Evaluation of High Resolution Melting analysis as an alternate tool to screen for risk alleles associated with small kidneys in Indian newborns

<p>Abstract</p> <p>Background</p> <p>Single nucleotide polymorphisms (SNPs) are the most common forms of sequence variations in the human genome. They contribute to the human phenotypic spectrum and are associated with variations in response to pathogens, drugs and vaccines. Recently, SNPs in three human genes involved in kidney development (<it>RET</it>, <it>PAX2 </it>and <it>ALDH1A2</it>) have been reported to be associated with variation in renal size and function. These known SNPs could potentially be used in the clinic as markers for identifying babies who may have smaller kidneys and permit close follow up for early detection of hypertension and acquired renal dysfunction. The aim of this study was to evaluate the use of High Resolution Melting technique (HRM) as a tool for detecting the known SNPs in these three genes in comparison to sequencing which is the gold standard.</p> <p>Methods</p> <p>High resolution melting analysis was performed on 75 DNA samples that were previously sequenced for the known polymorphisms in <it>RET </it>(rs1800860), <it>PAX2 </it>(rs11190688) and <it>ALDH1A2 </it>(rs7169289) genes. The SNPs were G > A transitions in <it>RET </it>and <it>PAX2 </it>and A > G in <it>ALDH1A2 </it>gene. A blinded assessment was performed on these samples for evaluation of the HRM technique as compared to sequencing.</p> <p>Results</p> <p>Each variant had a unique melt curve profile that was reproducible. The shift in melting temperature (Tm) allowed visual discrimination between the homozygous alleles (major and minor) in all three genes. The shape of the melting curve as compared to the major allele homozygous curve allowed the identification of the heterozygotes in each of the three SNPs. For validation, HRM was performed on 25 samples for each of the three SNPs. The results were compared with the sequencing results and 100% correct identification of the samples was obtained for <it>RET</it>, <it>PAX2</it>, and <it>ALDA1H2 </it>gene.</p> <p>Conclusion</p> <p>High Resolution Melting analysis is a simple, rapid and cost effective technique that could be used in a large population to identify babies with the risk alleles. These high risk children could be followed up for early detection of hypertension and acquired renal dysfunction.</p

'I have the confidence to ask': thickening agency among adolescent girls in Karnataka, South India.

Gender norms serve to normalise gender inequalities and constrain girls' agency. This paper examines how girls' agency, along a continuum, is influenced by the interplay between constraining and enabling influences in the girls' environments. We analyse data from a qualitative study nested within a cluster randomised evaluation of Samata, a multi-layered programme supporting adolescent girls to stay in school and delay marriage in Karnataka, South India. Specifically, we compare agency among 22 girls from intervention communities and 9 girls in control communities using data from the final round of interviews in a qualitative cohort. Using the concept of 'thin' and 'thick' agency on a continuum, we identified shocks like mothers' death or illness, poverty stress, gender norms and poor school performance as thinning influences. Good school examination results; norms in support of education; established educational aspirations; supportive parents, siblings and teachers; and strategic government and Samata resources enabled thicker agency. The intervention programme's effect increased in parallel to the gradient from thin to thicker agency among girls in progressively supportive family contexts. Engagement with the programme was however selective; families adhering to harmful gender norms were not receptive to outreach. In line with diffusion theory, late adopters required additional peer encouragement to change norms

Epigenome erosion and SOX10 drive neural crest phenotypic mimicry in triple-negative breast cancer

Intratumoral heterogeneity is caused by genomic instability and phenotypic plasticity, but how these features co-evolve remains unclear. SOX10 is a neural crest stem cell (NCSC) specifier and candidate mediator of phenotypic plasticity in cancer. We investigated its relevance in breast cancer by immunophenotyping 21 normal breast and 1860 tumour samples. Nuclear SOX10 was detected in normal mammary luminal progenitor cells, the histogenic origin of most TNBCs. In tumours, nuclear SOX10 was almost exclusive to TNBC, and predicted poorer outcome amongst cross-sectional (p = 0.0015, hazard ratio 2.02, n = 224) and metaplastic (p = 0.04, n = 66) cases. To understand SOX10’s influence over the transcriptome during the transition from normal to malignant states, we performed a systems-level analysis of co-expression data, de-noising the networks with an eigen-decomposition method. This identified a core module in SOX10’s normal mammary epithelial network that becomes rewired to NCSC genes in TNBC. Crucially, this reprogramming was proportional to genome-wide promoter methylation loss, particularly at lineage-specifying CpG-island shores. We propose that the progressive, genome-wide methylation loss in TNBC simulates more primitive epigenome architecture, making cells vulnerable to SOX10-driven reprogramming. This study demonstrates potential utility for SOX10 as a prognostic biomarker in TNBC and provides new insights about developmental phenotypic mimicry—a major contributor to intratumoral heterogeneity

Phytochemical Analysis, Antifungal and Antioxidant Activity of Leaf and Fruit of Zizyphus xylopyrus (Retz.) Willd. (Rhamnaceae)

In the present study, phytochemical screening, content of total phenolics, antifungal and antioxidant activity of leaf and fruit extract of Zizyphus xylopyrus (Retz.) Willd. (Rhamnaceae) was investigated in vitro. The leaf and fruit extracts were obtained by maceration process using methanol as solvent. The extracts were screened for phytochemicals by standard tests. Flavonoids, saponins, terpenoids, tannins and phenols were found in both extracts. Antifungal activity of extracts was evaluated by poisoned food technique. Leaf extract suppressed the growth of test fungi to high extent when compared to fruit extract. Antioxidant activity was determined by DPPH radical scavenging, ABTS radical scavenging and Ferric reducing assays. Both extracts exhibited radical scavenging and ferric reducing activity in dose dependent manner with marked activity exhibited by leaf extract. Folin-Ciocalteau reagent method was employed to estimate total phenolic content of extracts. Phenolic content was slightly higher in leaf extract. A positive correlation was observed between phenolic content and antioxidant activity. The plant can be used against phytopathogenic fungi and to prevent and control oxidative damage. Further studies are to be carried out to isolate active principles from the plant and to determine their bioactivities

Deep Learning-Based Signal Detection for Rate-Splitting Multiple Access Under Generalized Gaussian Noise

In this paper, we propose a long short-term memory-based deep learning (DL) architecture for signal detection in uplink and downlink rate-splitting multiple access systems with multi-carrier modulation, over Nakagami-m fading and generalized Gaussian noise (GGN). The proposed DL detector completely eliminates the need for the use of successive interference cancellation (SIC), which suffers from disadvantages such as error propagation. In an orthogonal frequency division multiplexing setting, we show that the proposed DL detector outperforms the standard SIC receivers such as the least squares detector and the minimum mean-squared error receiver, and attains the performance of the optimal maximum likelihood detector, in terms of the symbol error rate (SER). Furthermore, we study the effects of the shaping parameter of GGN, hyperparameters of the DL network such as batch size and learning rate on the SER performance

Surgical management of an endodontic retreatment failure of a mandibular first molar

One of the common endodontic iatrogenic mishaps is the extrusion of obturation material which has a negative effect on the long-term prognosis of the tooth. Surgical endodontics has enabled us to save teeth with persistent infections and extrusions when orthograde treatment has been unsuccessful. Apicoectomy of the molars is not frequently performed even though its success rate can reach that of anteriors and premolars. This case report describes the orthograde and surgical management of a mandibular first molar with external root resorption, instrument separation in the canal, and extruded obturation material, which had been unresponsive to endodontic retreatment

Clinico-Laboratory Profile of Autoimmune Encephalitis Amongst Children at Tertiary Care Hospital

Introduction: The causes of encephalitis in children are numerous and is often thought to be mediated by infections, commonly viral. Children with acute encephalitis often undergo extensive testing for infectious etiologies without discovery of a causative agent. In the past 10 years growing number of non-infectious causes like autoimmune encephalitis have been identified making them one of the important causes of encephalitis in children. Hence, we planned this study.Aims and objectives of The Study: To study the clinical and laboratory profile of children with autoimmune encephalitis.Material and methods: This prospective observational study on clinical and laboratory profiles of children with autoimmune encephalitis was conducted from October 2018 to April 2020 at Indira Gandhi Institute of child health, Bengaluru. Children 1year to 18years of age attending a tertiary care super specialty children hospital who fulfil the diagnostic criteria of possible autoimmune encephalitis were considered. History clinical examination and laboratory investigations were carried out with a pre-designed proforma after obtaining the consent from the care givers. Study population were treated with the first, second- and third-line immunotherapy and followed up for 6 months.Results: Out of 30 cases of suspected autoimmune encephalitis, 9 (30%) were serologically confirmed anti-NMDAR encephalitis and 21(70%) are seronegative. 15(50%) were males and 15 (50%) were females. Clinical symptoms were insidious in onset in 23(76%) children. Mean age of onset of symptoms was 6 y. various clinical features seen were seizures 24 (80%), movement disorders 15(50%), speech disturbances 19(63%) and psychiatric symptoms 10 (33%). sleep disturbances 7(23%) and autonomic dysfunction 3(10%). One child had preceding herpes simplex viral (HSV) encephalitis. MRI Brain was abnormal in 14(46%), cerebral atrophy in 2(14%), cystic lesion in temporal horn in 1(7%), diffusion restriction in bilateral frontal lobe in 1(7%), hyperintense signal changes in thalamus in 2(14%) and periventricular white matter signal changes in 8(57%). EEG was abnormal in 16(53%). 13 (43%) had CSF lymphocytic pleocytosis. out of 30 cases of autoimmune encephalitis 12 cases responded to iv methylprednisolone,8 cases each to IVIG and rituximab. Out of 30 cases 4 required mechanical ventilation among which 2cases were seropositive and 2 cases were seronegative AIE. Among 30 cases, 25 cases (83%) improved among which 17 cases were seronegative, and 8cases were seropositive. 3 cases left against medical advice and two children expired due to ventilator associated pneumonia each being seropositive and seronegative. Cases were followed up for 6 months. Mean duration of hospital stay was found to be 15 days. Screening for tumors was done in all and was found to be negative. Early diagnosis and initiation of immunosuppressive therapy has shown that 16(53%) cases had partial improvement, 9(30%) cases had full improvement. Hence timely initiation of immunosuppressive therapy has shown reduction in severity of the disease and improves the overall outcome of the disease. Conclusion: Clinical history and examination play an important role in diagnosis of autoimmune encephalitis. laboratory evidence of CSF analysis, autoantibody panel in serum and CSF, MRI brain as imaging modality further strengthens the diagnosis of autoimmune encephalitis. Early diagnosis and timely intervention is essential for long term outcome. However, it is never too late for diagnosing and treating this entity

Neurofibromatosis type 1 with unusual oral manifestations

Neurofibromatosis (NF) is a genetically transmitted autosomal dominant disorder with variable penetrance and about 50% of cases representing new mutations. It is progressive in nature and one of its unique feature is the diversity of clinical expression from one patent to another and even within a family. The disease is often characterized by complex and multicellular neurofibroma. It may also lead to different complications throughout the life of an affected individual. We report a case of NF involving alveolus of maxilla and mandible causing expansion of the buccal and lingual cortical plates in a 10-year-old gir

Effect of epidural clonidine on characteristics of spinal anaesthesia in patients undergoing gynaecological surgeries: A clinical study

Background and Aims: Combined spinal–epidural (CSE) anaesthesia is being increasingly used for effective post-operative analgesia. This study was designed to evaluate the effect of epidural clonidine on characteristics of spinal anaesthesia for gynaecological surgeries. Methods: This was a prospective randomised, double-blind, controlled study involving sixty patients belonging to American Society of Anesthesiologists Physical Status I and II who underwent gynaecological surgeries were randomly divided into clonidine (C) group and saline (S) group of thirty each. All patients received CSE anaesthesia. Ten minutes before subarachnoid block (SAB), Group C received clonidine 150 μg diluted to 5 ml in normal saline (NS) and Group S received NS epidurally. Hyperbaric bupivacaine (15 mg) was administered intrathecally for both groups after epidural injection. Sensory and motor block characteristics, analgesia, sedation and haemodynamics were observed. Statistical analysis was performed using appropriate tests. Results: Epidural clonidine produced faster onset (37.83 ± 8.58 s in Group C compared to 50.33 ± 8.80 s in Group S, P = 0.001) and prolonged duration of sensory block (241.17±18.65 minutes in group C compared to 150.33±19.16 minutes in group S, P = 0.001). Time for two segment regression of sensory block was193.67 ± 19.82 min in Group C and 109.33 ± 18.56 min Group S (P < 0.001). The duration of analgesia was 299.00 ± 43.38 min in Group C and 152.50 ± 21.04 min in Group S (P < 0.001). Haemodynamics and sedation scores were comparable between two groups. Conclusion: Administration of clonidine epidurally, 10 min before SAB, caused early onset and prolonged duration of motor blockade and analgesia, without any significant post-operative complication