RESUMO
PURPOSE: To conduct a comparative effectiveness analysis between robot-assisted radical cystectomy (RARC) and open approach (ORC). MATERIALS AND METHODS: A retrospective cohort study was conducted involving all patients undergoing radical cystectomy and urinary diversion for invasive bladder cancer at our institution from 2010 to 2018. Of a total 296 patients, we matched ORC and RARC cases based on age, BMI, Charlson comorbidity index, pathological TN staging of the tumor, prior radiotherapy, and type of diversion. The perioperative complications and oncological outcomes were compared. RESULTS: Eighty-nine patients were matched in the ORC and RARC groups. The median operative time was longer in RARC group (430 min) than that of ORC group (372 min) (p = 0.03); however, the median estimated blood loss (EBL) was significantly lower in RARC group (500 ml) than that of ORC (700 ml) (p < 0.0001). The median length of hospital stay (LOS) was significantly reduced in the RARC group (7 days) compared to the ORC group (8 days) (p = 0.02). There were no significant differences between both groups in 30- and 90-day postoperative complications (p = 0.3 and p = 0.2, respectively). A total of 68 deaths (38.2%) were observed, of which 36 (40.4%) were in ORC group while 32 (36%) were in RARC group (p = 0.5). The results were comparable in both groups regarding 5-year survival rate and cancer-specific survival (p = 0.3 and p = 0.1, respectively). CONCLUSION: RARC showed better perioperative outcomes in the form of less EBL and shortened LOS compared to ORC group. However, both RARC and ORC provide similar postoperative oncologic control, in terms of similar positive surgical margins, cancer-specific rates, and 5-year survival rates.
Assuntos
Procedimentos Cirúrgicos Robóticos , Neoplasias da Bexiga Urinária , Humanos , Cistectomia , Pontuação de Propensão , Estudos Retrospectivos , Neoplasias da Bexiga Urinária/cirurgiaRESUMO
[Figure: see text].
Assuntos
Átrios do Coração/inervação , Nó Sinoatrial/inervação , Neurônios Adrenérgicos/fisiologia , Animais , Nó Atrioventricular/inervação , Nó Atrioventricular/fisiologia , Sistema Nervoso Autônomo/anatomia & histologia , Sistema Nervoso Autônomo/fisiologia , Biomarcadores/análise , Neurônios Colinérgicos/fisiologia , Vasos Coronários/anatomia & histologia , Feminino , Gânglios Autônomos/anatomia & histologia , Humanos , Masculino , Ilustração Médica , Contração Miocárdica/fisiologia , Fenótipo , Nó Sinoatrial/fisiologia , Suínos , Porco Miniatura , Sinapses/fisiologia , Função Ventricular Esquerda/fisiologia , Proteínas Vesiculares de Transporte de Acetilcolina/análiseRESUMO
PURPOSE: To report the overall effect of ERAS protocol implementation in patients undergoing radical cystectomy and its impact on the length of hospital stay (LOS) and surgical outcomes considering their comorbid conditions. METHODS: Retrospective cohort study including 296 patients (146 non-ERAS patients vs. 150 ERAS patients) undergoing radical cystectomy and urinary diversion from 2010 to 2018. Age-adjusted Charlson Comorbidity Index (ACCI) score eight was set as cut off value between low-risk and high-risk patients. The primary outcome was LOS. Secondary outcomes were time to bowel movements, tolerance of regular diet, the incidence of postoperative ileus, postoperative complications, and 30- and 90-day readmission rates. RESULTS: A higher comorbidity burden was identified in the ERAS group compared to non-ERAS patients (p = 0.04). Median (IQR) LOS for non-ERAS was group 8(4) and 8(5) for ERAS group (p = 0.07). ERAS group demonstrated shorter time to resume bowel movements as well as time to tolerance of regular diet (p = 0.007, p = 0.023, respectively). Low-risk patients managed by the ERAS protocol demonstrated a significantly shortened gastrointestinal (GIT) recovery time (p = 0.001) as well as a reduction of LOS (p = 0.04). No significant reduction of LOS was identified for patients with higher comorbidity when placed on the ERAS protocol (p = 0.65). There were no significant differences in postoperative complications or readmission rates between groups. CONCLUSION: ERAS protocol implementation following radical cystectomy showed significant improvements in GIT recovery, nevertheless, it did not result in a decrease in LOS or readmission rates. Low-risk patients appeared to derive more benefit from ERAS protocol implementation than high-risk patients.
Assuntos
Cistectomia , Recuperação Pós-Cirúrgica Melhorada , Idoso , Estudos de Coortes , Cistectomia/métodos , Feminino , Humanos , Tempo de Internação , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/epidemiologia , Estudos Retrospectivos , Resultado do TratamentoRESUMO
OBJECTIVE: To study the effect of alvimopan and the Enhanced Recovery After Surgery protocol on length of hospital stay in patients undergoing radical cystectomy. METHODS: Our retrospective study involved 296 consecutive patients undergoing radical cystectomy for bladder cancer at our institution from 2010 through 2018. Patients were grouped according to three stages of the Enhanced Recovery After Surgery protocol implementation: (i) pre-Enhanced Recovery After Surgery (group A; n = 146); (ii) pre-alvimopan Enhanced Recovery After Surgery (group B; n = 102); and (iii) Enhanced Recovery After Surgery plus alvimopan (group C; n = 48). The primary outcome was the length of hospital stay. Secondary outcomes were time to first bowel movement, time to tolerate a regular diet, the incidence of postoperative ileus, postoperative complications and 30-day readmission rate. RESULTS: Group C showed a significantly shorter median length of hospital stay (7 days, P = 0.003), shorter gastrointestinal recovery time (4 days, P = 0.018) and a lower rate of postoperative ileus (14.6%, P = 0.005). The reduction in length of hospital stay, gastrointestinal recovery time and a lower rate of postoperative ileus was significant after controlling for other confounders on multivariable regression analysis. With the open approach, group C showed a significantly shorter length of hospital stay and gastrointestinal recovery time (P = 0.005, P = 0.001, respectively); however, in robotic cohorts, no significant differences were observed. There was no difference among groups in the 30-day readmission rate or postoperative complications. CONCLUSIONS: Patients undergoing radical cystectomy and managed by an Enhanced Recovery After Surgery protocol experience a significantly shorter length of hospital stay when receiving alvimopan as part of the protocol. Patients seem to derive the optimum benefits of alvimopan when it is used with an open approach; however, these benefits become less obvious with the robotic approach.
Assuntos
Cistectomia , Recuperação Pós-Cirúrgica Melhorada , Cistectomia/efeitos adversos , Fármacos Gastrointestinais , Humanos , Tempo de Internação , Piperidinas , Complicações Pós-Operatórias/epidemiologia , Complicações Pós-Operatórias/etiologia , Complicações Pós-Operatórias/prevenção & controle , Estudos RetrospectivosRESUMO
The prevalence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) quickly reached pandemic proportions, and knowledge about this virus and coronavirus disease 2019 (COVID-19) has expanded rapidly. This review focuses primarily on mechanisms that contribute to acute cardiac injury and dysfunction, which are common in patients with severe disease. The etiology of cardiac injury is multifactorial, and the extent is likely enhanced by preexisting cardiovascular disease. Disruption of homeostatic mechanisms secondary to pulmonary pathology ranks high on the list, and there is growing evidence that direct infection of cardiac cells can occur. Angiotensin-converting enzyme 2 (ACE2) plays a central role in COVID-19 and is a necessary receptor for viral entry into human cells. ACE2 normally not only eliminates angiotensin II (Ang II) by converting it to Ang-(1-7) but also elicits a beneficial response profile counteracting that of Ang II. Molecular analyses of single nuclei from human hearts have shown that ACE2 is most highly expressed by pericytes. Given the important roles that pericytes have in the microvasculature, infection of these cells could compromise myocardial supply to meet metabolic demand. Furthermore, ACE2 activity is crucial for opposing adverse effects of locally generated Ang II, so virus-mediated internalization of ACE2 could exacerbate pathology by this mechanism. While the role of cardiac pericytes in acute heart injury by SARS-CoV-2 requires investigation, expression of ACE2 by these cells has broader implications for cardiac pathophysiology.
Assuntos
Betacoronavirus/patogenicidade , Infecções por Coronavirus/enzimologia , Cardiopatias/enzimologia , Peptidil Dipeptidase A/metabolismo , Pericitos/enzimologia , Pneumonia Viral/enzimologia , Internalização do Vírus , Enzima de Conversão de Angiotensina 2 , Animais , COVID-19 , Infecções por Coronavirus/virologia , Cardiopatias/fisiopatologia , Cardiopatias/virologia , Interações Hospedeiro-Patógeno , Humanos , Pandemias , Pericitos/virologia , Pneumonia Viral/virologia , SARS-CoV-2RESUMO
PURPOSE: We assessed the effect of enhanced recovery after surgery protocol related fluid restriction on kidney function and the incidence of postoperative acute kidney injury and 3-month kidney function. MATERIALS AND METHODS: In a retrospectively collected, single institution cohort we studied 296 consecutive patients (146 pre-enhanced recovery after surgery vs 150 enhanced recovery after surgery) who underwent radical cystectomy from 2010 to 2018. The primary outcome was the incidence of postoperative acute kidney injury. Secondary outcomes were length of hospital stay, time to bowel movements, time to tolerate regular diet, postoperative complications and 30-day readmission rate. Study limitations include its retrospective design and relatively modest sample size. RESULTS: We observed an increased rate of postoperative acute kidney injury in patients on the enhanced recovery after surgery protocol (42.7% vs 30.1%, OR 1.725, p=0.025). On multivariate analysis enhanced recovery after surgery protocol remained a significant predictor of acute kidney injury even when controlling for other covariates including baseline kidney function (OR 1.8, 95% CI 1.04-3.30, p=0.036). Patients with postoperative acute kidney injury demonstrated significantly higher odds of stage 3 chronic kidney disease at 3 months even after controlling for baseline renal function (OR 2.5, 95% CI 1.3-4.9, p=0.016). CONCLUSIONS: Use of an enhanced recovery after surgery protocol following radical cystectomy was associated with a higher risk of postoperative acute kidney injury in patients who had baseline chronic kidney disease which could be related to the restricted perioperative fluid management mandated by enhanced recovery after surgery. Use of the enhanced recovery after surgery protocol did not impact the length of hospital stay or readmission rates.
Assuntos
Injúria Renal Aguda/epidemiologia , Cistectomia/efeitos adversos , Recuperação Pós-Cirúrgica Melhorada/normas , Complicações Pós-Operatórias/epidemiologia , Insuficiência Renal Crônica/epidemiologia , Injúria Renal Aguda/etiologia , Injúria Renal Aguda/fisiopatologia , Idoso , Ingestão de Líquidos/fisiologia , Feminino , Humanos , Incidência , Rim/fisiopatologia , Tempo de Internação/estatística & dados numéricos , Masculino , Pessoa de Meia-Idade , Readmissão do Paciente/estatística & dados numéricos , Complicações Pós-Operatórias/etiologia , Complicações Pós-Operatórias/fisiopatologia , Insuficiência Renal Crônica/complicações , Insuficiência Renal Crônica/fisiopatologia , Estudos Retrospectivos , Fatores de Risco , Neoplasias da Bexiga Urinária/complicações , Neoplasias da Bexiga Urinária/cirurgia , Equilíbrio Hidroeletrolítico/fisiologiaRESUMO
PURPOSE: Skeletal muscle and fat mass indexes have emerged as easily obtained, objective and useful tools to assess susceptibility to unfavorable postoperative outcomes. We examined the association between skeletal muscle and fat mass indexes, and the discharge disposition after radical cystectomy. MATERIALS AND METHODS: In a retrospectively collected, single institution cohort we studied patients who underwent radical cystectomy with pelvic lymphadenectomy of primary, nonmetastatic muscle invasive bladder cancer between 2009 and 2015. Included patients had undergone adequate axial computerized tomography at the L3 level within 90 days prior to surgery. Skeletal muscle and fat mass indexes were measured on preoperative computerized tomography and relationships to the outcomes of interest were analyzed. Multivariable logistic regression analysis was performed to assess the effect of the skeletal muscle and fat mass indexes on the discharge disposition while controlling for age, comorbidities, complications and previous neoadjuvant chemotherapy. RESULTS: A total of 136 patients met study inclusion criteria. The median skeletal muscle index among women and men in our study cohort was 36.4 and 47.6 cm2/m2, respectively. On multivariable logistic regression analysis a decreased skeletal muscle index (OR 0.94, 95% CI 0.90-0.98) and an increased fat mass index (OR 1.24, 95% CI 1.04-1.48) were associated with greater odds of discharge to a facility. Higher fat mass-to-skeletal muscle [corrected] index ratios were also associated with greater odds of discharge to a facility (OR 1.69, 95% CI 1.22-2.44). Study limitations include the retrospective design and unknown confounders. CONCLUSIONS: Low skeletal muscle and high fat compositions are independent predictors of discharge to a facility after radical cystectomy of nonmetastatic muscle invasive bladder cancer.
Assuntos
Tecido Adiposo/diagnóstico por imagem , Cistectomia , Músculo Esquelético/diagnóstico por imagem , Alta do Paciente/estatística & dados numéricos , Tomografia Computadorizada por Raios X , Neoplasias da Bexiga Urinária/cirurgia , Idoso , Comorbidade , Feminino , Humanos , Tempo de Internação/estatística & dados numéricos , Excisão de Linfonodo , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica , Estadiamento de Neoplasias , Estudos Retrospectivos , Neoplasias da Bexiga Urinária/patologiaRESUMO
BACKGROUND: Frequent premature ventricular contractions (PVCs) can cause a reversible reduction in systolic function. Most studies use 24-hour ambulatory electrocardiograms (AECGs) to assess PVC burden; however, PVC counts vary across 24-hour periods. We hypothesized that extended AECG monitoring would better identify clinically significant ectopy. METHODS: All 14-day AECGs performed at the San Francisco Veterans Affairs Medical Center between 2012 and 2015 (N = 694) were reviewed, and individuals with PVC counts ≥1.0% of total heartbeats were included (N = 101). Daily PVC counts and the range of these values across 24-hour periods were assessed. Median time for these ranges to cross clinically significant thresholds (PVCs ≥ 10%, 15%, or 20% of total heartbeats) was determined. RESULTS: Median PVC burden was 2.6% of total heartbeats (interquartile range [IQR]: 1.6-5.4%) and the median range across 24-hour periods was 3.6% (IQR: 2.0-9.1%). Individual ranges of daily PVC burden crossed thresholds of 10%, 15%, and 20% of total heartbeats in 26.7%, 16.8%, and 6.9% of patients, respectively. Median time to detecting an individual's maximum PVC burden was 6 days (IQR: 2-11 days). While 75% of those who reached the 20% threshold did so on day one of monitoring, only 53% of those reaching the 10% threshold did similarly, with a continually increasing yield throughout the 14-day monitoring period. CONCLUSIONS: PVC burden varies widely from day-to-day. While most patients with PVC burdens ≥20% were detected with 24 hours of monitoring, extended monitoring nearly doubled the identification of those reaching the 10% threshold.
Assuntos
Eletrocardiografia Ambulatorial/métodos , Complexos Ventriculares Prematuros/diagnóstico , Idoso , Feminino , Humanos , Masculino , Fatores de TempoRESUMO
Octopamine (OA) is a major neurotransmitter that has not been studied in the Pacific white shrimp, Litopenaeus vannamei. Therefore, we investigated changes in OA levels, its distribution in regions of the central nervous system (CNS) and ovary during the ovarian maturation cycle, as well as its possible role in regulating ovarian maturation. OA exhibited the highest concentration in the brain and thoracic ganglia at ovarian stage II, and then declined to the lowest concentration at ovarian stages III and IV. In the cerebral ganglia, OA-immunoreactivity (OA-ir) was present in neurons of clusters 6, 17, the anterior and posterior medial protocerebral, olfactory, antenna II, and tegumentary neuropils. In the circumesophageal, subesophageal, thoracic ganglia and abdominal ganglia, OA-ir was detected in several neuropils, neurons and fibers. The high level of intensity in OA immunostaining was observed in early developmental stage of oocyte by comparison with low level of OA-ir in late stages of oocyte development. Functionally, OA-injected female shrimps at doses of 2.5×10(-7) and 2.5×10(-6)mol/shrimp, showed significantly decreased gonado-somatic indices, oocyte diameters, and hemolymph vitellogenin levels, compared with control groups. This study showed changes of OA in the CNS and ovary reaching the highest level in early ovarian stages and declining in late stages, and it decreased hemolymph vitellogenin levels, suggesting significant involvement of OA in female reproduction in this species.
Assuntos
Sistema Nervoso Central/metabolismo , Octopamina/metabolismo , Ovário/crescimento & desenvolvimento , Ovário/metabolismo , Penaeidae , Animais , Feminino , Hemolinfa/química , Hemolinfa/metabolismo , Humanos , Neurônios/citologia , Neurônios/metabolismo , Oogênese/fisiologia , Penaeidae/crescimento & desenvolvimento , Penaeidae/metabolismo , Vitelogeninas/análise , Vitelogeninas/metabolismoRESUMO
Prostaglandins (PGs) are important bioactive mediators for many physiological functions. In some decapod crustaceans, prostaglandin E2 (PGE2) has been detected in reproductive organs, and may play a role in the control of ovarian maturation. However, in the freshwater prawn, Macrobrachium rosenbergii, the presences of PGE2 and key enzymes for PGE2 biosynthesis, as well as its effects on ovarian maturation have not yet been investigated. In this study we reported the presence of PGE2, cyclooxygenase1 (COX1) and prostaglandin E synthase (PGES) in the ovarian tissues of M. rosenbergii, using immunohistochemistry. Intense immunoreactivities of PGE2 (PGE2-ir), COX1 (Cox1-ir) and PGES (PGES-ir) were detected in previtellogenic oocytes (Oc1 and Oc2), while the immunoreactivities were absent in the late vitellogenic oocytes (Oc4). This finding supports the hypothesis that the PGE2 biosynthesis occurs in the ovary of this prawn. To ascertain this finding we used LC-MS/MS to quantitate PGE2 concentrations during ovarian developmental cycle. The levels of PGE2 were significantly higher in the early ovarian stages (St I and II) than in the late stages (St III and IV). Moreover, we found that administration of PGE2 stimulated the ovarian maturation in this species by shortening the length of the ovarian cycle, increasing ovarian-somatic index, oocyte proliferation, and vitellogenin (Vg) level in the hemolymph.
Assuntos
Dinoprostona/metabolismo , Oócitos/citologia , Ovário/citologia , Ovário/metabolismo , Palaemonidae/crescimento & desenvolvimento , Palaemonidae/metabolismo , Vitelogeninas/metabolismo , Animais , Proliferação de Células , Ciclo-Oxigenase 1/metabolismo , Feminino , Água Doce , Hemolinfa/metabolismo , Imuno-Histoquímica , Oxirredutases Intramoleculares/metabolismo , Oócitos/metabolismo , Prostaglandina-E Sintases , Espectrometria de Massas em TandemRESUMO
The coronary circulation plays a crucial role in balancing myocardial perfusion and oxygen demand to prevent myocardial ischemia. Extravascular compressive forces, coronary perfusion pressure, and microvascular resistance are involved to regulate coronary blood flow throughout the cardiac cycle. Autoregulation of the coronary blood flow through dynamic adjustment of microvascular resistance is maintained by complex interactions among mechanical, endothelial, metabolic, neural, and hormonal mechanisms. This review focuses on the neural mechanism. Anatomy and physiology of the coronary arterial innervation have been extensively investigated using animal models. However, findings in the animal heart have limited applicability to the human heart as cardiac innervation is generally highly variable among species. So far, limited data are available on the human coronary artery innervation, rendering multiple questions unresolved. Recently, the clinical entity of ischemia with non-obstructive coronary arteries has been proposed, characterized by microvascular dysfunction involving abnormal vasoconstriction and impaired vasodilation. Thus, measurement of microvascular resistance has become a standard diagnostic for patients without significant stenosis in the epicardial coronary arteries. Neural mechanism is likely to play a pivotal role, supported by the efficacy of cardiac sympathetic denervation to control symptoms in patients with angina. Therefore, understanding the coronary artery innervation and control of microvascular resistance of the human heart is increasingly important for cardiologists for diagnosis and to select appropriate therapeutic options. Advancement in this field can lead to innovations in diagnostic and therapeutic approaches for coronary artery diseases.
Assuntos
Circulação Coronária , Vasos Coronários , Resistência Vascular , Humanos , Vasos Coronários/inervação , Resistência Vascular/fisiologia , Circulação Coronária/fisiologia , Animais , Microcirculação , Doença da Artéria Coronariana/fisiopatologia , Isquemia Miocárdica/fisiopatologiaRESUMO
Cardiac control is mediated via nested-feedback reflex control networks involving the intrinsic cardiac ganglia, intra-thoracic extra-cardiac ganglia, spinal cord, brainstem, and higher centers. This control system is optimized to respond to normal physiologic stressors; however, it can be catastrophically disrupted by pathologic events such as myocardial ischemia. In fact, it is now recognized that cardiac disease progression reflects the dynamic interplay between adverse remodeling of the cardiac substrate coupled with autonomic dysregulation. With advances in understanding of this network dynamic in normal and pathologic states, neuroscience-based neuromodulation therapies can be devised for the management of acute and chronic cardiac pathologies.
Assuntos
Coração , Humanos , Coração/fisiologia , Coração/fisiopatologia , Sistema Nervoso Autônomo/fisiopatologia , Sistema Nervoso Autônomo/fisiologia , Cardiopatias/fisiopatologiaRESUMO
Detailed study of non-failing human hearts rejected for transplantation provides a unique opportunity to perform structural analyses across microscopic and macroscopic scales. These techniques include tissue clearing (modified immunolabeling-enabled three-dimensional (3D) imaging of solvent-cleared organs) and immunohistochemical staining. Mesoscopic examination procedures include stereoscopic dissection and micro-computed tomographic (CT) scanning. Macroscopic examination procedures include gross dissection, photography (including anaglyphs and photogrammetry), CT, and 3D printing of the physically or virtually dissected or whole heart. Before macroscopic examination, pressure-perfusion fixation may be performed to maintain the 3D architecture and physiologically relevant morphology of the heart. The application of these techniques in combination to study the human heart is unique and crucial in understanding the relationship between distinct anatomic features such as coronary vasculature and myocardial innervation in the context of the 3D architecture of the heart. This protocol describes the methodologies in detail and includes representative results to illustrate progress in the research of human cardiac anatomy.
Assuntos
Coração , Imageamento Tridimensional , Humanos , Coração/anatomia & histologia , Coração/diagnóstico por imagem , Imageamento Tridimensional/métodos , Microtomografia por Raio-X/métodos , Dissecação/métodos , Impressão TridimensionalRESUMO
Objective: Radical cystectomy is a complex lengthy procedure associated with postoperative morbidity. We aimed to assess the operative time (OT) in patients undergoing radical cystectomy and its impact on 90-day postoperative complications and readmission rates. Methods: The retrospective cohort study included 296 patients undergoing radical cystectomy and urinary diversion from May 2010 to December 2018 in our institution. The OT of 369 min was set as a cutoff value between short and long OT groups. The primary outcome was 90-day postoperative complication rates. Secondary outcomes were gastrointestinal recovery time, length of hospital stay, and 90-day readmission rates. Results: The overall incidence of 90-day postoperative complications was 79.7% where 43.2% representing low-grade complications according to the Clavien-Dindo classification (Grade 1 and Grade 2), and 36.5% representing high-grade complications (Grade≥3). Gastrointestinal tract and infectious complications are the most common complications in our data set (45.9% and 45.6%, respectively). On multivariable analysis, prolonged OT was significantly associated with odds of high-grade complications (odds ratio 2.340, 95% confidence interval 1.288-4.250, p=0.005). After propensity score-matched analysis, a higher incidence of major complications was identified in the long OT group 55 (51.4%) compared to 35 (32.7%) in the short OT group (p=0.006). A shorter gastrointestinal tract recovery time was noticed in the short OT group (p=0.009). Prolonged OT was associated with a higher 90-day readmission rate on univariate and multivariate analyses (p<0.001, p=0.001, respectively). Conclusion: Prolonged OT (>369 min) is associated with an increased risk of postoperative complications and readmission rates. The perception of potential postoperative complications requires careful monitoring of these patients which could translate into better operative outcomes.
RESUMO
We present a detailed histological description of the central nervous system (CNS: brain, subesophageal ganglion, thoracic ganglia, abdominal ganglia) of the blue crab, Portunus pelagicus. Because the presence of gonadotropin-releasing hormone (GnRH) in crustaceans has been disputed, we examine the presence and localization of a GnRH-like peptide in the CNS of the blue crab by using antibodies against lamprey GnRH (lGnRH)-III, octopus GnRH (octGnRH) and tunicate GnRH (tGnRH)-I. These antibodies showed no cross-reactivity with red-pigment-concentrating hormone, adipokinetic hormone, or corazonin. In the brain, strong lGnRH-III immunoreactivity (-ir) was detected in small (7-17 µm diameter) neurons of clusters 8, 9 and 10, in medium-sized (21-36 µm diameter) neurons of clusters 6, 7 and 11 and in the anterior and posterior median protocerebral neuropils, olfactory neuropil, median and lateral antenna I neuropils, tegumentary neuropil and antenna II neuropil. In the subesophageal ganglion, lGnRH-III-ir was detected in medium-sized neurons and in the subesophageal neuropil. In the thoracic and abdominal ganglia, lGnRH-III-ir was detected in medium-sized and small neurons and in the neuropils. OctGnRH-ir was observed in neurons of the same clusters with moderate staining, particularly in the deutocerebrum, whereas tGnRH-I-ir was only detected in medium-sized neurons of cluster 11 in the brain. Thus, anti-lGnRH-III shows greater immunoreactivity in the crab CNS than anti-octGnRH and anti-tGnRH-I. Moreover, our functional bioassay demonstrates that only lGnRH-III has significant stimulatory effects on ovarian growth and maturation. We therefore conclude that, although the true identity of the crab GnRH eludes us, crabs possess a putative GnRH hormone similar to lGnRH-III. The identification and characterization of this molecule is part of our ongoing research.
Assuntos
Proteínas de Artrópodes/metabolismo , Braquiúros/metabolismo , Sistema Nervoso Central/metabolismo , Hormônio Liberador de Gonadotropina/metabolismo , Peptídeos/metabolismo , Animais , Antenas de Artrópodes/citologia , Antenas de Artrópodes/metabolismo , Braquiúros/citologia , Sistema Nervoso Central/citologia , Neurônios/citologia , Neurônios/metabolismo , Neurópilo/citologia , Neurópilo/metabolismoAssuntos
Remodelamento Atrial , Infarto do Miocárdio , Animais , Arritmias Cardíacas , Cães , Ventrículos do Coração , NeurôniosRESUMO
Red pigment concentrating hormone (RPCH) is a member of the chromatophorotropic hormones and, in crustaceans, it is synthesized in the eyestalk. We have isolated a full-length cDNA for a RPCH preprohormone gene (Scyol-RPCH) from the eyestalks of female mud crabs, Scylla olivacea. The open reading frame consists of 642 nucleotides, and encodes a deduced 108 amino acid precursor protein, which includes a signal peptide, the RPCH (pQLNFSPGWamide), and an associated peptide. We show that the mud crab RPCH peptide exhibits 100% identity with 15 other decapods. Expression of Scyol-RPCH within adult mud crab takes place in the eyestalk, brain, and ventral nerve cord, comprising subesophageal ganglion, thoracic ganglion, and abdominal ganglion. In situ hybridization demonstrates specific expression within neuronal clusters 1, 2, 3, and 4 of the eyestalk X-organ, clusters 6, 8, 9, 10, and 17 of the brain, and in neuronal clusters of the ventral nerve cord. We found that administration of 5-HT up-regulates RPCH gene expression in the eyestalk, suggesting that RPCH may play a role as a downstream hormone of 5-HT.
Assuntos
Braquiúros/metabolismo , Hormônios de Invertebrado/biossíntese , Oligopeptídeos/biossíntese , Precursores de Proteínas/biossíntese , Ácido Pirrolidonocarboxílico/análogos & derivados , Serotonina/farmacologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Braquiúros/genética , Feminino , Expressão Gênica/efeitos dos fármacos , Hormônios de Invertebrado/genética , Masculino , Dados de Sequência Molecular , Oligopeptídeos/genética , Filogenia , Precursores de Proteínas/genética , Alinhamento de Sequência , Distribuição TecidualRESUMO
Neurotransmitters and neurohormones are agents that control gonad maturation in decapod crustaceans. Of these, serotonin (5-HT) and dopamine (DA) are neurotransmitters with known antagonist roles in female reproduction, whilst gonadotropin-releasing hormones (GnRHs) and corazonin (Crz) are neurohormones that exercise both positive and negative controls in some invertebrates. However, the effects of these agents on the androgenic gland (AG), which controls testicular maturation and male sex development in decapods, via insulin-like androgenic gland hormone (IAG), are unknown. Therefore, we set out to assay the effects of 5-HT, DA, l-GnRH-III, oct-GnRH and Crz, on the AG of small male Macrobrachium rosenbergii (Mr), using histological studies, a BrdU proliferative cell assay, immunofluorescence of Mr-IAG, and ELISA of Mr-IAG. The results showed stimulatory effects by 5-HT and l-GnRH-III through significant increases in AG size, proliferation of AG cells, and Mr-IAG production (P<0.05). In contrast, DA and Crz caused inhibitory effects on the AG through significant decreases in AG size, proliferation of AG cells, and Mr-IAG production (P<0.05). Moreover, the prawns treated with Crz died before day 16 of the experimental period. We propose that 5-HT and certain GnRHs can be now used to stimulate reproduction in male M. rosenbergii, as they induce increases in AG and testicular size, IAG production, and spermatogenesis. The mechanisms by which these occur are part of our on-going research.