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Abstract Background Integrated management of hemicellulosic fraction and its economical transformation to value-added products is the key driver towards sustainable lignocellulosic biorefineries. In this aspect, microbial cell factories are harnessed for the sustainable production of commercially viable biochemicals by valorising C5 and C6 sugars generated from agro-industrial waste. However, in the terrestrial ecosystem, microbial systems can efficiently consume glucose. On the contrary, pentose sugars are less preferred carbon source as most of the microbes lack metabolic pathway for their utilization. The effective utilization of both pentose and hexose sugars is key for economical biorefinery. Results Bioprospecting the food waste and selective enrichment on xylose-rich medium led to screening and isolation of yeast which was phylogenetically identified as Pichia fermentans. The newly isolated xylose assimilating yeast was explored for xylitol production. The wild type strain robustly grew on xylose and produced xylitol with > 40% conversion yield. Chemical mutagenesis of isolated yeast with ethyl methanesulphonate (EMS) yielded seven mutants. The mutant obtained after 15 min EMS exposure, exhibited best xylose bioconversion efficiency. This mutant under shake flask conditions produced maximum xylitol titer and yield of 34.0 g/L and 0.68 g/g, respectively. However, under the same conditions, the control wild type strain accumulated 27.0 g/L xylitol with a conversion yield of 0.45 g/g. Improved performance of the mutant was attributed to 34.6% activity enhancement in xylose reductase with simultaneous reduction of xylitol dehydrogenase activity by 22.9%. Later, the culture medium was optimized using statistical design and validated at shake flask and bioreactor level. Bioreactor studies affirmed the competence of the mutant for xylitol accumulation. The xylitol titer and yield obtained with pure xylose were 98.9 g/L and 0.67 g/g, respectively. In comparison, xylitol produced using non-detoxified xylose rich pre-hydrolysate from sugarcane bagasse was 79.0 g/L with an overall yield of 0.54 g/g. Conclusion This study demonstrates the potential of newly isolated P. fermentans in successfully valorising the hemicellulosic fraction for the sustainable xylitol production.

Cancer cells frequently harbour genetic aberrations that protect them from programmed cell death. Here, the authors show in non-small cell lung cancer that the anti-apoptotic gene MCL-1 is subject to copy number gains and that deletion of MCL-1 reduces tumour formation.

Establishment of a physiologically relevant human hepatocyte‐like cell system for in vitro translational research has been hampered by the limited availability of cell models that accurately reflect human biology and the pathophysiology of human disease. Here we report a robust, reproducible, and scalable protocol for the generation of hepatic organoids from human induced pluripotent stem cells (hiPSCs) using short exposure to nonengineered matrices. These hepatic organoids follow defined stages of hepatic development and express higher levels of early (hepatocyte nuclear factor 4A [HNF4A], prospero‐related homeobox 1 [PROX1]) and mature hepatic and metabolic markers (albumin, asialoglycoprotein receptor 1 [ASGR1], CCAAT/enhancer binding protein α [C/EBPα]) than two‐dimensional (2D) hepatocyte‐like cells (HLCs) at day 20 of differentiation. We used this model to explore the biology of the pleiotropic TRIB1 (Tribbles‐1) gene associated with a number of metabolic traits, including nonalcoholic fatty liver disease and plasma lipids. We used genome editing to delete the TRIB1 gene in hiPSCs and compared TRIB1‐deleted iPSC‐HLCs to isogenic iPSC‐HLCs under both 2D culture and three‐dimensional (3D) organoid conditions. Under conventional 2D culture conditions, TRIB1‐deficient HLCs showed maturation defects, with decreased expression of late‐stage hepatic and lipogenesis markers. In contrast, when cultured as 3D hepatic organoids, the differentiation defects were rescued, and a clear lipid‐related phenotype was noted in the TRIB1‐deficient induced pluripotent stem cell HLCs. Conclusion: This work supports the potential of genome‐edited hiPSC‐derived hepatic 3D organoids in exploring human hepatocyte biology, including the functional interrogation of genes identified through human genetic investigation.

Abstract Background Xylitol is a commercially important chemical with multiple applications in the food and pharmaceutical industries. According to the US Department of Energy, xylitol is one of the top twelve platform chemicals that can be produced from biomass. The chemical method for xylitol synthesis is however, expensive and energy intensive. In contrast, the biological route using microbial cell factories offers a potential cost-effective alternative process. The bioprocess occurs under ambient conditions and makes use of biocatalysts and biomass which can be sourced from renewable carbon originating from a variety of cheap waste feedstocks. Result In this study, biotransformation of xylose to xylitol was investigated using Yarrowia lipolytica, an oleaginous yeast which was firstly grown on a glycerol/glucose for screening of co-substrate, followed by media optimisation in shake flask, scale up in bioreactor and downstream processing of xylitol. A two-step medium optimization was employed using central composite design and artificial neural network coupled with genetic algorithm. The yeast amassed a concentration of 53.2 g/L xylitol using pure glycerol (PG) and xylose with a bioconversion yield of 0.97 g/g. Similar results were obtained when PG was substituted with crude glycerol (CG) from the biodiesel industry (titer: 50.5 g/L; yield: 0.92 g/g). Even when xylose from sugarcane bagasse hydrolysate was used as opposed to pure xylose, a xylitol yield of 0.54 g/g was achieved. Xylitol was successfully crystallized from PG/xylose and CG/xylose fermentation broths with a recovery of 39.5 and 35.3%, respectively. Conclusion To the best of the author’s knowledge, this study demonstrates for the first time the potential of using Y. lipolytica as a microbial cell factory for xylitol synthesis from inexpensive feedstocks. The results obtained are competitive with other xylitol producing organisms.

During the past decade, novel Deep Learning (DL) algorithms, workloads and hardware have been developed to tackle a wide range of problems. Despite the advances in workload and hardware ecosystems, the programming methodology of DL systems is stagnant. DL workloads leverage either highly-optimized, yet platform-specific and inflexible kernels from DL libraries, or in the case of novel operators, reference implementations are built via DL framework primitives with underwhelming performance. This work introduces the Tensor Processing Primitives (TPP), a programming abstraction striving for efficient, portable implementation of DL workloads with high-productivity. TPPs define a compact, yet versatile set of 2D-tensor operators [or a virtual Tensor Instruction Set Architecture (ISA)], which subsequently can be utilized as building-blocks to construct complex operators on high-dimensional tensors. The TPP specification is platform-agnostic, thus, code expressed via TPPs is portable, whereas the TPP implementation is highly-optimized and platform-specific. We demonstrate the efficacy and viability of our approach using standalone kernels and end-to-end DL & High Performance Computing (HPC) workloads expressed entirely via TPPs that outperform state-of-the-art implementations on multiple platforms.

Introduction: Identification of the nature of lymph nodes is one of the most important parameters for diagnosis in any oncological setting. The lymph nodes help in staging and this directly affects the prognostic outcome and treatment decision. Ultrasound is the preferred imaging method for distinguishing between benign and malignant superficial lymph nodes given ease of availability and its low cost. However, there is no reliable and definitive sonographic criterion in ultrasound to categorise lymph nodes as benign or malignant. Aim: To evaluate the role of sonography and sonoelastography in characterisation of cervical lymphadenopathy and to associate the imaging findings with cytopathology. Materials and Methods: This was a prospective cohort study, conducted over a period of one year in Teerthanker Mahaveer Medical College and Research Centre, Moradabad, Uttar Pradesh, India, constituting a total of 60 patients with enlarged cervical lymph nodes and evaluated on B-mode ultrasound and elastography in the present study. The ultrasonography was performed using either Acuson S3000 ultrasound system (Siemens Medical Solutions, Mountain View, CA) with a 4 to 9 MHz linear transducer for conventional sonography and Acoustic Radiation Force Impulse (ARFI) imaging. Results of radiological findings were tabulated, evaluated and collated with pathological findings. Results: ARFI imaging displayed a sensitivity and specificity of 80% and 97.5% respectively with a Positive Predictive Value (PPV) and Negative Predictive Value (NPV) of 94.1% and 90.7% respectively in distinguishing malignant from benign cervical lymphadenopathy with the diagnostic accuracy being 91.67%. Using Area Under the Receiver Operating Characteristic (ROC) Curve (AUC) of 0.925, a cut-off value for shear wave velocity of 2.98 m/s was obtained. Conclusion: ARFI is a novel technique which can be used to characterise the nature of cervical lymph nodes. Present study demonstrated high diagnostic accuracy (91.67%) in comparison with histopathological findings, ARFI elastography may help in distinction of benign from malignant nodes in adjunct to the gray scale.

Background and aims Secondary Prevention by Structured Semi-Interactive Stroke Prevention Package in India (SPRINT India) is a complex intervention delivered across 30 centres in 16 Indian states. The study delivers intervention in 12 languages in the form of workbook, videos and short messaging using cellular device and internet as a tool. Objectives of process evaluation are to assess whether trial was implemented as planned (fidelity and dose); whether, how and why the intervention is effective by looking at the stakeholders’ experiences (effectiveness); to assess reach of intervention in population (reach); how intervention fits into treatment plan to cause behavioural change when adopted (adoption); usefulness for target population and bring behaviour change (maintenance). Methods SPRINT India process evaluation is a prospective, multicentre study conducted with mixed-methods approach. Sample size of centres and stakeholders will be selected by maximum variance purposive sampling strategy. Centres will be stratified primarily for representing the 11 regional language in which the intervention is delivered. Qualitative data will comprise of interviews of patients, care givers and health professionals at these centres using semi structured interview guide. Quantitative data will comprise of all the randomised patients. Process evaluation framework is based on Realist and RE-AIM evaluation models presented according to Medical Research Council’s guidance. The four sections of the framework are context, trial implementation, mechanisms of impact and trial outcomes. Results Interviews of approximately 100 stakeholders and focus group interviews of health professionals and SPRINT India study central coordinating staff will be conducted. Analysis will be done using triangulation methodology. It will incorporate use of both quantitative and qualitative data, data collection techniques, data sources, evaluation models, stakeholders and researchers. Conclusions Process evaluation will identify efficacious factors in intervention package and consolidate use of secondary stroke semi-interactive stroke prevention package into practice and policy to prevent recurrent stroke.

Defects in the development of the ocular lens can cause congenital cataracts. To understand the various etiologies of congenital cataracts, it is important to characterize the genes linked to this developmental defect and to define their downstream pathways that are relevant to lens biology and pathology. Deficiency or alteration of several RNA-binding proteins, including the conserved RBP Celf1 (CUGBP Elav-like family member 1), has been described to cause lens defects and early onset cataracts in animal models and/or humans. Celf1 is involved in various aspects of post-transcriptional gene expression control, including regulation of mRNA stability/decay, alternative splicing and translation. Celf1 germline knockout mice and lens conditional knockout (Celf1cKO) mice develop fully penetrant cataracts in early postnatal stages. To define the genome-level changes in RNA transcripts that result from Celf1 deficiency, we performed high-throughput RNA-sequencing of Celf1cKO mouse lenses at postnatal day (P) 0. Celf1cKO lenses exhibit 987 differentially expressed genes (DEGs) at cut-offs of >1.0 log2 counts per million (CPM), ≥±0.58 log2 fold-change and Celf1cKO lenses. The DEGs were subjected to various downstream analyses including iSyTE lens enriched-expression, presence in Cat-map, and gene ontology (GO) and representation of regulatory pathways. Further, a comparative analysis was done with previously generated microarray datasets on Celf1cKO lenses P0 and P6. Together, these analyses validated and prioritized several key genes mis-expressed in Celf1cKO lenses that are relevant to lens biology, including known cataract-linked genes (e.g., Cryab, Cryba2, Cryba4, Crybb1, Crybb2, Cryga, Crygb, Crygc, Crygd, Cryge, Crygf, Dnase2b, Bfsp1, Gja3, Pxdn, Sparc, Tdrd7, etc.) as well as novel candidates (e.g., Ell2 and Prdm16). Together, these data have defined the alterations in lens transcriptome caused by Celf1 deficiency, in turn uncovering downstream genes and pathways (e.g., structural constituents of eye lenses, lens fiber cell differentiation, etc.) associated with lens development and early-onset cataracts.

Lens epithelial explants are comprised of lens epithelial cells cultured in vitro on their native basement membrane, the lens capsule. Biologists have used lens epithelial explants to study many different cellular processes including lens fiber cell differentiation. In these studies, fiber differentiation is typically measured by cellular elongation and the expression of a few proteins characteristically expressed by lens fiber cells in situ. Chromatin and RNA was collected from lens epithelial explants cultured in either un-supplemented media or media containing 50% bovine vitreous humor for one or five days. Chromatin for ATAC-sequencing and RNA for RNA-sequencing was prepared from explants to assess regions of accessible chromatin and to quantitatively measure gene expression, respectively. Vitreous humor increased chromatin accessibility in promoter regions of genes associated with fiber differentiation and, surprisingly, an immune response, and this was associated with increased transcript levels for these genes. In contrast, vitreous had little effect on the accessibility of the genes highly expressed in the lens epithelium despite dramatic reductions in their mRNA transcripts. An unbiased analysis of differentially accessible regions revealed an enrichment of cis-regulatory motifs for RUNX, SOX and TEAD transcription factors that may drive differential gene expression in response to vitreous.

Hydroponic sweet basil (O. basilicum L.) farming uses a recirculating nutrient solution that may spread waterborne microbial contamination including algae. Plasma, the fourth state of matter, generates antimicrobial reactive oxygen and nitrogen species when exposed to water. The objective of this work was to study the effect of plasma-treated water-based nutrient solution on plant growth and in reduction of algae. Basil plants were grown in isolated ebb and flow hydroponic systems (under monitored environmental conditions) using nutrient solution (NS) and plasma-activated nutrient solution (PANS) with two separate treatments: the same irrigation solutions were used in the growth cycle (Treatment 1: NST1 and PANST1 once at the beginning growth cycle) and new irrigation solutions at every week of the growth cycle (Treatment 2: NST2 and PANST2). The plant growth parameters (height, fresh and dry weight, number of branches and nodes, root length, leaf index), quality parameters (color, texture, aroma, and tissue nutrients concentration), and algae concentrations were measured. Compared to NST1, plants grown on PANST1 were significantly taller (up to 12%), had a higher fresh mass (up to 29%) and dry mass (up to 45%), and had a higher greenness value (up to 28%). Algae growth was significantly reduced in the PANST2 reservoir (up to 24%) compared to the NST2 reservoir. It was confirmed that Treatment 1 significantly improved the yield, morphology, and quality of sweet basil plants, while Treatment 2 was best suited to decreasing algae concentration in the hydroponic environment. This preliminary study indicated that PANS could improve the quality and growth of sweet basil in hydroponic farming while controlling the algae growth in the growing environment.

GWAS was applied to the antinutritive compounds vicine and convicine (V, C) in winter faba bean. V, C and V + C data for 189 inbred lines (five environments) were predicted by NIRS. These lines do not carry the strong “vc-“ allele (locus VC1). Lab data for 646 samples enabled our NIRS calibration, which performed well for V and V + C yet poor for C. Heritability was high (0.911; 0.868) for V and V + C and lower for C (0.737). From the 2542 mapped SNPs, 47 were significantly associated with V and one with V + C. Four SNPs mapped near to the VC1 locus and were significant for V. Seemingly, non-“vc-“ alleles at that locus contributed to V variation. Marker-assisted breeding with this germplasm can reduce the V + C content to about 0.44%, compared to the current lowest line with 0.55%. Further research will show inasmuch this can serve agronomy and breeding.

Introduction: Breast carcinoma demands attention as it causes high morbidity and mortality. It is important to recognise benign lesions to distinguish them from in situ and invasive breast cancer and to assess a patient’s risk of developing breast cancer, so that the most appropriate treatment modality for each case can be established. The p63 has been characterised as a reliable marker of myoepithelial cells of lactiferous duct. It is exclusively expressed in myoepithelial cells of normal breast tissue. Hence, p63 can be of great help in the differential diagnosis involving benign lesions. Also, p63 may aid in distinguishing benign from malignant lesions. Aim: To study the Immunohistochemistry (IHC) expression of p63 in benign and malignant breast lesions. Materials and Methods: The prospective study was conducted on 76 breast specimens for a period of one year, from 1st December 2018 to 30th November 2019 in the Department of Pathology, Bhagat Phool Singh, Government Medical College for Women, Khanpur Kalan, Sonepat, Haryana, India. IHC assessment for p63 nuclear protein was performed. The intensity of p63 expression was evaluated as continuous positive, discontinuous positive and negative. The extent was scored on the basis of percentage of positive cells and assigned a score of negative (0%), 1 (

Background: Lack of compliance to medication and uncontrolled risk factors are associated with increased risk of recurrent stroke and acute coronary syndrome in patients with recent stroke. Multimodal patient education may be a strategy to improve the compliance to medication and early adoption of nonpharmacological measures to reduce the vascular risk factor burden in patients with stroke. We thus aim to develop multilingual short messaging services (SMS), print, and audio-visual secondary stroke prevention patient education package. The efficacy of the package will be tested in a randomized control trial to prevent major cardiovascular and cerebrovascular events. Methods: In the formative stage, intervention materials (SMS, video, and workbook) were developed. In the acceptability stage, the package was independently assessed and modified by the stakeholders involved in the stroke patient care and local language experts. The modified stroke prevention package was tested for implementation issues (implementation stage). Results: Sixty-nine SMS, six videos, and workbook with 11 chapters with 15 activities were developed in English language with a mean ± SD SMOG index of 9.1 ± 0.4. A total of 355 stakeholders including patients (24.8%), caregivers (24.8%), doctors (10.4%), nurses (14.1%), local language experts (2.8%), physiotherapists (13.2%), and research coordinators (9.8%) participated in 10 acceptability stage meetings. The mean Patient Education Material Assessment Tool understandability score in all languages for SMS, video scripts, and workbook was 95.2 ± 2.6%, 95.2 ± 4.4%, and 95.3 ± 3.6%, respectively. The patients [n = 20, mean age of 70.3 ± 10.6 years and median interquartile range (IQR) baseline NIHSS 1 (0–3)] or the research coordinators (n = 2) noted no implementation issues at the end of 1 month. Conclusion: An implementable complex multilingual patient education material could be developed in a stepwise manner. The efficacy of the package to prevent major adverse cardiovascular events is being tested in the SPRINT INDIA study.

Background: Vasculitic neuropathies are an uncommon heterogeneous group of nerve disorders characterized by inflammation of the vasa nervorum, which may be either systemic vasculitic neuropathy (SVN) or non-SVN (NSVN). Settings and Design: This is a retrospective, observational study. Materials and Methods: Nerve biopsy-proven cases of vasculitic neuropathy (VN) from January 2011 to December 2017 were included in the study. Results: Twenty-five percent (40/156) patients had vasculitis on biopsy, 62.5% were male and the mean age was 52.8 ± 14.9 years. The median (interquartile range) duration of symptoms before diagnosis was 75 (345) days. Thirty (75%) patients had NSVN and 10 (25%) patients had SVN. The clinical pattern of peripheral nervous system involvement was as follows: 20 (50%) had polyneuropathy, 13 (32.5%) had mononeuritis multiplex, and 7 (32.5%) had polyradiculoneuropathy. Twenty-three (57.5%) patients received immunomodulatory therapy. Conclusion: VN has a wide spectrum of clinical presentation. High index of suspicion followed by nerve biopsy is needed for diagnosis. Early treatment may improve prognosis in this group of patients.

Usually, innovative products came to market with no standardization in their characteristics, resulting in frequent incompatibilities among similar products from different manufacturers. Thus, it is important to have standards available to industry as soon as possible. However, if standards are developed too early, maybe products finally do not succeed in the market and standardization efforts are lost, or maybe the technology continues evolving before it achieves some stability. If standards are developed too late, then probably their impact and usefulness for industry is much more limited. Basing in the adoption curve for innovative products, we formulated a proposal about the right moment to develop new standards for such products. To facilitate comparisons, we have defined an adequate common metric scale. This proposal has been verified by the analysis of some cases of innovative products, in which we have reviewed the adoption process and the starting of the standardization activity.

Collision tumour is defined as the presence of two distinct tumours in the same organ without any histological intermixing. Such tumours involving ovaries are extremely rare. We are presenting report of two cases of ovarian collision tumour. Our first case revealed features of mature cystic teratoma with mucinous cystadenoma. The second case showed components of both mature cystic teratoma and serous cystadenoma. Correct diagnosis of collision tumour will guide the surgeon towards proper treatment and favorable prognosis of patient.

Ewing sarcoma (ES) is an uncommon mesenchymal neoplasm that typically develops as a bone mass, although up to 30% arise in extraskeletal sites. ES of the gastrointestinal (GI) and hepatobiliary tract is rare and may be misdiagnosed as other, more common neoplasms that occur in these sites. However, the correct classification of extraskeletal ES is important for timely clinical management and prognostication. We reviewed our experience of ES in the GI and hepatobiliary tract in order to further highlight the clinicopathologic features of these neoplasms and document the potential for misdiagnosis in this setting. The archives and consultation files of 6 academic institutions were retrospectively queried for cases of ES occurring in the GI and hepatobiliary tract. The histologic slides and ancillary studies were reviewed and clinical data were retrieved for each case through the electronic medical records, when available. Twenty-three patients with ES in the GI and/or hepatobiliary tract were identified from 2000 to 2022. Of these, 11 were women and 12 were men with a median age of 38 years (range, 2 to 64). Tumor locations included the pancreas (n=5), liver (n=2), stomach (n=3), colorectum (n=3), and small intestine (n=5), as well as tumors involving multiple organs, pelvis and retroperitoneum (n=5). Tumor size varied between 2 cm and 18 cm. Twenty were primary and 3 were metastases. Of the 23 cases, only 17% were initially diagnosed as ES. The most common misdiagnoses involved various forms of neuroendocrine neoplasia due to expression of synaptophysin and other neuroendocrine markers (22%). A wide variety of diagnoses including GI stromal tumor was considered due to aberrant CD117 expression (4%). The diagnosis of ES was ultimately confirmed by detection of the EWSR1 rearrangement in 22 cases. The remaining case was diagnosed using traditional immunohistochemistry. Follow-up information was available in 20 cases, with follow-up time varying between 2 and 256 months. Six patients with follow-up died of disease between 6 and 60 months following initial presentation. Our data indicate ES in the GI and hepatobiliary tract is commonly misdiagnosed leading to a delay in therapy. In light of the attendant therapeutic and prognostic implications, ES should be considered in the differential diagnosis of any GI or hepatobiliary tumor with epithelioid and/or small round cell morphology.

ImportanceMost research to understand postacute sequelae of SARS-CoV-2 infection (PASC), or long COVID, has focused on adults, with less known about this complex condition in children. Research is needed to characterize pediatric PASC to enable studies of underlying mechanisms that will guide future treatment.ObjectiveTo identify the most common prolonged symptoms experienced by children (aged 6 to 17 years) after SARS-CoV-2 infection, how these symptoms differ by age (school-age [6-11 years] vs adolescents [12-17 years]), how they cluster into distinct phenotypes, and what symptoms in combination could be used as an empirically derived index to assist researchers to study the likely presence of PASC.Design, setting, and participantsMulticenter longitudinal observational cohort study with participants recruited from more than 60 US health care and community settings between March 2022 and December 2023, including school-age children and adolescents with and without SARS-CoV-2 infection history.ExposureSARS-CoV-2 infection.Main outcomes and measuresPASC and 89 prolonged symptoms across 9 symptom domains.ResultsA total of 898 school-age children (751 with previous SARS-CoV-2 infection [referred to as infected] and 147 without [referred to as uninfected]; mean age, 8.6 years; 49% female; 11% were Black or African American, 34% were Hispanic, Latino, or Spanish, and 60% were White) and 4469 adolescents (3109 infected and 1360 uninfected; mean age, 14.8 years; 48% female; 13% were Black or African American, 21% were Hispanic, Latino, or Spanish, and 73% were White) were included. Median time between first infection and symptom survey was 506 days for school-age children and 556 days for adolescents. In models adjusted for sex and race and ethnicity, 14 symptoms in both school-age children and adolescents were more common in those with SARS-CoV-2 infection history compared with those without infection history, with 4 additional symptoms in school-age children only and 3 in adolescents only. These symptoms affected almost every organ system. Combinations of symptoms most associated with infection history were identified to form a PASC research index for each age group; these indices correlated with poorer overall health and quality of life. The index emphasizes neurocognitive, pain, and gastrointestinal symptoms in school-age children but change or loss in smell or taste, pain, and fatigue/malaise-related symptoms in adolescents. Clustering analyses identified 4 PASC symptom phenotypes in school-age children and 3 in adolescents.Conclusions and relevanceThis study developed research indices for characterizing PASC in children and adolescents. Symptom patterns were similar but distinguishable between the 2 groups, highlighting the importance of characterizing PASC separately for these age ranges.

Author(s): Virinder Kumar Bansal (corresponding author) [1]; Krishna Asuri [1]; Deepti Singh [1]; Keshav Agarwal [2]; Raghunandan Dixit [1]; Om Prakash [1]; Subodh Kumar [1]; Rajeshwari Subramaniam [3]; Rashmi Ramachandran [...], Introduction: Laparoscopic transperitoneal adrenalectomy was first described by Gagner M et al. Here, we present our experience of more than two decades of laparoscopic adrenalectomy performed in a single surgical unit at a tertiary care centre. Patients and Methods: A prospectively collected database of patients undergoing laparoscopic adrenalectomy from December 1994 to May 2020 was analysed retrospectively. The demographic profile, details of clinical workup and laboratory parameters were recorded in a pre-structured pro forma. Functional workup and anatomical imaging were performed for all the patients. Patients were taken up for surgery after adequate pre-operative optimisation using a multidisciplinary approach. All the patients were operated by a single surgical team of experienced laparoscopic surgeons. Results: A total of 158 patients underwent laparoscopic transperitoneal adrenalectomy. The majority patients were females (64.5). The median tumour size was 5 cm (range, 1-18 cm). The diagnosis in the majority of the patients was pheochromocytoma (56.3). The mean operative time was 80 min (range: 45-210 min). The most common complication was bleeding in 6 (3.7) patients, which required laparotomy. The median duration of post-operative hospital stay was 3 days (range: 1-13). There was no 30-day mortality. The mean follow-up period was 15 months (range: 6-72 months), during which two patients developed local recurrence. Conclusion: The advantages of laparoscopic surgery are well established and have been extensively explored for the management of adrenal lesions. A multidisciplinary approach to management, consisting of endocrinologists, surgeons and anaesthesiologists is preferred. Pre-operative evaluation, optimisation and accurate selection of patients are crucial for successful laparoscopic adrenalectomy. Keywords: Adrenalectomy, laparoscopic, pheochromocytoma

Mutations/deficiency of TDRD7, encoding a tudor domain protein involved in post-transcriptional gene expression control, causes early onset cataract in humans. While Tdrd7 is implicated in the control of key lens mRNAs, the impact of Tdrd7 deficiency on microRNAs (miRNAs) and how this contributes to transcriptome misexpression and to cataracts, is undefined. We address this critical knowledge-gap by investigating Tdrd7-targeted knockout (Tdrd7-/-) mice that exhibit fully penetrant juvenile cataracts. We performed Affymetrix miRNA 3.0 microarray analysis on Tdrd7-/- mouse lenses at postnatal day (P) 4, a stage preceding cataract formation. This analysis identifies 22 miRNAs [14 over-expressed (miR-15a, miR-19a, miR-138, miR-328, miR-339, miR-345, miR-378b, miR-384, miR-467a, miR-1224, miR-1935, miR-1946a, miR-3102, miR-3107), 8 reduced (let-7b, miR-34c, miR-298, miR-382, miR-409, miR-1198, miR-1947, miR-3092)] to be significantly misexpressed (fold-change ≥ ± 1.2, p-value < 0.05) in Tdrd7-/- lenses. To understand how these misexpressed miRNAs impact Tdrd7-/- cataract, we predicted their mRNA targets and examined their misexpression upon Tdrd7-deficiency by performing comparative transcriptomics analysis on P4 and P30 Tdrd7-/- lens. To prioritize these target mRNAs, we used various stringency filters (e.g., fold-change in Tdrd7-/- lens, iSyTE-based lens-enriched expression) and identified 98 reduced and 89 elevated mRNA targets for overexpressed and reduced miRNAs, respectively, which were classified as “top-priority” “high-priority,” and “promising” candidates. For Tdrd7-/- lens overexpressed miRNAs, this approach identified 18 top-priority reduced target mRNAs: Alad, Ankrd46, Ceacam10, Dgat2, Ednrb, H2-Eb1, Klhl22, Lin7a, Loxl1, Lpin1, Npc1, Olfm1, Ppm1e, Ppp1r1a, Rgs8, Shisa4, Snx22 and Wnk2. Majority of these targets were also altered in other gene-specific perturbation mouse models (e.g., Brg1, E2f1/E2f2/E2f3, Foxe3, Hsf4, Klf4, Mafg/Mafk, Notch) of lens defects/cataract, suggesting their importance to lens biology. Gene ontology (GO) provided further insight into their relevance to lens pathology. For example, the Tdrd7-deficient lens capsule defect may be explained by reduced mRNA targets (e.g., Col4a3, Loxl1, Timp2, Timp3) associated with “basement membrane”. GO analysis also identified new genes (e.g., Casz1, Rasgrp1) recently linked to lens biology/pathology. Together, these analyses define a new Tdrd7-downstream miRNA-mRNA network, in turn, uncovering several new mRNA targets and their associated pathways relevant to lens biology and offering molecular insights into the pathology of congenital cataract.