LAG3 ectodomain structure reveals functional interfaces for ligand and antibody recognition.

The immune checkpoint receptor lymphocyte activation gene 3 protein (LAG3) inhibits T cell function upon binding to major histocompatibility complex class II (MHC class II) or fibrinogen-like protein 1 (FGL1). Despite the emergence of LAG3 as a target for next-generation immunotherapies, we have little information describing the molecular structure of the LAG3 protein or how it engages cellular ligands. Here we determined the structures of human and murine LAG3 ectodomains, revealing a dimeric assembly mediated by Ig domain 2. Epitope mapping indicates that a potent LAG3 antagonist antibody blocks interactions with MHC class II and FGL1 by binding to a flexible 'loop 2' region in LAG3 domain 1. We also defined the LAG3-FGL1 interface by mapping mutations onto structures of LAG3 and FGL1 and established that FGL1 cross-linking induces the formation of higher-order LAG3 oligomers. These insights can guide LAG3-based drug development and implicate ligand-mediated LAG3 clustering as a mechanism for disrupting T cell activation.

Recycling of polypropylene by supercritical carbon dioxide for extraction of contaminants from beverage cups. A comparison with polyethylene terephthalate and polylactic acid.

BACKGROUND: EU policies towards a circular economy address plastic packaging as one of the significant concerns and sets ambitious recycling targets. Polyolefins (POs) cannot be recycled for food contact using conventional polyethylene terephthalate (PET) recycling approaches. Thermal degradation prevents the use of high temperatures and, consequently, decontamination of POs may be insufficient when using lower temperatures. Polypropylene (PP) beverage cups were decontaminated using supercritical fluid extraction with carbon dioxide (scCO2 ). Decontamination efficiencies (DEs) of selected markers were determined in challenge tests following European Food Safety Authority guidelines. The effects of time (10-60 min) for PET, polylactic acid (PLA), and PP and temperature (60-80 °C) for PP were studied at constant pressure. The physical properties, sensorial properties, and overall migration of treated scCO2 PP were analysed and compared with virgin PP. RESULTS: PP showed the highest average DE, and PET the lowest, for all the surrogates and in all time conditions. A relative increase in the DE with the increase in process time, particularly for PET and to some extent for PLA, was seen. For PP, no significant impact of time and temperature was observed under the conditions tested. The DE of volatile surrogates was higher than that of semi-volatiles. Results indicate that the scCO2 treatment did not affect the physical and sensorial properties, nor the overall migration of PP, although it contributes to a considerable reduction in extractable n < C24 alkanes. CONCLUSIONS: Results indicate that scCO2 can be used to decontaminate post-consumption PP beverage cups with higher DEs than those for PET and PLA, applying mild processing conditions. © 2022 Society of Chemical Industry.

Cancer Drug Development of Carbonic Anhydrase Inhibitors beyond the Active Site.

Carbonic anhydrases (CAs) catalyze the reversible hydration of carbon dioxide to produce bicarbonate and a proton. Multiple CA isoforms are implicated in a range of diseases, including cancer. In solid tumors, continuously dividing cells create hypoxic conditions that eventually lead to an acidic microenvironment. Hypoxic tumor cells have different mechanisms in place to regulate and adjust the surrounding microenvironment for survival. These mechanisms include expression of CA isoform IX (CA IX) and XII (CA XII). These enzymes help maintain a physiological intracellular pH while simultaneously contributing to an acidic extracellular pH, leading to tumor cell survival. Expression of CA IX and CA XII has also been shown to promote tumor cell invasion and metastasis. This review discusses the characteristics of CA IX and CA XII, their mechanism of action, and validates their prospective use as anticancer targets. We discuss the current status of small inhibitors that target these isoforms, both classical and non-classical, and their future design in order to obtain isoform-specificity for CA IX and CA XII. Biologics, such as monoclonal antibodies, monoclonal-radionuclide conjugated chimeric antibodies, and antibody-small molecule conjugates are also discussed.

Unveiling the profound influence of sucralose on metabolism and its role in shaping obesity trends.

Artificial sweeteners, prominently exemplified by sucralose, have become pervasive in contemporary diets, prompting intriguing questions about their impact on metabolism and their potential role in the unfolding trends of obesity. Covering topics from its discovery to analytical methods for detection and determination in food samples, the manuscript scrutinizes the metabolic effects of sucralose. Notably, the association between sucralose intake and obesity is examined, challenging the conventional belief of its role in weight management. The document comprehensively examines in vivo studies, revealing sucralose's implications on insulin resistance, gut microbiota, and metabolic syndrome, providing a nuanced comprehension of its impact on human health. Additionally, it explores sucralose's effects on glucose and lipid metabolism, blood pressure, and cardiovascular health, underscoring its possible involvement in malignancy development. The review concludes with a call for increased public awareness, education, and updated dietary guidelines to help individuals make informed choices about sweetener consumption. The future perspectives section highlights the need for longitudinal studies, exploring alternative sweeteners, and refining acceptable daily intake limits to ensure public health recommendations align with evolving regulatory guidelines. Overall, the manuscript provides a comprehensive overview of sucralose's multifaceted impact on health, urging further research and a balanced perspective on sweetener consumption.

Short term biodistribution and in vivo toxicity assessment of intravenously injected pristine graphene oxide nanoflakes in SD rats.

The present study aimed to elucidate the short term biodistribution of nano sized graphene oxide (GO) along with the toxicological assessment under in-vivo condition with an intent to analyse the toxic effects of sudden accidental exposure of GO The synthesised GO was characterized using UV-Visible spectroscopy, XRD, FTIR, Raman spectroscopy, TGA and DLS. The morphological imaging was performed using SEM, TEM and AFM. With a lateral size of less than 300 nm, these nanoparticles exhibit significant organ barrier permeability of up to 20%. Upon acute exposure to 10 mg/kg dose of ICG-tagged GO nanoflakes through intravenous route, various organs such as kidney, spleen and liver were observed, and the nanoparticles predominantly accumulated in the liver upon 24 h of exposure. Upon confirming the accumulation of these particles in liver through IVIS imaging, our next attempt was to analyse various biochemical and serum parameters. An elevation in various serum parameters such as ALT, AST, Creatinine and Bilirubin was observed. Similarly, in the case of biochemical parameters tested in liver homogenates, an increase in NO, Catalase, GSH, SOD, ROS, LPO, GR, GPx, and GST was observed. This study highlights the potential toxicological risk associated with GO exposure which must be taken into account for any risk analysis associated with GO based consumer products and the occupational hazards.

Handling drug-target selectivity: A study on ureido containing Carbonic Anhydrase inhibitors.

Here we report the synthesis of a series of taurine substituted sulfonamide derivatives 1-29 having the ureido moiety installed at the tail section as selective inhibitors of the tumor associated human (h) Carbonic Anhydrase (CA; EC 4.2.1.1) IX and XII. The series was deeply investigated for their kinetic features which demonstrated a strong dependence on the ureido moiety. High resolution X-ray crystallographic investigation on selected ligand adducts complexed with hCA II and hCA IX-mimic revealed a strong correlation between the ureido moiety and the amino acid residues Q92 and Q67 in both the hCA II and hCA IX-mimic, contributing to highly stabilized ligand-protein complex.

Inhibition of Carbonic Anhydrase Using SLC-149: Support for a Noncatalytic Function of CAIX in Breast Cancer.

Carbonic anhydrase IX (CAIX) is considered a target for therapeutic intervention in solid tumors. In this study, the efficacy of the inhibitor, 4-(3-(2,4-difluorophenyl)-oxoimidazolidin-1-yl)benzenesulfonamide (SLC-149), is evaluated on CAIX and a CAIX-mimic. We show that SLC-149 is a better inhibitor than acetazolamide against CAIX. Binding of SLC-149 thermally stabilizes CAIX-mimic at lower concentrations compared to that of CAII. Structural examinations of SLC-149 bound to CAIX-mimic and CAII explain binding preferences. In cell culture, SLC-149 is a more effective inhibitor of CAIX activity in a triple-negative breast cancer cell line than previously studied sulfonamide inhibitors. SLC-149 is also a better inhibitor of activity in cells expressing CAIX versus CAXII. However, SLC-149 has little effect on cytotoxicity, and high concentrations are required to inhibit cell growth, migration, and invasion. These data support the hypothesis that CAIX activity, shown to be important in regulating extracellular pH, does not underlie its ability to control cell growth.

Physico-chemical and biological treatment strategies for converting municipal wastewater and its residue to resources.

An increase in urbanization and industrialization has not only contributed to an improvement in the lifestyle of people, but it has also contributed to a surge in the generation of wastewater. To date, conventional physico-chemical and biological treatment methods are widely used for the treatment of wastewater. However, the efficient operation of these systems require substantial operation and maintenance costs, and the application of novel technologies for the treatment and disposal of sludge/residues. This review paper focuses on the application of different treatment options such as chemical, catalyst-based, thermochemical and biological processes for wastewater or sludge treatment and membrane-based technologies (i.e. pressure-driven and non-pressure driven) for the separation of the recovered products from wastewater and its residues. As evident from the literature, a wide variety of treatment and resource recovery options are possible, both from wastewater and its residues; however, the lack of planning and selecting the most appropriate design (treatment train) to scale up from pilot to the field scale has limited its practical application. The economic feasibility of the selected technologies was critically analyzed and the future research prospects of resource recovery from wastewater have been outlined in this review.

"The unpredictable brain tumor".

Histiocytosis is a group of rare diseases with vast imaging findings, few of which are distinctive and characteristic that help to differentiate each one of them. Therefore, typical imaging appearances must be recognized to include the possibility in the differential diagnosis, whenever considered pertinent. Hereby, we present one such unique case of histiocytosis in a 26-year-old female, which involved intertwined and overlapping features of radiological findings.

Combating Climate Change-induced Heat Stress: Assessing Cool Roofs and Its Impact on the Indoor Ambient Temperature of the Households in the Urban Slums of Ahmedabad.

BACKGROUND: The rising global temperature and frequent heatwaves are the adverse effects of climate change. The causalities and ill impacts of the heat stress were higher among the slum dwellers because of the vulnerable household structures, which were made by heat-trapping materials like tin sheets, cement sheet (asbestos), plastic, and tarpaulin. The houses are not only dwellings but also a source of livelihood for many slum dwellers as they are involved in home-based work. The increase in the temperature of more than 40°C severely affects health and increases energy expenditures. OBJECTIVE: The present study conducted to identify the efficient cool roof technologies that reduce indoor temperature of the households and improve the heat resilience of dwellings located in the urban slums of Ahmedabad. METHODOLOGY: The performances of cool roof interventions were compared with the nonintervention - roof types, namely, tin, asbestos/cement sheet, and concrete. Relative humidity/temperature data loggers (Lascar EL-USB-2-LCD, Sweden) were used to measure the indoor ambient temperature and humidity. The questionnaire-based survey also has been conducted to understand the socioeconomic status and the perceptions related to roofing and health. RESULTS: The results revealed that selected cool roof technologies including Thermocol insulation, solar reflective white paint on the outer surface of the roof, and Modroof are effectively reducing the indoor temperature as compared to the nonintervention roofing. CONCLUSION: Cool roof technologies have a wider scope as number of informal settlements are increasing across the cities in India and other developing countries. The governments may not able to provide proper housing to all these inhabitants due to various reasons including the land tenure of the habitats. Validated cool roof technologies can be promoted as these structures are not requires legal sanctions and easily dismantled and installed in multiple places and safeguards the investment of urban poor.

PET-CT upstaging of unilateral operable breast cancer and its correlation with molecular subtypes.

CONTEXT: Prognosis and survival rates for breast cancer vary greatly depending on the cancer stage of the patient. Instead of a step-by-step approach using multiple investigations, we can get all the information about the metastatic load of the disease in PET-CT imaging by one single investigation. There is also a correlation between prognosis, FDG uptake, and molecular subtype of breast cancer (Luminal A, Luminal B, Human epidermal growth factor receptor 2 (HER2) positive and Triple-negative). Pre-treatment baseline PET-CT scan was done in 156 unilateral early and operable breast cancer patients from November 2017 to April 2019 in our prospective observational study. AIMS: To evaluate the utility of PET-CT in staging and upstaging of early and operable breast cancer by detection of unsuspected lymph nodes and distant organ metastases.To determine the prognostic association between SUVmax of the primary breast lesion in the upstaged cases and the molecular subtypes. RESULTS: Thus, PET-CT can serve as one-stop imaging in unilateral operable early breast cancer patients for upstaging and prognostication based on the correlation of SUVmax with molecular subtypes of breast lesions in patients who will surely benefit from whole-body imaging.Out of 156 patients, approximately 27 patients were upstaged after pre-treatment PET CT.Six patients were upstaged to stage IIIC and 21 patients were upstaged to IV.Regional nodes like internal mammary and supraclavicular nodes were detected in 7 patients and 5 patients, respectively, out of 156 patients.Non-regional distant nodes and organ metastases were detected in 11 and 18 patients out of 156 patients.Most common molecular subtype detected in the upstaged cases in our study was Luminal A (13 patients) followed by Triple negative (6), Luminal B (3) and HER2-neu-positive subtypes (1). CONCLUSIONS: FDG PET-CT is a substantial modality to provide information on regional, non regional lymph nodes and distant metastases in early operable breast cancer.It helps in evaluating the whole body metastatic burden in a single sitting, therefore, reducing the need for multiple investigations.SUVmax association of the index lesion with molecular subtype in the FDG PET scanning can serve as a prognostication factor in operable early breast cancer patients.

The unforeseen orbital tumor.

Inflammatory myofibroblastic tumor is a rare group of neoplasms showing a mixture of spindle-shaped myofibroblasts or fibroblasts and a variable amount of inflammatory cells (eosinophils, plasma cells, and lymphocytes). They are not usually included in the differential diagnosis of nodules and masses because of their rarity, therefore, remaining an underdiagnosed entity. We report one such rare case in a 3-year-old female.

Status and determinants of birth preparedness and complication readiness in a rural block of Haryana.

INTRODUCTION: Pregnancy and motherhood is a physiological phenomenon. However, approximately 830 women die from preventable causes related to pregnancy and childbirth every day. Birth preparedness and complication readiness (BPACR) improves preventive behavior and improves knowledge of mothers about danger signs. OBJECTIVE: The objective of the study is to assess the status and sociodemographic determinants of BPACR among the women who have experienced motherhood recently. MATERIALS AND METHODS: The study was carried out among 200 women in a rural block of Haryana over a period of 6 months. The tool used to collect data was adapted from survey tools of Johns Hopkins Program for International Education in Gynaecology and Obstetrics/Maternal and Neonatal Health Program. RESULTS: BPACR index came out to be 66.93 and 58.5% women were well prepared for BPACR. Education and occupation of participants, education of participant's husband, socioeconomic status, and caste were found to be significantly associated with BPACR. CONCLUSION: BPACR is a comprehensive strategy to ensure safer pregnancy and motherhood. Providing educational and skill acquisition opportunities for rural women for their empowerment and increasing their role in decision-making are imperative in order to improve BPACR and promote utilization of skilled attendants at every delivery.

Development of fisetin-loaded folate functionalized pluronic micelles for breast cancer targeting.

The natural flavonoid fisetin (FS) has shown anticancer properties but its in-vivo administration remains challenging due to its poor aqueous solubility. The aim of the study was to develop FS loaded pluronic127 (PF)-folic acid (FA) conjugated micelles (FS-PF-FA) by the way of increasing solubility, bioavailability and active targetability of FS shall increase its therapeutic efficacy. FA-conjugated PF was prepared by carbodiimide crosslinker chemistry. FS-PF-FA micelles were prepared by thin-film hydration method and evaluated in comparison with free FS and FS loaded PF micelles (FS-PF). The smooth surfaces with spherical in shape of FS-PF-PF micelles displayed smaller in size (103.2 ± 6.1 nm), good encapsulation efficiency (82.50 ± 1.78%), zeta potential (-26.7 ± 0.44 mV) and sustained FS release. Bioavailability of FS from FS-PF-PF micelles was increased by 6-fold with long circulation time, slower plasma elimination and no sign of tissue toxicity as compared to free FS. Further, the FS-PF-FA micelles demonstrated active targeting effect on folate overexpressed human breast cancer MCF-7 cells. The concentration of the drug needed for growth inhibition of 50% of cells in a designed time period (GI50) was 14.3 ± 1.2 µg/ml for FS while it was greatly decreased to 9.8 ± 0.78 µg/ml, i.e. a 31.46% decrease for the FS-PF. Furthermore, the GI50 value for FS-PF-FA was 4.9 ± 0.4 µg/ml, i.e. a 65.737% decrease compared to FS and 50% decrease compare to FS-PF. The results indicate that the FS-PF-FA micelles have the potential to be applied for targeting anticancer drug delivery.

Protective Role of Melatonin in Streptozotocin Induced Pancreatic Damages in Diabetic Wistar Rat.

BACKGROUND AND OBJECTIVE: Hyperglycemia is a representative hallmark and risk factor for diabetes and is closely linked to diabetes associated complications. The aim of the present study was to evaluate the therapeutic potential of exogenous melatonin against the streptozotocin induced pancreatic damages in rats. MATERIALS AND METHODS: Streptozotocin was injected for consecutive 6 days. Diabetes was confirmed by blood glucose measurement after 72 h and on 7th day after injection. Animals having blood glucose level above 250 mg dL-1 were considered as diabetic and were administered exogenous melatonin for 4 weeks. Animals were euthanized after last dose, pancreas were dissected out, weighed and fixed in Bouin's fixative for histology and further tissues were kept at -20°C for biochemistry. RESULTS: Diabetic rats displayed significant increase in lipid peroxidation, but pancreatic weight index, antioxidant system (GSH, SOD and CAT) showed decrease. Melatonin treatment to diabetic rats restored the alteration in physiological and biochemical markers. Results were supported by the histopathological observations, STZ treated pancreas showed damage in islets of langerhans, while as melatonin treated diabetic rats recovered the cellular architecture which inturn normalize the function of the pancreas. CONCLUSION: Therefore, melatonin might be considered as a molecule to protect the pancreatic damages.

4-Hydroxy-3-nitro-5-ureido-benzenesulfonamides Selectively Target the Tumor-Associated Carbonic Anhydrase Isoforms IX and XII Showing Hypoxia-Enhanced Antiproliferative Profiles.

Human carbonic anhydrases (CA, EC, 4.2.1.1) IX and XII are overexpressed in cancer cells as adaptive response to hypoxia and acidic conditions characteristic of many tumors. In addition, hypoxia facilitates the activity of specific oxido-reductases that may be exploited to selectively activate bioreductive prodrugs. Here, new selective CA IX/XII inhibitors, as analogues of the antitumor phase II drug SLC-0111 are described, namely ureido-substituted benzenesulfonamides appended with a nitro-aromatic moiety to yield an antiproliferative action increased by hypoxia. These compounds were screened for the inhibition of the ubiquitous hCA I/II and the target hCA IX/XII. Six X-ray crystallographies with CA II and IX/mimic allowed for the rationalization of the compounds inhibitory activity. The effects of some such compounds on the viability of HT-29, MDA-MB-231, and PC-3 human cancer cell lines in both normoxic and hypoxic conditions were examined, providing the initiation toward the development of hypoxia-activated antitumor CAIs.

Maternal Near-Miss Audit: Lessons to Be Learnt.

Mother and child constitute a large, vulnerable, and a priority group as the risk is involved with childbearing in women and of growth and development in children. For every woman who dies from pregnancy or childbirth-related causes, it is estimated that twenty more suffer from pregnancy-related illness or experience other severe complications. These women who nearly escape death are categorized under "near miss" which has been defined as "a woman who nearly died but survived a complication that occurred during pregnancy, childbirth or within 42 days of termination of pregnancy." Maternal near-miss audits give us an opportunity to study the cases which were almost similar to those where maternal deaths happened; thus, their review may give concrete evidence of reasons/deficiencies in health care leading to severe complications and even grave consequences as maternal deaths. Near-miss audits will allow the care of critically ill women to be analyzed, deficiencies in the provision of care to be identified, and comparison within and between institutions and, ultimately, improve the quality of obstetric care and further reduce maternal morbidity and mortality.