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Pulmonary hypertension and hereditary hemorrhagic telangiectasia (HHT) have an association mediated by activin A receptor type II-like 1 (ACVRL1) gene pathogenic variants. A 30-year-old woman was previously admitted to a hospital due to lung hemorrhage, and was diagnosed with pulmonary hypertension, but stopped follow-up visits. At 48 years of age, she was admitted to our hospital and was diagnosed with HHT. Genetic testing revealed an ACVRL1 pathogenic variant. After the initiation of pulmonary vasodilator treatment, the patient's mean pulmonary artery pressure started to decrease from 43 mmHg, declining to 37 mmHg when she was 58 years of age. This is the first report describing the 28-year follow-up of an HHT and pulmonary hypertension patient with an ACVRL1 mutation.

Background: Hypertension (HTN) causes vascular injury identified by endothelial dysfunction, vascular stiffening, and remodeling, which contributes to kidney damage leading to chronic kidney disease (CKD). MicroRNAs (miRNAs) repress/degrade target mRNAs. Their role in vascular injury in HTN remains unclear. We aimed to identify differentially expressed (DE) miRNAs in gluteal subcutaneous arteries of patients with HTN associated or not with CKD to shed light on the pathophysiological molecular mechanisms. Methods: Normotensive subjects and patients with HTN associated or not with CKD grades 3-4 were studied (n=15-16). Small arteries were isolated from gluteal subcutaneous biopsies, RNA extracted and small and total RNA sequencing performed by Illumina HiSeq-2500. DE genes were identified with a P 1.3. Top 3 DE miRNAs ( P 2, mean read count number (MRCN) >3,000 in all groups and having predicted mRNA targets) were selected for validation by reverse transcription-quantitative PCR (RT-qPCR). The mRNA targets of the top selected miRNA were predicted by TargetScan with P 1.5 and MRCN>150 and the top 9 targets were validated by RT-qPCR using gain- and loss-of-function in human aortic endothelial cells (HAECs). Gene ontology enrichment analysis (GOEA) was done in Cytoscape. Results: DE miRNAs and mRNAs were identified uniquely associated with HTN (miRNAs: 10, mRNAs: 68), CKD (miRNAs: 68, mRNAs: 395), and in both groups (miRNAs: 2, mRNAs: 32). miR-338-3p presented the best correlation between RNA sequencing and RT-qPCR (R 2 =0.328, P PTPRS , FC: 0.80±0.08 vs 1.00±0.00) and glutathione peroxidase 3 ( GPX3 , FC: 0.88±0.04 vs 1.00±0.00) were down-regulated in HAECs transfected with miR-338-3p mimics ( P GPX3 with oxidative stress detoxification ( q PTPRS with the immune system, neuronal system and developmental process ( q Conclusion: Down-regulated miR-338-3p in gluteal subcutaneous small arteries of hypertensive patients with CKD targets PTPRS and GPX3 that may play a role in vascular injury in HTN.

The basolateral amygdala (BLA) is a critical brain region for cocaine-memory reconsolidation. Corticotropin-releasing factor receptor type 1 (CRFR1) is densely expressed in the BLA, and CRFR1 stimulation can activate intra-cellular signaling cascades that mediate memory reconsolidation. Hence, we tested the hypothesis that BLA CRFR1 stimulation is necessary and sufficient for cocaine-memory reconsolidation. Using an instrumental model of drug relapse, male and female Sprague-Dawley rats received cocaine self-administration training in a distinct environmental context over 10 days followed by extinction training in a different context over 7 days. Next, rats were re-exposed to the cocaine-paired context for 15 min to initiate cocaine-memory retrieval and destabilization. Immediately or 6 h after this session, the rats received bilateral vehicle, antalarmin (CRFR1 antagonist; 500 ng/hemisphere), or corticotropin-releasing factor (CRF; 0.2, 30 or 500 ng/hemisphere) infusions into the BLA. Resulting changes in drug context-induced cocaine seeking (index of context-cocaine memory strength) were assessed three days later. Female rats self-administered more cocaine infusions and exhibited more extinction responding than males. Intra-BLA antalarmin treatment immediately after memory retrieval (i.e., when cocaine memories were labile), but not 6 h later (i.e., after memory reconsolidation), attenuated drug context-induced cocaine seeking at test independent of sex, relative to vehicle. Conversely, intra-BLA CRF treatment increased this behavior selectively in females, in a U-shaped dose-dependent fashion. In control experiments, a high (behaviorally ineffective) dose of CRF treatment did not reduce BLA CRFR1 cell-surface expression in females. Thus, BLA CRFR1 signaling is necessary and sufficient, in a sex-dependent manner, for regulating cocaine-memory strength.

Background Angiotensin II (Ang II ) can cause hypertension and tissue impairment via AGTR 1 (Ang II receptor type 1), particularly in renal proximal tubule cells, and can cause DNA damage in renal cells via nicotinamide adenine dinucleotide phosphate oxidase. BubR1 (budding uninhibited by benzimidazole‐related 1) is a multifaceted kinase that functions as a mitotic checkpoint. BubR1 expression can be induced by Ang II in smooth muscle cells in vitro, but the relationship between systemic BubR1 expression and the Ang II response is unclear. Methods and Results Twenty 24‐week‐old male BubR1 low‐expression mice (BubR1 L/L mice) and age‐matched BubR1 +/+ mice were used in this study. We investigated how Ang II stimulation affects BubR1 L/L mice. The elevated systolic blood pressure caused by Ang II stimulation in BubR1 +/+ mice was significantly attenuated in BubR1 L/L mice. Additionally, an attenuated level of Ang II –induced perivascular fibrosis was observed in the kidneys of BubR1 L/L mice. Immunohistochemistry revealed that the overexpression of AGTR 1 induced by Ang II stimulation was repressed in BubR1 L/L mice. We evaluated AGTR 1 and Nox‐4 (nicotinamide adenine dinucleotide phosphate oxidase‐4) levels to determine the role of BubR1 in the Ang II response. Results from in vitro assays of renal proximal tubule cells suggest that treatment with small interfering RNA targeting BubR1 suppressed Ang II ‐induced overexpression of AGTR 1. Similarly, the upregulation in Nox4 and Jun N‐terminal kinase induced by Ang II administration was repressed by treatment with small interfering RNA targeting BubR1. Conclusions Ang II –induced hypertension is caused by AGTR 1 overexpression in the kidneys via the upregulation of BubR1 and Nox4.

Objective— Diabetic macular edema is a major cause of visual impairment. It is caused by blood-retinal barrier breakdown that leads to vascular hyperpermeability. Current therapeutic approaches consist of retinal photocoagulation or targeting VEGF (vascular endothelial growth factor) to limit vascular leakage. However, long-term intravitreal use of anti-VEGFs is associated with potential safety issues, and the identification of alternative regulators of vascular permeability may provide safer therapeutic options. The vascular specific BMP (bone morphogenetic protein) receptor ALK1 (activin-like kinase receptor type I) and its circulating ligand BMP9 have been shown to be potent vascular quiescence factors, but their role in the context of microvascular permeability associated with hyperglycemia has not been evaluated. Approach and Results— We investigated Alk1 signaling in hyperglycemic endothelial cells and assessed whether BMP9/Alk1 signaling could modulate vascular permeability. We show that high glucose concentrations impair Alk1 signaling, both in cultured endothelial cells and in a streptozotocin model of mouse diabetes mellitus. We observed that Alk1 signaling participates in the maintenance of vascular barrier function, as Alk1 haploinsufficiency worsens the vascular leakage observed in diabetic mice. Conversely, sustained delivery of BMP9 by adenoviral vectors significantly decreased the loss of retinal barrier function in diabetic mice. Mechanistically, we demonstrate that Alk1 signaling prevents VEGF-induced phosphorylation of VE-cadherin and induces the expression of occludin, thus strengthening vascular barrier functions. Conclusions— From these data, we suggest that by preventing retinal vascular permeability, BMP9 could serve as a novel therapeutic agent for diabetic macular edema.

INTRODUCTION AND OBJECTIVES:Psychological stress exacerbates bladder dysfunction such as overactive bladder (OAB) and bladder pain syndrome/interstitial cystitis (BPS/IC). We previously reported that bombesin (BB), a stress-related neuropeptide, centrally induces facilitation of the rat micturition reflex. Brain BB is reported to activate hypothalamo-pituitary-adrenal axis and sympathetic nerves, as representative responses to stressful conditions, via brain corticotropin-releasing factor (CRF), another stress-related neuropeptide. Therefore, we examined whether brain CRF is involved in the BB-induced frequent urination in rats.METHODS:(1) In urethane anesthetized (0.8 g/kg, ip) male Sprague-Dawley (SD) rats (300-400 g), a catheter was inserted into the bladder to perform cystometry (CMG, 12 ml/h saline infusion). Astressin (a non-selective CRF receptor antagonist, 1 or 3 nmol/rat) was intracerebroventricularly (icv) pretreated 60 min before BB administration (0.03 nmol/rat, icv). CMG was started 60 min b...

Numerous epidemiological studies have evaluated the association between transforming growth factor beta receptor type 1 ( TGFBR1) polymorphisms and the risk of cancer; however, the results remain inconclusive and controversial. To determine the association between breast cancer risk and the *6A polymorphism of the TGFBR1 gene, a case-control study of 280 breast cancer patients and 280 controls was performed in Iranian women. Our study demonstrates that women who carry the TGFBR1*6A allele are at lower risk of developing breast cancer. The highest protection against breast cancer was observed in 6A/6A homozygotes (OR = 0.32, p = 0.04). A lower frequency of the TGFBR1*6A allele in breast cancer patients may be an important genetic determinant that contributes to a lower risk of breast cancer in Iranian women. The results also showed that the allelic length of TGFBR1 polymorphisms had no significant association with the age at onset or the grade of disease, nor with the expression of progesterone and estrogen receptors and HER2.

Non-synonymous GRK4 variants, R65L, A142V and A486V, are associated with essential hypertension in diverse populations. This study replicated the association of GRK4 variants, including GRK4(142V), with human essential hypertension in a Japanese population (n=588; hypertensive, n=486 normotensive controls) and determined whether the presence of GRK4 variants predicted the blood pressure (BP) response to angiotensin receptor blockers (ARBs) in patients with essential hypertension. We analyzed 829 patients and compared the response to ARBs between individuals with no GRK4 variants (n=136) and those with variants at one or any of the three loci (n=693). Carriers of hGRK4(142V) had a greater decrease in systolic BP in response to ARBs than non-carrier hypertensive patients. By contrast, those with variants only at GRK4(486V) were less likely to achieve the BP goal in response to an ARB than those with no variants. These studies showed for the first time the association between GRK4(142V) and a larger decrease in BP with ARBs in hypertensive patients.

Inositol 1,4,5-trisphosphate receptor type II (InsP3R-II) is the most prevalent isoform of the InsP3R in hepatocytes and is concentrated under the canalicular membrane, where it plays an important role in bile secretion. We hypothesized that altered calcium (Ca2+) signaling may be involved in metabolic dysfunction, as InsP3R-mediated Ca2+signals have been implicated in the regulation of hepatic glucose homeostasis. Here, we find that InsP3R-II, but not InsP3R-I, is reduced in the livers of obese mice. In our investigation of the functional consequences of InsP3R-II deficiency, we found that organic anion secretion at the canalicular membrane and Ca2+signals were impaired. However, mice lacking InsP3R-II showed no deficits in energy balance, glucose production, glucose tolerance, or susceptibility to hepatic steatosis. Thus, our results suggest that reduced InsP3R-II expression is not sufficient to account for any disruptions in metabolic homeostasis that are observed in mouse models of obesity. We conclude that metabolic homeostasis is maintained independently of InsP3R-II. Loss of InsP3R-II does impair secretion of bile components; therefore, we suggest that conditions of obesity would lead to a decrease in this Ca2+-sensitive process.

LY3022859 is an anti-TGFβRII IgG1 monoclonal antibody that inhibits receptor-mediated signaling activation. The primary objective of this phase I study was to determine a phase II dose in patients with advanced solid tumors. Secondary objectives were to assess safety and pharmacokinetics (PK). LY3022859 was infused intravenously (IV) at 1.25 mg/kg over 1 h every 2 weeks (Q2W) (cohort 1A) and at flat doses of 12.5 mg (cohort 1B) and 25 mg (cohort 2) over 3 h Q2W. Fourteen patients were enrolled in cohorts 1A (n = 2), 1B (n = 5), and 2 (n = 7). DLTs were experienced by both patients in cohort 1A (infusion-related reaction) and 2 patients in cohort 2 (cytokine release syndrome and infusion-related reaction). No MTD was determined. At the 25 mg dose level (cohort 2), after fifth infusion, LY3022859 had a short t1/2 (4.37-7.80 h) and rapid clearance (CLss, 0.412 L/h). Exposure increased twofold (from 28.5 to 60.2 μg·h/mL) with increase in dose from 12.5 to 25 mg. No accumulation was observed after repeat administration. The MTD for LY3022859 was not determined. Dose escalation beyond 25 mg was considered unsafe due to worsening symptoms (uncontrolled cytokine release) despite prophylaxis (corticosteroids and antihistamines). clinicaltrials.gov Identifier: NCT01646203.

Renin-angiotensin system inhibitors (RASi) aim to reduce the negative effects of Angiotensin (Ang) II, either by decreasing its production or by inhibiting its binding to the Ang II receptor Type 1 (AT1R) using angiotensin-converting enzyme inhibitors (ACEi) or Ang receptor blockers (ARB)

Transforming growth factor-â (TGF-â) diduga berperan pada preeklamsia. Reseptor TGF-â tipe II (TâR-II) dihasilkan dari transkripsi gen TGF-â receptor type II (TGFBR2). Polimorfisme gen TGFBR2 pada basa C1167T dapat menyebabkan hipoksia yang menginduksi iskemia serta meningkatkan produksi solubel endoglin (sEng) dan vascular cell adhesion molecule-1 (VCAM-1). Tujuan penelitian ini untuk mengetahui korelasi polimorfisme gen TGFBR2 pada basa C1167T dengan kadar sEng dan VCAM-1 ibu preeklamsia. Subjek adalah ibu preeklamsia usia kehamilan 28–42 minggu dan kehamilan normal sebagai kontrol, masing-masing 120 orang. Penelitian dilakukan di Rumah Sakit Hasan Sadikin, Bandung, September 2008–Mei 2009. Sampel berupa darah vena, pemeriksaan polimorfisme dilakukan dengan DNA Wizard® genomic DNA purification, kadar sEng dan VCAM-1 dengan imunoesai. Hasil penelitian menunjukkan polimorfisme CT pada kelompok preeklamsia 92 (76,7%) dan kontrol 70 (58,3%) {p

The levels of several soluble cytokine receptors in body fluids of healthy individuals change with age. Clinical application of the measurement of the serum soluble interleukin‐1 receptor type I (sIL‐1RI) level depends critically on the samples used as the controls. At present, there is no information regarding the levels of serum sIL‐1RI in healthy subjects. The purpose of this study is to reveal the age‐related changes that occur in the serum sIL‐1RIlevels of healthy individuals. We determined the serum sIL‐1RI levels of healthy Japanese children using ELISA. The serum sIL‐1RI level of children (0–14 years) was significantly higher than that of adults (more than 15 years) (P=0.0138, n=90). Thus, it is recommended that when the serum sIL‐1RI level of patients is evaluated, it should be compared against age‐matched controls. J. Clin. Lab. Anal. 23:175–178, 2009. © 2009 Wiley‐Liss, Inc.

An involvement of pituitary adenylate cyclase activating polypeptide-specific receptor type 1 (PAC1-R) in behavioral performance of retrieval of memory upon the stimulation of perception of sensory modality was investigated in mice. Eighteen heterozygous (-/+) transgenic mutant PAC1-R-deficient mice ((-/+) mice) and 18 wild-type (+/+) mice of littermates ((+/+) mice) as a control were used. No homozygous (-/-) transgenic mutant PAC1-R-deficit mouse was bred in our colony. It was observed that; (1) changing a single alley in the multiple mazes interfered with retrieval of memory in both (-/+) mice and (+/+) mice, and they made considerable errors; (2) a rotation of the multiple mazes for 180 degrees in relation to the geomagnetic field made also considerable errors, but in only (+/+) mice and not in (-/+) mice; and (3) an exposure for perception of fear signals made also considerable errors, but in only (+/+) mice and not in (-/+) mice. It is inferred that PAC1-R may play a gating role in passage permitting through perception of geomagnetic orientation and fear signals, but not perception of visual orientation, and it regulates behavioral performance in retrieval of memory in mice.

A 20-year-old fit male soldier presented on two separate occasions 16 months apart with severe, symptomatic hyponatraemia and a clinical and biochemical picture consistent with the syndrome of inappropriate secretion of antidiuretic hormone (SIADH). In the intervening period, repeated plasma sodium values were in the reference range. Intensive investigation failed to reveal a cause for SIADH that was initially considered idiopathic. The description of a family comprising several adults with intermittent or water load induced-hyponatraemia associated with an activating mutation in the arginine vasopressin (AVP) receptor type 2 (AVPR2) raised the question of whether our patient could have a similar ‘nephrogenic syndrome of inappropriate antidiuresis’. Mutational screening of AVPR2 in our patient revealed a single missense mutation (R137C) in the second intracellular loop, which has been associated with constitutive activation of the AVPR2. In conclusion, adults with intermittent, severe hyponatraemia may have a constitutively activating mutation in the AVPR2 with resultant nephrogenic syndrome of inappropriate antidiuresis. Patients with idiopathic SIADH, particularly those with unmeasurable circulating AVP concentrations, should be considered for mutational screening of AVPR2.

Objective: There are some clues that measurement of triglyceride (TG) and high-density lipoprotein (HDL) and their correlation with tumor necrosis factor alpha (TNF-α), soluble TNF-α receptor type 1 and type 2 (sTNFR1, sTNFR2) in serum could be valuable in the assessment of disease activity of systemic lupus erythematosus (SLE) patients. Methods: In this cross-sectional study, fasting blood samples were obtained from 86 SLE patients referred to the Rheumatology Research Center of Tehran University in Shariati Hospital, Iran. Disease activity was determined by using the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI). TG and HDL obtained after overnight fasting were analysed by routine chemistry. Levels of circulating TNF-α, sTNFR1, and sTNFR2 were determined by enzyme-linked immunosorbent assay. Results: Triglyceride levels were associated with SLEDAI (r = 0.59, P 0.23) and (r = –0.17, P > 0.1), respectively. In multiple logistic regression models, the levels of TNF-α and HDL were omitted. Conclusion: Dyslipoproteinemia with high TG/low HDL levels correlates with disease activity in SLE, and enhanced activity in the TNF-α/sTNFR system. With results of this study and the same studies, serum levels of TG, HDL, TNF-α, sTNFR1, sTNFR2 are valuable markers for estimation of disease activity in SLE.

To investigate contribution of glucocorticoids to the maintenance of gastric mucosal integrity during stress we predominantly used ulcerogenic stress models. Using these models we demonstrated that glucocorticoids released in response to the ulcerogenic stimuli attenuated their harmful action on the gastric mucosa.In the present study we hypothesized that mild stressors does not damage the gastric mucosa due to gastroprotective action of glucocorticoids released in response to these stimuli. To verify the hypothesis the effects of normally non-ulcerogenic mild stimuli (15-30 min cold-restraint) on the gastric mucosa have been studied under the circumstances of inhibition ofthe hypothalamic-pituitary-adrenocortical (HPA) axis in rats. The HPA axis was inhibited by: 1) fast inhibitory action of NBI 27914, the selective antagonist of cortricotropin-releasing factor receptor type 1; 2) fast inhibitory action of metyrapone, inhibitor glucocoricoid synthesis; 3) delayed inhibitory action of a single pharmacolog...

Stress-induced inhibition of food intake is reportedly blocked by a selective corticotropin-releasing factor (CRF) type 1 receptor (CRF1) antagonist, suggesting the involvement of CRF1 in the inhibitory mechanism. CRF1 and CRF2 are considered to function in the inhibition of food intake by CRF-related peptides with different time courses.This study was designed to clarify whether CRF2 is also involved in stress-induced inhibition of food intake and to examine the relation of CRF1to CRF2 in the inhibitory mechanism.Antisauvagine-30 (AS-30), a selective CRF2 antagonist, and/or CRA1000, a selective CRF1 antagonist, were pre-administered intracerebroventricularly and intraperitoneally, respectively, to male Wistar rats deprived of food for 24 h before the animals were exposed to a 1-h period of stressors and food intake in 1 h after stress exposure was examined. The effect of both antagonists on locomotor activity was also examined.Pre-administration of 5-30 microg of AS-30 attenuated inhibition of food intake induced by restraint, electric footshock or emotional stress using a communication box. CRA1000 also attenuated the restraint-induced inhibition of food intake at doses of 5 and 10 mg/kg body weight. The reversal of restraint-induced inhibition of food intake by co-administration of AS-30 and CRA1000 was not larger than that by AS-30 or CRA1000 alone. Both antagonists did not affect locomotor activity.These results suggest that not only CRF1, but also CRF2, are involved in stress-induced inhibition of food intake, and that both subtypes of CRF receptor function probably in series in 1 h after stress exposure.

In this article, we review the recent findings regarding a new derivative of angiotensin-(1-7) [Ang-(1-7)], alamandine, and its receptor, the Mas-related G-coupled receptor type D (MrgD) with a special emphasis on its role and how it can be formed.Over the last decade, there have been significant conceptual changes regarding the understanding of the renin-angiotensin system (RAS). A cardioprotective axis has been elucidated by the discovery of the Mas receptor for the biologically active Ang-(1-7), and the angiotensin-converting enzyme 2 (ACE2) that coverts Ang II into Ang-(1-7). In addition, several components of the system, such as Ang-(1-12), Angiotensin A (Ang A) and the newly discovered peptide, alamandine, have been identified. Alamandine is generated by catalysis of Ang A via ACE2 or directly from Ang-(1-7).Alamandine is a vasoactive peptide with similar protective actions as Ang-(1-7) that acts through the MrgD and may represent another important counter-regulatory mechanism within the RAS.

The effects of conditioned fear on the release of noradrenaline in the hypothalamic paraventricular nucleus (PVN) and the involvement of corticotropin-releasing factor (CRF) receptor type 1 (CRFR1) in conditioned fear-induced changes in noradrenaline release were examined by intracerebral microdialysis in rats. Conditioned fear was produced by placing animals into a box where they had previously been exposed to a 5-min period of electric footshock, 135 min prior to the start of experiment. Conditioned fear for 20 min produced a significant increase in the release of noradrenaline in the PVN. Intraperitoneal preadministration of a selective nonpeptidic CRFR1 antagonist, CRA1000, completely blocked the conditioned fear-induced release of noradrenaline. These results suggest that CRFR1 is involved in the release of noradrenaline in the hypothalamic PVN induced by conditioned fear.

The receptor type mediating the positive inotropic effect of histamine was examined in left atria from neonatal guinea pigs. The positive inotropic effect of histamine, as well as its action potential prolonging effect, was antagonized by ranitidine, but not by chlorpheniramine or thiperamide. The positive inotropic effect was enhanced by isobutylmethylxantine. Receptor binding studies revealed the presence of both H1 and H2 receptor types. These results suggest that the positive inotropic effect of histamine in the neonatal guinea-pig atrium is mediated by H2 receptors.

The possible effect of different corticotropin-releasing factor receptor (CRFR) antagonists (alpha-helical CRF, CP-154,526 and AS-30) on the maintenance and reactivation of morphine-conditioned place preference (CPP) induced by morphine or footshock stress, respectively, were investigated in rats. The results show that morphine-induced maintenance of CPP was not affected by pretreatment with any CRFR antagonists. However, morphine-induced the reactivation of CPP was significantly attenuated by pre-administration of 10 microg alpha-helical CRF (i.c.v.). The maintenance of morphine CPP could be induced by repeated footshock and this effect was significantly attenuated by pretreatment of 10 microg alpha-helical CRF (i.c.v.) and 10 mg CP-154,526 (i.p.). Furthermore, following a 28-day extinction of morphine CPP, a single footshock could again elicit the reactivation of place preference that was blocked by pretreatment with 10 microg alpha-helical CRF (i.c.v.) and 1 or 10 mg CP-154,526 (i.p.). The present study demonstrates that CRFR type 1, but not CRFR type 2, mediates the stress-induced maintenance and reactivation of morphine CPP. These findings suggest that CRFR type 1 antagonists might be of some value in the treatment and prevention of stress-induced relapse to drug dependence long after detoxification.

Expression of transforming growth factor (TGF)-β3 as well as cellular localization of TGF-β receptors has not been demonstrated in placenta. TGF-β receptor type I (RI) and type II (RII) are required to transmit TGF-β signals, therefore the determination of cells expressing both receptors in concert is necessary to identify target cells for TGF-β. We investigated presence and localization of TGF-βs and their receptors (RI, RII) in human term placenta using reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry. RT-PCR showed that messenger RNA for TGF-β1, -β2, -β3, and RI and RII is present in human term placenta. We found intense staining for all TGF-β isoform and receptor proteins in the syncytiotrophoblastic layer, chorionic plate, and in cells of the extravillous trophoblasts using immunohistochemistry. The simultaneous expression of ligands and their receptors support the hypothesis that TGF-β may play an important role in regulating growth, differentiation, and function of the human placenta.

The stress neurohormone corticotropin releasing factor (CRF) activates at least two receptor types. Expression of corticotropin releasing factor receptor type II (CRF2) has been demonstrated in the hypothalamic ventromedial nucleus (VMH) of the adult and developing rat, but the physiological functions of VMH-CRF2 have not been elucidated. The VMH has been documented as an important participant in the regulation of food intake and its interactions with the hypothalamic-pituitary-adrenal axis and circadian rhythms. Regulation of VMH-CRF2 may thus play a role in the interplay of physiological alterations in metabolic state with the neuroendocrine and anorexic effects of CRF. This study determined the regulation of CRF2-mRNA expression in infant rats by the physiological consequences of maternal deprivation, i.e., fasting and stress. Using in situ hybridization, maternally deprived pups had an average 62% reduction of VMH-CRF2-mRNA levels compared with stress-free controls. Maternal deprivation also resulted in elevated plasma corticosterone levels (3.8 +/- 0.3 vs. 1.3 +/- 0.1 microg/dl) and an average 5.7% body weight loss. This study demonstrates that maternal deprivation, via fasting and HPA activation, leads to a dramatic decrease of CRF2-mRNA levels in the VMH. These results are consistent with a role for CRF2 activation in mediating some of the complex interactions of CRF (or urocortin) with regulation of food intake in the developing rat.

e13125Background: Human epidermal growth receptor type 2 (HER2) overexpression/amplification is associated with the efficacy of HER2 targeted therapies in breast and gastric cancers, where candidates for its targeted therapies are selected by immunohistochemistry (IHC) and/or fluorescence in situ hybridization (FISH) testing. However, the HER2 status in colorectal cancer (CRC) has not been fully investigated to date. CancerPlex is a comprehensive next-generation sequencing-based test that evaluates 400-plus genes for mutations and copy number changes, including HER2. In the present study, we aimed to clarify the association between HER2 status and HER2 gene alteration detected with CancerPlex in CRC. Methods: This retrospective study included 100 patients with Stage I-IV CRC, who were evaluated HER2 status and HER2 gene alteration. IHC for HER2 was classified as 4 grades according to the immunoreactivity. The patients with IHC2+ were further tested with FISH. IHC3+/IHC2+ and FISH positive patients were de...

Germline mutations of BMPR2 gene ( bone morphogenetic protein receptor type 2 ), or more rarely of ACVRL1 ( activin A receptor type II-li

Effects of subjects’ taste sensitivity (expressed as taste detection threshold), tastant quality and taste transduction mechanism on pulsation-induced taste enhancement were tested. Taste intensities of pulsatile MSG and NaCl stimuli at pulsation periods below, at and above individual taste fusion periods (TFP in seconds) were compared to taste intensities of a continuous reference of the same net tastant concentration and quality. In line with results previously reported for sucrose, pulsation-induced taste enhancement peaked around TFP for both MSG and NaCl and did not require perception of tastant pulsation. TFP and pulsation effects were independent of the taste transduction mechanism (G-protein-coupled receptor for MSG versus ion-channel for NaCl). The absence of a relation between TFP and taste sensitivity suggests that temporal gustatory resolution and taste sensitivity are not necessarily influenced by the same factors. The results support earlier findings that early stages of taste transduction are involved in pulsation-induced taste enhancement. Pulsation-induced taste enhancement is determined by the pulsation rate (i.e. TFP) which is longer for MSG than NaCl. This is probably due to the tastant-specific interaction with the receptor rather than the taste transduction mechanism (G-protein-coupled receptor versus ion-channel) involved.

We report on a patient with familial bicuspid aortic valve disease operated on for proximal aortic aneurysm. The surgery was complicated by intraoperative aortic dissection. Multi-generational genetic analysis demonstrated a mutation in the transforming growth factor-beta receptor type II gene. This case confirms the clinical hypothesis that the proximal aortic disease has a genetic origin in some bicuspid aortic valve patients.

The aim of our investigation was to analyze the pattern of interleukin-1 (IL-1) family compounds: IL-1 beta, IL-1 receptor accessory protein (Acp), IL-1 receptor antagonist (IL-1Ra) and IL-1 receptor type II (IL-1RII) in the serum and cerebrospinal fluid (CSF) from 67 multiple sclerosis (MS) patients and 31 controls. We found significantly elevated CSF levels of IL-1 beta, IL-1Ra and Acp in MS patients compared to controls (p=0.001), while IL-1 beta and Acp were significantly elevated in MS sera (p=0.001). IL-1Ra and/or IL-1 RII increased in sera of all 10 investigated patients after the steroid treatment for relapse. Our findings suggest the important beneficial role of the induction of IL-1 RII and IL-1Ra in MS.

P0320 TARGETING TGF-BETA I WITH THE TRANSFORMING GROWTH FACTOR RECEPTOR TYPE I KINASE INHIBITOR, LY2157299, MODULATES STEMNESS-RELATED BIOMARKERS IN HEPATOCELLULAR CARCINOMA B. Rani, F. Dituri, Y. Cao, U. Engstrom, L. Lupo, S. Dooley, A. Moustakas, G. Giannelli. Department of Biomedical Sciences and Human Oncology, University of Bari, Bari, Italy; Biomedical Center, Ludwig Institute for Cancer Research, Science for Life Laboratory, Uppsala, Sweden; Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy; Molecular Hepatology-Alcohol associated diseases, Department of medicine II, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany; Department of Medical Biochemistry and Microbiology, and Ludwig Institute for Cancer Research, Science for Life Laboratory, Box 581 Biomedical Center, Uppsala, Sweden E-mail: bhavna.rani@uniba.it

Targeting Tgf-Beta I With The Transforming Growth Factor Receptor Type I Kinase Inhibitor, Ly2157299, Modulates Stemness-Related Biomarkers In Hepatocellular Carcinoma

Appropriate N-terminus modification can result in somatostatin (SRIF) octapeptide analogs that are both more potent and more selective in vitro for the human SRIF receptor type 2 (hsst2). In addition, these modifications can improve the duration of action and bioavailability of SRIF analogs following parenteral administration, as shown by both pharmacological and distribution studies in vivo with BIM-23190 and BIM-23197 in the rat.

Recently, we described a splice variant of the human natriuretic peptide receptor type B (NPR-Bi) in human proximal tubule cells [immortalized human kidney epithelial cells (IHKE-1) that lacks a functional guanylate cyclase domain (Hirsch JR, Meyer M, Mägert HJ, Forssmann WG, Mollerup S, Herter P, Weber G, Cermak R, Ankorina-Stark I, Schlatter E, and Kruhøffer M. J Am Soc Nephrol 10: 472–480, 1999). Its signaling pathway does not include cGMP, cAMP, or Ca2+but leads to inhibition of K+channels. In patch-clamp experiments, effects of tyrosine kinase receptor blockers on C-type natriuretic peptide (CNP)-mediated depolarizations of membrane voltages ( Vm) of IHKE-1 cells were tested. The epidermal growth factor (EGF) receptor blocker genistein (10 μM) abolished the effect of CNP (0.2 ± 0.4 mV, n = 7), and comparable results were obtained with 10 μM daidzein ( n = 8). Aminogenistein (10 μM, n = 5) and tyrphostin AG1295 (10 μM, n = 5) had no significant effects. EGF (1 nM) hyperpolarized cells by –5.3 ± 0.8 mV ( n = 5). This effect was completely blocked by genistein or daidzein. The Cl–channel blocker NPPB (10 μM, n = 5) inhibited the EGF-mediated hyperpolarization. mRNA expression of NPR-B and NPR-Bi shows reversed patterns along the human nephron. NPR-B is highly expressed in glomeruli and proximal tubules, whereas NPR-Bi shows strong signals in the distal nephron. Expression of NPR-Bi in the cortical collecting duct was also confirmed with immunohistochemistry. In other human tissues, NPR-Bi shows strongest expression in pancreas and lung, whereas in the heart and liver NPR-B is the dominating receptor. In conclusion, CNP inhibits an apical K+channel in IHKE-1 cells independently of cGMP and so far this effect can only be blocked by genistein and daidzein. Tyrosine phosphorylation might be the missing link in the signaling pathway of CNP/NPR-Bi.

The involvement of 5-HT1Breceptors in the regulation of vigilance states was assessed by investigating the spontaneous sleep–waking cycles and the effects of 5-HT receptor ligands on sleep in knock-out (5-HT1B−/−) mice that do not express this receptor type. Both 5-HT1B−/− and wild-type 129/Sv mice exhibited a clear-cut diurnal sleep–wakefulness rhythm, but knock-out animals were characterized by higher amounts of paradoxical sleep and lower amounts of slow-wave sleep during the light phase and by a lack of paradoxical sleep rebound after deprivation. In wild-type mice, the 5-HT1Bagonists CP 94253 (1–10 mg/kg, i.p.) and RU 24969 (0.25–2.0 mg/kg, i.p.) induced a dose-dependent reduction of paradoxical sleep during the 2–6 hr after injection, whereas the 5-HT1B/1Dantagonist GR 127935 (0.1–1.0 mg/kg, i.p.) enhanced paradoxical sleep. In addition, pretreatment with GR 127935, but not with the 5-HT1Aantagonist WAY 100635, prevented the effects of both 5-HT1Bagonists. In contrast, none of the 5-HT1Breceptor ligands, at the same doses as those used in wild-type mice, had any effect on sleep in 5-HT1B−/− mutants. Finally, the 5-HT1Aagonist 8-OH-DPAT (0.2–1.2 mg/kg, s.c.) induced in both strains a reduction in the amount of paradoxical sleep. Altogether, these data indicate that 5-HT1Breceptors participate in the regulation of paradoxical sleep in the mouse.

We investigated whether corticotropin-releasing factor (CRF) receptor type 1 (CRFR1) is involved in emotional stress-induced inhibition of food intake and behavioral changes in rats. The inhibition of food intake and increase in locomotor activity induced by emotional stress using a communication box were reversed by both intracerebroventricular injection of α-helical CRF (9–41), a non-selective CRF receptor antagonist, and intraperitoneal injection of a selective non-peptidic CRFR1 antagonist. These results suggest that CRFR1 mediates at least in part the emotional stress-induced inhibition of feeding behavior and increase in locomotor activity.

The mechanisms by which interferon-beta-1b (IFNbeta-1b) acts in the treatment of patients with multiple sclerosis (MS) are not completely known. A total of 10 MS patients were treated with 8 million units of IFNbeta-1b every other day. Compared to baseline and control group the expression of TGFbeta-1-mRNA by PBMC was persistently increased at week 6, month 3 and month 6 (por = 0.04), that of the TGFbeta-1 receptor type II from day 5 up to month 6 (p0.01). The mRNA and protein expression of tumor necrosis factor-alpha (TNFalpha)-receptor (55 kDa) was only temporarily elevated at the beginning of the therapy. Serum levels of sVCAM were increased during the whole time of treatment (p0.01). The CD8CD38 lymphocyte subpopulation was continuously elevated from day 5 up to month 6 (p0.01). No persistently significant changes were demonstrable concerning the percentage of total CD4, CD8, CD19 or in CD4 subpopulations (CD4CD29, CD4CD45RA). The present data suggest that IFNbeta-1b induces the expression of TGFbeta-1- and TGFbeta-R-II-mRNA by PBMC and increases levels of sVCAM-1 and of circulating activated CD8 cells (CD8CD38) in serum. These might be other mechanisms by which IFNbeta-1b mediates its positive effects in the treatment of MS patients.

Plasma concentrations of soluble interleukin (IL)-1 receptor type II, IL-10, and IL-13 were measured in 42 patients with clinically defined sepsis during a 3-day follow-up and in 7 healthy humans after intravenous injection of endotoxin (2 ng/kg). Levels of soluble IL-1 receptor type II were persistently elevated in patients with sepsis than in healthy controls and higher in nonsurviving patients (n = 22) than in surviving patients (n = 20) at all time points. IL-10 was found in the circulation of 81% of patients with sepsis, while it was not detectable in normal plasma. During follow-up, IL-10 remained invariably high only in nonsurviving patients, while it significantly decreased in survivors. Endotoxin induced IL-10, while soluble IL-1 receptor type II remained unchanged. IL-13 remained undetectable in the vast majority of patients and was not induced by endotoxin. Enhanced IL-13 production does not seem to be part of an inducible host defense mechanism during sepsis.