ORAI1 channel gating and selectivity is differentially altered by natural mutations in the first or third transmembrane domain.

KEY POINTS: Gain-of-function mutations in the highly selective Ca2+ channel ORAI1 cause tubular aggregate myopathy (TAM) characterized by muscular pain, weakness and cramping. TAM-associated mutations in ORAI1 first and third transmembrane domain facilitate channel opening by STIM1, causing constitutive Ca2+ influx and increasing the currents evoked by Ca2+ store depletion. Mutation V107M additionally decreases the channel selectivity for Ca2+ ions and its inhibition by acidic pH, while mutation T184M does not alter the channel sensitivity to pH or to reactive oxygen species. The ORAI blocker GSK-7975A prevents the constitutive activity of TAM-associated channels and might be used in therapy for patients suffering from TAM. ABSTRACT: Skeletal muscle differentiation relies on store-operated Ca2+ entry (SOCE) mediated by STIM proteins linking the depletion of endoplasmic/sarcoplasmic reticulum Ca2+ stores to the activation of membrane Ca2+ -permeable ORAI channels. Gain-of-function mutations in STIM1 or ORAI1 isoforms cause tubular aggregate myopathy (TAM), a skeletal muscle disorder with muscular pain, weakness and cramping. Here, we characterize two overactive ORAI1 mutants from patients with TAM: V107M and T184M, located in the first and third transmembrane domain of the channel. When ectopically expressed in HEK-293T cells or human primary myoblasts, the mutated channels increased basal and store-operated Ca2+ entry. The constitutive activity of V107M, L138F, T184M and P245L mutants was prevented by low concentrations of GSK-7975A while the G98S mutant was resistant to inhibition. Electrophysiological recordings confirmed ORAI1-V107M constitutive activity and revealed larger STIM1-gated V107M- and T184M-mediated currents with conserved fast and slow Ca2+ -dependent inactivation. Mutation V107M altered the channel selectivity for Ca2+ ions and conferred resistance to acidic inhibition. Ca2+ imaging and molecular dynamics simulations showed a preserved sensitivity of T184M to the negative regulation by reactive oxygen species. Both mutants were able to mediate SOCE in Stim1-/- /Stim2-/- mouse embryonic fibroblasts expressing the binding-deficient STIM1-F394H mutant, indicating a higher sensitivity for STIM1-mediated gating, with ORAI1-T184M gain-of-function being strictly dependent on STIM1. These findings provide new insights into the permeation and regulatory properties of ORAI1 mutants that might translate into therapies against diseases with gain-of-function mutations in ORAI1.

A cross-sectional retrospective study comparing handwritten operation notes with electronic operation notes.

INTRODUCTION: Electronically completed medical notes have been shown to be superior in legibility and completeness to handwritten ones. Despite this, surgeons continue to use handwritten operation notes. This paper aims to compare the quality of handwritten versus electronic operation notes. METHODS: This is a retrospective cross-sectional single-centre study done at Darent Valley Hospital, a district general hospital at Dartford, UK. We looked at 405 operation notes of patients who had general surgery procedures between 1 January 2020 to 31 January 2021 checking for legibility and completeness of operation note criteria as given by the Royal College of Surgeons of England's Good Surgical Practice. Data were collated using an app that populates comparison criteria in an Excel sheet and were analysed using SPSS (Statistical Package for the Social Sciences). The results are presented in bar graphs and frequency table. RESULTS: In 17 out of the 18 criteria in RCS England's Good Surgical Practice, electronic notes were better completed than handwritten ones (p<0.001). Signature as a criterion had comparable level of completeness in both handwritten and electronic notes, 95% versus 91% respectively. There was 8.3% illegibility in the handwritten note and none in the electronic ones. CONCLUSIONS: Electronic notes are far better completed than handwritten notes in 17 out of the 18 criteria of a good operation note by RCS England. The difference between both forms of notes is far too much; we propose a complete shift in practice from handwritten to electronic format.

Corrigendum to "WCN23-1242 Genetic interaction between XBP1 and Pkd1 modulates ADPKD progression" [Kidney International Reports Volume 8, Issue 3, Supplement, March 2023, Page S250].

[This corrects the article DOI: 10.1016/j.ekir.2023.02.564.].

Unsuspected pheochromocytoma multisystem crisis: a fatal outcome in a young male patient.

Pheochromocytoma is a great mimicker and has varied presentation. It can present as medical emergency with hypertensive emergencies, acute cardiac event, neurological manifestations, systemic inflammatory response syndrome, acute respiratory distress syndrome, and metabolic emergencies. Here we report a young individual who after a bout of exercise developed breathlessness and rapidly developed multiorgan failure which was fatal and post mortem examination revealed pheochromocytoma in right adrenal gland. Pheochromocytoma multisystem crisis is discussed.

Sonocatalytic recovery of ceria from graphite and inhibition of graphite erosion by ionic liquid based platinum nanocatalyst.

Use of ultrasound as an intensified non-destructive decontamination technique for processing graphite limits its reusability beyond a few number of decontamination cycles due to the exfoliation of graphite due to cavitation effects. The current work establishes that the use of platinum nanoparticles in the leachant reduces the erosion of graphite substrate due to cavitation. It presents an improved way of sonochemical recovery of ceria using a mixture of nitric acid, formic acid and hydrazinium nitrate in the presence of platinum nanoparticles and ionic liquid. The platinum nanoparticles catalyst in ionic liquid prevented the generation of the carbon residue due to the combined effect of denitration and reduced sonication. The presence of the catalyst showed a fivefold increase in dissolution kinetics of ceria as well as absence of graphite erosion, facilitating better chances of graphite recycling than the decontamination without the catalyst. The catalytic approach offers a better recycle strategy for graphite with reduced exfoliation and NOx generation due to denitration, making it a more sustainable decontamination process. Since ceria is used as a surrogate for plutonium oxide, the results can be extended to decontaminate such deposits clearly establishing the utility of the presented results in the nuclear industry.

Short- and long-term outcome of laparostomy following intra-abdominal sepsis.

AIM: This study reports the short- and long-term outcomes of laparostomy for intra-abdominal sepsis. METHOD: Twenty-nine sequential patients with intra-abdominal sepsis treated with a laparostomy over 6 years were included. RESULTS: The median age of the patients was 51 years, postoperative intensive care unit stay was 8 days, postoperative length of hospital stay was 87 days and follow up was 2 years. The expected mortality of 25% was insignificantly different from the observed mortality of 33% (P = 0.35). Seven per cent of patients required percutaneous drainage of intra-abdominal collections. An enterocutaneous fistula developed in 31% of all patients and in 15% of those treated with vacuum dressings. Component-separation fascial reconstruction was successful and uncomplicated in 83% of recipients compared with 25% of mesh repairs. CONCLUSION: Laparostomy does not significantly reduce mortality from the expected rate and commits the patient to a prolonged recovery with a high risk of enterocutaneous fistulation. Component-separation fascial reconstruction has a better outcome than mesh repair.

Efficacy of postharvest sodium nitroprusside application to extend storability by regulating physico-chemical quality of pear fruit.

Quality loss in pear fruit during storage reduces its marketability for long run. To increase its storability, the efficacy of postharvest dip treatment donor sodium nitroprusside (SNP) 0.000, 0.001, 0.002 and 0.003 mol L-1 were investigated on pear fruit cv. Patharnakh under storage conditions (low temperature 0-1 °C and relative humidity (90-95%). SNP effectively lowered fruit mass loss, retained colour and higher firmness, suppressed browning and respiration rate and sustained soluble solids content, titratable acidity, total phenol content and ascorbic acid thus conserved the fruit quality for longer period. SNP treatments suppressed the activity of polyphenol oxidase and increased activity of superoxide dismutase enzyme. Additionally, the SNP treated fruit exhibited lesser activities of fruit softening enzymes like pectin methylesterase, polygalacturonase and cellulase. Among all, 0.002 mol L-1 SNP concentration was superior to lengthen storability and sensory quality of pear up to 60 d under cold storage.

Sonochemical recovery of uranium from nanosilica-based sorbent and its biohybrid.

Use of nanomaterials to remove uranium by adsorption from nuclear wastewater is widely applied, though not much work is focused on the recovery of uranium from the sorbents. The present work reports the recovery of adsorbed uranium from the microstructures of silica nanoparticles (SiO2M) and its functionalized biohybrid (fBHM), synthesized with Streptococcus lactis cells and SiO2M, intensified using ultrasound. Effects of temperature, concentration of leachant (nitric acid), sonic intensity, and operating frequency on the recovery as well as kinetics of recovery were thoroughly studied. A comparison with the silent operation demonstrated five and two fold increase due to the use of ultrasound under optimum conditions in the dissolution from SiO2M and fBHM respectively. Results of the subsequent adsorption studies using both the sorbents after sonochemical desorption have also been presented with an aim of checking the efficacy of reusing the adsorbent back in wastewater treatment. The SiO2M and fBHM adsorbed 69% and 67% of uranium respectively in the second cycle. The adsorption capacity of fBHM was found to reduce from 92% in the first cycle to 67% due to loss of adsorption sites in the acid treatment. Recovery and reuse of both the nuclear material and the sorbent (with some make up or activation) would ensure an effective nuclear remediation technique, catering to UN's Sustainable Development Goals.

Cavitation-assisted decontamination of yttria from graphite of different densities.

Yttria coated graphite crucibles are widely used to handle molten refractory and radioactive metals like uranium and plutonium. However, the coated layer suffers damages like cracking and peeling off owing to thermal cycles. As a result, removal of the yttria layer from the graphite surface is essential to ensure reuse of the crucible and minimization of radioactive waste. The present work investigates intensified dissolution of yttria from the coated graphite samples using ultrasound as a non-destructive decontamination technique to recycle the graphite substrate. The optimum conditions established for maximum dissolution were 8 M as acid strength, frequency of 30 kHz, temperature of 45 °C and power density of 8 W cm-2 that resulted in maximum dissolution of 52% in 30 min. Use of an oxidant H2O2 to the acid, did not yield any improvement in the dissolution kinetics, instead, increased oxidation of the graphite substrate was observed, leading to the anomalous weight gain of the graphite substrate despite surface erosion. Effect of ultrasound on the dissolution was pronounced, with almost a threefold increase compared to dissolution performed under silent conditions. Rates of dissolution of yttria from the substrate of different densities and pore size distribution were also studied. The dissolution was slowest from graphite of density 1.82 g cm-3 as the pore size distribution was conducive to accommodate the yttria particles. The dissolution in nitric acid followed ash layer diffusion controlled kinetics. The study has demonstrated the efficacy of application of ultrasound for accelerated decontamination of graphite substrates.

Intensified dissolution of uranium from graphite substrate using ultrasound.

Decontamination of graphite structural elements and recovery of uranium is crucial for waste minimization and recycle of nuclear fuel elements. Feasibility of intensified dissolution of uranium-impregnated graphite substrate using ultrasound has been studied with objective of establishing the effect of operating parameters and the kinetics of sonocatalytic dissolution of uranium in nitric acid. The effect of operating frequency and acoustic intensity as well as the acid concentration and temperature on the dissolution of metal has been elucidated. It was observed that at lower acid concentrations (6 M-8 M), the dissolution ratio increases by 15% on increasing the bath temperature from 45 to 70 °C. At higher acid concentration (>10 M), the increase was only around 5-7% for a similar change in temperature. With 12 M HNO3, pitting was also observed on the graphite surface along with erosion due to high local reaction rates in the presence of ultrasound. For higher frequency of applied ultrasound, lower dissolution rate of uranium was observed though it also leads to high rates of erosion of the substrate. It was thus established that suitable optimization of frequency is required based on the nature of the substrate and the choice of recycling it. The dissolution rate was also demonstrated to increase with acoustic intensity till it reaches to the maximum at the observed optimum (1.2 W/cm2 at 33 kHz). Comparison with silent conditions revealed that enhanced rate was obtained due to the use of ultrasound under optimum conditions. The work has demonstrated the effective application of ultrasound for intensifying the extent of dissolution of metal.

Safety and probiotic evaluation of two Bacillus strains producing antioxidant compounds.

Bacillus species are becoming increasingly relevant for use as probiotics or feed additives where their heat stability can ensure survival in the food matrix or enable long-term storage at ambient temperature. Some Bacillus species are pigmented and in this study, we have examined two strains, one Bacillus pumilus (pigmented red) and the other Bacillus megaterium (pigmented yellow) for their safety for potential use in humans as dietary supplements. In addition, we have set out to determine if they might confer any potential health benefits. Both strains produce C30 carotenoids while the B. pumilus strain also produced large quantities of riboflavin equivalent to genetically modified Bacillus strains and most probably contributing to this strain's pigmentation. Riboflavin's and carotenoids are antioxidants, and we have evaluated the ability of vegetative cells and/or spores to influence populations of Faecalibacterium prausnitzii in the colon of mice. While both strains increased levels of F. prausnitzii, spores of the B. pumilus strain produced a significant increase in F. prausnitzii levels. If found to be reproducible in humans such an effect might, potentially, confer health benefits particularly for those suffering from inflammatory bowel disease.

A case of venous gangrene, treated successfully with thrombolytic therapy and skin grafting.

Venous gangrene is a rare condition. We report a case of venous gangrene, treated successfully with thrombolytic therapy and subsequently by skin grafting.

Proopiomelanocortin-derived peptides are synthesized and released by human keratinocytes.

Proopiomelanocortin (POMC), the precursor for melanotropic, corticotropic, and opioid peptides such as alpha-melanocyte-stimulating hormone (alpha MSH), ACTH, and other related peptides, was originally identified as a product of the pituitary gland. However, recent evidence shows that POMC products can also be produced by nonpituitary tissues. Because keratinocytes, the major constituent of the epidermis exhibit the capacity to release a variety of proinflammatory and immunomodulatory mediators, the present study was performed to investigate whether human keratinocytes are able to produce POMC-derived peptides. Supernatants of human normal keratinocytes and an epidermal carcinoma cell line (A431) contained significant levels of immunoreactive alpha MSH and ACTH. Upon immuneprecipitation and size-exclusion chromatography, keratinocyte-derived alpha MSH exhibited a molecular mass of approximately 1 kD and was biologically active as demonstrated in a tyrosinase bioassay. Northern blot analysis revealed the expression of POMC-specific transcripts (1.3 kb) in both normal keratinocytes and A431 cells. The production of alpha MSH and ACTH could be significantly upregulated both at the protein and mRNA level upon treatment with phorbol myristate acetate, ultraviolet light, or interleukin 1. These data provide first evidence that human keratinocytes produce POMC-derived peptides such as alpha MSH and ACTH. Because POMC-derived peptides recently have been recognized as potent immunomodulatory mediators, their presence in the epidermis may have a major impact on the skin immune system.

Chest pain, dynamic ECG changes and coronary artery disease.

BACKGROUND: Chest pain is a common emergency. Some patients have dynamic ECG changes with chest pain. Purpose of study was to find out the incidence of coronary artery disease (CAD) in patients presenting with chest pain and dynamic ECG changes. MATERIAL AND RESULTS: Seventy two patients presenting with chest pain and dynamic ECG changes were subjected to coronary angiography. Thirty nine patients had typical angina/unstable angina (USA). Twenty seven were males and 12 were females. Mean age was 57.28 +/- 8.91 years. 36 (92.3%) of these were found to have CAD. Only three patients of these had normal coronaries. Thirty three patients had atypical chest pain. Twenty two were females and 11 were males. Mean age was 45.15 +/- 10.18 years. Only two (6%) of these patients had CAD. Thirty one (94%) patients had normal coronaries. CONCLUSION: History of typical angina with dynamic ECG changes is highly suggestive of CAD as a cause of chest pain. Patients with atypical chest pain with dynamic ECG changes have low likelihood of CAD. Young patients and females may have dynamic ECG changes without CAD.

Separation of nuclear isomers for cancer therapeutic radionuclides based on nuclear decay after-effects.

177Lu has sprung as a promising radionuclide for targeted therapy. The low soft tissue penetration of its ß- emission results in very efficient energy deposition in small-size tumours. Because of this, 177Lu is used in the treatment of neuroendocrine tumours and is also clinically approved for prostate cancer therapy. In this work, we report a separation method that achieves the challenging separation of the physically and chemically identical nuclear isomers, 177mLu and 177Lu. The separation method combines the nuclear after-effects of the nuclear decay, the use of a very stable chemical complex and a chromatographic separation. Based on this separation concept, a new type of radionuclide generator has been devised, in which the parent and the daughter radionuclides are the same elements. The 177mLu/177Lu radionuclide generator provides a new production route for the therapeutic radionuclide 177Lu and can bring significant growth in the research and development of 177Lu based pharmaceuticals.

Study of 7 Cases of Giant Cell Tumor of Soft Tissue.

BACKGROUND: Primary giant cell tumour of soft tissues is a distinct but uncommon group of neoplasms morphologically identical to osseous giant cell tumor. METHODS: 7 patients with painless growing soft tissue mass, having no attachment to underlying bone, were identified in a four years retrospective study from two zonal hospitals of armed forces. Histopathology of these lesions revealed admixture of multinucleated giant cell with mononuclear cells. All patients were treated by surgical resection and followed up for recurrence. Results : There were 5 male and 2 female patients in the age group of 18 to 56 years. All lesions were superficial, circumscribed and involved extremities except one. Histologic transition between benign and malignant lesion was present in only one of the 7 patients that recurred after three months of surgery for which she had to be operated again. 2 of our 7 cases were lost in follow up. CONCLUSION: Primary giant cell tumour of soft tissues usually present as a painless mass and needs to be differentiated from other giant cell rich soft tissue tumors. Benign clinical course is expected if the lesion is excised adequately. Its biological behaviour to have low malignant potential is recognized; but this cannot be predicted and metastasis does occur rarely.

Engaging private providers and Ayurvedic practitioners in Bilaspur, India: did it increase TB case detection?

To find 'missing' tuberculosis (TB) cases, in November 2014 we trained private practitioners (PPs) and Ayurvedic practitioners (APs; Indian system of medicine) in Bilaspur district, India, to identify patients with presumptive TB and refer them to sputum microscopy centres. To reinforce this training, we sent weekly text message reminders during January-March 2015. All 50 APs and 23 of 29 PPs participated. The number of patients with presumptive TB referred by the PPs and APs increased from 38 (January-March 2014) to 104 (January-March 2015), and the number of smear-positive TB patients diagnosed increased from 5 to 16, a 220% increase. While the intervention increased the number of referrals, it did not impact case detection at district level, due to the short duration of the intervention and the non-dominant private sector.

Dans le but de trouver des cas de tuberculose (TB) « manquants ¼, nous avons formé des praticiens privés (PP) et des praticiens ayurvédiques (AP ; médecine indienne) du district de Bilaspur, Inde, en novembre 2014 afin d'identifier des patients avec une TB présumée et de les référer dans des centres de microscopie de crachats. Pour renforcer cette formation, nous avons envoyé des messages sous forme de texto chaque semaine comme aide-mémoire de janvier à mars 2015. Tous les 50 AP et 23 des 29 PP ont participé. Le nombre de patients présumés TB référés par les PP et AP a augmenté de 38 (janvier­mars 2014) à 104 (janvier­mars 2015) et le nombre de patients à frottis de TB positifs diagnostiqués a augmenté de 5 à 16 (augmentation de 220%). Si l'intervention a augmenté la référence de patients au laboratoire, elle n'a pas eu d'impact sur la détection des cas au niveau du district à cause de sa durée brève et du fait que le secteur privé n'est pas dominant dans la région.

Con el propósito de detectar los casos de tuberculosis (TB) que 'se pasan por alto', se llevó a cabo una capacitación de los médicos del sector privado (PP) y los médicos del sistema Ayurveda (AP) de medicina tradicional de la India; la formación, realizada en el distrito de Bilaspur en noviembre del 2014, buscó mejorar el reconocimiento de los pacientes con presunción clínica de TB y su remisión a los centros de examen microscópico del esputo. Con el objeto de reforzar la capacitación, se enviaron mensajes recordatorios por SMS de enero a marzo del 2015. Participaron en el estudio todos los 50 AP y 23 de los 29 PP del distrito. El cociente del número de pacientes con presunción de TB remitidos por los PP y los AP de 38 (entre enero y marzo del 2014) a 104 (entre enero y marzo del 2015) y el número de casos diagnosticados de TB con baciloscopia positiva aumentó de cinco a 16 (incremento del 220%). Si bien la intervención aumentó el número de remisiones, la detección de casos a escala del distrito no se modificó, debido a la corta duración de la intervención y a que el sector privado no es predominante en este distrito.

The B-cell tumor promoter Bcl-3 suppresses inflammation-associated colon tumorigenesis in epithelial cells.

Bcl-3 is an atypical member of the inhibitor of kappa light polypeptide gene enhancer in B-cells (IκB) family. It associates with p50/nuclear factor-κB1 (NF-κB1) and p52/NF-κB2 homodimers in nuclei where it modulates transcription in a context-dependent manner. A subset of B-cell tumors exhibits recurrent translocations of Bcl-3, resulting in overexpression. Elevated expression without translocations is also observed in various B-cell lymphomas and even some solid tumors. Here we investigated the role of Bcl-3 in azoxymethane/dextran sulfate sodium (AOM/DSS)-induced colon tumors, a mouse model for colitis-associated colorectal cancers in humans. Contrary to expectations, Bcl-3 suppressed colorectal tumor formation: Bcl-3-deficient mice were relatively protected from DSS-induced epithelial damage and developed more polyps after AOM/DSS treatment, although polyp size was unaffected. DSS-challenged mutant mice exhibited increased recruitment of myeloid-derived suppressor cells, consistent with protection of the epithelium. Loss of Bcl-3 in intestinal epithelial cells was sufficient to increase tumorigenesis. The added tumor burden in mutant mice was dependent on tumor necrosis factor-α (TNFα), a tumorigenic, NF-κB-mediated signaling pathway that was dampened by Bcl-3. These findings reveal a tumor-suppressive role for Bcl-3 in this inflammation-associated cancer model. Bcl-3 thus functions as a tumor promoter or suppressor, depending on the cellular and environmental context.

Macrophage-derived angiogenesis factors.

A majority of angiogenic factors has been shown to be produced by macrophages. This review will give a concise description of their biochemical nature, their isolation from macrophages and their angiogenic activity. Among the factors with mitogenic effects on endothelial cells are basic fibroblast growth factor (bFGF), transforming growth factor-alpha (TGF-alpha) and very probably insulin-like growth factor-1 (IGF-1). Other secretory products such as angiotropin and human angiogenic factor (HAF) are nonmitogenic but promote angiogenesis by inducing migration of endothelial cells. Prostaglandins, platelet-derived growth factor (PDGF), granulocyte-macrophage- and granulocyte-colony stimulating factor (GM-CSF, G-CSF), interleukin 6 (IL-6) and angiotensin converting enzyme (ACE) have also been shown to be angiogenic, but their mode of action is still to be clearly defined. As the extracellular matrix appears to be involved in the control of angiogenesis, macrophage-derived factors that can alter this structure via degradation or via the clotting system will also be discussed. Tumor necrosis factor alpha (TNF-alpha), interleukin 1 (IL-1) and transforming growth factor-beta (TGF-beta) have complex actions on endothelial cells, and can partially inhibit angiogenesis. Among the factors which solely inhibit neovascularization are the interferons. As it is not known whether all of these factors play a role in angiogenesis in vivo attempts to detect them in situ during the course of neovascularization will be described. Finally macrophages will be discussed as cells that may not be mandatory for each phase of the angiogenic process but whose angiogenic capabilities are comprehensive and unsurpassed by any other cell.