G1 Treatment Research Articles

GPR30 activation improves memory and facilitates DHPG-induced LTD in the hippocampal CA3 of middle-aged mice

Reduced estrogen levels and decreased expression of related receptors are typical cerebral features of aging. The G protein-coupled estrogen receptor 1 (GPER1, also known as GPR30) is considered a novel therapeutic target for neurodegenerative diseases. In this study, we demonstrated that hippocampal GPR30 expression was reduced in middle-aged mice compared with young adult mice. GPR30 agonist G1 improved both fear and spatial memory in both male and female middle-aged mice, but not in young adult mice, which were blocked by the GPR30 antagonist G15. Interestingly, a group I metabotropic glutamate receptor (mGluR) agonist, 3,5-dihydroxyphenylglycine (DHPG)-induced long-term depression (LTD) in mossy fiber-cornu ammonis 3 (MF-CA3) synapses but not Schaffer collateral-CA1 (SC-CA1) synapses was facilitated in brain slices from G1-treated middle-aged mice. Long-term potentiation (LTP) in SC-CA1 synapses was not affected in slices from G1-treated mice. The effects of GPR30 activation on memory and DHPG-LTD in MF-CA3 synapses were further confirmed by viral expression of GPR30 in the CA3. The regulation of hippocampal synaptic plasticity by G1 treatment might be related to brain-derived neurotrophic factor (BDNF)-tropomyosin receptor kinase B (TrkB) signaling, as G15 also blocked G1-induced activation of the BDNF-TrkB pathway. Moreover, we found that DHPG triggered GluA internalization in slices from G1-treated mice but not control mice. Pharmacological experiments showed that G1-mediated facilitation of DHPG-induced LTD in MF-CA3 synapses was dependent on protein kinase B (Akt), mammalian target of rapamycin (mTor), and TrkB signaling. In conclusion, our results indicate that GPR30 activation improves memory in middle-aged mice, likely through facilitating synaptic plasticity in the CA3. This study provides novel evidence that GPR30 activation can improve memory in middle-aged animals.

Simple heat profiles and biogeochemical patterns for analysis the influence on soil microbial community of plastic-greenhouse and open field condition

The diversity of the soil microbial community is remarkably sensitive to different cropping techniques. To examine the effects of cultivation in a greenhouse on the soil microbial activity and diversity, we collected soil samples from vegetable crops continuously grown in a plastic greenhouse (G) and an open field (N). Heat profiling was obtained using microcalorimeter in the presence of different carbon sources. Power time curves were recorded after glucose and ammonium sulphate supplementation. The total heat release QT (Jg-1), maximum heat flow Pmax (μW) and growth rate constant k (h-1) are higher in G treatment group than the in N treatment group. On the other hand, the time (tmax) to reach the peak heat flow is lower in G treatment group compared to N treatment group, as is cell specific heat rate JQ/N (J cell-1). Sample G1 has the highest Pmax (1246.07 μW) and the lowest tmax (11.86 h); in contrast, sample N5 has the lowest Pmax (411.03 μW) and the highest tmax (21.78 h). The microbial community in the soil was estimated from phospholipid fatty acid (PLFA) biomarkers analysis and a viable count, followed by a subsequent determination of the physicochemical parameters. The total average PLFA concentration range is found as 129.76 - 142.54 nmol g-1. Both thermodynamics properties and PLFA concentration was are significantly greater in G1, G2 and G3 treatment than in the other two (N5 & N6) management systems (P < 0.05). Combination of both thermodynamic and PLFA profiling determines that the microbial population is higher in the soil from the greenhouse fertilization than from the open field cultivation. This novel approach could be applicable to a better understanding of (i) the changes in the soil microbial community in green house and open field cultivation practices, and (ii) the relationship between physicochemical properties and soil microbial community’s diversity. Moreover, this would be a notable technique for the decision maker to choose the potential cultivation method for sustainable green-food development program.

GPER activation is effective in protecting against inflammation-induced nigral dopaminergic loss and motor function impairment

Increasing evidence suggest that excessive inflammatory responses from overactivated microglia play a critical role in Parkinson’s disease (PD), contributing to, or exacerbating, nigral dopaminergic (DA) degeneration. Recent results from our group and others demonstrated that selective activation of G protein-coupled estrogen receptor (GPER) with the agonist G1 can protect DA neurons from 1-methyl-4-phenylpyridinium (MPP+) and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) neurotoxins. However, it is not known whether modulation of microglial responses is one of the mechanisms by which G1 exerts its DA neuroprotective effects. We analyzed, in the N9 microglial cell line, the effect of G1 on microglial activation induced by lipopolysaccharide (LPS) exposure. The results revealed that G1 significantly decrease phagocytic activity, expression of inducible nitric oxide synthase (iNOS) and release of nitric oxide (NO) induced by LPS. To determine the relevance of this anti-inflammatory effect to the protection of nigral DA cells, the effect of G1 was analyzed in male mice injected unilaterally in the substantia nigra (SN) with LPS. Although G1 treatment did not decrease LPS-induced increase of ionized calcium binding adaptor molecule 1 (iba-1) positive cells it significantly reduced interleukin-1beta (IL-1β), cluster of differentiation 68 (CD68) and iNOS mRNA levels, and totally inhibited nigral DA cell loss and, as a consequence, protected the motor function. In summary, our findings demonstrated that the G1 agonist is able to modulate microglial responses and to protect DA neurons and motor functions against a lesion induced by an inflammatory insult. Since G1 lacks the feminizing effects associated with agonists of the classical estrogen receptors (ERs), the use of G1 to selectively activate the GPER may be a promising strategy for the development of new therapeutics for the treatment of PD and other neuroinflammatory diseases.

Estrogen stabilizes hypoxia-inducible factor 1α through G protein-coupled estrogen receptor 1 in eutopic endometrium of endometriosis

To investigate whether G protein-coupled estrogen receptor (GPER, also known as GPR30 and GPER1) stabilizes hypoxia-inducible factor 1α (HIF-1α) in eutopic endometrium (EuEM) of endometriosis. Immunohistochemical analysis and experimental invitro study. University hospital. Patients with or without endometriosis. The EuEM and normal control endometrium (CoEM) were obtained by curettage. Primary cultured endometrial stromal cells (ESCs) were treated with 17β-E2, G1, or G15. The EuEM and CoEM were collected for immunohistochemistry. Western blot, polymerase chain reaction, ELISA, and dual luciferase experiments were used to detect expression of GPER, HIF-1α, vascular endothelial growth factor (VEGF), and matrix metalloproteinase 9 (MMP9) in ESCs. Estradiol and G1 were used as agonists of GPER, G15 as an antagonist. Migration of ESCs and endothelial tube formation of human umbilical vein endothelial cells cultured in medium collected from ESCs were measured. Protein levels of GPER and HIF-1α were higher in EuEM than in CoEM. Protein levels of HIF-1α but not HIF-1α mRNA levels increased concurrently with GPER after E2 and G1 treatment. Furthermore, expression and activity of VEGF and MMP9 increased under E2 and G1 stimulation. However, these effects disappeared when GPER was blocked. G protein-coupled estrogen receptor stabilizes HIF-1α and thus promotes HIF-1α-induced VEGF and MMP9 in ESCs, which play critical roles in endometriosis.

Activation of GPER ameliorates experimental pulmonary hypertension in male rats

RationalePulmonary hypertension (PH) is characterized by pulmonary vascular remodeling that leads to pulmonary congestion, uncompensated right-ventricle (RV) failure, and premature death. Preclinical studies have demonstrated that the G protein-coupled estrogen receptor (GPER) is cardioprotective in male rats and that its activation elicits vascular relaxation in rats of either sex. ObjectivesTo study the effects of GPER on the cardiopulmonary system by the administration of its selective agonist G1 in male rats with monocrotaline (MCT)-induced PH. MethodsRats received a single intraperitoneal injection of MCT (60mg/kg) for PH induction. Experimental groups were as follows: control, MCT+vehicle, and MCT+G1 (400μg/kg/daysubcutaneous). Animals (n=5pergroup) were treated with vehicle or G1 for 14days after disease onset. Measurements and Main ResultsActivation of GPER attenuated exercise intolerance and reduced RV overload in PH rats. Rats with PH exhibited echocardiographic alterations, such as reduced pulmonary flow, RV hypertrophy, and left-ventricle dysfunction, by the end of protocol. G1 treatment reversed these PH-related abnormalities of cardiopulmonary function and structure, in part by promoting pulmonary endothelial nitric oxide synthesis, Ca2+ handling regulation and reduction of inflammation in cardiomyocytes, and a decrease of collagen deposition by acting in pulmonary and cardiac fibroblasts. ConclusionsG1 was effective to reverse PH-induced RV dysfunction and exercise intolerance in male rats, a finding that have important implications for ongoing clinical evaluation of new cardioprotective and vasodilator drugs for the treatment of the disease.

Oestrogens Stimulate Proliferation in Colorectal Cancer via GPER and the Hippo signalling pathway

The concentration of circulating oestrogens is associated with the incidence and outcomes of colorectal cancer (CRC). Both the physiological and pathological effects of oestrogens are mediated by oestrogen receptors. This project aims to investigate the potential role of G protein-coupled oestrogen receptor 1 (GPER) as an oestrogen-induced mediator in CRC proliferation. To achieve this, colorectal adenoma and carcinoma cell lines were examined for protein expression of oestrogen receptors. Immunoblotting did not report ERa and ERb expression, although GPER was detected. Following this, the GPER-associated gene connective tissue growth factor (CTGF) expression was measured after I7β-estradiol (E2) and GPER agonist (G1) treatment. qRT-PCR results showed no significant increase in CTGF expression levels, 24 and 48 hours after treatment. In addition, GPER interaction with the Hippo pathway in CRC was examined by treating cells with E2 and G1 for 0, 15, 30 minutes, 1, and 2 hours. Alterations in P-Y API expression were unclear after treatment in the cell lines examined. However, addition of GPER antagonist, G15, resulted in significant inhibition in YAP1 phosphorylation in HCT116 cells, 15 and 30 minutes of treatment. Consequently, our data supports that oestrogens and G1 treatment leads to increase in YAP phosphorylation and nucleus-cytoplasmic shuttling via GPER stimulation. YAP1 knock-down studies and pharmacological inhibition followed by proliferation assays established that this early metabolic effect translates into increased cellular proliferation. Collectively, our data propose a novel oestrogen-driven pro-proliferative pathway via GPER through Hippo pathway's key downstream effector, YAP1, in CRC. Further studies are required to 'reveal the detailed signalling cascade by which GPER mediates the increased YAP1 phosphorylation.

Abstract P600: Activation Of GPER Ameliorates Pulmonary Hypertension In Female Rats

Introduction: Pulmonary hypertension (PH) is primarily a disease of women (female-to-male ratio 4:1) and is associated with cardiac dysfunction. Aim: The activation of GPER by its agonist G1 was evaluated in monocrotaline (MCT)-induced PH rats. Methods: Depletion of estrogen was induced by bilateral oophorectomy (OVX; n = 18) in female Wistar rats (12 wks old). Experimental groups were: SHAM or OVX that received i.p. injection of MCT (60 mg/kg) for PH induction followed by administration of vehicle or G1 (400 μg/kg/day s.c.) for 14 days (n=7 per group). Hemodynamic parameters were determined by echocardiography. The effects of G1 in the maintenance of exercise capacity was investigated using a treadmill test. Results: MCT injection and estrogen loss led to a significant decrease in pulmonary acceleration time (PAT) and an increase in RV free wall thickness and MCT-related changes were attenuated by treatment with G1 ( P < 0.05; Table 1). Right ventricular systolic pressure (RVSP) was higher in MCT-injected rats and the magnitude of this increase in OVX group was significantly higher than that in SHAM group. G1 normalized RVSP in both SHAM and OVX rats (Table 1). G1 treatment reversed altered expression of SERCA2a and phospholamban proteins in the RV (Table 1). Interaction between estrogen loss and MCT also reduced treadmill time to exhaustion in PH rats ( P < 0.05), and chronic administration of G1 restored the exercise capacity ( P < 0.05). Conclusion: G1 reversed PH-related cardiopulmonary dysfunction and exercise intolerance in female rats, a finding that may have important implications for the ongoing clinical evaluation of new drugs for the treatment of the disease in aging females.

Abstract 267: Activation of a Novel Estrogen Receptor Ameliorates Experimental Pulmonary Hypertension in Male Rats

Introduction: Pulmonary arterial hypertension (PAH) is characterized by pulmonary vascular remodeling that leads to uncompensated RV failure, and premature death. Preclinical studies have demonstrated that activation of the G protein-coupled estrogen receptor (GPER) is cardioprotective in male rats, and that a selective agonist G1, elicits vascular relaxation in rats of either sex. This work investigated the effects of G1 in male rats with monocrotaline (MCT)-induced PAH. Methods and Results: Male Wistar rats received a single intraperitoneal injection of MCT (60 mg/kg) for PAH induction. Experimental groups were: control, MCT + vehicle, and MCT + G1 (400 μg/kg/day s.c.). Animals were treated with vehicle or G1 for 14 days after the onset of disease (n = 5 per group). Treadmill test and transthoracic echocardiography were performed to access exercise capacity and cardiac function, respectively. RV systolic pressure (RVSP) and mean arterial pressure (MAP) were measured. Time to exhaustion in the treadmill (s) was reduced from 1042.0 ± 66.4 to 188.0 ± 53.1 (MCT + vehicle) and recovered to 809.4 ± 61.5 after G1 treatment. Pulmonary acceleration time (PAT) (ms) was reduced from 44.7 ± 1.4 to 24.7 ± 1.2 in MCT + vehicle group and restored to 41.8 ± 1.0 in MCT + G1 group. RVSP (mmHg) was increased from 24.6 ± 0.6 to 41.1 ± 1.4 (MCT + vehicle) and was reduced to 27.5 ± 0.7 (MCT + G1). MAP (mmHg) was reduced from 100.9 ± 1.1 to 77.1 ± 2.4 (MCT + vehicle), indicating HF induced by the RV dysfunction, and G1 normalized it to 92.4 ± 1.4. G1 decreased pulmonary vascular remodeling and normalized endothelial nitric oxide enzyme levels in lungs from PAH rats. PLB/SERCA2a ratio increased, as a sign of contractility dysfunction, in PAH rats and tumor necrosis factor alpha upregulation was significantly correlated with these Ca 2+ handling proteins impairment. Activation of GPER beneficially reduced PLB/SERCA2a ratio and TNF-α levels in RV tissue of MCT-induced PAH rats. Conclusions: G1 was effective to reverse PAH-induced RV dysfunction and exercise intolerance in male rats, a finding that may have important implications for ongoing clinical evaluation of new drugs for the treatment of the disease.

Activation of GPR30 improves exercise capacity and skeletal muscle strength in senescent female Fischer344 × Brown Norway rats

The molecular mechanisms of muscle weakness and sarcopenia in postmenopausal women are largely unknown. To determine the effect of a new estrogen receptor, GPR30, in the maintenance of exercise capacity and skeletal muscle function in females, the selective GPR30 agonist, G1 (100 μg/kg/day), or vehicle (V, soybean oil) was administered subcutaneously daily (n = 7 per group) to ovariectomized (OVX) 27-month-old Fischer 344 × Brown Norway (F344BN) female rats. Following 8 weeks of treatment, the exercise capacity (treadmill walk time to exhaustion) was reduced in OVX vs. sham rats (5.1 ± 1.4 vs. 11.0 ± 0.9 min, P < 0.05), and chronic G1 restored exercise capacity (12.9 ± 1.2 min; P < 0.05 vs. OVX-V). Similarly, the peak twitch of electrically stimulated soleus muscles was decreased by 22% in OVX vs. sham rats (P < 0.05), and G1 attenuated this decline (P < 0.05). Western blot analysis showed that chronic G1 treatment attenuated OVX-associated decreases in heat shock protein (HSP) 90, HSP70, and HSP27 expressions. In vitro studies using the L6 myoblast cell line demonstrated that G1 increased mRNA levels of HSPs in cultured cells. Collectively, these data demonstrate that the activation of GPR30 mitigates the adverse effects of estrogen loss on exercise capacity and skeletal muscle contractile function in old F344BN rats. The protective effects of GPR30 might be through its upregulation of heat shock proteins in skeletal muscle.

NADPH-Diaphorase Colocalizes with GPER and Is Modulated by the GPER Agonist G1 in the Supraoptic and Paraventricular Nuclei of Ovariectomized Female Rats

Nitric oxide is produced in the brain by the neuronal nitric oxide synthase (nNOS) and carries out a wide range of functions by acting as a neurotransmitter-like molecule. Gonadal hormones are involved in the regulation of the brain nitrergic system. We have previously demonstrated that estradiol, via classical estrogen receptors (ERs), regulates NOS activity in the supraoptic (SON) and paraventricular (PVN) nuclei of the hypothalamus, acting through both ERα and ERβ. Magnocellular and parvocellular neurons in the SON and PVN also express the G protein-coupled ER (GPER). In this study, we have assessed whether GPER is also involved in the regulation of nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase in the SON and PVN. Adult female ovariectomized rats were treated with G1, a selective GPER agonist, or with G1 in combination with G15, a selective GPER antagonist. G1 treatment decreased NADPH-diaphorase expression in the SON and in all PVN subnuclei. The treatment with G1 + G15 effectively rescued the G1-dependent decrease in NADPH-diaphorase expression in both brain regions. In addition, the activation of extracellular signal-regulated kinase (ERK) 1/2, one of the kinases involved in the GPER-dependent intracellular signaling pathway and in NOS phosphorylation, was assessed in the same brain nuclei. Treatment with G1 significantly decreased the number of p-ERK 1/2-positive cells in the SON and PVN, while the treatment with G1 + G15 significantly recovered its number to control values. These findings suggest that the activation of GPER in the SON and PVN inhibits the phosphorylation of ERK 1/2, which induces a decrease in NADPH-diaphorase expression.

GPER inhibits diabetes-mediated RhoA activation to prevent vascular endothelial dysfunction

The effect of estrogen receptors on diabetes-induced vascular dysfunction is critical, but ambiguous. Individuals with diabetic vascular disease may require estrogen receptor-specific targeted therapy in the future. The G protein-coupled estrogen receptor (GPER) has beneficial effects on vascular function. However, its fundamental mechanisms are unclear. The RhoA/Rho-kinase pathway contributes to diabetic vascular complications, whereas estrogen can suppress Rho-kinase function. Thus, we assumed that GPER inhibits diabetes-mediated RhoA activation to prevent vascular dysfunction. We further investigated the underlying mechanisms involved in this process. Vascular endothelial cells and ex vivo cultured ovariectomized (OVX) C57BL/6 mouse aortae were treated with high glucose (HG) alone or in combination with GPER agonist (G1). G1 treatment was also administered to OVX db/db mice for 8 weeks. An ex-vivo isovolumic myograph was used to analyze the endothelium-dependent vasodilation and endothelium-independent contraction of mouse aortae. Apoptosis, oxidative stress, and inflammation were attenuated in G1-pretreated vascular endothelial cells. G1 significantly decreased the phosphorylation of inhibitory endothelial nitric oxide (NO) synthase residue threonine 495 (eNOS Thr495), inhibited RhoA expression, and increased NO production. Additionally, G1 rescued the impaired endothelium-dependent relaxation and inhibited RhoA activation in the thoracic aorta of OVX db/db mice and ex-vivo cultured OVX C57BL/6 mouse aortae treated with HG. Estrogens acting via GPER could protect vascular endothelium, and GPER activation might elicit ERα-independent effect to inhibit RhoA/Rho-kinase pathway. Additionally, GPER activation might reduce vascular smooth muscle contraction by inhibiting RhoA activation. Thus, the results of the present study suggest a new therapeutic paradigm for end-stage vascular dysfunction by inhibiting RhoA/Rho-kinase pathway via GPER activation.

DAMPAK APLIKASI HERBISIDA IPA GLIFOSAT DALAM SISTEM TANPA OLAH TANAH (TOT) TERHADAP TANAH DAN TANAMAN PADI SAWAH

Weed is one of the important constraints in rice production, and therefore its effective control measure should be considered that the production can be optimized. The application of no tillage by using herbicide is assumed more effective and efficient than conventional one. However, herbicide which is used as contionusly can influence residue in soil, plant and rice. The objectives of the research were to analyze the effect of no tillage and its combination of IPA glyphosate herbicide concentration levels to rice productivity and to analyze IPA glyphosate herbicide residue in soil, straw and rice. The study was conducted in the field by using IPA glyphosate herbicide with five treatments, namely maximum tillage (Gm), no herbicide spraying (G0), glyphosate herbicide doses 1.5 l ha -1 (G1), 3 l ha -1 (G2) and 4.5 l ha -1 (G3). The analysis of glyphosate residue was done in soil, straw and rice samples by using HPLC (high performance liquid chromatography) method. The research results showed that Gm and G1 treatments had highest with rice yield average were 938 g m -2 and 728 g m -2 , respectively. Gm treatmen more profitable with R / C ratio of 1.84 with a profit of Rp 13.714 million, but using more labor than G1 treatment. Thereby, no-tillage treatment (G1) could be done by using glyphosate herbicide in doses 1.5 l ha -1 , economically. Glyphosate contained in soil samples, straw, and rice proved that using of gyphosate intensively could have negative impacts on soil microbial activity, plant resistance to plant diseases and residue of glyphosate carried by plants. Glyphosate residue concentration was highest found on rice sample in treatments G3 was 0.272 mg kg -1 . These glyphosate residual values on rice was highest than maximum residue limit which was decided by Indonesia government (0.1 mg kg -1 ). Glyphosate residues contained in food is not within safe limits if taken every day and can cause adverse effects to human health. Keywords: glyphosate herbicide, maximum residue limits, paddy field, weed control

Abstract 351: Late-life Activation of GPR30 by G1 Reverses Diastolic Dysfunction in Senescent Fischer344 x Brown Norway Female Rats

Introduction: The aged heart is characterized by impaired lusitropy and increased stiffness, and no proven pharmacologic interventions exist that limit these manifestations of left ventricular diastolic dysfunction (LVDD). G1, a selective agonist to the estrogen receptor GPR30, attenuated the effects of estrogen (E2) loss on LVDD in a hypertensive model. To extend prior studies, we determined the cardioprotective potential of chronic GPR30 activation in the female Fischer344 x Brown Norway (F344BN) rat, a normotensive aging model. Methods and Results: Bilateral oophorectomy (OVX) or sham-operations were performed in middle- (Mid) (18 months) and Old-aged rats (27 months). At 28 months of age, Old rats were randomized to daily G1 (100 μg/kg/day, s.c.) or vehicle (VEH; soybean oil) injections (n=7 rats/group). Following 8 weeks of treatment, left ventricle (LV) mass was greater in Old vs. Mid-aged rats, independent of E2 or G1, while systolic function and LV wall thicknesses were similar among groups. Myocardial relaxation (e’) was reduced (P&lt;0.001) and LV filling pressure (E/e’) was increased (P&lt;0.01) in Old vs. Mid-aged rats, and this was exacerbated by OVX (P&lt;0.01) and reversed by late-life G1 treatment. Systolic blood pressure (SBP) increased by 20% after mid-life OVX, whereas late-life E2 loss had no effect on SBP, irrespective of G1. Cardiac chymase expression was increased in E2 deficient hearts determined by Western blot and G1 modestly limited this E2/age effect. Conclusion: Our data indicate that late-life G1 treatment reverses the detrimental effects of advanced age and E2 deficiency on diastolic function, possibly via interactions with the noncanonical cardiac renin angiotensin system.

Osteoprotective effects of estrogen membrane receptor GPR30 in ovariectomized rats

G protein-coupled estrogen receptor 30 (GPR30) is expressed in bone tissue. However, little is known regarding the function of GPR30 in postmenopausal osteoporosis. In this study, we examined the effects of GPR30 on ovariectomy (OVX)-induced osteoporosis in rats, including the effects on proliferation, differentiation, and expression of proteins in osteoblasts. Administration of G1 (35μg/kg, ip, 3 times/week for 6 weeks), a specific agonist of GPR30, prevented OVX-induced increase in bone turnover rate, decrease in bone mineral content and bone mineral density, damage to bone structure, and aggravation of bone biomechanical properties. In addition, G1 did not affect uterine weight in the OVX rats. Osteoblasts isolated from calvarias from newborn rats were used to explore the underlying mechanisms. G1 (150pM) promoted proliferation and differentiation of osteoblasts through a positive feedback of GPR30, which then activated the PI3K-Akt, ERK, and CREB pathways. G15 (750pM), a specific antagonist of GPR30, reversed the above effects initiated by G1 treatment. In conclusion, activation of GPR30 protected bones against osteoporosis in OVX rats and exerted no untoward effect on the uterus. We suggest that GPR30 can be used as an effective therapeutic target for the prevention and treatment of postmenopausal osteoporosis.

Dampak Aplikasi Herbisida IPA Glifosat dalam Sistem Tanpa Olah Tanah (TOT) Terhadap Tanah dan Tanaman Padi Sawah

Weed is one of the important constraints in rice production, and therefore its effective control measure should be considered that the production can be optimized. The application of no tillage by using herbicide is assumed more effective and efficient than conventional one. However, herbicide which is used as contionusly can influence residue in soil, plant and rice. The objectives of the research were to analyze the effect of no tillage and its combination of IPA glyphosate herbicide concentration levels to rice productivity and to analyze IPA glyphosate herbicide residue in soil, straw and rice. The study was conducted in the field by using IPA glyphosate herbicide with five treatments, namely maximum tillage (Gm), no herbicide spraying (G0), glyphosate herbicide doses 1.5 l ha -1 (G1), 3 l ha -1 (G2) and 4.5 l ha -1 (G3). The analysis of glyphosate residue was done in soil, straw and rice samples by using HPLC (high performance liquid chromatography) method. The research results showed that Gm and G1 treatments had highest with rice yield average were 938 g m -2 and 728 g m -2 , respectively. Gm treatmen more profitable with R / C ratio of 1.84 with a profit of Rp 13.714 million, but using more labor than G1 treatment. Thereby, no-tillage treatment (G1) could be done by using glyphosate herbicide in doses 1.5 l ha -1 , economically. Glyphosate contained in soil samples, straw, and rice proved that using of gyphosate intensively could have negative impacts on soil microbial activity, plant resistance to plant diseases and residue of glyphosate carried by plants. Glyphosate residue concentration was highest found on rice sample in treatments G3 was 0.272 mg kg -1 . These glyphosate residual values on rice was highest than maximum residue limit which was decided by Indonesia government (0.1 mg kg -1 ). Glyphosate residues contained in food is not within safe limits if taken every day and can cause adverse effects to human health. Keywords: glyphosate herbicide, maximum residue limits, paddy field, weed control

Effects of Long-Term Treatment with Estradiol and Estrogen Receptor Subtype Agonists on Serotonergic Function in Ovariectomized Rats

Acute estradiol treatment was reported to slow the clearance of serotonin via activation of estrogen receptors (ER)β and/or GPR30 and to block the ability of a selective serotonin reuptake inhibitor (SSRI) to slow serotonin clearance via activation of ERα. In this study, the behavioral consequences of longer-term treatments with estradiol or ER subtype-selective agonists and/or an SSRI were examined in the forced swim test (FST). Ovariectomized rats were administered the following for 2 weeks: estradiol, ERβ agonist (diarylpropionitrile, DPN), GPR30 agonist (G1), ERα agonist (PPT), and/or the SSRI sertraline. Similar to sertraline, longer-term treatment with estradiol, DPN or G1 induced an antidepressant-like effect. By contrast, PPT did not, even though it blocked the antidepressant-like effect of sertraline. Uterus weights, used as a peripheral measure of estrogenic activity, were increased by estradiol and PPT but not DPN or G1 treatment. A second part of this study investigated, using Western blot analyses in homogenates from hippocampus, whether these behavioral effects are accompanied by changes in the activation of specific signaling pathways and/or TrkB. Estradiol and G1 increased phosphorylation of Akt, ERK and TrkB. These effects were similar to those obtained after treatment with sertraline. Treatment with DPN increased phosphorylation of ERK and TrkB, but it did not alter that of Akt. Treatment with PPT increased phosphorylation of Akt and ERK without altering that of TrkB. In conclusion, activation of at least TrkB and possibly ERK may be involved in the antidepressant-like effect of estradiol, ERβ and GPR30 agonists whereas Akt activation may not be necessary.

GPER1-mediated immunomodulation and neuroprotection in the myenteric plexus of a mouse model of Parkinson's disease

Lewy pathology affects the gastrointestinal tract in Parkinson's disease (PD) and recent reports suggest a link between the disorder and gut inflammation. In this study, we investigated enteric neuroprotection and macrophage immunomodulation by 17β-estradiol (E2) and the G protein-coupled estrogen receptor 1 (GPER1) in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse PD model. We found that both E2 and the GPER1 agonist G1 are protective against the loss of dopamine myenteric neurons and inhibited enteric macrophage infiltration in MPTP-treated mice. Coadministration of GPER1 antagonist G15, while completely blocking the neuroprotective and anti-inflammatory effects of G1 also partially prevented those of E2. Interestingly, we found that E2 and G1 treatments could directly alter MPTP-mediated immune responses independently from neurodegenerative processes. Analyses of monocyte/macrophage NF-κB and iNOS activation and FACs immunophenotype indicated that 1-methyl-4-phenylpyridinium (MPP+) treatment induces a strong immune response in monocytes, comparable to that of canonical challenge by lipopolysaccharide. In these cells, G1 and E2 treatment are equally potent in promoting a shift toward an anti-inflammatory “M2” immunophenotype reducing MPP+-induced NF-κB and iNOS activation. Moreover, G15 also antagonized the immunomodulatory effects of G1 in MPP+-treated macrophages. Together these data provide the first evidence for the role of GPER1 in enteric immunomodulation and neuroprotection. Considering increasing recognition for myenteric pathology as an early biomarker for PD, these findings provide a valuable contribution for better understanding and targeting of future therapeutic strategies.

Activation of GPR30 inhibits cardiac fibroblast proliferation.

The incidence of left ventricular diastolic dysfunction significantly increases in postmenopausal women suggesting the association between estrogen loss and diastolic dysfunction. The in vivo activation of G protein-coupled estrogen receptor (GPR30) attenuates the adverse effects of estrogen loss on cardiac fibrosis and diastolic dysfunction in mRen2.Lewis rats. This study was designed to address the effects of GPR30 on cardiac fibroblast proliferation in rats. The expression of GPR30 in cardiac fibroblasts isolated from adult Sprague-Dawley rats was confirmed by RT-PCR, Western blot analysis, and immunofluorescence staining. Results from BrdU incorporation assays, cell counting, carboxyfluorescein diacetate succinimidyl ester labeling in conjunction with flow cytometry, and Ki-67 staining showed that treatment with G1, a specific agonist of GPR30, inhibited cardiac fibroblast proliferation in a dose-dependent manner, which was associated with decreases in CDK1 and cyclin B1 protein expressions. In the GPR30-KO cells, BrdU incorporation, and CDK1 and cyclin B1 expressions significantly increased when compared to GPR30-intact cells. G1 had no effect on BrdU incorporation, CDK1 and cyclin B1 mRNA levels in GPR30-KO cells. In vivo studies showed increases in CDK1 and cyclin B1 mRNA levels, Ki-67-positive cells, and the immunohistochemistry staining of vimentin, a fibroblast marker, in the left ventricles from ovariectomized mRen2.Lewis rats versus hearts from ovary-intact littermates; 2 weeks of G1 treatment attenuated these adverse effects of estrogen loss. This study demonstrates that GPR30 is expressed in rat cardiac fibroblasts, and activation of GPR30 limits proliferation of these cells likely via suppression of the cell cycle proteins, cyclin B1, and CDK1.

Immunogenicity and safety of measles–mumps–rubella vaccine delivered by disposable-syringe jet injector in healthy Brazilian infants: A randomized non-inferiority study

This study aimed to determine if immunogenicity to measles–mumps–rubella vaccine delivered to infants via a disposable-syringe jet injector (DSJI) was non-inferior to that administered by needle and syringe (NS). Vaccination safety was evaluated, as were the use, performance, and acceptability of each delivery method. The DSJI was the PharmaJet® 2009 generation-1 device (G1) and the vaccine was measles–mumps–rubella vaccine from Bio-Manguinhos. Five hundred eighty-two healthy Brazilian infants were randomized to receive vaccine via G1 or NS. Seroconversion rates against measles and mumps viruses in the G1 treatment group did not meet non-inferiority criteria when compared with the NS group; however, responses in the G1 group to rubella virus were non-inferior to those of NS vaccinees. Most adverse events were mild or moderate. Crying after injection was more frequent in the NS group, and local skin reactions were more common in the G1 group. Five serious adverse events were judged causally unrelated to treatment and all resolved. Parents/guardians expressed a strong preference for G1 over NS for their children. Vaccinators found the G1 easy to use but noted incomplete vaccine delivery in some cases. Although the G1 has been superseded by an updated device, our results are important for the continued improvement and evaluation of DSJIs, which have the potential to overcome many of the challenges and risks associated with needle-based injections worldwide. Recommendations for future DSJI clinical studies include rigorous training of vaccinators, quantitative measurement of wetness on the skin following injection, and regular monitoring of device and vaccinator performance.

Influence of furostanol glycosides treatments on strawberry (Fragaria × ananassa Duch.) growth and photosynthetic characteristics under drought condition

Water availability is one of the major limiting factors for plant growth and productivity. In recent years, efforts to search for specific and practical approaches to improve the adaptability of plants to this environmental condition have been made. In this study we have examined the influence of two furostanol glycoside application on the capacity for acclimation to drought of two strawberry (Fragaria×ananassa Duch.) cultivars, Real and Magic, grown in greenhouse conditions. Two regimes of irrigation were used: well-watered (80% WHC) and drought stress (50% WHC) for one month. Two furostanol glycoside aqueous solutions (G1 and G2) were applied by foliar spraying during late afternoon at 3 days intervals on 5 mature plants for each variant. Changes in plant growth and photosynthesis (gas exchange and chlorophyll fluorescence measurements) were studied.In drought conditions, glycoside application induced a decrease in leaf size and increase in roots length, resulting in significant increases of root/shoot ratio. The two glycoside treatments alleviated the effect of drought and improved photosynthetic rate (A) and water use efficiency (A/E) in both cultivars. In addition, G1 treatment reduced the severity of drought effect by increasing the stomata responsiveness thus maintaining a higher relative water content (RWC), while G2 treatment led to a lower decrease of stomatal conductance (gs) and transpiration rate (E) and a significant decrease in internal CO2 (Ci) concentration compared to untreated plants subjected to stress. The chlorophyll fluorescence measurements indicate that furostanol glycoside treatments mitigated the reduction of photosystem II efficiency (ΦPS II) and electron transport rate (ETR) caused by water stress, compared to untreated plants. Moreover, under drought stress, treated plants recorded higher values of non-photochemical processes (NPQ) than untreated ones. One can safely assume that glycoside treatments induced an enhancement of non-photochemical processes as well as an alleviation of the drought-induced alteration in PS II by increasing the efficiency of light utilization and dissipation of excitation energy in the PSII antennae.