Level Rise Research Articles

Impact of two human milk oligosaccharides and lactose on the faecal microbiome of infants with probable cow's milk allergy.

Cow's milk protein allergy (CMPA) in infancy is associated with intestinal microbial dysbiosis, characterised by low Bifidobacteriaceae levels. The present study aimed to investigate the impact of two human milk oligosaccharides (HMO), lactose (L), and their combination on the faecal microbiome and metabolome of infants with CMPA. Stool samples of 12 term infants with probable CMPA (mean age 4.3 months) were analysed using a validated intestinal fermentation assay (SIFR® technology). For each substrate (i.e. HMO (2'-fucosyllactose [2'-FL] and lacto-N-neotetraose [LNnT]), L and HMO + L), taxonomic microbiome characterisation and untargeted metabolite profiling were performed at multiple timepoints. At baseline, the tested faecal microbiota overall displayed low abundances of Bifidobacteriaceae. Fermentation with either HMO or lactose significantly enriched Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium pseudocatenulatum and, for HMO + L, also Bifidobacterium bifidum. The increase in HMO-utilising bifidobacteria was associated with a significant rise in levels of short-chain fatty acids, aromatic lactic acids and N-acetylated amino acids, with additive effects being observed for HMO + L. The above data suggest that the combination of 2'-FL, LNnT and lactose helps to alleviate the previously reported CMPA-associated intestinal bacterial dysbiosis and induces the production of several beneficial metabolites. The clinical significance of these findings for infants with CMPA requires further investigation.

Dexmedetomidine leads to the amelioration of stress response to surgery.

To determine the impact of intravenous Dexmedetomidine (Dex) administered perioperatively on stress response markers in patients undergoing ENT surgeries. This randomized interventional study was conducted at POFs Hospital and NUMS affiliated WMC, Wah Cantt, in collaboration with IIMC Rawalpindi, Pakistan, from October 2021 to April 2022. One hundred patients aged between 15-60 years, after satisfying the inclusion standards were randomly divided into two groups C and D. Group-C (n=50) received normal saline in addition to the standard anesthesia protocol. The intervention Group-D (n=50) was administered 1µg/kg dexmedetomidine hydrochloride intravenously over 10 minutes just before the induction followed 0.5μg/kg/hr as maintenance dose till the end of surgery. Serum inflammatory biomarkers (interleukins-6, TNF-α and cortisol) were measured in blood samples in both groups, taken at 0 (T0), 30 minutes(T1), and two hours (T2) time intervals and analyzed by using Enzyme-linked immunosorbent assay (ELISA). Data was statistically analyzed using SPSS 23. The patients receiving Dex showed marked decrease in serum levels of cortisol, TNF- α and interleukins-6, from 139.73 to10.18, 99.51 to 0.96 and 85.09 to 0.96 respectively. Comparison between C and D groups revealed significant differences (p≤0.05) in these serum biomarkers. In the present study, intravenous Dex suppressed the intraoperative rise in levels of cytokine secretion and enhanced smooth recovery with no incidence of nausea and vomiting. These effects could be attributed to the anti-inflammatory effects of dexmedetomidine.

Effective Utilization of Pre Engineering building (PEB) construction for Development of Rapid Healthcare facilitates in Medical Emergency

World is developing at a faster pace in all spheres of technology. The parameters whichdetermine the development have changed a lot in the recent past and will keep changing in future too.Rapid rise in levels of education, high rates of technological innovations and applications, ever faster andcheaper communication that dissolves physical and social barriers both within countries andinternationally, greater availability and easier access to information, and the further opening up of globalmarkets are the set of catalytic forces that is accelerating the speed of social change throughout theworld.Pre Engiering building is one of the best application of fast track construction Pre-Engineered Buildingconcept involves the steel building systems which are predesigned and prefabricated .The presentconstruction methodology calls for the best aesthetic look, high quality fast construction, costeffective innovative touch. One has to think for alternative construction system like pre-engineeredsteel buildings. In recent years, the introduction of Pre Engineered Building (PEB) concept in the designof structures has helped in optimizing design. In this report the detail explore of pre engineering as afast tack construction’s application discuss . As the world battles the ongoing COVID-19 pandemic, ithas become very clear that societies depend on functioning healthcare facilities. The sufficientavailability of hospital beds, healthcare staff, protective gear or ventilators can make all the differencebetween a situation that is still manageable and a severe crisis. for this infrastructure facilities should beprepare in short time ,For this EPS Pannel and UPVC panels will be use for rapid construction ofhealthcare facilities

Arginyltransferase 1 (ATE1)-mediated proteasomal degradation of viral haemagglutinin protein: a unique host defence mechanism.

The extensive protein production in virus-infected cells can disrupt protein homeostasis and activate various proteolytic pathways. These pathways utilize post-translational modifications (PTMs) to drive the ubiquitin-mediated proteasomal degradation of surplus proteins. Protein arginylation is the least explored PTM facilitated by arginyltransferase 1 (ATE1) enzyme. Several studies have provided evidence supporting its importance in multiple physiological processes, including ageing, stress, nerve regeneration, actin formation and embryo development. However, its function in viral pathogenesis is still unexplored. The present work utilizes Newcastle disease virus (NDV) as a model to establish the role of the ATE1 enzyme and its activity in pathogenesis. Our data indicate a rise in levels of N-arginylated cellular proteins in the infected cells. Here, we also explore the haemagglutinin-neuraminidase (HN) protein of NDV as a presumable target for arginylation. The data indicate that the administration of Arg amplifies the arginylation process, resulting in reduced stability of the HN protein. ATE1 enzyme activity inhibition and gene expression knockdown studies were also conducted to analyse modulation in HN protein levels, which further substantiated the findings. Moreover, we also observed Arg addition and probable ubiquitin modification to the HN protein, indicating engagement of the proteasomal degradation machinery. Lastly, we concluded that the enhanced levels of the ATE1 enzyme could transfer the Arg residue to the N-terminus of the HN protein, ultimately driving its proteasomal degradation.

Special Aspects of Cholesterol Metabolism in Women.

Cardiovascular diseases due to arteriosclerosis are the most common causes of death and disability in both men and women. Hypercholesterolemia, a treatable risk factor, is often detected after a delay in women, and then inadequately treated. It is, therefore, important to know the sex-specific aspects of cholesterol metabolism and to address them specifically. We conducted a selective literature search in PubMed with particular attention to current guidelines. In the population as a whole, the age-associated rise in serum cholesterol levels occurs approximately 10 years later in women than in men. Women are exposed to a higher cholesterol load than men at the beginning of their lives, and especially after menopause. This is correlated with a later, but nonetheless clinically relevant rise in the incidence of myocardial infarction in older women. Because women's LDL cholesterol and lipoprotein(a) levels rise after menopause, their lipid profiles should be re-evaluated at this time. Moreover, conditions that are specific to women such as polycystic ovary syndrome, contraception, and especially the phases of life-such as planning to become pregnant, pregnancy, and breastfeeding-need to be considered for both diagnostic and therapeutic purposes. Sex-specific differences and cholesterolassociated risks are particularly pronounced in women with familial hypercholesterolemia (prevalence 1:250). Lowering high cholesterol levels, especially in postmenopausal women, may prevent the development of cardiovascular diseases.

The role of pantothenic acid in alleviating hypoxia-induced liver injury of sub-adult grass carp (Ctenopharyngodon idellus): Possible mechanisms and implication

In aquaculture, hypoxia poses a significant threat to fish growth and survival. Despite this, there is limited research on the impact of pantothenic acid (PA) in enhancing the anti-hypoxia stress capability of fish. The primary objective of this study was to explore the effects and potential mechanisms of PA in alleviating hypoxia-induced liver injury in sub-adult grass carp. To achieve this, six experimental diets containing varying levels of PA (0.08, 11.95, 24.95, 37.95, 50.95, and 63.95 mg/kg) were formulated and administered to grass carp (initial weight 696.22 ± 2.45 g) for 63 days. Following the dietary intervention, the fish were segregated into hypoxia and control groups and treated for 96 h. The findings revealed that optimal PA levels could enhance the growth performance of sub-adult grass carp and ameliorate the adverse impacts of hypoxia. First, optimal PA levels reduced serum levels of glutamate-oxaloacetate transaminase (GOT), glutamate-pyruvate transaminase (GPT), and lactate dehydrogenase (LDH), as well as mitigated hepatocyte vacuolization, improved liver and mitochondrial morphology, thereby alleviating hypoxia-induced liver injury. Furthermore, optimal PA levels were associated with increased contents of serum cortisol, glucose, liver glycogen, pyruvate, and the activities of Pantothenate kinase 2 (PanK2) and coenzyme A (CoA), while liver lactate content and LDH activity were reduced, indicating improvements in energy metabolism and PA utilization, potentially mediated by the down-regulation of hypoxia-inducible factor-1α (HIF-1α) mRNA level. Moreover, optimal PA levels decreased reactive oxygen species (ROS) and malondialdehyde (MDA) levels, enhanced liver antioxidant capacity, down-regulated autophagy-related gene mRNA levels, and up-regulated autophagy receptor protein 62 (p62) protein level, suggesting a reduction in oxidative damage and autophagy. Finally, a rise in levels of mitochondrial quality control-related mRNAs such as mitofusin 1/2 (MFN1/2), dynamin-related protein-1 (DRP-1), and nuclear respiratory factor 1/2 (Nrf-1/2) was observed, together with increment in the protein levels of PTEN induced putative kinase 1 (PINK1), BCL-2 interacting protein 3 (BNIP3) and FUN14 domain-containing protein 1 (FUNDC1). These alterations hinted at the role of PA in preserving mitochondrial homeostasis. In conclusion, PA exhibited a protective role against hypoxia-induced liver injury by enhancing liver energy metabolism, bolstering antioxidant defenses, suppressing autophagy, and maintaining mitochondrial homeostasis. Through an assessment of percentage weight gain (PWG) and liver ROS level, the requirements of PA for promoting growth and resilience to hypoxia-induced liver injury in sub-adult grass carp were determined to be 37.44 mg/kg and 42.57 mg/kg, respectively.

Sea−level rise and arsenic−rich soils: A toxic relationship

In the United States, dangerously high arsenic (As) levels have been found in drinking water wells in more than 25 states, potentially exposing 2.1 million people to drinking water high in As; a known carcinogen. The anticipated sea−level rise (SLR) is expected to alter soil biogeochemical and hydrological conditions, potentially impacting their ability to sequester As. In our study of coastal Wilmington, DE, an area projected to experience a 1 −meter SLR by 2100, we examined the spatial distribution, speciation, and release possibilities of As due to SLR. To understand the complex dynamics at play, we employed a comprehensive approach, including bulk and micro X−ray absorption spectroscopy measurements, hydrological pattern evaluation, and macroscopic stirred−flow experiments. Our results suggest that introducing reducing and saline conditions can increase As release in both river water and seawater inundation scenarios, most likely due to ionic competition and the dissolution of As−bearing Fe/Mn oxides. Regardless of the salinity source, the released As concentrations consistently exceeded the EPA threshold for drinking water. Our results provide valuable insights for developing appropriate remedial and management strategies for this site and numerous others facing similar environmental challenges. Environmental ImplicationWith nearly two hundred million individuals living within coastal flood plains and with two million square kilometers of land and one trillion dollars' worth of assets lying less than 1 m above current sea level, sea–level rise (SLR) is one of the significant socio–economic threats associated with global warming. Arsenic is a prevalent contaminant in coastal areas impacted by industrial activities, many of which are susceptible to being impacted by SLR. This study examines SLR's impact on arsenic fate and speciation in a densely populated coastline in Wilmington, DE, expecting 1 meter of SLR by 2100.

To Compare the Efficacy of Sub-Pecto-Interfascial Block with Erector Spinae Plane Block in Postoperative Pain Management in Patients Undergoing Cardiac Surgery

Objectives: Pain following sternotomy has always been an issue of major concern for anesthesiologists. The incidence of pain is as high as 49% at rest following coronary artery bypass grafting. We planned to utilize the sub-pecto-interfascial plane (SIP) block and erector spinae plane (ESP) block to determine its efficacy and quality of analgesia as compared to conventional intravenous analgesia. Material and Methods: After the Institutional Ethics Committee’s approval, we recruited 105 patients and randomized them into three groups. Group 1 received conventional analgesia, group 2 SIP, and group 3 ESP block. Group 2 and Group 3 received allocated blocks after induction of balanced general anesthesia under ultrasound guidance. With high-frequency linear probe (13 MHz) planes identified, a mixture of injection ropivacaine (0.375%) and dexmedetomidine (1.1 mcg/mL) was deposited. In group 2, six injections of 6 mL each at the 2nd, 4th, and 6th intercostal spaces in the bilateral parasternal region. In group 3, two injections of 20 mL of the above-mentioned mixture on each side above the transverse process of the T5 vertebra under the erector spinae muscle plane were injected. All patients were monitored throughout the procedures, and their vitals were recorded. Heart rate, systolic blood pressure, and diastolic blood pressure were noted at baseline, at skin incision, at sternotomy, and 30 min post-extubation. Patients were followed for total fentanyl consumption, pain score (numerical rating scale [NRS] score), and peak inspiratory flow rate, which were noted at extubation and 2, 4, 6, 8, 12, and 24 hours post-extubation. Levels of C-reactive protein (CRP), cortisol, and prolactin were done at baseline and 24 h after surgery. Extubation time, time of first oral intake, and total length of intensive care unit (ICU) stay were also compared between the groups. Results: Total fentanyl consumption was significantly lower (P < 0.005) in group 2 and group 3 compared to group 1. Patient receiving blocks were significantly lower NRS score compared to the control group. Spirometry has shown improved results in block groups catering to early discharge from ICU. Rise in levels of CRP, cortisol, and prolactin were much higher in group 1 as compared to group 2 and group 3. Conclusion: The application of ESP and SIP blocks is far superior than conventional parenteral analgesics in terms of pain score, total opioid’s consumption, hemodynamic maintenance, spirometry efforts, and length of ICU stays.

Gauged and historical abrupt wave front floods (‘walls of water’) in Pennine rivers, northern England

AbstractExtremely rapid rates of rise in level and discharge in a subset of flash floods (‘abrupt wave front floods’, AWF) are separate hazards from peak level. Such flood events are investigated for Pennine catchments in northern England using both gauged and historical information. Gauged level and flow digital records at 15‐min intervals provide recent data. Historical information for 122 AWF events is extracted from a chronology of flash floods for Britain. Historical AWF events are mapped and found to occur on every major Pennine catchment; catchment descriptors are derived as a basis for assessing catchment vulnerability. We discuss the disputed origin of AWF. Using gauged data, we contrast the rising limb of AWF and ‘normal’ floods. We investigate time series of historical AWF, noting a puzzling peak in the late 19th century. Current rainfall and river monitoring does not provide a reliable basis for understanding AWF processes or for operational response and we suggest improvements. Similarly, current models for design flood estimation and forecasting do not generate the observed rapid increase in level in AWF floods.

Correlation of serum biomarker (LDH, CRP, IL-8) levels in pre-malignant and malignant lesions of head and neck

A precancerous lesion is often defined as morphologically altered tissue during which the occurrence of carcinoma is more likely than its normal counterpart. Head and neck cancers include cancers of the mouth and therefore the larynx, also as some rarer cancers like cancer of the sinuses, salivary glands, nose, and tympanic cavity. Head and neck cancers are often described as consistent with the sort of cell they begin with. A biomarker could be defined and interpreted as biological molecule which is mostly found in body fluids, blood or even tissues that indicates abnormalities of our body. Here we've taken three biomarkers (LDH, CRP, IL-8) to gauge their level rise in pre-malignant and malignant lesions which are present in our head and neck. Patients attending ENT OPD with primary symptoms indicating premalignant and malignant lesions in the region of head and neck had been included for the analytical study. Complete ENT examination followed by biopsy of the suspicious lesion and blood sample collection was done. A complete of 30 patients were included in each group of premalignant and malignant lesions. No mean serum rise of serum LDH was noted in premalignant and malignant lesions. A mean serum rise of serum CRP was noted in premalignant lesions but was not noted in malignant lesions. However, both premalignant and malignant lesions noted a mean serum rise of IL-8. From a future perspective serum evaluation of biomarkers can become a replacement diagnostic modality for the early evaluation of premalignant lesions and malignant lesions and for monitoring the prognosis of an equivalent.

Синоптические условия повышения уровня рек на юге Иркутской области и Забайкалья в июле 2023 года

The article discusses natural and anthropogenic factors of rising river levels and associated environmental consequences, existing probabilistic estimates of the amount of floods using various hydrological and climate models. Orographic, climatic and circulation features of the occurrence of floods in the Irkutsk region are identified. A study was carried out of the circulation and weather factors of increasing the intensity and duration of precipitation in early July 2023, which was accompanied by an increase in river levels in the south of the Irkutsk region and the Baikal region. The dynamics of atmospheric processes near the Earth's surface and at the levels of isobaric surfaces AT850 hPa, AT-700 hPa, AT-500 hPa and AT-300 hPa, reflecting the conditions of surface and highaltitude cyclo- and frontogenesis, are analyzed. The contribution of advective flows of heat and moisture, mesojets in the middle troposphere and upper tropospheric jet currents in increasing the amount and duration of atmospheric precipitation is highlighted. A significant contribution to the increase in precipitation and the rise in river levels was made by moist air masses moving in the lower troposphere from the water area of the lake. Baikal, in the middle troposphere from the Yellow Sea. The time of passage of the flood and the height of the rise in the level of individual watercourses as a result of the rains were estimated.

Lake level dynamics in the centre of the East European Plain during the Holocene

Based on the lithostratigraphic and geomorphological approach, the study of lake level fluctuations in Nero and Seliger over the past 15,000 years was conducted. Common features of level changes were revealed: deep regression at the end of the late glacial period and early Holocene, intensive rise in level in the early-middle Holocene, stabilization (with a slight upward trend) in the late Holocene. Similar features in level changes were revealed in the history of a number of lakes in Germany and Scandinavia. A connection was found between lake level fluctuations and the intensity of fluvial processes in the center of the East European Plain.

Horizontal Integration Strategies and Competitiveness of Firms in EPZ Kenya

Global challenges have given a rise in levels of competitiveness among industries that produce similar or related goods and services. One possible solution to this is the use of horizontal integration strategy to ensure efficient production. This study therefore intended to determine the influence of horizontal integration strategy on the competitiveness of EPZ firms. The study was guided by two theories, namely; the resource-based view theory and the transaction economic cost theory. The study adopted the use of a questionnaire. Both quantitative and qualitative approaches were used. Descriptive technique, specifically survey was used in the research design. The target population was all the 137 EPZ firms within Kenya and the sample was drawn from this population. The stratified random sampling technique was used. The EPZ firms are situated within the 74 gazetted zones. The quantitative data that was collected was analysed using the Statistical Package for Social Scientists. Further, the significance level of each independent variable was tested against the dependent variable at 95% level of confidence using ANOVA, regression and correlation techniques. The findings of this study indicated that horizontal integration had a positive and statistically significant relationship with the competitiveness of firms in EPZ Kenya. This study recommends the inclusion of horizontal integration in the strategic plans of the different EPZ firms. This will subsequently lead to an increase in employment opportunities, real-value added growth, and foreign direct investment (KNBS, 2022). In conclusion, from the analysis of findings, horizontal integration is significant for competitiveness of firms in EPZ Kenya.

Energy Efficient Design for a Conventional Building using BIM Tools

Nowadays, energy has become a key player in the AEC industry because of the increase in global warming and energy crisis. Due to global warming the consumption of energy has increased enormously in buildings. The no. of buildings have grown in tandem with rise in levels of urbanization in developing & developed countries, which in return rises the bar for energy consumption. The improved energy performance of the building is significantly influenced by both interior and exterior building factors. It is preferred to perform energy analysis in the design stage of the building. The current paper is the talks about the energy model by improving the design of a conventional buildings, an energy – efficient model is been created which uses less energy and reduces its carbon footprint. Energy analysis using BIM can be used to optimize the building’s design and identify opportunities for improvement, such as implementing energy-saving technologies, improving insulation and air tightness, and reducing energy loss through window frames, it’s glazing type and also redesigning the other parameters including HVAC types, Window shades, change in orientation etc. In the earlier, the conventional model to produce energy through renewable sources like solar and wind, and incorporate them into the building’s design. Thus, obtained results after analysis can be used to take design decisions and ensure the building meets energy efficiency standards and sustainability goals. The use of BIM in the design and analysis of energy-efficient buildings can help support the transition to a low-carbon, sustainable future.

Impact of feeding diets with enhanced vegetable protein content and presence of umami taste-stimulating additive on gastrointestinal amino acid sensing and feed intake regulation in rainbow trout

The regulation of feed intake in fish is dependent upon different neuroendocrine and metabolic mechanisms including amino acid sensing in the gastrointestinal tract (GIT). However, there is little information regarding the impact of diets on such mechanisms. Therefore, in this study, we fed rainbow trout (Oncorhynchus mykiss) with 3 diets: a diet with high content of fishmeal and low content of soybean protein concentrate (SPC) (HF), a diet with a reduced content of fishmeal and high content of SPC (LF), and the LF diet supplemented with an umami taste-stimulating additive (LFU). Fish were fed ad libitum once a day for 4 weeks, with no significant differences being registered in feed intake among groups. At the end of the feeding trial, we collected samples of different areas of the GIT (stomach, proximal and distal intestine) and hypothalamus at different times: after 48 h of fasting (time 0), and 1 h, 4 h, and 24 h after feeding. We evaluated the activity of pepsin in the stomach and trypsin and chymotrypsin in the proximal intestine, as well as mRNA abundance of transcripts encoding amino acid transporters and taste receptors, intracellular signalling molecules, and hormones. Moreover, we assessed the hypothalamic mRNA abundance of neuropeptides involved in feed intake regulation. Feeding rainbow trout with LF did not result in marked alterations in parameters related to digestive function and amino acid sensing in the rainbow trout GIT, nor in the expression of gastrointestinal hormones (except cck) and hypothalamic neuropeptides. In contrast, supplementation of the LF diet with an umami taste-stimulating additive resulted in a general improvement of digestive or absorptive function (increased protein, dry matter and energy digestibility, and earlier peak in plasma amino acid levels) and activation of gut-brain axis mechanisms involved in feed intake regulation through the transcriptional activation of amino acid transporters, taste receptors, signalling molecules, and hormones. These results demonstrate that the dietary inclusion of umami receptor stimulants has the potential to improve fish physiological responses to the rise in levels of vegetable protein in the diet.

Numerical simulation of tsunami in the system of sevastopol bays

Within the framework of numerical simulation, a study was made of the penetration of tsunami waves into the system of Sevastopol bays. The non-linear SWASH hydrodynamic model was used to simulate the tsunami propagation. To determine the boundary conditions on the liquid boundary of the computational domain, using the Black Sea tsunami model, the level fluctuations near Sevastopol in the region of depths of 90 m were calculated during the passage of tsunami waves from three potential tsunami foci caused by underwater earthquakes of magnitude 7. It was found that in because of tsunami penetration into the bays of Sevastopol from the nearest focus, the rise in sea level in the tops of the bays could reach 1–2 m. The maximum amplitudes of level fluctuations were received in Pesochnaya and Karantinnaya bays, where they reached 2 m. In the Sevastopol Bay, the level rises were about 0.5–1 m. The most intense fluctuations were observed in the first 3–3.5 hours of the tsunami action. It is shown that the coastal zone of Sevastopol is protected from waves coming from distant foci by Cape Сhersones. Numerical experiments have shown that the protective piers at the entrance to the Sevastopol Bay do not have a significant effect on the sea level fluctuations caused by the tsunami inside the bay.

FRI538 Discovery And Characterization Of Orally Bioavailable Nonpeptide Thyroid Stimulating Hormone Receptor (TSHR) Antagonists For The Treatment Of Graves’ Disease

Abstract Disclosure: M.A. Fowler: Employee; Self; Crinetics Pharmaceuticals. Stock Owner; Self; Crinetics Pharmaceuticals. C. Regan: Employee; Self; Crinetics Pharmaceuticals. Stock Owner; Self; Crinetics Pharmaceuticals. J. Zhao: Employee; Self; Crinetics Pharmaceuticals. Stock Owner; Self; Crinetics Pharmaceuticals. E. Coutinho: Employee; Self; Crinetics Pharmaceuticals. Stock Owner; Self; Crinetics Pharmaceuticals. B. Fleck: Employee; Self; Crinetics Pharmaceuticals. Stock Owner; Self; Crinetics Pharmaceuticals. A.A. Castellanos: Employee; Self; Crinetics Pharmaceuticals. E. Muller: Employee; Self; Crinetics Pharmaceuticals. M. Johns: Employee; Self; Crinetics Pharmaceuticals. Stock Owner; Self; Crinetics Pharmaceuticals. Y. Tang: Employee; Self; Crinetics Pharmaceuticals. Stock Owner; Self; Crinetics Pharmaceuticals. E. Sturchler: Employee; Self; Crinetics Pharmaceuticals. Stock Owner; Self; Crinetics Pharmaceuticals. M. Chen: Employee; Self; Crinetics Pharmaceuticals. Stock Owner; Self; Crinetics Pharmaceuticals. K. Retting: Employee; Self; Crinetics Pharmaceuticals. Stock Owner; Self; Crinetics Pharmaceuticals. D. Dalvie: Employee; Self; Crinetics Pharmaceuticals. Stock Owner; Self; Crinetics Pharmaceuticals. S.F. Betz: Employee; Self; Crinetics Pharmaceuticals. Stock Owner; Self; Crinetics Pharmaceuticals. S. Markison: Employee; Self; Crinetics Pharmaceuticals. Stock Owner; Self; Crinetics Pharmaceuticals. Graves’ disease is an autoimmune condition that affects approximately 1 in 100 people in the United States and 2-3% of the population worldwide. It is characterized by the production of autoantibodies against TSHR, and the pathology of Graves’ disease is driven by TSHR stimulatory antibodies (TSAb) that result in heightened activation of TSHR. This overstimulation results in hyperthyroidism due to excessive production of thyroid hormones. Approximately 30% of Graves’ disease patients also develop thyroid eye disease (TED or Graves’ orbitopathy) due to overactivation of TSHR in orbital fibroblasts leading to excessive production of hyaluronic acid, adipogenesis, cytokine production, and fibrosis. This can cause a myriad of debilitating symptoms including pain, swelling, blurry vision, diplopia, and proptosis. Several treatments for Graves’ hyperthyroidism are available including anti-thyroid drugs, radioactive iodine (RAI), and surgery. RAI and surgery are definitive treatments for Graves’ hyperthyroidism, but often result in hypothyroidism. In addition, none of the current treatments for Graves’ hyperthyroidism are effective in treating TED and, in some cases, such as with RAI, worsen the condition. Blocking TSHR activation directly via a TSHR antagonist may provide an important new therapeutic mechanism to treat patients with Graves’ disease that would effectively treat both the hyperthyroidism and TED. We have identified several potent and orally bioavailable nonpeptide allosteric antagonists with acceptable drug-like properties. One analog, TSHRant-1, demonstrated potent negative allosteric modulator activity at both the human and rat TSHR. To evaluate the in vivo pharmacodynamics of TSHant-1, we developed a rat model of hyperthyroidism. In this model, subcutaneous administration of the TSAb, M22, to female rats resulted in a robust and long-lasting rise in levels of the thyroid hormone thyroxin (T4). Oral administration of TSHRant-1 dose-dependently suppressed M22-stimulated T4, providing evidence that a nonpeptide allosteric antagonist of TSHR may serve as an effective treatment for Graves’ disease and associated orbitopathy (TED). Presentation: Friday, June 16, 2023

OR19-01 Srd5a1 Undergoes Distinct Changes in Methylation and Is Differentially Expressed During Pre-Pubertal Development in the Ovary and Hypothalamus

Abstract Disclosure: B. Bar-Sadeh: None. P. Melamed: None. The steroidogenic enzyme, 5α-reductase-1, is responsible for conversion of testosterone to dihydrotestosterone, and production of other 5α-reduced steroids and their derivatives, including in the brain. We reported previously that Srd5a1 levels are reduced in mice hypothalamic preoptic area (POA) and ovaries following early-life immune stress, which appeared to underlie delayed puberty onset and faster depletion of the ovarian reserve. This drop in ovarian Srd5a1 expression was concomitant with hypermethylation at two CpGs in its first intron, and also at the orthologous site in buccal DNA of women who experienced childhood immune challenge1. We thus hypothesized that this region of Srd5a1 acts as a transcriptional enhancer and is regulated by DNA methylation. Here we examined Srd5a1 expression and methylation at these CpGs across normal mouse development and in cultured cells. Ovarian Srd5a1 increased 8-fold and CpG methylation decreased by as much as 75% between PND10-30. During this time estradiol (E2) levels rise, and E2 exposure increased Srd5a1 mRNA levels in primary ovarian cells and KK-1 cell line. Chromatin immunoprecipitation (ChIP) confirmed binding of estrogen receptor to the locus of the CpGs, and identified an adjacent region strongly enriched with H3K4me1, characteristic of transcriptional enhancers. In the POA, in sharp contrast, Srd5a1 mRNA levels decreased by 70% between PND7-10 and then stayed quite constant, with little correlation to CpG methylation levels. The neonatal drop in POA Srd5a1 coincides with an increase in glucocorticoid (GC) levels, and we had seen lower Srd5a1 expression in mice following early-life stress, we thus treated GT1-7 hypothalamic neuronal cells with dexamethasone. This treatment reduced Srd5a1 expression, and ChIP confirmed that the GC receptor binds the enhancer locus. Srd5a1 expression was 12-fold higher in the ovarian KK1 cells compared to the neuronal GT1-7 cells, in accordance with much higher methylation levels in the latter. We thus used these cells to investigate the role of this CpG methylation, using gRNAs targeting dCas9-TET1 for demethylation in GT1-7 cells, or dCas9-DNMT3 for inducing methylation in KK1 cells. In GT1-7 cells, methylation at the first CpG dropped >50%, though was unaltered at the second CpG, and no effect was seen on Srd5a1 expression or its response to E2. In contrast, methylation in KK1 cells increased at the second CpG only, by more than 2.5 fold, and this abolished the response to E2. These distinct changes in Srd5a1 expression during development, together with our findings in cultured cells suggest a cell-specific GC-activated repression of Srd5a1 in the POA, and its E2-activated stimulation in the ovaries regulated by methylation at one of these CpGs. Reference: 1. Bar-Sadeh, B., Amichai, O.E., Pnueli, L., Begum, K., Leeman, G., Emes, R.D., Stöger, R., Bentley, G.R., and Melamed, P. (2022). BMC Biol. 20, 11. Presentation Date: Saturday, June 17, 2023

Photoperiod and UV light influence secondary metabolites of Hyoscyamus niger callus induced from leaves

Conclusion: Light significantly affected the concentration of several secondary metabolism chemicals. Despite this, UV light showed that the majority of secondary metabolites of N. niger had high significance when exposed to UV light for 120 min. This included a rise in levels of all compounds except for atropine, which showed a significant increase after only 60 minutes of UV exposure.

Bempedoic Acid: An Emerging Therapy for Uncontrolled Low-Density Lipoprotein (LDL) Cholesterol

Atherosclerotic cardiovascular disease (ASCVD) is a silent epidemic, which is progressing relentlessly across the globe. Developing countries such as India have a high prevalence of dyslipidemia and consequently a huge burden of coronary artery disease (CAD) and ASCVD. Low-density lipoprotein is regarded as the primary culprit in the genesis of ASCVD, and statins are the first line therapy for LDL-C lowering. Statin therapy has unequivocally demonstrated the benefit of lowering LDL-C in patients across the spectrum of CAD and ASCVD. Muscle symptoms and worsening of glycemic homeostasis could be challenges with statin therapy, especially with the use of high doses. A large fraction of patients are also unable to achieve their LDL goals with statins alone in clinical practice. Moreover, LDL-C goals have become aggressive over years, necessitating a combination of lipid lowering therapies. PCSK-9 inhibitors and Inclisiran have emerged as robust and safe lipid-lowering agents, but parenteral administration and high cost precludes their widespread use. Bempedoic acid is a novel lipid-lowering agent working upstream of statins by inhibiting the enzyme ATP citrate lyase (ACL). The drug produces an average LDL lowering of 22-28% in statin-naïve patients and 17-18% when given to preexisting statin users. Because skeletal muscles lack the ACL enzyme, there is minimal risk of muscle-related symptoms. In combination with ezetimibe, the drug synergistically reduced LDL-C by 39%. Moreover, the drug has no adverse effect on glycemic parameters and lowers hsCRP (inflammation) like statin. The series of four randomized CLEAR trials, involving >4000 patients, have shown consistent LDL lowering across the spectrum of ASCVD patients with or without background therapy. The large and only cardiovascular outcome trial of the drug (CLEAR Outcomes) has recently demonstrated a 13% reduction of MACE at 40 months. Rise in levels of uric acid (four times) and acute gout (three times) are more common compared to placebo with the drug, owing to competitive renal transportation by OAT 2. In a nutshell, Bempedoic acid represents a value addition to the inventory of dyslipidemia management.