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Background: Prosthetic valve implantation requires postoperative prophylactic anticoagulation to preclude thrombotic events. The aim of this review is to assess the role of anticoagulation therapy in the management of valve replacement patients. Methodology: Literature from PubMed, Embase, Medline and Google Scholar were searched using the terms "valvular heart disease", "anticoagulant", "mechanical heart valve", "bioprosthesis", "bridging", "Vitamin K antagonist (VKA)", and "acenocoumarol". A committee comprising leading cardiothoracic surgeons from India was convened to review the literature and suggest key practice points. Results: Prosthetic valve implantation requires postoperative prophylactic anticoagulation to preclude thrombotic events. A paramount risk of thromboembolic events is observed during the first three months after surgery for both mechanical and bioprosthetic devices. The VKA therapy with individualized target international normalized ratio (INR) is recommended in patients after prosthetic valve replacement. Therapies for the management of prosthetic valve complications should be based on the type of complications. Special care is mandated in distinguished individuals and those with various co-morbidities. Conclusion: In patients with prosthetic valve replacement, anticoagulant therapy with VKA seems to be an effective option. The role for non-VKA oral anticoagulants in the setting of prosthetic valve replacement has yet to be established. Furthermore, whether the novel oral anticoagulants are safe and efficacious in patients after placement of a bioprosthetic valve remains unanswered.
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Anticoagulantes/uso terapêutico , Implante de Prótese de Valva Cardíaca , Complicações Pós-Operatórias/prevenção & controle , Tromboembolia/prevenção & controle , Próteses Valvulares Cardíacas , Hemorragia/induzido quimicamente , Hemorragia/terapia , Humanos , Coeficiente Internacional Normatizado , Medição de RiscoRESUMO
This work provides an electrodeposition-based methodology for synthesizing multicomponent nanowires containing Ag, Co and Ni atoms. Nanowire morphology was obtained by using an anodic alumina membrane with cylindrical pores of â¼ 200-nm diameter. Structural, compositional and magnetic characterization revealed that the as-synthesized nanowires adopted a core-shell microstructure. The core (axial region) contained pure Ag phase volumes with a plate-like morphology oriented perpendicular to the nanowire axis. The shell (peripheral region) contained pure Ag nanoparticles along with superparamagnetic Co and Ni rich clusters.
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This report provides information about an electrodeposition-based two-step synthesis methodology for producing core-shell Ag-(Ni-O) nanowires and their detailed structural and compositional characterization using electron microscopy technique. Nanowires were produced by employing anodic alumina templates with a pore diameter of 200 nm. In the first step of the synthesis process, nanocrystalline Ni-O was electrodeposited in a controlled manner such that it heterogeneously nucleated and grew only on the template pore walls without filling the pores from bottom upwards. This alumina template with pore walls coated with Ni-O was then utilized as a template during the electrodeposition of Ag in the second step. Electrodeposited Ag filled the template pores to finally produce Ag-(Ni-O) core-shell nanowires with an overall diameter of 200 nm.
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The genetic diversity among 28 pea (Pisum sativum L.) genotypes was analyzed using 32 simple sequence repeat markers. A total of 44 polymorphic bands, with an average of 2.1 bands per primer, were obtained. The polymorphism information content ranged from 0.657 to 0.309 with an average of 0.493. The variation in genetic diversity among these cultivars ranged from 0.11 to 0.73. Cluster analysis based on Jaccard's similarity coefficient using the unweighted pair-group method with arithmetic mean (UPGMA) revealed 2 distinct clusters, I and II, comprising 6 and 22 genotypes, respectively. Cluster II was further differentiated into 2 subclusters, IIA and IIB, with 12 and 10 genotypes, respectively. Principal component (PC) analysis revealed results similar to those of UPGMA. The first, second, and third PCs contributed 21.6, 16.1, and 14.0% of the variation, respectively; cumulative variation of the first 3 PCs was 51.7%.
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Variação Genética , Repetições de Microssatélites/genética , Pisum sativum/genética , Marcadores Genéticos , Genótipo , Filogenia , Análise de Componente PrincipalRESUMO
The brown planthopper, Nilaparvata lugens (Stål) is a major sucking insect pest of rice. This insect has long been considered as migratory; however, its route in India is still unknown. Hence, to find out its migration route genetic diversity, genetic structure and gene flow of 16 N. lugens populations from major rice growing regions of India was studied based on mitochondrial cytochrome oxidase I (COI). The results revealed a high genetic homogeneity among the populations on the basis of genetic diversity statistics and neutrality tests. There was a prevalence of a single major haplotype across the country. No spatial relevance was found with the genetic structure of the populations indicating presence of excessive gene flow among them. Extensive gene flow among populations was also confirmed with the presence of higher number of immigrants in North, Central, and East India. To further clarify the migration sources, 48 h air-mass reverse trajectory was performed for Varanasi just aftermath of cyclones Amphan and Yaas, which disclosed Eastern/Northeastern states along with Bangladesh and Myanmar as the possible source areas. Overall, the results revealed a single panmictic homogeneous population of N. lugens in India with extensive gene flow as a consequence of their migration. These findings will help in better forecasting enabling efficient regional management of this important rice pest. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03337-6.
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The combination activities of temephos, fenthion and petroleum ether extract of Solanum xanthocarpum were observed for their larvicidal activities against Culex quinquefasciatus. The combination of temephos and S. xanthocarpum was studied at ratios of 1:1, 1:2, and 1:4. Similar ratios were also used for the combination of fenthion and S. xanthocarpum. The temephos/plant extract combination acted antagonistically. The combination of fenthion and plant extract acted synergistically against the target organisms at a ratio of 1:1, which showed the best results of: LC50 0.0144 and 0.0056 ppm and LC90 0.0958 and 0.0209 ppm at 24 and 48 hours, respectively. The present study will be helpful in developing a commercial formulation for effective vector management.
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Culex/efeitos dos fármacos , Filariose/prevenção & controle , Insetos Vetores/efeitos dos fármacos , Inseticidas/toxicidade , Larva/efeitos dos fármacos , Controle de Mosquitos/métodos , Extratos Vegetais/toxicidade , Solanum/toxicidade , Alcanos/toxicidade , Animais , Fention/toxicidade , Filariose/parasitologia , Inseticidas/química , Solanum/química , Temefós/toxicidade , Testes de Toxicidade AgudaRESUMO
Malaria control in developing countries is based largely on vector eradication by the use of mosquito larvicides which is an ideal method for controlling mosquito and the related epidemics. On account of ecohazardous nature, nontarget specificity of chemical insecticides and evidences of developing resistance against them in the exposed species, currently, importance of secondary plant metabolites has been acknowledged. Insecticides of plant origin are environmentally safe, degradable, and target specific. In view of this fact, the present work highlights the larvicidal property of extracts of Amaranthus oleracea and Euphorbia hirta against the third instar larvae of Anopheles stephensi, the urban malaria vector. LC(50) values for the carbon tetrachloride fraction of A. oleracea against larvae are 17,768.00 and 13,780.00 ppm after 24 and 48 h of exposure accordingly. For the methanol extract of the same, LC(50) values are 15,541.00 and 10,174.00 ppm after 24 and 48 h of exposure. In the case of petroleum ether extract, LC(50) values after 24 and 48 h of exposure are 848.75 and 311.50 ppm. LC(50) values for carbon tetrachloride extracts of E. hirta against the larvae are 11,063.00 and 10,922.00 ppm after 24 and 48 h of exposure, respectively. For methanol extract of the same extract, the LC(50) values are 19,280.00 and 18,476.00 ppm after 24 and 48 h of exposure. In the case of petroleum ether extract, LC(50) values after a 24- and 48-h exposure period are 9,693.90 and 7,752.80 ppm. The results obtained for petroleum extracts of A. oleracea are encouraging and there are probabilities that the active principle contained in this extract may be more effective than its crude form and may serve as ecofriendly mosquito larvicide.
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Amaranthus/química , Anopheles/efeitos dos fármacos , Euphorbia/química , Inseticidas/farmacologia , Extratos Vegetais/farmacologia , Animais , Vetores de Doenças , Índia , Inseticidas/isolamento & purificação , Extratos Vegetais/isolamento & purificação , Análise de Sobrevida , Fatores de TempoRESUMO
In view of the recently increased interest in developing plant-based insecticides as an alternative to chemical insecticides, this study was undertaken to assess the larvicidal potential of the various fruit wall extracts of Momordica charantia (cucurbitaceae) against two species of mosquito vectors, Anophels stephensi and Culex quinquefasciatus. Among the extracts tested, petroleum ether (LC(50) = 27.60; 17.22 ppm and 41.36; 15.62 ppm) extract was found more effective than carbon tetrachloride (LC(50) = 49.58; 16.15 ppm and 80.61; 27.64 ppm) and methanol (LC(50) = 142.82; 95.98 ppm and 1,057.49; 579.93 ppm) extracts towards anopheline and culicine larvae after 24 and 48 h of exposure respectively. Thus, all fruit wall extracts of M. charantia are toxic to both the larval species. M. charantia may, therefore, act as an effective biolarvicide against mosquitoes in the future.
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Anopheles/efeitos dos fármacos , Culex/efeitos dos fármacos , Frutas/química , Inseticidas/isolamento & purificação , Inseticidas/farmacologia , Momordica charantia/química , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Animais , Larva/efeitos dos fármacos , Análise de SobrevidaRESUMO
In search of a natural larvicide, petroleum ether, carbon tetrachloride, and methanol extracts of Azadirachta indica fruits and seed extracts of bitter gourd (Momordica charantia) and castor (Ricinus communis) were tested for larvicidal activity against Culex quinquefasciatus. Among the extracts tested, the methanol extract of Az. indica was observed the most potent with LC(50) at 74.04 and 58.52 ppm and LC(-90) at 201.83 and 171.70 ppm as compared to methanol extract of M. charantia with LC(50) at 101.18 and 93.58 ppm and LC(90) at 322.81 and 302.62 ppm carbon tetrachloride extract of R. communis with LC(50) at 144.11 and 92.44 ppm and LC(90) at 432.42 and 352.89 ppm after 24 and 48 h, respectively. The methanol extract of Az. indica exhibited potential results and can be exploited as a preferred natural larvicide for the control of filarial vector, Cx. quinquefasciatus.
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Azadirachta/química , Culex/efeitos dos fármacos , Inseticidas/farmacologia , Momordica charantia/química , Extratos Vegetais/farmacologia , Ricinus communis/química , Animais , Frutas/química , Inseticidas/isolamento & purificação , Larva/efeitos dos fármacos , Dose Letal Mediana , Extratos Vegetais/isolamento & purificação , Sementes/química , Análise de SobrevidaRESUMO
PURPOSE: Valvular heart disease (VHD) patients after prosthetic valve implantation are at risk of thromboembolic events. Follow-up care of patients with prosthetic valve has a paramount role in reducing the morbidity and mortality. Currently, in India, there is quintessential need to stream line the follow-up care of prosthetic valve patients. This mandates the development of a consensus guideline for the antithrombotic therapy in VHD patients post prosthetic valve implantation. METHODS: A national level panel was constituted comprising 13 leading cardio care experts in India who thoroughly reviewed the up to date literature, formulated the recommendations, and developed the consensus document. Later on, extensive discussions were held on this draft and the recommendations in 8 regional meetings involving 79 additional experts from the cardio care in India, to arrive at a consensus. The final consensus document is developed relying on the available evidence and/or majority consensus from all the meetings. RESULTS: The panel recommended vitamin K antagonist (VKA) therapy with individualized target international normalized ratio (INR) in VHD patients after prosthetic valve implantation. The panel opined that management of prosthetic valve complications should be personalized on the basis of type of complications. In addition, the panel recommends to distinguish individuals with various co-morbidities and attend them appropriately. CONCLUSIONS: Anticoagulant therapy with VKA seems to be an effective option post prosthetic valve implantation in VHD patients. However, the role for non-VKA oral therapy in prosthetic valve patients and the safety and efficacy of novel oral anticoagulants in patients with bioprosthetic valve need to be studied extensively.
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Larvicidal potential of petroleum ether, carbon tetrachloride and methanol extracts of Aloe barbadensis and Cannabis sativa has been investigated against Culex quinquefasciatus. Among the extracts examined, Carbon tetrachloride extract (Cte) of Aloe barbadensis was the most effective with LC50 values of 15.31 and 11.01 ppm after 24 and 48 hr of exposure, respectively followed by pertoleum ether extract (Pee) of A barbadensis, Cte of C. sativa, methanol extract (Mee) of A. barbadensis, methanol and petroleum ether of C. saliva, LC, being 25.97, 88.51, 144.44, 160.78 and 294.42 ppm affer 24hr and 16.60, 68.69, 108.38, 71.71 and 73.32 ppm after 48 hr of post treatment, respectively. Cte of both the plants exhibits potential larvicidal activity and can be used as ecofriendly alternative in the management of the filariasis vector, Culex quinquefasciatus.
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Aloe/química , Cannabis/química , Culex/efeitos dos fármacos , Insetos Vetores/efeitos dos fármacos , Extratos Vegetais/toxicidade , Animais , Culex/crescimento & desenvolvimento , Filariose , Insetos Vetores/crescimento & desenvolvimento , Larva/efeitos dos fármacos , Dose Letal Mediana , Folhas de Planta/químicaRESUMO
With a goal of minimal application of environmentally hazardous chemical insecticides, the larvicidal activity of cypermethrin was studied alone and in combination with the root extract of Solanum xanthocarpum against anopheline larvae. Petroleum ether extract was observed to be the most toxic, with LC,, of 1.41 and 0.93 ppm and LC90 of 16.94 and 8.48 ppm at 24 and 48 hours after application, respectively, followed by carbon tetrachloride and methanol extracts. The values for cypermethrin were an LC50 of 0.0369 ppm after 24 hours and 0.0096 ppm after 48 hours and LC90 of 0.0142 and 0.0091 ppm after 24 and 48 hours, respectively. The ratios of cypermethrin and petroleum ether extracts tested were 1:1, 1:2 and 1:4. Of the various ratios tested, the cypermethrin and petroleum ether extract ratio of 1:1 was observed to be more efficient than the other combinations. From the individual efficacy of each constituent, synergism was noted. This is an ideal ecofriendly approach for the control of malaria vector, Anopheles stephensi.
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Anopheles/efeitos dos fármacos , Insetos Vetores/efeitos dos fármacos , Inseticidas/toxicidade , Larva/efeitos dos fármacos , Malária/prevenção & controle , Controle de Mosquitos/métodos , Extratos Vegetais/toxicidade , Piretrinas/toxicidade , Solanum/toxicidade , Alcanos/toxicidade , Animais , Anopheles/crescimento & desenvolvimento , Bioensaio , Tetracloreto de Carbono/toxicidade , Combinação de Medicamentos , Malária/parasitologia , Extratos Vegetais/química , Raízes de Plantas/química , Raízes de Plantas/toxicidade , Análise de Regressão , Testes de Toxicidade AgudaRESUMO
Larvicidal potential of petroleum ether (Pee), carbon tetrachloride (Cte) and methanol extract (Mee) of Artemisia annua, Chenopodium album and Sonchus oleraceus was observed against malaria vector, Anopheles stephensi Liston. The Pee of A. annua with LC50 16.85 ppm after 24 h and 11.45 ppm after 48 h of treatment was found most effective, followed by Cte of A. annua and Ch. album, Pee of Ch. album and Mee of A. annua. However, no significant larvicidal activity was observed in Mee of Ch. album and all the three extracts of S. oleraceous. The Pee of A. annua was further investigated for its effect on the metamorphosis and the development of the malaria vector. It influenced the early life cycle of An. stephensi by reducing the percentage of hatching, larval, pupal and adult emergence and also lengthening the larval and pupal periods. The growth index was also reduced significantly. As the extract has remarkable effect on the metamorphosis and high larvicidal potential, it could, therefore, be used as an effective biocontrol agent against the highly nuisant malaria vector.
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Anopheles/efeitos dos fármacos , Insetos Vetores/efeitos dos fármacos , Larva/efeitos dos fármacos , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Alcanos/farmacologia , Animais , Artemisia/química , Tetracloreto de Carbono/farmacologia , Chenopodium album/química , Relação Dose-Resposta a Droga , Metanol/farmacologia , Controle de Mosquitos/métodos , Sonchus/químicaRESUMO
Spermatogenesis is a complex process that is both remarkable and enigmatic. While circulating hormones clearly play an important role in initiating and regulating the process, the Sertoli cell barrier prevents most substances from entering the seminiferous tubule compartment and directly influencing germ cell development. Therefore, the testis cannot rely solely upon the delivery of circulating hormones, nutrients, and growth factors, but must independently produce its own regulatory substances. A rapidly increasing number of testicular factors that appear to modulate spermatogenesis in a paracrine fashion are now being identified. These discoveries are beginning to contribute to our understanding of the intricate network of testicular cell-cell interactions that control male reproduction.
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DBA/2 mice infected with lethal dosages of Friend virus complex (FVC) can be 100% cured by split-dose total body irradiation (TBI) at 150 cGy, an effect associated with the restoration of the cellular immunity which is compromised by the virus. The exact mechanism underlying the curative effect is unknown, but it may involve the interferon (IFN) system and interleukin-2 (IL-2) production. Initial studies indicated that TBI did not directly inactivate the virus, suggesting that irradiation either acted on the target cells for virus replication or on other cells mediating the effect. We have now examined the effect of this relatively low dose TBI on replication, transcription, and protein expression of the Friend virus. Northern blot analysis revealed that in FVC infected mice treated with curative low dose TBI, no spleen focus-forming virus (SFFV)-specific mRNA species were detected. Southern blot analysis revealed that a 6.0 kb SFFV fragment could be detected in infected, untreated spleen cells, but not in cells from FVC-infected mice treated with TBI, or in uninfected spleen cells. Western blot analysis revealed that the SFFV envelope glycoprotein was expressed in the spleen cells from untreated FVC infected mice, but not in the cells from TBI treated FVC infected mice. These results, consistent with our previous findings of greatly reduced spleen focus forming units in mice with FVC which had been treated with this regimen of TBI, suggest the possibility of using such treatments in other retroviral associated disorders.
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DNA Viral/efeitos da radiação , Vírus da Leucemia Murina de Friend/efeitos da radiação , Leucemia Experimental/radioterapia , RNA Mensageiro/efeitos da radiação , RNA Viral/efeitos da radiação , Proteínas dos Retroviridae/metabolismo , Vírus Formadores de Foco no Baço/efeitos da radiação , Irradiação Corporal Total , Animais , Northern Blotting , Southern Blotting , Western Blotting , Relação Dose-Resposta a Droga , Feminino , Vírus da Leucemia Murina de Friend/genética , Leucemia Experimental/genética , Leucemia Experimental/microbiologia , Camundongos , Camundongos Endogâmicos , RNA Viral/genética , Proteínas dos Retroviridae/genética , Vírus Formadores de Foco no Baço/genéticaRESUMO
Ethanolic and acetone extracts of Nerium indicum and Thuja orientelis have been studied against III instar larvae of Anopheles stephensi and Culex quinquefasciatus. Ethanolic extract of N. indicum is found more effective than its acetone extract against anopheline larvae with LC50 values of 185.99 and 148.05 ppm for former and 229.28 and 149.43 ppm for the later after 24 and 48 hrs of exposure. The acetone extract with LC50 values of 209.00 and 155.97 ppm is more effective in case of culicine larvae than its ethanolic extract with LC50 494.07 and 194.49 ppm after 24 and 48 hours of treatment. Ethanolic extract of T. orientelis is more effective against both the larval species with LC50 values of 13.10 and 9.02 ppm after 24 and 48 hours for anopheline and 22.74 and 16.72 ppm against culicine larvae. The acetone extract showed LC50 values of 200.87 and 127.53 ppm against anopheline and 69.03 and 51.14 ppm against culicine larvae. Thus ethanolic extract of T. orientelis is an ideal potential larvicide for both types of mosquito larvae.
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Anopheles/efeitos dos fármacos , Culex/efeitos dos fármacos , Insetos Vetores/efeitos dos fármacos , Inseticidas/intoxicação , Nerium/química , Thuja/química , Acetona , Animais , Anopheles/metabolismo , Culex/metabolismo , Etanol , Insetos Vetores/metabolismo , Larva/efeitos dos fármacos , Larva/metabolismo , Dose Letal Mediana , Extratos Vegetais/intoxicaçãoRESUMO
Mosquito larvicidal activity of crude carbon-tetra-chloride, methanol and petroleum ether extracts of Solanum xanthocarpum fruits was examined against Anopheles stephensi and Culex quinquefasciatus. Among the extracts tested, carbon-tetra-chloride extract was the most effective with LC50 values of 5.11 ppm after 24 hours and 1.27 ppm after 48 hours of treatment against An. stephensi. In the case of Cx. quinquefasciatus the petroleum ether extract was observed as most toxic with LC50 values of 62.62 ppm after 24 hours and 59.45 ppm after 48 hours of exposure period respectively. It is, therefore, suggested that S. xanthocarpum can be applied as an ideal potential larvicide against An. stephensi and Cx. quinquefasciatus.
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Culicidae/efeitos dos fármacos , Frutas/química , Controle de Insetos/métodos , Extratos Vegetais/toxicidade , Solanum/química , Alcanos , Animais , Tetracloreto de Carbono , Larva/efeitos dos fármacos , Dose Letal Mediana , Metanol , Fatores de TempoRESUMO
A GH-releasing hormone (GHRH) messenger RNA (mRNA) has been identified in hypothalamus, placenta, and testicular germ cells. The GHRH mRNA produced by spermatogenic cells is approximately 1700 nucleotides in length, whereas GHRH transcripts in hypothalamus and placenta are 750 nucleotides. To correlate the structure of testicular GHRH mRNA with cell type-specific expression, we determined its sequence. A GHRH clone isolated from a rat testicular complementary DNA library was found to be identical in the coding sequence to hypothalamic GHRH. Rapid amplification of complementary DNA ends analysis of the 5'-end of germ cell GHRH mRNA and comparison with the genomic sequence revealed that GHRH transcription in testis initiates approximately 700 basepairs 5' to transcription initiation in placenta and 10.7 kilobasepairs 5' to that in hypothalamus. Reverse transcription-polymerase chain reaction analysis of germ cell RNA using primers from testicular exons 1 and 4 demonstrated that part of the placental exon 1 sequence is contained in some testicular GHRH transcripts, as an extra exon, between testicular exon 1 and the common exon 2. This was confirmed by a Northern blot of testicular mRNA using a testicular exon 1 probe. The 5'-flanking region of the testicular GHRH gene was analyzed and found to contain a TATA-like motif and sequences homologous to spermatogenic-specific cis-acting elements. Southern blot analysis of rat liver DNA suggested that just one GHRH gene is present in rat. These results indicate that both alternative transcription initiation and splicing of the GHRH gene exist in rat testicular germ cells.
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Hormônio Liberador de Hormônio do Crescimento/genética , Espermatogênese/genética , Processamento Alternativo , Sequência de Aminoácidos , Animais , Sequência de Bases , Southern Blotting , Éxons , Amplificação de Genes , Hormônio Liberador de Hormônio do Crescimento/química , Hipotálamo/metabolismo , Masculino , Dados de Sequência Molecular , Placenta/metabolismo , Reação em Cadeia da Polimerase , RNA Mensageiro/biossíntese , RNA Mensageiro/química , Ratos , Ratos Sprague-Dawley , Espermatozoides/metabolismo , Testículo/metabolismoRESUMO
A GHRH-like mRNA and peptide (t-GHRH) have been detected in rat and human testis. In rat, t-GHRH mRNA is localized to developing spermatogenic cells. We predicted that the most likely target cell of t-GHRH action would be the Sertoli cell. To test this prediction, we evaluated GHRH action on Sertoli cell function. Rat GHRH at a concentration of 10 nM or 100 nM stimulated cAMP production 2-fold over control levels after a 30 min incubation. This stimulation was obliterated by preincubation with a 10-fold excess of the GHRH antagonist (N-Ac-Tyr1, D-Arg2)-GRF(1-29)-NH2. The effect of treatment with [His1,Nle27]GHRH(1-32)-NH2, a GHRH analog, on Sertoli cell mRNAs was also assessed. Treatment with the analog significantly increased levels of c-fos and steel factor (the product of the Steel gene, also termed SCF) mRNAs above controls, but had no effect on sulfated glycoprotein-2 mRNA. We conclude that GHRH acts via adenylate cyclase to modulate specific Sertoli cell products, possibly as part of a network of local interacting factors controlling Sertoli and germ cell function.
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Hormônio Liberador de Hormônio do Crescimento/farmacologia , Células de Sertoli/efeitos dos fármacos , Adenilil Ciclases/metabolismo , Animais , Células Cultivadas , AMP Cíclico/biossíntese , Expressão Gênica/efeitos dos fármacos , Genes fos/genética , Hormônio Liberador de Hormônio do Crescimento/antagonistas & inibidores , Fatores de Crescimento de Células Hematopoéticas/genética , Masculino , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Células de Sertoli/fisiologia , Fator de Células-TroncoRESUMO
GH-releasing hormone (GHRH)-like mRNA and immunoreactivity (t-GHRH-li) are present in the testes of rats and humans. To learn more about the physiology of t-GHRH-li mRNA, we performed a series of experiments that disrupted various testicular functions. We employed ethylene dimethanesulfonate, a Leydig cell toxin, to assess the effects of Leydig cell ablation on t-GHRH-li mRNA and protein levels in prepubertal and postpubertal male rats. The ethylene dimethanesulfonate treatment resulted in decreases in serum testosterone, but had no effect on t-GHRH-li mRNA or peptide levels. To assess the effect of GHRH on Leydig cell steroidogenesis, Leydig cells were isolated by Percoll gradient centrifugation and cultured in the presence or absence of hCG, GHRH, or hCG plus GHRH. GHRH had no effect on steroidogenesis by Leydig cells, either alone or in combination with hCG. To localize t-GHRH-li mRNA within rat testis, in situ hybridization analysis was performed on testicular sections from normal rats, using a [35S]GHRH riboprobe. Grains were detected in spermatogenic cells with the antisense probe, whereas none was detected with the sense strand (control) probe. To verify these results, Northern blot analysis of RNA from separated testicular cells was performed. t-GHRH-li mRNA was detected in spermatocytes and round spermatids and to a lesser extent in Sertoli cells, but not in elongating spermatids, Leydig cells, or peritubular myoid cells. t-GHRH-li mRNA was also not found in epididymis. Since our experiments localized t-GHRH-li mRNA to spermatogenic cells, methoxyacetic acid (MAA), a pachytene spermatocyte toxin, was administered to postpubertal rats to determine whether t-GHRH-li is expressed primarily in pachytene spermatocytes. MAA treatment caused a decrease in testicular weight, which gradually returned to control levels by 42 days. Serum FSH levels in the treated animals fluctuated over the course of the experiment, while those in control animals remained steady. However, there was no difference in testicular GHRH-li mRNA levels between control and treated animals at any treatment time. Insulin-like growth factor-I and -II mRNA levels were also unaltered by the MAA treatment. We conclude from these results that t-GHRH-li is synthesized in early spermatogenic cells, but not in mature sperm, and that testicular GHRH-li does not play a major role in steroidogenesis by the Leydig cell.