18 results on '"Rocha JN"'
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2. Opsonization but not pretreatment of equine macrophages with hyperimmune plasma nonspecifically enhances phagocytosis and intracellular killing of Rhodococcus equi.
- Author
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Harvey AB, Bordin AI, Rocha JN, Bray JM, and Cohen ND
- Subjects
- Animals, Antibodies, Bacterial, Horses, Macrophages, Phagocytosis, Actinomycetales Infections veterinary, Horse Diseases, Rhodococcus, Rhodococcus equi
- Abstract
Background: Evidence regarding the efficacy of equine hyperimmune plasma to prevent pneumonia in foals caused by Rhodococcus equi is limited and conflicting., Hypothesis: Opsonization with R. equi-specific hyperimmune plasma (HIP) will significantly increase phagocytosis and decrease intracellular replication of R. equi by alveolar macrophages (AMs) compared to normal plasma (NP)., Animals: Fifteen adult Quarter Horses were used to collect bronchoalveolar lavage cells., Methods: In the first experiment, AMs from 9 horses were pretreated (incubated) with either HIP, NP, or media only (control) and then infected with nonopsonized R. equi. In a second experiment, AMs from 6 horses were infected with R. equi either opsonized with HIP or opsonized with NP. For both experiments, AMs were lysed at 0 and 48 hours and the number of viable R. equi quantified by culture were compared among groups using linear mixed-effects modeling with significance set at P < .05., Results: Opsonization with either HIP or NP increased phagocytosis by AMs (P < .0001) and decreased intracellular survival of organisms in AMs (P < .0001). Pretreating AMs with either HIP or NP without opsonizing R. equi had no effects on phagocytosis or intracellular replication., Conclusions and Clinical Importance: Opsonizing R. equi with either NP or HIP decreases intracellular survival of organisms in AMs, but the effect does not appear to be enhanced by using HIP. Mechanisms other than effects on AMs must explain any clinical benefits of using HIP over NP to decrease the incidence of R. equi pneumonia in foals., (© 2020 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals LLC. on behalf of the American College of Veterinary Internal Medicine.)
- Published
- 2021
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3. Investigating accessibility in Latin American science museums and centers.
- Author
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Rocha JN, Massarani L, Abreu WV, Inacio LGB, and Molenzani AO
- Abstract
Science museums are powerful places not only for science communication, informal science teaching, and leisure but also for promoting science and technology in an equitable, diverse, and inclusive mode. The paper presents an overview of accessibility in Latin American science museums and centers, based on a questionnaire comprised of open and closed questions in Portuguese and Spanish. The answers received from directors and other personnel responsible for 109 institutions in twelve countries were analyzed using an accessibility indicator tool. The findings suggest that these institutions generally offer some physical accessibility resources and fewer communicational and attitudinal accessibility resources. Data also show that there is an absence of institutional practices that might underpin any endeavor to take into consideration the inclusion of people with disabilities. More funding must be made available for practices and research in this area. We also recommend that persons with disabilities play a greater role as protagonists and professionals and likewise as a research public in the realm of science communication and at the region's science museums and centers.
- Published
- 2020
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4. Activity Mapping of Children in Play Using Multivariate Analysis of Movement Events.
- Author
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Rocha JN, Barnes CM, Rees P, Clark CT, Stratton G, and Summers HD
- Subjects
- Accelerometry instrumentation, Ankle, Child, Child, Preschool, Female, Humans, Male, Multivariate Analysis, Principal Component Analysis, Time and Motion Studies, Child Behavior physiology, Exercise physiology, Movement physiology, Play and Playthings
- Abstract
Purpose: (i) To develop an automated measurement technique for the assessment of both the form and intensity of physical activity undertaken by children during play. (ii) To profile the varying activity across a cohort of children using a multivariate analysis of their movement patterns., Methods: Ankle-worn accelerometers were used to record 40 min of activity during a school recess, for 24 children over five consecutive days. Activity events of 1.1 s duration were identified within the acceleration time trace and compared with a reference motif, consisting of a single walking stride acceleration trace, obtained on a treadmill operating at a speed of 4 km h. Dynamic time warping of motif and activity events provided metrics of comparative movement duration and intensity, which formed the data set for multivariate mapping of the cohort activity using a principal component analysis (PCA)., Results: The two-dimensional PCA plot provided clear differentiation of children displaying diverse activity profiles and clustering of those with similar movement patterns. The first component of the PCA correlated to the integrated intensity of movement over the 40-min period, whereas the second component informed on the temporal phasing of activity., Conclusions: By defining movement events and then quantifying them by reference to a motion-standard, meaningful assessment of highly varied activity within free play can be obtained. This allows detailed profiling of individual children's activity and provides an insight on social aspects of play through identification of matched activity time profiles for children participating in conjoined play.
- Published
- 2020
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5. Transfusion With 2 L of Hyperimmune Plasma is Superior to Transfusion of 1 L or Less for Protecting Foals Against Subclinical Pneumonia Attributed to Rhodococcus equi.
- Author
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Kahn SK, Blodgett GP, Canaday NM, Bevevino KE, Rocha JN, Bordin AI, and Cohen ND
- Subjects
- Animals, Horses, Pneumonia, Bacterial prevention & control, Retrospective Studies, Actinomycetales Infections veterinary, Horse Diseases prevention & control, Pneumonia, Bacterial veterinary, Rhodococcus equi
- Abstract
Transfusing foals with Rhodococcus equi hyperimmune plasma (REHIP) is a standard practice at many horse-breeding farms to help prevent R. equi pneumonia. At many large breeding farms, pneumonia is most commonly recognized as subclinical based on thoracic ultrasonography findings. The efficacy of REHIP transfusion and the impact of the volume of plasma transfused for reducing the cumulative incidence of subclinical R. equi pneumonia are unknown. A retrospective cohort study was conducted among foals born and residing through weaning at a large breeding farm. Foals were transfused with either 0 L (n = 2 foals), 1 L (n = 85 foals), or 2 L (n = 62 foals) of REHIP within 36 hours of birth. Volume transfused was principally based on intended use of the foals. All foals at the ranch were routinely screened using thoracic ultrasonography at 5, 7, and 9 weeks of age to detect subclinical pneumonia attributed to R. equi based on farm history. The proportion of the foals receiving < 1 L REHIP that developed subclinical pneumonia (32%; 26/82) was significantly (P = .0068; chi-squared test) greater than that among foals transfused with 2 L of REHIP (12%; 8/68). Despite the important limitations of this observational study, it provides evidence supporting the need for well-designed clinical trials to evaluate the impact of the use and dose of REHIP for preventing subclinical pneumonia. Reducing the incidence of subclinical pneumonia is important because reducing antibiotic treatment of subclinical cases will decrease selection pressure for antimicrobial resistance in R. equi., (Copyright © 2019 Elsevier Inc. All rights reserved.)
- Published
- 2019
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6. In vitro evaluation of complement deposition and opsonophagocytic killing of Rhodococcus equi mediated by poly-N-acetyl glucosamine hyperimmune plasma compared to commercial plasma products.
- Author
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Folmar CN, Cywes-Bentley C, Bordin AI, Rocha JN, Bray JM, Kahn SK, Schuckert AE, Pier GB, and Cohen ND
- Subjects
- Actinomycetales Infections immunology, Animals, Antibodies, Bacterial blood, Complement C1 immunology, Female, Horse Diseases immunology, Horse Diseases microbiology, Horses immunology, Male, Neutrophils, Plasma immunology, Acetylglucosamine immunology, Actinomycetales Infections veterinary, Rhodococcus equi immunology
- Abstract
Background: The bacterium Rhodococcus equi can cause severe pneumonia in foals. The absence of a licensed vaccine and limited effectiveness of commercial R. equi hyperimmune plasma (RE-HIP) create a great need for improved prevention of this disease., Hypothesis: Plasma hyperimmune to the capsular polysaccharide poly-N-acetyl glucosamine (PNAG) would be significantly more effective than RE-HIP at mediating complement deposition and opsonophagocytic killing (OPK) of R. equi., Animals: Venipuncture was performed on 9 Quarter Horses., Methods: The ability of the following plasma sources to mediate complement component 1 (C1) deposition onto either PNAG or R. equi was determined by ELISA: (1) PNAG hyperimmune plasma (PNAG-HIP), (2) RE-HIP, and (3) standard non-hyperimmune commercial plasma (SP). For OPK, each plasma type was combined with R. equi, equine complement, and neutrophils isolated from horses (n = 9); after 4 hours, the number of R. equi in each well was determined by quantitative culture. Data were analyzed using linear mixed-effects regression with significance set at P < .05., Results: The PNAG-HIP and RE-HIP were able to deposit significantly (P < .05) more complement onto their respective targets than the other plasmas. The mean proportional survival of R. equi opsonized with PNAG-HIP was significantly (P < .05) less (14.7%) than that for SP (51.1%) or RE-HIP (42.2%)., Conclusions and Clinical Importance: Plasma hyperimmune to PNAG is superior to RE-HIP for opsonizing and killing R. equi in vitro. Comparison of these 2 plasmas in field trials is warranted because of the reported incomplete effectiveness of RE-HIP., (© 2019 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.)
- Published
- 2019
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7. PNAG-specific equine IgG 1 mediates significantly greater opsonization and killing of Prescottella equi (formerly Rhodococcus equi) than does IgG 4/7 .
- Author
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Rocha JN, Dangott LJ, Mwangi W, Alaniz RC, Bordin AI, Cywes-Bentley C, Lawhon SD, Pillai SD, Bray JM, Pier GB, and Cohen ND
- Subjects
- Actinomycetales Infections immunology, Actinomycetales Infections prevention & control, Age Factors, Animals, Animals, Newborn, Antibodies, Bacterial classification, Antibodies, Bacterial immunology, Horse Diseases immunology, Horse Diseases prevention & control, Horses immunology, Immunoglobulin G classification, Opsonin Proteins, Pneumonia, Bacterial immunology, Pneumonia, Bacterial prevention & control, Acetylglucosamine immunology, Actinomycetales Infections veterinary, Antibodies, Bacterial blood, Complement C1 immunology, Immunoglobulin G blood, Phagocytosis, Rhodococcus equi immunology
- Abstract
Prescottella equi (formerly Rhodococcus equi) is a facultative intracellular bacterial pathogen that causes severe pneumonia in foals 1-6 months of age, whereas adult horses are highly resistant to infection. We have shown that vaccinating pregnant mares against the conserved surface polysaccharide capsule, β-1 → 6-linked poly-N-acetyl glucosamine (PNAG), elicits opsonic killing antibody that transfers via colostrum to foals and protects them against experimental infection with virulent. R. equi. We hypothesized that equine IgG
1 might be more important than IgG4/7 for mediating protection against R. equi infection in foals. To test this hypothesis, we compared complement component 1 (C1) deposition and polymorphonuclear cell-mediated opsonophagocytic killing (OPK) mediated by IgG1 or IgG4/7 enriched from either PNAG hyperimmune plasma (HIP) or standard plasma. Subclasses IgG1 and IgG4/7 from PNAG HIP and standard plasma were precipitated onto a diethylaminoethyl ion exchange column, then further enriched using a protein G Sepharose column. We determined C1 deposition by enzyme-linked immunosorbent assay (ELISA) and estimated OPK by quantitative microbiologic culture. Anti-PNAG IgG1 deposited significantly (P < 0.05) more C1 onto PNAG than did IgG4/7 from PNAG HIP or subclasses IgG1 and IgG4/7 from standard plasma. In addition, IgG1 from PNAG HIP mediated significantly (P < 0.05) greater OPK than IgG4/7 from PNAG HIP or IgG1 and IgG4/7 from standard plasma. Our findings indicate that anti-PNAG IgG1 is a correlate of protection against R. equi in foals, which has important implications for understanding the immunopathogenesis of R. equi pneumonia, and as a tool for assessing vaccine efficacy and effectiveness when challenge is not feasible., (Copyright © 2019 Elsevier Ltd. All rights reserved.)- Published
- 2019
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8. Antibody to Poly-N-acetyl glucosamine provides protection against intracellular pathogens: Mechanism of action and validation in horse foals challenged with Rhodococcus equi.
- Author
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Cywes-Bentley C, Rocha JN, Bordin AI, Vinacur M, Rehman S, Zaidi TS, Meyer M, Anthony S, Lambert M, Vlock DR, Giguère S, Cohen ND, and Pier GB
- Subjects
- Animals, Animals, Newborn, Horses, Rhodococcus equi, Acetylglucosamine immunology, Actinomycetales Infections immunology, Antibodies, Bacterial immunology, Antigens, Bacterial immunology, Bacterial Vaccines immunology
- Abstract
Immune correlates of protection against intracellular bacterial pathogens are largely thought to be cell-mediated, although a reasonable amount of data supports a role for antibody-mediated protection. To define a role for antibody-mediated immunity against an intracellular pathogen, Rhodococcus equi, that causes granulomatous pneumonia in horse foals, we devised and tested an experimental system relying solely on antibody-mediated protection against this host-specific etiologic agent. Immunity was induced by vaccinating pregnant mares 6 and 3 weeks prior to predicted parturition with a conjugate vaccine targeting the highly conserved microbial surface polysaccharide, poly-N-acetyl glucosamine (PNAG). We ascertained antibody was transferred to foals via colostrum, the only means for foals to acquire maternal antibody. Horses lack transplacental antibody transfer. Next, a randomized, controlled, blinded challenge was conducted by inoculating at ~4 weeks of age ~10(6) cfu of R. equi via intrabronchial challenge. Eleven of 12 (91%) foals born to immune mares did not develop clinical R. equi pneumonia, whereas 6 of 7 (86%) foals born to unvaccinated controls developed pneumonia (P = 0.0017). In a confirmatory passive immunization study, infusion of PNAG-hyperimmune plasma protected 100% of 5 foals against R. equi pneumonia whereas all 4 recipients of normal horse plasma developed clinical disease (P = 0.0079). Antibodies to PNAG mediated killing of extracellular and intracellular R. equi and other intracellular pathogens. Killing of intracellular organisms depended on antibody recognition of surface expression of PNAG on infected cells, along with complement deposition and PMN-assisted lysis of infected macrophages. Peripheral blood mononuclear cells from immune and protected foals released higher levels of interferon-γ in response to PNAG compared to controls, indicating vaccination also induced an antibody-dependent cellular release of this critical immune cytokine. Overall, antibody-mediated opsonic killing and interferon-γ release in response to PNAG may protect against diseases caused by intracellular bacterial pathogens., Competing Interests: Gerald B. Pier is an inventor of intellectual properties [human monoclonal antibody to PNAG and PNAG vaccines] that are licensed by Brigham and Women’s Hospital to Alopexx Vaccine, LLC, and Alopexx Pharmaceuticals, LLC, entities in which GBP also holds equity. As an inventor of intellectual properties, GBP also has the right to receive a share of licensing-related income (royalties, fees) through Brigham and Women’s Hospital from Alopexx Pharmaceuticals, LLC, and Alopexx Vaccine, LLC. GBP’s interests were reviewed and are managed by the Brigham and Women’s Hospital and Partners Healthcare in accordance with their conflict of interest policies. Colette Cywes-Bentley is an inventor of intellectual properties [use of human monoclonal antibody to PNAG and use of PNAG vaccines] that are licensed by Brigham and Women’s Hospital to Alopexx Pharmaceuticals, LLC. As an inventor of intellectual properties, CC-B also has the right to receive a share of licensing-related income (royalties, fees) through Brigham and Women’s Hospital from Alopexx Pharmaceuticals, LLC. Noah D. Cohen has received an unrestricted gift to the EIDL from Alopexx Vaccines, LLC. Daniel Vlock, holds an equity share and potential royalty income from Alopexx Vaccines, LLC for vaccines to PNAG and monoclonal antibody to PNAG from Alopexx Pharmaceuticals, LLC. Mark Meyer holds minority equity shares of Mg Biologics, Inc. Sarah Anthony and McKenzie Lambert are employees of Mg Biologics, Inc.
- Published
- 2018
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9. Cystometric analysis of the transplanted bladder.
- Author
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Rocha JN
- Subjects
- Animals, Female, Models, Animal, Pressure, Rats, Wistar, Reference Values, Reproducibility of Results, Time Factors, Transplantation, Autologous, Treatment Outcome, Urinary Bladder pathology, Urination physiology, Urinary Bladder physiopathology, Urinary Bladder transplantation
- Abstract
Objective: cystometric evaluation of the bladder after autotransplant and isogeneic transplant in female rats., Material and Methods: two groups were constituted: (A) bladder autotransplant with two subgroups: R1 - (control) and R2 - (bladder transplant); (B) isogeneic bladder transplant with three subgroups; T1 - (control); T2-T3, two subgroups observed for 30 and 60 days after transplant, respectively. All animals underwent cystometric evaluation. Afterwards, the bladders were removed for histological study., Results: the transplanted bladders did not show significant changes in filling/storage and emptying/micturition functions after 30 and 60 days of evolution. Upon macroscopical evaluation, there was good revascularization and the tissue was well preserved. Cystometry results: did not show significant differences in the micturition pressure in subgroups T2-T3, but did between subgroups R1-R2, T1-T2, and T1-T3. Significant differences were verified in the micturition interval between T1-T3, T2-T3, but not between R1-R2, T1-T2. There was significant difference in the micturition duration between T1-T3 but not between R1-R2, T1-T2 and T2-T3. No fistula was noted on the suture site nor leakage of urine in the abdominal cavity or signs of necrosis or retraction were observed., Conclusions: transplant of the bladder was shown to be a viable procedure. The results indicate that there was structural and functional regeneration of transplanted bladders, and these results indicate that it is possible that vascular endothelium growth and neurogenesis factors are involved and activated in the process of the preservation or survival of the transplanted organ., (Copyright® by the International Brazilian Journal of Urology.)
- Published
- 2017
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10. Extracellular adenosine 5'-triphosphate concentrations changes in rat spinal cord associated with the activation of urinary bladder afferents. A microdialysis study.
- Author
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Rocha JN
- Subjects
- Adenosine Triphosphate pharmacology, Animals, Female, Luminescent Measurements, Neurons, Afferent drug effects, Neurons, Afferent metabolism, Rats, Rats, Sprague-Dawley, Spinal Cord drug effects, Urinary Bladder drug effects, Adenosine Triphosphate analysis, Microdialysis methods, Neurons, Afferent physiology, Spinal Cord chemistry, Urinary Bladder innervation, Visceral Afferents
- Abstract
Objective: To determine adenosine 5'-triphosphate levels in the interstice of spinal cord L6-S1 segment, under basal conditions or during mechanical and chemical activation of urinary bladder afferents., Methods: A microdialysis probe was transversally implanted in the dorsal half of spinal cord L6-S1 segment in female rats. Microdialysate was collected at 15 minutes intervals during 135 minutes, in anesthetized animals. Adenosine 5'-triphosphate concentrations were determined with a bioluminescent assay. In one group of animals (n=7) microdialysate samples were obtained with an empty bladder during a 10-minutes bladder distension to 20 or 40cmH2O with either saline, saline with acetic acid or saline with capsaicin. In another group of animals (n=6) bladder distention was performed and the microdialysis solution contained the ectonucleotidase inhibitor ARL 67156., Results: Basal extracellular adenosine triphosphate levels were 110.9±35.34fmol/15 minutes, (mean±SEM, n=13), and bladder distention was associated with a significant increase in adenosine 5'-triphosphate levels which was not observed after bladder distention with saline solution containing capsaicin (10µM). Microdialysis with solution containing ARL 67156 (1mM) was associated with significantly higher extracellular adenosine 5'-triphosphate levels and no further increase in adenosine 5'-triphosphate was observed during bladder distension., Conclusion: Adenosine 5'-triphosphate was present in the interstice of L6-S1 spinal cord segments, was degraded by ectonucleotidase, and its concentration increased following the activation of bladder mechanosensitive but not of the chemosensitive afferents fibers. Adenosine 5'-triphosphate may originate either from the central endings of bladder mechanosensitive primary afferent neurons, or most likely from intrinsic spinal neurons, or glial cells and its release appears to be modulated by capsaicin activated bladder primary afferent or by adenosine 5'-triphosphate itself., Objetivo: Determinar as concentrações extracelulares do 5'-trifosfato de adenosina no interstício dos segmentos medulares L6-S1, em condições basais ou durante a ativação mecânica e química das fibras aferentes vesicais., Métodos: Um cateter de microdiálise foi implantado no sentido transversal na parte dorsal da medula espinal, entre os segmentos L6-S1 de ratas. O microdialisado foi coletado em intervalos de 15 minutos, durante 135 minutos, com os animais anestesiados. A concentração de 5'-trifosfato de adenosina nas amostras foi determinada mediante ensaio de bioluminescência. Em um grupo de animais (n=7), as amostras de microdialisado foram obtidas com a bexiga vazia, com distensão da bexiga para volume de 20 ou 40cmH2O, com solução salina, solução salina com ácido acético, ou solução salina com capsaicina. Em outro grupo (n=6), foi realizada com a bexiga distendida, e a solução para microdiálise continha o inibidor de ectonucleotidase ARL 67156., Resultados: Os níveis extracelulares de trifosfato de adenosina no início do estudo foram 110,9±35,36fmol/15 minutos (média±EPM, n=13), e a distensão da bexiga causou um aumento nos níveis de 5'-trifosfato de adenosina, o que não foi observado após a distensão da bexiga com solução salina contendo capsaicina (10µM). A microdiálise com solução contendo ARL 67156 (1mM) foi associada com significante aumento dos níveis de trifosfato de adenosina extracelular, e nenhum aumento do trifosfato de adenosina foi observado durante a distensão da bexiga., Conclusão: O 5'-trifosfato de adenosina está presente no interstício do segmento L6-S1 da medula espinal, é degradado por ectonucleotidases, e sua concentração aumentou com a ativação das fibras aferentes mecanossensíveis da bexiga, mas não das quimiossensíveis. O 5'-trifosfato de adenosina pode ter sido liberado das terminações centrais dos neurônios aferentes primários mecanossensíveis ou, mais provavelmente, de neurônios espinais intrínsecos, ou ainda de células gliais. Sua liberação parece ser modulada por fibras aferentes primárias da bexiga ativadas pela capsaicina ou pelo próprio 5'-trifosfato de adenosina., Competing Interests: none.
- Published
- 2016
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11. Effect of S-methyl-l-thiocitrulline dihydrochloride on rat micturition reflex.
- Author
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Rocha JN
- Subjects
- Anesthetics, Intravenous, Animals, Arginine pharmacology, Citrulline pharmacology, Female, Injections, Spinal, Muscle Contraction drug effects, Muscle Contraction physiology, Pressure, Rats, Wistar, Reference Values, Thiourea pharmacology, Time Factors, Urethane pharmacology, Urethra physiology, Urinary Bladder physiology, Urination physiology, Citrulline analogs & derivatives, Enzyme Inhibitors pharmacology, Nitric Oxide Synthase Type I pharmacology, Thiourea analogs & derivatives, Urethra drug effects, Urinary Bladder drug effects, Urination drug effects
- Abstract
Objective: To evaluate the effect of neuronal nitric oxide synthase on the striated urethral sphincter and the urinary bladder., Materials and Methods: A coaxial catheter was implanted in the proximal urethra and another one in the bladder of female rats, which were anesthetized with subcutaneous injection of urethane. The urethral pressure with saline continuous infusion and bladder isovolumetric pressure were simultaneously recorded. Two groups of rats were formed. In group I, an intrathecal catheter was implanted on the day of the experiment at the L6-S1 level of the spinal cord; in group II, an intracerebroventricular cannula was placed 5-6 days before the experiment., Results: It was verified that the group treated with S-methyl-L-thio-citrulline, via intrathecal pathway, showed complete or partial inhibition of the urethral sphincter relaxation and total inhibition of the micturition reflexes. The urethral sphincter and the detrusor functions were recovered after L-Arginine administration. When S-methyl-Lthio-citrulline was administered via intracerebroventricular injection, there was a significant increase of urethral sphincter tonus while preserving the sphincter relaxation and the detrusor contractions, at similar levels as before the use of the drugs. Nevertheless there was normalization of the urethral tonus when L-Arginine was applied., Conclusions: The results indicate that, in female rats anaesthetized with urethane, the nNOS inhibitor administrated through the intrathecal route inhibits urethral sphincter relaxation, while intracerebroventricular injection increases the sphincter tonus, without changing bladder function. These changes were reverted by L-Arginine administration. These findings suggest that the urethral sphincter and detrusor muscle function is modulated by nitric oxide., Competing Interests: Conflicts of Interest: None declared., (Copyright® by the International Brazilian Journal of Urology.)
- Published
- 2016
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12. Oral Administration of Electron-Beam Inactivated Rhodococcus equi Failed to Protect Foals against Intrabronchial Infection with Live, Virulent R. equi.
- Author
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Rocha JN, Cohen ND, Bordin AI, Brake CN, Giguère S, Coleman MC, Alaniz RC, Lawhon SD, Mwangi W, and Pillai SD
- Subjects
- Actinomycetales Infections diagnostic imaging, Administration, Oral, Animals, Antibody Formation immunology, Antibody Specificity immunology, Horse Diseases diagnostic imaging, Horses, Immunity, Cellular, Immunoglobulin G metabolism, Interferon-gamma biosynthesis, Leukocytes, Mononuclear metabolism, Nose immunology, Treatment Outcome, Ultrasonography, Vaccination veterinary, Virulence, Actinomycetales Infections veterinary, Bronchi microbiology, Electrons, Horse Diseases immunology, Rhodococcus equi physiology
- Abstract
There is currently no licensed vaccine that protects foals against Rhodococcus equi-induced pneumonia. Oral administration of live, virulent R. equi to neonatal foals has been demonstrated to protect against subsequent intrabronchial challenge with virulent R. equi. Electron beam (eBeam)-inactivated R. equi are structurally intact and have been demonstrated to be immunogenic when administered orally to neonatal foals. Thus, we investigated whether eBeam inactivated R. equi could protect foals against developing pneumonia after experimental infection with live, virulent R. equi. Foals (n = 8) were vaccinated by gavaging with eBeam-inactivated R. equi at ages 2, 7, and 14 days, or gavaged with equal volume of saline solution (n = 4), and subsequently infected intrabronchially with live, virulent R. equi at age 21 days. The proportion of vaccinated foals that developed pneumonia following challenge was similar among the vaccinated (7/8; 88%) and unvaccinated foals (3/4; 75%). This vaccination regimen did not appear to be strongly immunogenic in foals. Alternative dosing regimens or routes of administration need further investigation and may prove to be immunogenic and protective.
- Published
- 2016
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13. Use of Liposomal Gentamicin for Treatment of 5 Foals with Experimentally Induced Rhodococcus equi Pneumonia.
- Author
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Cohen ND, Giguère S, Burton AJ, Rocha JN, Berghaus LJ, Brake CN, Bordin AI, and Coleman MC
- Subjects
- Actinomycetales Infections microbiology, Animals, Anti-Bacterial Agents administration & dosage, Anti-Bacterial Agents therapeutic use, Clarithromycin administration & dosage, Clarithromycin therapeutic use, Gentamicins administration & dosage, Horse Diseases drug therapy, Horses, Rifampin administration & dosage, Rifampin therapeutic use, Actinomycetales Infections veterinary, Gentamicins therapeutic use, Horse Diseases microbiology, Liposomes, Rhodococcus equi
- Abstract
Background: Adverse effects of, and bacterial resistance to, macrolides used to treat Rhodococcus equi infections have prompted search for clinically effective alternative antimicrobials. Liposomal gentamicin (LG) is effective against R. equi in vitro and decreases tissue concentrations of R. equi in experimentally infected mice. Effectiveness of LG treatment of foals with R. equi pneumonia, however, has not been described., Hypothesis: Liposomal gentamicin is safe and effective for treating foals with R. equi pneumonia., Animals: Ten foals with experimentally induced R. equi pneumonia., Methods: Pilot treatment trial. Foals with pneumonia induced by intrabronchial instillation of R. equi were randomly allocated to receive either clarithromycin combined with rifampin (CLR + RIF) PO or LG IV, and followed by daily physical examinations and weekly thoracic ultrasonography and serum creatinine concentration determinations until the resolution of clinical signs. Treatment success was defined as the resolution of clinical signs and ultrasonographically identified pulmonary abscesses., Results: All 10 foals were successfully treated. Two of 5 foals treated with LG developed azotemia within 1 week; LG was discontinued and treatment switched to CLR + RIF for these foals. None of the CLR + RIF treated foals developed azotemia., Conclusions and Clinical Importance: Liposomal gentamicin IV can be effective for treatment of R. equi pneumonia, but nephrotoxicity indicates that an alternative dosing interval or route (such as nebulization) will be needed before LG is adequately safe for clinical use. Larger comparative trials will be needed to evaluate the relative efficacy of a safer LG dosage regimen., (Copyright © 2015 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.)
- Published
- 2016
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14. Functional and biochemical characteristics of urinary bladder muscarinic receptors in long-term alloxan diabetic rats.
- Author
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Rocha JN
- Subjects
- Alloxan administration & dosage, Animals, Bethanechol administration & dosage, Diabetes Mellitus, Experimental chemically induced, Disease Models, Animal, Dose-Response Relationship, Drug, Male, Muscle Contraction drug effects, Muscle Contraction physiology, N-Methylscopolamine administration & dosage, Rats, Wistar, Receptors, Muscarinic drug effects, Time Factors, Urinary Bladder drug effects, Urinary Bladder physiopathology, Urination drug effects, Urination physiology, Diabetes Mellitus, Experimental metabolism, Receptors, Muscarinic metabolism, Urinary Bladder metabolism
- Abstract
Objective: To re-examine the function of the urinary bladder in vivoas well as to determine the functional and biochemical characteristics of bladder muscarinic receptors in long-term alloxan-induced diabetes rats., Methods: Two-month-old male Wistar rats were injected with alloxan and the animals showing blood glucose levels >300mg/dL together with age-paired untreated animals were kept for 11 months. Body weight, bladder weight, blood glucose, and urinary volume over a period of 24 hours were determined in both groups of animals. A voiding cystometry in conscious control and diabetic rats was performed to determine maximal micturition pressure, micturition contraction interval and duration as well as voided and post-voiding residual volume. In addition, concentration-response curves for bethanechol in isolated bladder strips, as well as [3H]-N methyl-scopolamine binding site characteristics in bladder homogenates were determined., Results: Mean bladder weight was 162.5±21.2mg versus 290±37.9mg in control and treated animals, respectively (p<0.05). Micturition contraction amplitude (34.6±4.7mmHg versus 49.6±2.5mmHg), duration (14.5±1.7 seconds versus 23.33±4.6 seconds) and interval (87.5±17.02 seconds versus 281.11±20.24 seconds) were significantly greater in alloxan diabetic rats. Voided urine volume per micturition contraction was also significantly higher in diabetic animals. However the post-voiding residual volume was not statistically different. Bethanechol potency (EC50 3µM versus 5µM) and maximal effect (31.2±5.9g/g versus 36.1±6.8g/g) in isolated bladder strips as well as number (169±4fmol/mg versus 176±3fmol/mg protein) and affinity (0.69±0.1nM versus 0.57±0.1nM) of bladder muscarinic receptors were also not statistically different., Conclusion: Bladder function in vivo is altered in chronic alloxan-induced diabetes rats without changes in functional and biochemical characteristics of bladder muscarinic receptors.
- Published
- 2015
- Full Text
- View/download PDF
15. Nitric oxide metabolites in the lumbosacral spinal cord interstice and cerebrospinal fluid in female rats with acute cyclophosphamide-induced cystitis. An in vivo microdialysis study.
- Author
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Rocha JN and Ballejo G
- Subjects
- Animals, Cyclophosphamide, Cystitis metabolism, Cystitis pathology, Female, Lumbosacral Region, Luminescence, Microdialysis, Nitrates cerebrospinal fluid, Nitrates metabolism, Nitric Oxide cerebrospinal fluid, Nitric Oxide Synthase antagonists & inhibitors, Nitrites cerebrospinal fluid, Nitrites metabolism, Rats, Rats, Wistar, Reference Values, Spinal Cord metabolism, Time Factors, Urinary Bladder metabolism, Urinary Bladder pathology, omega-N-Methylarginine pharmacology, Cystitis chemically induced, Nitric Oxide metabolism, Spinal Cord chemistry
- Abstract
Objective: To determine the concentration of nitrate/nitrite in the cerebrospinal fluid and in the dorsal horn interstice of the L6-S1 spinal cord boundary in rats with or without cystitis induced by cyclophosphamide., Methods: All experiments were conducted using Wistar female rats. A microdialysis probe was implanted in the subarachnoid space or in the spinal cord tissue at the L6-S1 segments (confirmed histologically). Two days later, the microdialysis probe was perfused with artificial cerebrospinal fluid, containing or not NG-monomethyl-L-arginine. Samples were collected every 15 minutes and kept at -20ºC. Nitrite/nitrate concentrations were determined by chemiluminescence., Results: In normal animals, the mean values of nitrite/nitrate concentrations in the first microdialysate sample of the cerebrospinal fluid and of the spinal cord interstice were similar (482.5±90.2pmol/75µL, n=20, and 505.7±11.5pmol/75µL, n=6, respectively), whereas, in the samples from rats with cystitis, these values were significantly greater (955.5±66.3pmol/75µL, n=8, and 926.5±131.7pmol/75µL, n=11, respectively). In both groups, NG-monomethyl-L- arginine caused a significant reduction in the nitrite/nitrate concentration. Interestingly, the maximal reduction of nitrite/nitrate concentration caused by NG-monomethyl-L- arginine was no greater than 30% of the initial values., Conclusions: These results constitute the first demonstration that nitrite/nitrate concentrations in the cerebrospinal fluid and spinal cord interstice are elevated between 20- and 22 hours after cyclophosphamide-induced cystitis, and indicate that cystitis is associated with changes in the production of nitric oxide in the spinal cord segments, where most primary bladder afferents end.
- Published
- 2013
- Full Text
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16. Communication between organ donor families and recipients: a definitely controversial subject.
- Author
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Ono VC, Ramalho FS, Rocha JN, Martins AC, Ramalho LN, Zucoloto S, Martinelli AL, and Castro-E-Silva O
- Subjects
- Adult, Aged, Communication, Female, Humans, Income, Male, Middle Aged, Waiting Lists, Family, Interpersonal Relations, Kidney Transplantation psychology, Tissue Donors psychology
- Abstract
In transplant centers, few topics are more controversial than communication between organ donor families (ODF) and recipients (RE). The Organ Procurement Organizations and transplant centers have felt obliged to protect the confidentiality and interests of ODF and RE. However, some authors have reported favorable effects of contact between ODF and RE. This study sought to investigate the current situation of the communication between ODF and RE from the viewpoint of transplanted patients (n = 50) and waiting transplant patients (n = 50) at a Brazilian University Hospital, ODF (n = 10), physicians from transplant centers (n = 50), as well as the opinion of the general population of a Brazilian city (n = 100). This work was developed as a survey whose questions related to the issue of communication between ODF and RE. The results showed that the majority of transplanted patients (82%) and patients awaiting transplant (60%) wanted to meet ODF to express their gratitude for receiving the organ. Likewise, ODF (67%) wanted to have a meeting with recipients, which allowed them to confirm the benefit of their donation. The general population was also favorable (66%) to ODF and RE communication. In contrast, the physicians (74%) were opposed to the ODF and RE contact. They affirmed that direct contact could lead to serious emotional conflicts or attempts of material involvement. One believes that decisions concerning the contact between ODF and RE would have to be determined by the involved parties. The transplant team could analyze the requests case by case, but ODF and RE must have the right to make the final decision.
- Published
- 2008
- Full Text
- View/download PDF
17. Change in muscarinic modulation of transmitter release in the rat urinary bladder after spinal cord injury.
- Author
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Somogyi GT, Zernova GV, Yoshiyama M, Rocha JN, Smith CP, and de Groat WC
- Subjects
- Animals, Atropine pharmacology, Electric Stimulation, Female, Muscarinic Antagonists pharmacology, Piperidines pharmacology, Pirenzepine pharmacology, Rats, Rats, Sprague-Dawley, Receptor, Muscarinic M3, Receptors, Muscarinic drug effects, Urinary Bladder, Neurogenic metabolism, Neurotransmitter Agents metabolism, Receptors, Muscarinic metabolism, Spinal Cord Injuries metabolism, Urinary Bladder metabolism
- Abstract
Muscarinic facilitation of 14C-ACh release from post-ganglionic parasympathetic nerve terminals was studied in bladder strips prepared from spinal intact (SI) and spinal cord transected (SCT) rats. The spinal cord was transected at the lower thoracic spinal segments 3 weeks prior to the experiments. Using non-facilitatory stimulation (2 Hz) the release of ACh in spinal intact rats did not change in the presence of a non-specific muscarinic antagonist, atropine (100 nM), an M(1) specific antagonist (pirenzepine, 50 nM) or an M(1)-M(3) specific antagonist (4-DAMP, 5 nM). However, during a facilitatory stimulation paradigm (10 Hz or 40 Hz, 100 shocks) atropine and pirenzepine, but not 4-DAMP inhibited the release of ACh in bladders from spinal intact rats, indicating an M(1) receptor-mediated facilitation. In spinal cord transected rats, 2 Hz stimulation-induced release was significantly inhibited by atropine or 4-DAMP but not by pirenzepine indicating that a pre-junctional facilitatory mechanism mediated via M(3) muscarinic receptors could be induced by a non-facilitatory stimulation paradigm after spinal injury. In bladders of spinal cord transected rats, 10 Hz stimulation-evoked release of ACh was also inhibited by atropine and 4-DAMP (5 nM) but not by pirenzepine (50 nM). These results indicate that pre-junctional muscarinic receptors at cholinergic nerve endings in the bladder change after chronic spinal cord injury. It appears that low affinity M(1) muscarinic receptors are replaced by high affinity M(3) receptors. This change in modulation of ACh release may partly explain the bladder hyperactivity after chronic spinal cord injury.
- Published
- 2003
- Full Text
- View/download PDF
18. Modulation of urethral alpha-sympathetic by parasympathetic before and following bethanechol chloride injection.
- Author
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Suaid HJ, Rocha JN, Martins AC, Cologna AJ, and Tucci SJ
- Abstract
Introduction and Objectives: Chagas' disease causes specific parasympathetic denervation and in its digestive clinic form promotes also functional alterations in bladder. Thus, the aim was to investigate the existence of balance between sympathetic and parasympathetic systems in lower urinary tract, as occurs in other organs. We verified the urethral closing pressure before and following parasympathetic stimulus., Patients and Methods: For that, the urethral closure pressure was studied before and after the injection of 5 mg of bethanechol chloride subcutaneously in 28 voluntary female patients, divided into 4 groups. The constitution of theses groups was: A) normal control = 6 patients; B) Chagas' disease with positive serology only = 5 patients; C) Chagas' disease with cardiac disease = 6 patients, and D) Chagas' disease with digestive disease and vesical hyporeflexia = 11 patients. Urethral profilometry was performed through perfusion urethral catheter with a 6.5 ml/minute flow and a traction rate of 5 mm/minute., Results: Means and standard deviations for urethral closure pressure before bethanechol chloride were respectively: group A = 67.3 +/- 7.1; group B = 69.2 +/- 7.4; group C = 95.8 +/- 5.1; group D = 82.1 +/- 8.4. After bethanechol chloride they were: group A = 66.0 +/- 6.6; group B = 77.0 +/- 7.6; group C = 98.3 +/- 8.8; group D = 45.9 +/- 6.2. The Kruskal Wallis statistical test did not show statistically significance difference between groups A, B, C. However, it was statistically significant between groups C and D with p = 0.003. Wilcoxon test showed p = 0.001, only for values in group D before and following bethanechol chloride., Conclusions: Chagas' disease in its intestinal form seems to alter urethral function as well. Parasympathetic stimulation decreased urethral pressure, indicating potential modulation by the parasympathetic system over the sympathetic system.
- Published
- 2003
- Full Text
- View/download PDF
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