Prostanoid receptors in hyperfiltration-mediated glomerular injury: Novel agonists and antagonists reveal opposing roles for EP2 and EP4 receptors.

Increased fluid-flow shear stress (FFSS) contributes to hyperfiltration-induced podocyte and glomerular injury resulting in progression of chronic kidney disease (CKD). We reported that increased FFSS in vitro and in vivo upregulates PGE2 receptor EP2 (but not EP4 expression), COX2-PGE2 -EP2 axis, and EP2-linked Akt-GSK3ß-ß-catenin signaling pathway in podocytes. To understand and use the disparities between PGE2 receptors, specific agonists, and antagonists of EP2 and EP4 were used to assess phosphorylation of Akt, GSK3ß and ß-catenin in podocytes using Western blotting, glomerular filtration barrier function using in vitro albumin permeability (Palb ) assay, and mitigation of hyperfiltration-induced injury in unilaterally nephrectomized (UNX) mice at 1 and 6 months. Results show an increase in Palb by PGE2 , EP2 agonist (EP2AGO ) and EP4 antagonist (EP4ANT ), but not by EP2 antagonist (EP2ANT ) or EP4 agonist (EP4AGO ). Pretreatment with EP2ANT blocked the effect of PGE2 or EP2AGO on Palb . Modulation of EP2 and EP4 also induced opposite effects on phosphorylation of Akt and ß-Catenin. Individual agonists or antagonists of EP2 or EP4 did not induce significant improvement in albuminuria in UNX mice. However, treatment with a combination EP2ANT + EP4AGO for 1 or 6 months caused a robust decrease in albuminuria. EP2ANT + EP4AGO combination did not impact adaptive hypertrophy or increased serum creatinine. Observed differences between expression of EP2 and EP4 on the glomerular barrier highlight these receptors as potential targets for intervention. Safe and effective mitigating effect of EP2ANT + EP4AGO presents a novel opportunity to delay the progression of hyperfiltration-associated CKD as seen in transplant donors.

"Cock-walk" gait and "horseshoe moustache" sign on MRI in inherited hypermanganesemia.

Exogenous manganese (Mn) intoxication leads only to neurotoxicity, whereas inherited hypermanganesemia additionally can cause cirrhosis and polycythemia. We report two affected siblings in a family from South India with severe dysarthria, without dysphagia, generalized dystonia, and characteristic "cock-walk" gait which are clinical clues. Genetic study showed homozygous mutation in the first exon of solute carrier family 30 member 10 (SLC30A10) gene (c.134T>C) confirming the diagnosis of inherited hypermanganesaemia with dystonia 1 (HMNDYT1). Characteristic brain MRI finding is involvement of pontine tegmentum on T1 axial images (due to affliction of central tegmental tract [CTT]) with sparing of ventral pons giving rise to "horseshoe moustache" sign. Symmetric hyperintensities in dentate nucleus, globus pallidus, and putamen while relatively sparing caudate nucleus on T1 without signal intensity abnormalities on T2 images are highly suggestive of hypermanganesaemia. Axial diffusion tensor imaging confirmed the "horseshoe moustache" sign to be constituted by the affected CTT. Hypermanganesaemia-induced CTT involvement in T1 needs to be differentiated from the other more common pediatric causes of CTT affliction which are evident on T2 or diffusion weighted images. Identification is crucial as it is a treatable disorder of metal deposition amenable to chelation.

Conversion of No-Show Patients to Telehealth in a Primary Medicine Clinic.

No-shows in primary care clinics prevent patients from receiving essential care and decrease clinic productivity. The COVID-19 pandemic forced physicians to adjust to telemedicine as a necessary method to provide care. In this study no-show patients were converted to telehealth visits thereby allowing physicians to care for their patients and maintain hospital revenue. The most common reasons for "no-shows" were found to be forgetting appointments and transportation issues.

'Remediability Score' as a tool for selecting the most suitable remediation technology for contaminated sites.

Land gets contaminated due to different human activities like illegal industrial discharges, mining activities, and spillage of chemicals. The pollutants released through these activities can have prolonged effects on the environment and human health. Therefore, there is a need for immediate remediation of the contaminated site. Different remediation technologies are available for remediating a land contaminated with a particular contaminant. Selection of the most appropriate remediation technology among all those available would help in quick and efficient remediation of the land. There are multiple factors deciding the appropriateness of a technology for a given case. 'Remediability' is a term introduced in the scientific literature recently to describe the ease with which a contaminated land can be remediated and 'Remediability Score (RS)' quantifies remediability in a 0-100 scale. The higher the value of RS, the greater is the difficulty for remediation. The factors influencing remediability were presented before a group of experts in a Delphi process for assigning comparative importance. From the importance score obtained in a Saaty scale of 1 to 9, the weights of factors were calculated using Fuzzy AHP. The overall RS was calculated by taking the weighted impact of the factors after their magnitudes were normalized. RS for the various remediation technologies feasible for a contaminated site, Kuzhikandam, near Kochi, Kerala was calculated. The site was revealed to be contaminated with heavy metals and organic compounds in a field investigation. The most suitable technology for remediating the selected site was found to be vitrification with an RS of 22. The next best technology, stabilization/solidification, had an RS of 25. The study showed that RS can be a reliable matric for selecting the most suitable remediation technology for a contaminated site.

A Case of Leukemic Pleural Infiltration in the Blast Crisis Phase of Chronic Myeloid Leukemia: An Unusual Extramedullary Involvement.

The development of pleural effusion in chronic myeloid leukemia (CML) is not well-understood and rarely documented in literature. Extramedullary involvement (EMI), which occurs in about 10% of CML cases, typically affects lymph nodes and the spleen. Instances of extensive infiltration of leukemic cells into the pleura are infrequently reported in CML. Here, we report a case of 41-year-old man experiencing significant bilateral pleural effusion with leukemic infiltration during the blast crisis (BC) phase of refractory CML. Examination of the pleural fluid revealed cells with morphological characteristics of myeloblasts. Although very rare, pleural leukemic infiltration should be considered as a cause of pleural effusion in patients with CML, especially in the BC phase.

Comparison of Ultrasound Parameters and Clinical Parameters in Airway Assessment for Prediction of Difficult Laryngoscopy and Intubation: An Observational Study.

Background and objective The primary responsibility of the anesthesiologist is to provide adequate oxygenation and ventilation to the patient by securing the airway. Prediction of Cormack-Lehane (CL) grading preoperatively helps patients' airway management during anesthesia induction, particularly in difficult intubations. Our study aims to evaluate airway assessment modalities using ultrasound and conventional clinical screening methods for predicting difficult laryngoscopy and intubation. Materials and methods This prospective observational study was conducted on 100 patients aged between 18 and 70 years belonging to ASA classes I, II, and III scheduled for elective surgery requiring general anesthesia under endotracheal intubation was included in the study. Patients who needed rapid sequence induction and had a history of difficult intubation, obese patients with a body mass index (BMI) of more than 40, patients with notable swelling in the neck region (thyroid), pregnant patients, and patients with maxillofacial anomalies were excluded from the study. Clinical parameters such as body mass index, neck circumference, modified Mallampati grading, thyromental distance, and ultrasound parameters such as anterior neck soft tissue thickness at the level of the thyrohyoid membrane (ANS-TM) and anterior neck soft tissue thickness at the level of vocal cord (ANS-VC) were obtained preoperatively. After intubation, the CL grading was noted and categorized into two groups: easy (classes 1 and 2) and difficult (classes 3 and 4). Descriptive statistics included frequency and percentage for categorical variables and mean±standard deviation for continuous variables. The chi-square test was applied to find the relationship between easy and difficult laryngoscopy when compared with the outcome for categorical variables. A P value of less than 0.05 was considered significant throughout the study. The receiver operating characteristics curve (ROC curve) was used to determine the sensitivity and specificity to predict the outcomes. Results Ultrasound-guided measurements of ANS-TM and ANS-VC are independent predictors of difficult laryngoscopy compared with clinical screening tests. Of the two parameters, we found that ANS-TM has a better diagnostic value for predicting a difficult airway with an area under the ROC curve (AUC) of 91% compared with ANS-VC, which has an AUC of 84%. Of the clinical parameters, the modified Mallampati grading has an AUC of 81%, leading to better diagnostic value in the prediction of a difficult airway. Conclusion Our study demonstrated that ANS-TM and ANS-VC are independent predictors of a difficult airway. ANS-TM has a better correlation with CL grading. Clinical screening tests should be combined with ultrasound measurements to aid in the better prediction of difficult laryngoscopy.

Clinical Utility of Proband Only Clinical Exome Sequencing in Neurodevelopmental Disorders.

Chromosomal microarray is recommended as the first line of investigation in neurodevelopmental disorders (NDDs). However, advances in next-generation sequencing have unraveled more than 900 genes associated with NDDs, thus improving the genetic diagnosis. Therefore, this study was conducted to explore the utility of clinical exome sequencing (CES) in NDDs from a tertiary care centre in India. A retrospective observational analysis of 78 children with NDDs for whom CES was performed between 2017 and 2021 was conducted. The American College of Medical Genetics and Genomics (ACMG) criteria were used to classify the variants. The mean age was 5.8 ± 3.6 y, and 42 (53%) were male. Pathogenic, likely pathogenic, and variants of uncertain significance (VUS) were observed in 22 (28.2%), 10 (12.8%), and 26 (33.3%) patients, respectively, which included five copy number variants. The diagnostic yield for pathogenic and likely pathogenic variants in NDDs by CES was 41%, which was reasonably high.

A Case of Hypersexuality in a Patient Receiving Aripiprazole for Schizophrenia.

Aripiprazole is an antipsychotic with partial agonist activity at the dopamine D2 receptor which may lead to compulsive behaviors including hypersexuality. The case presented is of a 24-year-old male with schizophrenia who developed hypersexuality symptoms after initiation of aripiprazole. Cessation of the aripiprazole led to a reduction in hypersexual behaviors. Clinicians should be aware of compulsive behaviors including hypersexuality with aripiprazole use and consider discontinuation of aripiprazole treatment in patients suffering from hypersexuality.

Polyaza functionalized graphene oxide nanomaterial based sensor for Escherichia coli detection in water matrices.

Water quality is widely discussed owing to its significance in public health due to the inability to access clean water. Waterborne diseases account for the presence of pathogens like Escherichia coli (E. coli) in drinking water in the environmental community. Owing to the rapid increase of such bacterial microorganisms, a cost-effective sensor setup has been developed. Herein, we demonstrate the amine-functionalized graphene oxide (fGO) based 2D nanomaterial used to graft E. coli on its surface. The comparative analysis of the deposition of nanosheets on the glass substrate and PDMS was executed. The impedance variations of GO-based nanosensor at various concentrations of E. coli were performed and their potential difference was recorded. It was observed that the impedance changes inversely with the bacterial concentrations and was fed to the Arduino microcontroller. The experimental setup was standardized for the range of 0.01 Hz to 100 kHz. The obtained analog data was programmed with a microcontroller and the bacterial concentration in colony-forming units was displayed. The real-time analysis showsthe low-level detection of E. coli in aquatic environments. Experiments were conducted using the developed nanosensor to test the efficiency in complex water matrices and whose behavior changes with various physical, chemical, and environmental factors.

A mouse model of prenatal exposure to Interleukin-6 to study the developmental origin of health and disease.

Systemic inflammation in pregnant obese women is associated with 1.5- to 2-fold increase in serum Interleukin-6 (IL-6) and newborns with lower kidney/body weight ratio but the role of IL-6 in increased susceptibility to chronic kidney (CKD) in adult progeny is not known. Since IL-6 crosses the placental barrier, we administered recombinant IL-6 (10 pg/g) to pregnant mice starting at mid-gestation yielded newborns with lower body (p < 0.001) and kidney (p < 0.001) weights. Histomorphometry indicated decreased nephrogenic zone width (p = 0.039) with increased numbers of mature glomeruli (p = 0.002) and pre-tubular aggregates (p = 0.041). Accelerated maturation in IL-6 newborns was suggested by early expression of podocyte-specific protein podocin in glomeruli, increased 5-methyl-cytosine (LC-MS analysis for CpG DNA methylation) and altered expression of certain genes of cell-cycle and apoptosis (RT-qPCR array-analysis). Western blotting showed upregulated pJAK2/pSTAT3. Thus, treating dams with IL-6 as a surrogate provides newborns to study effects of maternal systemic inflammation on future susceptibility to CKD in adulthood.

Transcription Factor ß-Catenin Plays a Key Role in Fluid Flow Shear Stress-Mediated Glomerular Injury in Solitary Kidney.

Increased fluid flow shear stress (FFSS) in solitary kidney alters podocyte function in vivo. FFSS-treated cultured podocytes show upregulated AKT-GSK3ß-ß-catenin signaling. The present study was undertaken to confirm (i) the activation of ß-catenin signaling in podocytes in vivo using unilaterally nephrectomized (UNX) TOPGAL mice with the ß-galactosidase reporter gene for ß-catenin activation, (ii) ß-catenin translocation in FFSS-treated mouse podocytes, and (iii) ß-catenin signaling using publicly available data from UNX mice. The UNX of TOPGAL mice resulted in glomerular hypertrophy and increased the mesangial matrix consistent with hemodynamic adaptation. Uninephrectomized TOPGAL mice showed an increased ß-galactosidase expression at 4 weeks but not at 12 weeks, as assessed using immunofluorescence microscopy (p < 0.001 at 4 weeks; p = 0.16 at 12 weeks) and X-gal staining (p = 0.008 at 4 weeks; p = 0.65 at 12 weeks). Immunofluorescence microscopy showed a significant increase in phospho-ß-catenin (Ser552, p = 0.005) at 4 weeks but not at 12 weeks (p = 0.935) following UNX, and the levels of phospho-ß-catenin (Ser675) did not change. In vitro FFSS caused a sustained increase in the nuclear translocation of phospho-ß-catenin (Ser552) but not phospho-ß-catenin (Ser675) in podocytes. The bioinformatic analysis of the GEO dataset, #GSE53996, also identified ß-catenin as a key upstream regulator. We conclude that transcription factor ß-catenin mediates FFSS-induced podocyte (glomerular) injury in solitary kidney.

Unmasking the immune microecology of ductal carcinoma in situ with deep learning.

Despite increasing evidence supporting the clinical relevance of tumour infiltrating lymphocytes (TILs) in invasive breast cancer, TIL spatial variability within ductal carcinoma in situ (DCIS) samples and its association with progression are not well understood. To characterise tissue spatial architecture and the microenvironment of DCIS, we designed and validated a new deep learning pipeline, UNMaSk. Following automated detection of individual DCIS ducts using a new method IM-Net, we applied spatial tessellation to create virtual boundaries for each duct. To study local TIL infiltration for each duct, DRDIN was developed for mapping the distribution of TILs. In a dataset comprising grade 2-3 pure DCIS and DCIS adjacent to invasive cancer (adjacent DCIS), we found that pure DCIS cases had more TILs compared to adjacent DCIS. However, the colocalisation of TILs with DCIS ducts was significantly lower in pure DCIS compared to adjacent DCIS, which may suggest a more inflamed tissue ecology local to DCIS ducts in adjacent DCIS cases. Our study demonstrates that technological developments in deep convolutional neural networks and digital pathology can enable an automated morphological and microenvironmental analysis of DCIS, providing a new way to study differential immune ecology for individual ducts and identify new markers of progression.

MoNuSAC2020: A Multi-Organ Nuclei Segmentation and Classification Challenge.

Detecting various types of cells in and around the tumor matrix holds a special significance in characterizing the tumor micro-environment for cancer prognostication and research. Automating the tasks of detecting, segmenting, and classifying nuclei can free up the pathologists' time for higher value tasks and reduce errors due to fatigue and subjectivity. To encourage the computer vision research community to develop and test algorithms for these tasks, we prepared a large and diverse dataset of nucleus boundary annotations and class labels. The dataset has over 46,000 nuclei from 37 hospitals, 71 patients, four organs, and four nucleus types. We also organized a challenge around this dataset as a satellite event at the International Symposium on Biomedical Imaging (ISBI) in April 2020. The challenge saw a wide participation from across the world, and the top methods were able to match inter-human concordance for the challenge metric. In this paper, we summarize the dataset and the key findings of the challenge, including the commonalities and differences between the methods developed by various participants. We have released the MoNuSAC2020 dataset to the public.

Liability allocation in pollution involving multiple responsible parties.

World over, the instances where polluters are made to pay for the damages they have caused to the environment are on the rise. If more than one party is found responsible for pollution, our analyses of statutes of different countries, covering more than 57% of the world population and more than 45% of its land area, showed that liability is to be allocated among the involved parties on an equitable basis, often decided by the courts. Analyses of a number of court cases showed that the factors considered for allocation in such cases may be classified into two, technical and non-technical. It was also found that the legal liability of pollution was different from the technical liability in many cases due to the influence of non-technical factors that were very specific to the case. These non-technical factors often do not fall under the investigation purview of an environmental forensic expert who carries out technical investigations to find the responsible parties. Based on the analysis, it is suggested that the liability allocation be a two-stage process; the first stage being technical liability allocation done by the forensic expert and the second stage the final (legal) allocation by the court. It is also suggested that clear guidelines be prepared for technical liability allocation. There was also a felt need to quantify remediability to make the remediation liability allocation easier.

Paralytic rabies: An acute flaccid myelitis after inadequate post exposure prophylaxis.

Rabies has two distinct clinical syndromes, encephalitic (or 'furious') and paralytic (or 'dumb'). The paralytic form presents as acute flaccid myelitis and is more common in patients who received post-exposure anti-rabies vaccination without rabies immunoglobulins. We have recently had the opportunity to manage a middle-aged man presenting as 'dumb' paralytic rabies.

Draft Genome Sequence of Freshwater-Derived Streptomyces sp. Strain BPSDS2, Isolated from Damte Stream, Northeast India.

We report the draft genome sequence of Streptomyces sp. strain BPSDS2, isolated from freshwater sediments in Northeast India. The draft genome has a size of 8.27 Mb and 7,559 protein-coding sequences.

Draft Genome Sequence of Streptomyces thermocarboxydus BPSAC147, a Potentially Plant Growth-Promoting Endophytic Bacterium.

Streptomyces thermocarboxydus strain BPSAC147 is an endophytic actinobacterium isolated from root tissues of Rhynchotechum ellipticum in Mizoram, Northeast India. The strain showed potentially plant growth-promoting and biocontrol activities. The draft genome consists of 7.37 Mb and encodes 7,060 putative open reading frames.

Draft Genome Sequence of Plant Growth-Promoting Endophytic Microbacterium hydrothermale BPSAC84, Isolated from the Medicinal Plant Mirabilis jalapa.

Endophytic Microbacterium hydrothermale strain BPSAC84, which has antimicrobial potential, was isolated from root tissues of Mirabilis jalapa in Mizoram, Northeast India. The draft genome consists of 3.58 Mb and 3,444 protein-coding sequences.

Proliferation Tumour Marker Network (PTM-NET) for the identification of tumour region in Ki67 stained breast cancer whole slide images.

Uncontrolled proliferation is a hallmark of cancer and can be assessed by labelling breast tissue using immunohistochemistry for Ki67, a protein associated with cell proliferation. Accurate measurement of Ki67-positive tumour nuclei is of critical importance, but requires annotation of the tumour regions by a pathologist. This manual annotation process is highly subjective, time-consuming and subject to inter- and intra-annotator experience. To address this challenge, we have developed Proliferation Tumour Marker Network (PTM-NET), a deep learning model that objectively annotates the tumour regions in Ki67-labelled breast cancer digital pathology images using a convolution neural network. Our custom designed deep learning model was trained on 45 immunohistochemical Ki67-labelled whole slide images to classify tumour and non-tumour regions and was validated on 45 whole slide images from two different sources that were stained using different protocols. Our results show a Dice coefficient of 0.74, positive predictive value of 70% and negative predictive value of 88.3% against the manual ground truth annotation for the combined dataset. There were minimal differences between the images from different sources and the model was further tested in oestrogen receptor and progesterone receptor-labelled images. Finally, using an extension of the model, we could identify possible hotspot regions of high proliferation within the tumour. In the future, this approach could be useful in identifying tumour regions in biopsy samples and tissue microarray images.

Improved segmentation of cerebellar structures in children.

BACKGROUND: Consistent localization of cerebellar cortex in a standard coordinate system is important for functional studies and detection of anatomical alterations in studies of morphometry. To date, no pediatric cerebellar atlas is available. NEW METHOD: The probabilistic Cape Town Pediatric Cerebellar Atlas (CAPCA18) was constructed in the age-appropriate National Institute of Health Pediatric Database asymmetric template space using manual tracings of 16 cerebellar compartments in 18 healthy children (9-13 years) from Cape Town, South Africa. The individual atlases of the training subjects were also used to implement multi atlas label fusion using multi atlas majority voting (MAMV) and multi atlas generative model (MAGM) approaches. Segmentation accuracy in 14 test subjects was compared for each method to 'gold standard' manual tracings. RESULTS: Spatial overlap between manual tracings and CAPCA18 automated segmentation was 73% or higher for all lobules in both hemispheres, except VIIb and X. Automated segmentation using MAGM yielded the best segmentation accuracy over all lobules (mean Dice Similarity Coefficient 0.76; range 0.55-0.91; mean Hausdorff distance 0.9 mm; range 0.8-2.7 mm). COMPARISON WITH EXISTING METHODS: In all lobules, spatial overlap of CAPCA18 segmentations with manual tracings was similar or higher than those obtained with SUIT (spatially unbiased infra-tentorial template), providing additional evidence of the benefits of an age appropriate atlas. MAGM segmentation accuracy was comparable to values reported recently by Park et al. (Neuroimage 2014;95(1):217) in adults (across all lobules mean DSC=0.73, range 0.40-0.89). CONCLUSIONS: CAPCA18 and the associated multi-subject atlases of the training subjects yield improved segmentation of cerebellar structures in children.