPAE guidelines. RESULTS: During the study period, 8107 babies were screened for congenital hypothyroidism, 83 cases had confirmed disease and 47 had established etiology. There is a fair correlation between screening TSH and venous TSH (r = 0.7, p < 0.05). The estimated incidence of congenital hypothyroidism in present series is 1 in 97 cases. Out of the 83 cases, 36 (43.3%), 16 (19.2%) and 31 (37.3%) cases had screening TSH >20 µIU/mL, 6-20 µIU/mL and < 6 µIU/mL, respectively. Out of the 47 babies with screening TSH <20 µIU/mL, 23 (48.9%), 25 (53.1%) and 12 (25.5%) cases had prematurity, maternal thyroid disease and illness, respectively, as risk factors (some babies had more than one risk factor). Of the 83 cases, all of them had venous TSH >10 µIU/mL and five cases had low Free T4 (<1.1 ng/dl). Thus, none of the cases with congenital hypothyroidism were missed on the new ISPAE guidelines. CONCLUSION: The new ISPAE guidelines for neonatal screening for congenital hypothyroidism are very useful and applicable in Indian neonates.

Impact of maternal thyroid disease on neonatal thyroid status.

OBJECTIVES: Prevalence of Maternal and congenital hypothyroidism is on the rise. To present the thyroid stimulating hormone screening results in babies born to hypothyroid mothers and assess the burden, aetiology of hypothyroidism in these babies. METHODS: All antenatal mothers attending our hospital during the study period were enrolled into the study. Group I includes 249 term babies born to hypothyroid mothers and group II comprises 2154 newborns born to mothers who are euthyroid. Heel prick thyroid stimulating hormone was done for all newborns on day 3 for both groups. Confirmatory venous testing was done for all for babies in group I and screen positives belonging to group II. Evaluation and therapy done as per standard guidelines. RESULTS: Thyroid stimulating hormone values in the two groups are presented. There was significant correlation between peak maternal thyroid stimulating hormone and neonatal day 3 heel prick in group I (r=0.7, P<0.05). The prevalence of positive screening test in groups I and II was 3.8 and 1.03% (p<0.05) whereas corresponding values for confirmed disease was 4.3 and 0.6%, respectively (p<0.05). Aetiological evaluation revealed both transient hypothyroidism (33.3%) and permanent hypothyroidism (66.6%). CONCLUSION: 4.3% of babies born to hypothyroid mothers develop congenital hypothyroidism; aetiology being both transient and permanent. A venous test by 3 weeks is helpful in these babies to improve case identification.

LATCH Score as a Predictor of Exclusive Breastfeeding at 6 Weeks Postpartum: A Prospective Cohort Study.

INTRODUCTION: Assessment of breastfeeding efficiency by objective means is essential to increase the success of breastfeeding. AIMS AND OBJECTIVES: To study the utility of LATCH score in predicting exclusive breastfeeding rates at 6 weeks postpartum along with an optimum cutoff between January 2016 and June 2016. MATERIALS AND METHODS: One hundred mother-infant dyads born of vaginal delivery with gestational age ≥37 weeks and singleton births were included with LATCH scores assessed at birth and 48 hours/discharge and followed up at 6 weeks postpartum. RESULTS: Sixty-two of 100 (62%) infants were exclusively breastfeeding at 6 weeks. Mean LATCH scores at birth (7.17 ± 1.13 versus 4.26 ± 1.7, p = <0.0001) and 48 hours/discharge (9.22 ± 1.01 versus 5.78 ± 1.29, p = <0.0001) were higher in mothers who were breastfeeding at 6 weeks. LATCH score ≥6 at birth had a sensitivity 93.5% and specificity 65.78% and these mothers were 5.92 times more likely to be breastfeeding at 6 weeks. LATCH score ≥8 at 48 hours/discharge had a sensitivity 93.55% and specificity 92.1% with these mothers being 9.28 times more likely to be breastfeeding at 6 weeks postpartum. Receiver operating characteristic (ROC) for LATCH score at birth and exclusive breastfeeding (EBF) at 6 weeks had an area under curve (AUC) 0.915 (cutoff ≥5.5 sensitivity 93.5%, false positive rate-34.2%). ROC for LATCH score at 48 hours/discharge and EBF 6 weeks had an AUC 0.979 (cutoff ≥7.5 sensitivity 93.5%, false positive rate-7.9%). CONCLUSION: LATCH represents a simple, cost-effective tool capable of predicting breastfeeding as early as in the first 24 hours of life, especially in resource-limited settings.