Basal Concentrations Research Articles

Investigating serum concentration profiles of orally ingested short-chain fatty acid supplements.

Acetate, propionate, and butyrate are naturally-occurring short-chain fatty acids (SCFAs) derived from bacterial metabolism of dietary fibre and have been associated with numerous positive health outcomes. All three acids have been shown to offer unique physiological and metabolic effects and, therefore, could be targeted for co-ingestion as part of a nutritional/medicinal plan. However, a better understanding of the outcomes of supplementing in combination on circulating concentration profiles is necessary to confirm uptake efficacy. This study sought to investigate the acute circulating concentration profiles of acetate, propionate, and butyrate following oral supplementation. Three experimental trials were conducted including investigations to understand the impact of capsule coating on circulating concentration profiles, the effect of supplementation dose on uptake kinetics, and the outcome of a short, repeated, supplementation routine on circulating levels. Serum samples were analysed for SCFA content using a quantitative GC-MS assay. It was observed that an acid-resistant coated capsule caused a delayed and blunted blood concentration response, with the non-acid resistant trial displaying earlier and more intense peak serum concentrations. For dose comparison investigations, all SCFAs peaked within 60 min and returned to baseline concentrations by 120 min post-supplementation. A graded dose relationship was present for propionate and butyrate when considering the total circulating exposure across a 240 min monitoring period. In addition, a one-week, twice-daily, repeated supplementation protocol resulted in no changes in basal serum SCFA concentrations. Overall, these data indicate that acetate, propionate, and butyrate display relatively similar circulating concentration profiles following oral co-ingestion, adding knowledge to help inform supplementation strategies for future outcomes where acute elevation of circulating SCFAs is desired.

Effect of N-acetylcysteine on craving in substance use disorders (SUD): a meta-analysis of randomized controlled trials.

N-acetyl cysteine (NAC) appears promising as a treatment in patients with substance use disorder (SUD) as it helps rebalance glutamate levels in the central nervous system (CNS). Basal concentrations of glutamate are indeed reduced in SUD patients but increased during craving. We conducted a systematic review and meta-analysis of randomized controlled trials (RCTs). We assessed whether NAC reduce craving rating as compared to a placebo in SUD patients. Secondary outcomes were withdrawal symptoms (WS), side effects (SE) and drop-outs. Estimates are presented as standardized mean differences (SMD) or risk ratio (RR) with 95% confidence interval (CI). Eleven RCTs were included. NAC reduced craving rating (SMD -0.61 (-1.17, -0.06), p = 0.03, I2 = 85%), with no differences in the subgroup analysis according to the drug addiction (alcohol, cocaine, poly-drugs, amphetamine, nicotine) (p = 0.98). Among the secondary outcomes, for WS data showed no significant difference between groups (SMD -0.18 (-0.43, 0.08), p = 0.17); for SE no substantial difference was observed between the two treatment groups (RR = 1.06 (0.89-1.27), p = 0.52, I2 = 0%); for dropouts the results are in favor of the placebo but no statistically significant (RR 1.17 (0.85, 1.61), p = 0.34; I2 = 0%). NAC seem to reduce craving rating in SUD patients, but evidence is weak. More studies are needed to confirm this finding.

Zn2+ is essential for Ca2+ oscillations in mouse eggs.

Changes in the intracellular concentration of free calcium (Ca2+) underpin egg activation and initiation of development in animals and plants. In mammals, the Ca2+ release is periodical, known as Ca2+ oscillations, and mediated by the type 1 inositol 1,4,5-trisphosphate receptor (IP3R1). Another divalent cation, zinc (Zn2+), increases exponentially during oocyte maturation and is vital for meiotic transitions, arrests, and polyspermy prevention. It is unknown if these pivotal cations interplay during fertilization. Here, using mouse eggs, we showed that basal concentrations of labile Zn2+ are indispensable for sperm-initiated Ca2+ oscillations because Zn2+-deficient conditions induced by cell-permeable chelators abrogated Ca2+ responses evoked by fertilization and other physiological and pharmacological agonists. We also found that chemically or genetically generated eggs with lower levels of labile Zn2+ displayed reduced IP3R1 sensitivity and diminished ER Ca2+ leak despite the stable content of the stores and IP3R1 mass. Resupplying Zn2+ restarted Ca2+ oscillations, but excessive Zn2+ prevented and terminated them, hindering IP3R1 responsiveness. The findings suggest that a window of Zn2+ concentrations is required for Ca2+ responses and IP3R1 function in eggs, ensuring optimal response to fertilization and egg activation.

Zn2+ is essential for Ca2+ oscillations in mouse eggs

Changes in the intracellular concentration of free calcium (Ca2+) underpin egg activation and initiation of development in animals and plants. In mammals, the Ca2+ release is periodical, known as Ca2+ oscillations, and mediated by the type 1 inositol 1,4,5-trisphosphate receptor (IP3R1). Another divalent cation, zinc (Zn2+), increases exponentially during oocyte maturation and is vital for meiotic transitions, arrests, and polyspermy prevention. It is unknown if these pivotal cations interplay during fertilization. Here, using mouse eggs, we showed that basal concentrations of labile Zn2+ are indispensable for sperm-initiated Ca2+ oscillations because Zn2+-deficient conditions induced by cell-permeable chelators abrogated Ca2+ responses evoked by fertilization and other physiological and pharmacological agonists. We also found that chemically or genetically generated eggs with lower levels of labile Zn2+ displayed reduced IP3R1 sensitivity and diminished ER Ca2+ leak despite the stable content of the stores and IP3R1 mass. Resupplying Zn2+ restarted Ca2+ oscillations, but excessive Zn2+ prevented and terminated them, hindering IP3R1 responsiveness. The findings suggest that a window of Zn2+ concentrations is required for Ca2+ responses and IP3R1 function in eggs, ensuring optimal response to fertilization and egg activation.

Understanding the Structural Arrangement of Islets in Chronic Pancreatitis.

Islet transplantation has become an established method for the treatment of insulin-deficient diabetes such as type 1 and type 3C (pancreatogenic). An effective transplantation necessitates a thorough understanding of the islet architecture and related functions to improve engraftment outcomes. However, in chronic pancreatitis (CP), the structural and related functional information is inadequate. Hence, the present study is aimed to understand the cytoarchitecture of endocrine cells and their functional implications in CP with and without diabetes. Herein, a set of human pancreatic tissue specimens (normal, n=5 and CP, n=20) was collected and processed for islet isolation. Furthermore, immunohistochemistry was used to assess the vascular densities, cell mass, organization, and cell-cell interactions. The glucose-stimulated insulin release results revealed that in chronic pancreatitis without diabetes mellitus altered (CPNDA), at basal glucose concentration the insulin secretion was increased by 24.2%, whereas at high glucose concentration the insulin levels were reduced by 77.4%. The impaired insulin secretion may be caused by alterations in the cellular architecture of islets during CP progression, particularly in chronic pancreatitis with diabetes mellitus and CPNDA conditions. Based on the results, a deeper comprehension of islet architecture would be needed to enhance successful transplantation in CP patients: (J Histochem Cytochem XX.XXX-XXX, XXXX).

Key Drivers of Coagulation Factor Use in Von Willebrand Disease During Hospitalization: An Overview of the French BERHLINGO Cohort.

Von Willebrand disease (VWD) is the most common inherited bleeding disorder. However, studies of hospitalisation patterns with replacement treatment are scarce. The aim of this study was to investigate the current therapeutic management of VWD and determine the key drivers of coagulation factor uses in patients during hospitalisation. Hopscotch-WILL was a multi-centric retrospective study conducted over a 48-month period in any patients with VWD. The data were collected from the BERHLINGO Research Database and the French Hospital database. A total of 988 patients were included; 153 patients (15%) were hospitalised during 293 stays requiring treatment with von Willebrand factor (VWF) concentrates-pure or in association with Factor VIII (FVIII). Their median basal concentrations of VWF and FVIII were significantly lower than in untreated patients: VWF antigen < 30 IU/dL, VWF activity < 20 IU/dL and FVIII:C < 40 IU/dL. The median (interquartile range) concentrate consumption was similar between highly purified VWF or VWF combined with FVIII (72 [110] vs 57 [89] IU/kg/stay, p = 0.154). The use of VWF was highly heterogeneous by VWD type; type 3 had a particularly high impact on VWF consumption in non-surgical situations. The main admissions were for ear/nose/throat, hepato-gastroenterology, and trauma/orthopaedic conditions, besides gynaecological-obstetric causes in women. The use of VWF concentrates is mostly influenced by low basal levels of VWF and FVIII, but also by VWD type or the cause for hospitalisation. These results could inform future studies of newly released recombinant VWF.

Hypoglycemic Effect of Betulin in Rats With Experimental Diabetes

Betulin is natural pentacyclic triterpenoids lupane-structure, which shows a wide range of biological activity, including possibly hypoglycemic effects. Diabetes mellitus is a metabolic disease, accompanied by chronic hyperglycemia caused by absolute or relative insulin deficiency. The main effective drug in the treatment of type 1 diabetes is insulin injections. In the case of type 2 diabetes, it is an adjuvant therapy. To purpose of this study was to show hypoglycemic action of betulin in a model of experimentally induced diabetes in rats. For the induction of diabetes, alloxan at a dose of 170 mg/kg, i.p. was used. The hypoglycemic effect of betulin was evaluated using doses of 50 and 100 mg/kg/day for 30 days. The activity of glucose metabolism enzymes: liver glycogen phosphorylase, hexokinase and liver and skeletal muscle phosphofructokinase in rats, glucose-6-phosphate dehydrogenase (DG-6-P) of the liver was also evaluated. Betulin in a dose-dependent manner normalized of glycemia, increased the basal insulin concentration in the blood serum, increased the activity of carbohydrate metabolizing enzymes and on the gene expression profile of enzymes involved in carbohydrate metabolism. Betulin also significant improved the morphology of the pancreas. In particular, there was an increase in the area of the islands, their diameter, an increase in the number of β cells in the composition of the islands. We conclude that betulin has hypoglycemic properties, improving glycemia and carbohydrate metabolizing enzymes and pancreatic morphology.

Obesity and obesity-associated metabolic disease conditions in Connemara ponies in Ireland.

Equine obesity combined with insulin dysregulation (ID) is a major risk factor associated with laminitis. Some pony breeds appear to be at increased risk. However, little is known regarding the prevalence of obesity or hyperinsulinaemia as evidence of ID in Irish ponies. To investigate the prevalence of obesity and associated endocrine/metabolic disease conditions in Connemara ponies and to determine if hyperinsulinaemia in these ponies could be predicted by morphometric or metabolic markers. Cross-sectional study. The study population included registered Connemara ponies recruited through public and veterinary social media posts. Ponies underwent a physical examination and information on their management and clinical history was obtained via owner questionnaire. The body condition score (BCS) was measured using the Henneke system; cresty neck score (CNS) and regionalised adiposity were also assessed. Hyperinsulinaemia was confirmed by measuring serum basal insulin concentration (BIC) or insulin concentration after an oral sugar test (OST). Blood glucose and triglyceride concentrations were measured. Characteristics of hyperinsulinaemic and insulin-sensitive ponies were compared by logistic regression. Two hundred ponies were included; 59 ponies (29.5%) had a BCS ≥7, 58 (29.0%) had a CNS ≥2.5 and 135 (67.5%) had regionalised adiposity; 137 (68.5%) ponies had at least one of these abnormalities. Owner-reported history or clinical evidence of chronic laminitis was found in 92 ponies (46.0%). Hyperinsulinaemia was confirmed in 32 ponies (16.0%), including 23 of 91 (25.3%) detected by OST and 9 of 109 (8.3%) by BIC. Hypertriglyceridaemia was observed in 12 of 198 ponies (6.1%) ponies and hyperglycaemia in 11 of 197 ponies (5.6%) ponies. The odds of hyperinsulinaemia increased by a factor of 6.53 (95% confidence interval: 2.95, 15.21) when BCS was ≥7. The OST was not performed in all ponies. Increased adiposity, laminitis and metabolic derangements are prevalent in this native Irish pony breed.

Differential patterns of basal and naloxone-evoked dopamine efflux in the rat dorsal and ventral striatum following prolonged-intermittent exposure to morphine.

Hypodopaminergia in the ventral striatum is a putative neurobiological correlate of withdrawal in opioid-dependent individuals. This perspective stands in contrast to brain imaging studies with chronic opioid users showing that naloxone-enhanced dopamine (DA) release in the dorsal striatum is positively correlated with withdrawal aversion. Here, we examined regional differences in striatal DA function associated with opioid withdrawal in rats exposed to intermittent morphine injections for 31 days. Basal concentrations of DA were reduced (i.e., indicating a hypodopaminergic state) in the ventral striatum on Day 10 of morphine exposure, whereas a more prolonged period of morphine treatment was required to reveal hypodopaminergia in the dorsal striatum on Day 31. The ventral striatum consistently exhibited naloxone-induced transient reductions in DA below the hypodopaminergic basal levels, whereas morphine enhanced DA efflux. In the dorsal striatum, DA responsivity to naloxone shifted from a significant decrease on Day 10 to a notable increase above hypodopaminergic basal levels on Day 31, corroborating the findings in the human dorsal striatum. Unexpectedly, the magnitude of morphine-evoked increases in DA efflux on Day 31 was significantly blunted relative to values on Day 10. These findings indicate that prolonged-intermittent access to morphine results in a sustained hypodopaminergic state as reflected in basal levels in the striatum, which is accompanied by regional differences in DA responsivity to naloxone and morphine. Overall, our findings suggest that prolonging the duration of morphine exposure to 31 days is sufficient to reveal neuroadaptations that may underlie the transition from initial drug exposure to opioid dependence.

Are Proteins Such as MMP2, IGF1, IL-13, and IL-1ra Valuable as Markers of Fitness Status in Racehorses? A Pilot Study

In a recent study, in which more than a thousand racehorses were evaluated, three potential blood markers were selected. It was concluded that insulin-like growth factor 1 (IGF1), interleukin-1 receptor antagonist (IL-1ra), and matrix metalloproteinase 2 (MMP2) may enable the early detection of horses at risk of injuries. However, in other studies, it was suggested that cytokine concentrations indicate the athlete’s status better than mRNA expression in blood cells. Thus, the purpose of this study was to evaluate changes in IGF1, MMP-2, and novel markers such as interleukin-13 (IL-13) after exercise in horses at different fitness levels as well as after different intensities of exercise. ELISA tests were performed on thirty-one racehorses [n = 31], who were divided into an inexperienced [beginner] group [n = 20] and an experienced [advanced] group [n = 6]. In addition, differences between race and training session were evaluated to see the influence of different intensities of exercise. Blood samples were taken before and after exercise. The basal IGF1 concentration was lower in an inexperienced group (p &lt; 0.01) as well as IL-13 (p &lt; 0.05) in comparison to the experienced group. There were no differences between pre- and post-exercise samples, changes in multiples or between training, and racing exercises. In conclusion, the basal values of some cytokines may appear to be more beneficial in forecasting horse fitness level.

Effects of intravenous administration of ascorbic acid (vitamin C) on oxidative status in healthy adult horses.

Ascorbic acid (AA) is an antioxidant that might be beneficial for adjunctive treatment of sepsis in horses. The optimal dose and effects on oxidative status are unknown. Ascorbic acid administration will increase plasma AA concentrations and decrease determinants of reactive oxygen metabolites (dROM), basal and stimulant-induced intraerythrocytic reactive oxygen species (ROS) concentrations, and stimulant-induced neutrophil ROS production, and increase plasma antioxidant capacity (PAC) in a dose-dependent manner. Eight healthy horses. Randomized placebo-controlled crossover study. Each horse received 4 single-dose IV treatments including AA at 25, 50, and 100 mg/kg and saline (placebo) with each treatment separated by ≥1 week. Blood was collected at baseline, 2 and 6 hours for assessment of plasma dROM and PAC via photometer, intraerythrocytic ROS by flow cytometry, and stimulant-induced neutrophil ROS by a fluorometric assay. Plasma AA concentrations were measured by high-performance liquid chromatography/electrochemical detection. Ascorbic acid at 100 mg/kg resulted in decreased dROM 2 hours after treatment (P = .03, 95% CI 5.51-121.2, point estimate 63.3). There was no effect of AA on basal or stimulant-induced intraerythrocytic ROS (P = .88, 95% CI -0.156 to 0.081, point estimate -0.037; P = .93, 95% CI -0.123 to 0.112, point estimate -0.006, respectively), basal or stimulant-induced neutrophil ROS (P ≥ .12, 95% CI -644.9 to 56.2, point estimate -294.4), or PAC (P ≥ .64, 95% CI -1567 to 463.4, point estimate -552.0) at any dose or timepoint. Plasma AA concentrations increased in a dose-dependent manner. High-dose administration of AA might provide antioxidant benefits in horses.

Cytokines in predicting JAK inhibitors clinical efficacy for COVID-19

Janus kinases inhibitors (JAKi) have an immunosuppressive effect due to the normalization of levels of proinflammatory cytokines, are used in the treatment of cytokine storm in COVID-19. This fact makes it necessary to search a predictor of an efficacy of this small-molecules. The aim of the study. To evaluate the possibility of using cytokines in predicting the efficacy of JAKi in COVID-19. Materials and methods. We examined 86 patients with COVID-19: 50 men and 36 women. The age was 57,0; (50,0–65,0) years. The SARS-CoV-2 presence was confirmed by PCR. The control group consisted of 30 healthy donors matched by sex and age. The criterion for the efficacy of JAKi was recovery, in efficacy – switching to another drug, or death. The serum concentration of IL-1β, –6, –8, –17, TNF-α and GM–CSF by ELISA was estimates using Invitrogen test-systems (USA), according to the manufacturer’s instructions. Results. In COVID-19 patients who received baricitinib (BTC), compared with healthy individuals, basal (before JAKi starting) IL-6, TNF-α serum concentrations were increased; tofacitinib (TFT) – IL-6, ruxilitinib (RLT) – IL-6 and TNF-α. A decrease of IL-1β and GM–CSF serum concentration was noted before the start of TFT and RLT. Basal concentrations of IL-6 and TNF-α are most statistically strongly associated with the diagnosis COVID-19. After 5 days of treatment with BTC, there was an increase in IL-8 level compared to the basal, a decrease in the GM–CSF concentration and a slight increase IL-6 concentration relative to healthy donors (p &lt; 0.05 in all cases). Basal concentration of IL-17 (more than 13.9 pg/ml) indicated the possibility of successful RLT treatment (area under the ROC-curve = 0.99; AP = 99 %; DS = 80 %; OP+ = 5.0; OP– = 0.2; CI: 0.9–1.0). Conclusions. Basal serum concentration of cytokines may predict JAKi efficacy in COVID-19 patients.

261 Variations in Protein Deposition Might Be Associated with Insulin Resistance and Differences in Amino Acids Metabolism in Growing Gilts

Abstract Pigs with similar body weight (BW), same genetic background and sex vary in terms of protein deposition (PD) overgrowth. Insulin stimulates protein synthesis, and differences in PD could be associated with the variation among pigs in postprandial insulin secretion. Therefore, this study aimed to investigate if changes in preprandial and postprandial amino acids (AA) response and insulin resistance differ between pigs having Low or High PD. Seventy gilts (33 kg BW, ± 3.6; Alphagène) were assigned to a randomized complete block (BW) design and housed in the same pen equipped with automatic feeders. Pigs were fed ad libitum with a diet balanced to maximize PD. Dual X-Ray estimated body protein (BP) and lipid (BL) masses on d 1 and 21, and by difference, PD was calculated. On d 23, pigs were grouped in Low (157 g/d) and High (219 g/d) PD. Eight pigs per PD group were canulated in a jugular vein, and after 48h recovery receiving feed at 90% of the measured feed intake, they were fasted for eight hours to study preprandial metabolism (-5, -1 min before the meal). In sequence, all pigs received a 300 g meal and blood samples were collected at 10, 15, 20, 30, 40, 50, 60, 75, 90, 120, 150, 178, 178 min, relative to the meal, to measure AA, insulin and glucose concentrations. In addition, insulin resistance (HOMA-IR) was calculated based on fasting glucose and insulin concentrations. The AA response was modeled for each PD group based on the average group response using differential equations derived from the Erlang equation. Thus, the basal concentration, area under the curve (AUC) and AA flux could be compared among groups using the parameters obtained by the non-linear model. An F-test was used to compare the difference in the indexes between Low and High PD pigs. The basal concentration of methionine, valine and leucine were similar, whereas AUC was greater for Low PD pigs with no difference in flux, with the exception of methionine (P &amp;lt; 0.10; Table 1). Conversely, lysine (15%) and threonine (10%) basal concentrations were less in Low PD pigs. Lysine AUC was greater (8%), whereas others AA AUC were on average 27% less for High PD than for Low PD pigs. Lysine maximal concentration was 11% greater for High PD pigs and occurred later than for Low PD (1.15h vs 1.03h). Such differences in AA postprandial metabolism might be partially due to metabolic differences in response to anabolic hormones such as insulin. For instance, HOMA-IR differed (P &amp;lt; 0.05) among PD categories, with Low PD showing a 44% lower index for insulin resistance than high PD pigs. High PD pigs are less insulin-resistant and show differences in pre- and postprandial AA metabolism.

PSVIII-30 Impaired Glucose-Stimulated Insulin Secretion in Intrauterine Growth-Restricted Neonatal Lambs Is Improved By Daily Supplementation of ω-3 Polyunsaturated Fatty Acid Ca2+ Salts

Abstract Maternofetal heat stress during placental development induces fetal intrauterine growth restriction (IUGR), which in turn reduces birthweight. IUGR-born livestock exhibit deficits in insulin stimulus-secretion coupling and glucose metabolism that contribute to poor growth efficiency. Thus, our objective was to determine whether daily oral supplementation of Ca2+ salts of ω-3 polyunsaturated fatty acids (ω-3 PUFA) to IUGR-born neonatal lambs improves glucose-stimulated insulin secretion and other indicators of metabolic efficiency. Maternal heat stress (40°C, 35% humidity, THI = 85) from d 40 to 95 of gestation was used to induce IUGR. Lambs were weaned at birth and hand-reared on milk replacer. Beginning at birth, lambs received daily oral boluses of 0.5 mL molasses containing 0 or 0.42 g/kg bodyweight of ω-3 PUFA (Virtus Nutrition). All lambs were fitted with femoral catheters at d 24 and a square-wave hyperglycemic clamp study was performed on d 28 to assess basal (i.e., resting) and glucose-stimulated insulin secretion (GSIS). Blood glucose concentrations did not differ among IUGR lambs (n = 8), IUGR+ω-3 lambs (n = 7), and controls (n = 12), but by study design were increased (P &amp;lt; 0.05) 205% from basal during hyperglycemic periods. Basal plasma insulin concentrations did not differ among groups, but basal glucose-to-insulin ratios were greater (P &amp;lt; 0.05) for IUGR+ω-3 lambs than for controls or IUGR lambs. During hyperglycemia, plasma insulin concentrations were 53% less (P &amp;lt; 0.05) for IUGR lambs but only 30% less (P &amp;lt; 0.05) for IUGR+ω-3 lambs compared with controls. Hyperglycemic glucose-to-insulin ratios were greater (P &amp;lt; 0.05) for IUGR lambs (3.24 ± 0.59) compared with controls (1.10 ± 0.29) and were intermediate for IUGR+ω-3 lambs (2.08 ± 0.86). Blood lactate concentrations did not differ among groups or between periods, but blood pH was was greater (P &amp;lt; 0.05) for IUGR (7.448 ± 0.006) and IUGR+ω-3 lambs (7.439 ± 0.003) compared with controls (7.422 ± 0.004). Blood partial pressure of CO2 was less (P &amp;lt; 0.05) for IUGR lambs (43.0 ± 0.3 mmHg) than for controls (46.7 ± 0.9 mmHg) and was intermediate for IUGR+ω-3 lambs (44.0 ± 0.4 mmHg). Blood HCO3 concentrations and O2-bound hemoglobin were greater (P &amp;lt; 0.05) for IUGR and IUGR+ω-3 lambs than for controls. Blood Na+ tended to be less (P = 0.07) and blood K+ was less (P &amp;lt; 0.05) for IUGR and IUGR+ω-3 lambs than for controls. Blood Ca2+ was less (P &amp;lt; 0.05) for IUGR lambs but not for IUGR+ω-3 lambs than for controls. These findings demonstrate that daily oral supplementation of Ca2+ salts of ω-3 PUFA improved deficits in insulin stimulus-secretion coupling and several other metabolic indicators observed in IUGR-born offspring. However, it did not improve all electrolyte concentrations or acid-base balance of the blood.

Autophagic and apoptotic proteins in goat corpus luteum and the effect of Adiponectin/AdipoRon on luteal cell autophagy and apoptosis

The most abundant adipokine Adiponectin (APN) is present in ovaries. AdipoRon is a small molecule oral APN receptor agonist that binds and activates APN receptors. However, the function of APN/AdipoRon in regulation of luteal cell processes has not been elucidated. To investigate autophagic and apoptotic proteins in goat CLs and effects of APN/AdipoRon on goat luteal autophagy and apoptosis, goat CLs were collected during the early, mid and late luteal stages of the estrous cycle to evaluate autophagic and apoptotic protein patterns. LC3B, Beclin 1, Caspase-3 and Bax/Bcl-2 as well as p-AMPK were differentially abundant at different stages of CL development. All these proteins were primarily localized in large and small luteal steroidogenic cells. Then, isolated luteal steroidogenic cells were evaluated to ascertain the functions and mechanism of APN/AdipoRon in luteal autophagy and apoptosis. Treatment with AdipoRon (25 and 50 μM) and APN (1 μg/mL) for 48 h resulted in a decrease in cell viability and P4 level, increased autophagic and apoptotic proteins. Treatment with AdipoRon (25 μM) led to rapid and transient p-AMPK activation, with p-AMPK elevated at 30 min to 1 h with there being a return to a basal concentration at 2 h post-treatment. Moreover, treatment with AdipoRon led to an increase in autophagy by activating AMPK, which was markedly reduced with treatment with an AMPK inhibitor Compound C and siAMPK, however, abundances of apoptotic proteins were not affected by these treatments. In conclusion, autophagy and apoptosis are involved in the structural regression of goat CL. APN/AdipoRon led to a lesser cell viability and P4 concentration, and activated autophagy through induction of the AMPK while there was induction of apoptosis through an AMPK - independent pathway in goat luteal cells.

On Primary Adrenal Insufficiency with Normal Concentrations of Cortisol - Early Manifestation of Addison's Disease.

Primary adrenal insufficiency (AI) is an endocrine disorder in which hormones of the adrenal cortex are produced to an insufficient extent. Since receptors for adrenal steroids have a wide distribution, initial symptoms may be nonspecific. In particular, the lack of glucocorticoids can quickly lead to a life-threatening adrenal crisis. Therefore, current guidelines suggest applying a low threshold for testing and to rule out AI not before serum cortisol concentrations are higher than 500 nmol/l (18 μg/dl). To ease the diagnostic, determination of morning cortisol concentrations is increasingly used for making a diagnosis whereby values of>350 nmol/l are considered to safely rule out Addison's disease. Also, elevated corticotropin concentrations (>300 pg/ml) are indicative of primary AI when cortisol levels are below 140 nmol/l (5 μg/dl). However, approximately 10 percent of our patients with the final diagnosis of primary adrenal insufficiency would clearly have been missed for they presented with normal cortisol concentrations. Here, we present five such cases to support the view that normal to high basal concentrations of cortisol in the presence of clearly elevated corticotropin are indicative of primary adrenal insufficiency when the case history is suggestive of Addison's disease. In all cases, treatment with hydrocortisone had been started, after which the symptoms improved. Moreover, autoantibodies to the adrenal cortex had been present and all patients underwent a structured national education program to ensure that self-monitored dose adjustments could be made as needed.

Plasma endocannabinoids in cocaine dependence and their relation to cerebral metabotropic glutamate receptor 5 density

Animal models indicate that the endocannabinoid system (ECS) plays a modulatory role in stress and reward processing, both crucially impaired in addictive disorders. Preclinical findings showed endocannabinoid-modulated synaptic plasticity in reward brain networks linked to the metabotropic-glutamate-5 receptor (mGluR5), contributing to drug-reinforcing effects and drug-seeking behavior. Although animal models postulate a link between ECS and cocaine addiction, human translational studies are lacking. Here, we tested previous preclinical findings by investigating plasma endocannabinoids (eCBs) anandamide (AEA), 2-arachidonoylglycerol (2-AG), and the related N-acylethanolamines (NAEs) palmitoylethanolamide (PEA) and oleoylethanolamide (OEA), including their interaction with cerebral mGluR5, in chronic cocaine users (CU). We compared basal plasma concentrations between chronic CU (N = 103; 69 recreational CU and 34 dependent CU) and stimulant-naïve healthy controls (N = 92). Follow-up basal eCB/NAE plasma levels after 12 months were used for reliability and stability check (CU: N = 33; controls: N = 43). In an additional analysis using 11C-ABP688 positron emission tomography (PET) in a male subsample (CU: N = 18; controls: N = 16), we investigated the relationships between eCBs/NAEs and mGluR5 density in the brain. We found higher 2-AG plasma levels in dependent CU compared to controls and recreational CU. 2-AG levels were stable over time across all groups. In the PET-subsample, a positive association between 2-AG and mGluR5 brain density only in CU was found. Our results corroborate animal findings suggesting an alteration of the ECS in cocaine dependence and an association between peripheral 2-AG levels and cerebral mGluR5 in humans. Therefore, the ECS might be a promising pharmaco-therapeutic target for novel treatments of cocaine dependence.

Kinin B1 receptor modulates glucose homeostasis and physical exercise capacity by altering adrenal catecholamine synthesis and secretion

Our group has shown in several papers that kinin B1 receptor (B1R) is involved in metabolic adaptations, mediating glucose homeostasis and interfering in leptin and insulin signaling. Since catecholamines are involved with metabolism management, we sought to evaluate B1R role in catecholamine synthesis/secretion. Using B1R global knockout mice, we observed increased basal epinephrine content, accompanied by decreased hepatic glycogen content and increased glucosuria. When these mice were challenged with maximal intensity exercise, they showed decreased epinephrine and norepinephrine response, accompanied by disturbed glycemic responses to effort and poor performance. This phenotype was related to alterations in adrenal catecholamine synthesis: increased basal epinephrine concentration and reduced norepinephrine content in response to exercise, as well decreased gene expression and protein content of tyrosine hydroxylase and decreased gene expression of dopamine beta hydroxylase and kinin B2 receptor. We conclude that the global absence of B1R impairs catecholamine synthesis, interfering with glucose metabolism at rest and during maximal exercise.

BEVA primary care clinical guidelines: Diagnosis and management of equine pituitary pars intermedia dysfunction.

BEVA primary care clinical guidelines: Diagnosis and management of equine pituitary pars intermedia dysfunction.

THU044 The Subcellular Localization Of β-Catenin Is Associated With Tumor Size And The Surgical Remission Rate In Growth Hormone-producing Pituitary Adenomas

Abstract Disclosure: S. Kuwabara: None. H. Kameda: None. H. Nomoto: None. K. Cho: None. A. Nakamura: None. Y. Ishi: None. H. Motegi: None. F. Miki: None. H. Miyoshi: None. T. Atsumi: None. The Wnt/β-catenin pathway is the best understood Wnt signaling pathway and controls cell proliferation. In the presence of Wnt, the translocation of β-catenin from the cytoplasm to the nucleus leads to pathway activation. It has been reported that the nuclear accumulation of β-catenin is associated with Ki-67 expression in pituitary adenomas and that the invasiveness of growth hormone (GH)-producing pituitary adenomas is associated with β-catenin expression. We aimed to clarify the effects of the Wnt/β-catenin pathway in GH-producing pituitary adenomas. We performed immunofluorescence staining using antibodies against GH and β-catenin in pathological samples of pituitary adenoma from 62 patients with acromegaly who underwent pituitary surgery at our hospital between January 2008 and December 2021. We also obtained clinical data for the patients, including the results of endocrine testing, and evaluated the relationships of key indices with the intracellular localization of β-catenin. Samples from 26 cases (42%) showed strong punctate expression of β-catenin in the nuclei (dot pattern (DP) group), and samples from 36 cases (58%) showed β-catenin expression in the cytoplasm or cell membrane (non-DP group). There were no differences in the sex, age, GH concentration, or insulin-like growth factor I (IGF-1) SD scores of the two groups, but the DP group had larger tumors (diameters 18±8 vs. 13±7 mm) and a higher prevalence of macroadenomas (77% vs. 53%). In the DP group, a paradoxical preoperative GH response (an increase of ≥30% of the basal concentration) to 75-g oral glucose tolerance testing was more frequent (46 vs. 22%, p&amp;lt;0.05) and postoperative remission was less frequent (23% vs. 50%, p&amp;lt;0.05). Accordingly, the Wnt/β-catenin pathway appears to be more active in the DP group. The larger tumors and lower postoperative remission rate of the DP group suggest that the Wnt/β-catenin pathway may influence tumor growth. Wnt/β-catenin pathway could play a major role in GH-producing pituitary tumor growth, therefore this pathway would be a potential therapeutic target in the affected patients. Presentation: Thursday, June 15, 2023