Calling activity and vocal repertoire of hypsiboas prasinus (Anura, Hylidae), a treefrog from the Atlantic Forest of Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of mesenchymal stem cells on movement and urination of rats with spinal cord injury

Cell therapy has frequently been reported as a possible treatment for spinal trauma in humans and animals; however, without pharmacologically curative action on damage from the primary lesion. In this study, we evaluated the effect of administering human adipose-derived stem cells (hADSC) in rats after spinal cord injury. The hADSC were used between the third and fifth passages and a proportion of cells were transduced for screening in vivo after transplantation. Spinal cord injury was induced with a Fogarty catheter no. 3 inserted into the epidural space with a cuff located at T8 and filled with 80 mu L saline for 5 min. The control group A (n = 12) received culture medium (50 mu L) and group B (n = 12) received hADSC (1.2 x 10(6)) at 7 and 14 days post-injury, in the tail vein. Emptying of the bladder by massage was performed daily for 3 months. Evaluation of functional motor activity was performed daily until 3 months post-injury using the Basso-Beattie-Bresnahan scale. Subsequently, the animals were euthanized and histological analysis of the urinary bladder and spinal cord was performed. Bioluminescence analysis revealed hADSC at the application site and lungs. There was improvement of urinary bladder function in 83.3% animals in group B and 16.66% animals in group A. The analysis of functional motor activity and histology of the spinal cord and urinary bladder demonstrated no significant difference between groups A and B. The results indicate that transplanted hADSC improved urinary function via a telecrine mechanism, namely action at a distance.

Psychogenic dysphonia: diversity of clinical and vocal manifestations in a case series

Introduction: Psychogenic dysphonia is a functional disorder with variable clinical manifestations.Objective: To assess the clinical and vocal characteristics of patients with psychogenic dysphonia in a case series.Methods: The study included 28 adult patients with psychogenic dysphonia, evaluated at a University hospital in the last ten years. Assessed variables included gender, age, occupation, vocal symptoms, vocal characteristics, and videolaryngostroboscopic findings.Results: 28 patients (26 women and 2 men) were assessed. Their occupations included: housekeeper (n = 17), teacher (n = 4), salesclerk (n = 4), nurse (n = 1), retired (n = 1), and psychologist (n = 1). Sudden symptom onset was reported by 16 patients and progressive symptom onset was reported by 12; intermittent evolution was reported by 15; symptom duration longer than three months was reported by 21 patients. Videolaryngostroboscopy showed only functional disorders; no patient had structural lesions or changes in vocal fold mobility. Conversion aphonia, skeletal muscle tension, and intermittent voicing were the most frequent vocal emission manifestation forms.Conclusions: In this case series of patients with psychogenic dysphonia, the most frequent form of clinical presentation was conversion aphonia, followed by musculoskeletal tension and intermittent voicing. The clinical and vocal aspects of 28 patients with psychogenic dysphonia, as well as the particularities of each case, are discussed. (C) 2014 Associacao Brasileira de Otorrinolaringologia e Cirurgia Cervico-Facial. Published by Elsevier Editora Ltda. All rights reserved.

Aquecimento vocal para o canto erudito: teoria e prática

Vocal warm-ups are essential for the technical training of the singer, for good speech and performance, as well as essential for good vocal health and a long-lasting career. Objective: To analyze the theory and practice of the vocal warm-up in classical singing, from the perspective of different professionals in the voice field including: teachers, speech therapists and singers. Method: descriptive search, quantitative in nature, cross-sectional, with a questionnaire for voice teachers, singers and audiologists. The sample consisted of 165 subjects: 86 voice teachers, 64 speech pathologists, and 15 singers. Results: Teachers of Singing (97.7 %), singers (95.3 %) and students (93.3 %) use the vocal warm-up. Conclusion: From the results we can infer that most of the professionals surveyed point to the importance of the implementation of the vocal warm-up before a performance, and the strategy used by most voice instructors, speech therapists and singers is vocalization, i.e. aesthetic warm-up.

Voice disorders in teachers: clinical, videolaryngoscopical, and vocal aspects

Dysphonia is more prevalent in teachers than among the general population. The objective of this study was to analyze clinical, vocal, and videolaryngoscopical aspects in dysphonic teachers. Ninety dysphonic teachers were inquired about their voice, comorbidities, and work conditions. They underwent vocal auditory-perceptual evaluation (maximum phonation time and GRBASI scale), acoustic voice analysis, and videolaryngoscopy. The results were compared with a control group consisting of 90 dysphonic nonteachers, of similar gender and ages, and with professional activities excluding teaching and singing. In both groups, there were 85 women and five men (age range 31-50 years). In the controls, the majority of subjects worked in domestic activities, whereas the majority of teachers worked in primary (42.8%) and secondary school (37.7%). Teachers and controls reported, respectively: vocal abuse (76.7%; 37.8%), weekly hours of work between 21 and 40 years (72.2%; 80%), under 10 years of practice (36%; 23%), absenteeism (23%; 0%), sinonasal (66%; 20%) and gastroesophageal symptoms (44%; 22%), hoarseness (82%; 78%), throat clearing (70%; 62%), and phonatory effort (72%; 52%). In both groups, there were decreased values of maximum phonation time, impairment of the G parameter in the GRBASI scale (82%), decrease of F0 and increase of the rest of acoustic parameters. Nodules and laryngopharyngeal reflux were predominant in teachers; laryngopharyngeal reflux, polyps, and sulcus vocalis predominated in the controls. Vocal symptoms, comorbidities, and absenteeism were predominant among teachers. The vocal analyses were similar in both groups. Nodules and laryngopharyngeal reflux were predominant among teachers, whereas polyps, laryngopharyngeal reflux, and sulcus were predominant among controls.

Osteogenic potential of mesenchymal cells derived from canine umbilical cord matrix co-cultured with platelet-rich plasma and demineralized bone matrix

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Changes in T-cell phenotype and cytokines profile in maternal blood, cord blood and colostrum of diabetic mothers

The present study evaluated the cells and cytokine of maternal blood, cord blood and colostrum of diabetic mothers. The women evaluated were divided according to their body mass index (BMI) and glycemic status into non-diabetic (ND - N = 15), mild gestational hyperglycemic (MGH - N = 15), diabetes mellitus gestational (DMG - N = 13) and type-2 diabetes mellitus (DM2 - N = 15) groups. The subsets of cells and cytokine profile were determined by flow cytometry. Maternal blood from MGH group had increase percentage of CD3(+)T cells, and DM-2 group had decrease percentage of CD4(+) T cells. The cord blood from hyperglycemic groups showed lower percentage of CD3(+) T cells expressing CD45RO(+) and higher of CD4(+) T cells and CD4(+) T cells expressing CD45RA(+). In the colostrum, the CD4(+) T cells and CD4(+) T cells expressed CD45RA(+) increase in hyperglycemic groups. The DM2 group exhibited higher IL17 levels in maternal blood. IFN-γ was lower in cord blood from MGH and DMG groups with overweight/obese. Irrespective of the glycemic status, IL6 was higher in colostrum. The results obtained suggest that maternal hyperglycemia modifies the phenotypes of T cells and cytokines profile in maternal, cord blood and colostrum.

Development of a cryopreservation protocol for equine mesenchymal stem cells obtained from umbilical cord

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Experimental infection of suckling mice by subcutaneous inoculation with Oropouche virus

Oropouche virus, of the family Bunyaviridae, genus Orthobunyavirus, serogroup Simbu, is an important causative agent of arboviral febrile illness in Brazil. An estimated 500,000 cases of Oropouche fever have occurred in Brazil in the last 30 years, with recorded cases also in Panama, Peru, Suriname and Trinidad. We have developed an experimental model of Oropouche virus infection in neonatal BALB/c mouse by subcutaneous inoculation. The vast majority of infected animals developed disease on the 5th day post infection, characterized mainly by lethargy and paralysis, progressing to death within 10 days. Viral replication was documented in brain cells by in situ hybridization, immunohistochemistry and virus titration. Multi-step immunohistochemistry indicated neurons as the main target cells of OROV infection. Histopathology revealed glial reaction and astrocyte activation in the brain and spinal cord, with neuronal apoptosis. Spleen hyperplasia and mild meningitis were also found, without viable virus detected in liver and spleen. This is the first report of an experimental mouse model of OROV infection, with severe involvement of the central nervous system, and should become useful in pathogenesis studies, as well as in preclinical testing of therapeutic interventions for this emerging pathogen. (c) 2012 Elsevier B.V. All rights reserved.

Time-related effects of general functional training in spinal cord-injured rats

OBJECTIVES: This prospective, randomized, experimental study with rats aimed to investigate the influence of general treatment strategies on the motor recovery of Wistar rats with moderate contusive spinal cord injury. METHODS: A total of 51 Wistar rats were randomized into five groups: control, maze, ramp, runway, and sham (laminectomy only). The rats underwent spinal cord injury at the T9-T10 levels using the NYU-Impactor. Each group was trained for 12 minutes twice a week for two weeks before and five weeks after the spinal cord injury, except for the control group. Functional motor recovery was assessed with the Basso, Beattie, and Bresnahan Scale on the first postoperative day and then once a week for five weeks. The animals were euthanized, and the spinal cords were collected for histological analysis. RESULTS: Ramp and maze groups showed an earlier and greater functional improvement effect than the control and runway groups. However, over time, unexpectedly, all of the groups showed similar effects as the control group, with spontaneous recovery. There were no histological differences in the injured area between the trained and control groups. CONCLUSION: Short-term benefits can be associated with a specific training regime; however, the same training was ineffective at maintaining superior long-term recovery. These results might support new considerations before hospital discharge of patients with spinal cord injuries.

Multiple Intravenous Administrations of Human Umbilical Cord Blood Cells Benefit in a Mouse Model of ALS

Background: A promising therapeutic strategy for amyotrophic lateral sclerosis (ALS) is the use of cell-based therapies that can protect motor neurons and thereby retard disease progression. We recently showed that a single large dose (25x10(6) cells) of mononuclear cells from human umbilical cord blood (MNC hUCB) administered intravenously to pre-symptomatic G93A SOD1 mice is optimal in delaying disease progression and increasing lifespan. However, this single high cell dose is impractical for clinical use. The aim of the present pre-clinical translation study was therefore to evaluate the effects of multiple low dose systemic injections of MNC hUCB cell into G93A SOD1 mice at different disease stages. Methodology/Principal Findings: Mice received weekly intravenous injections of MNC hUCB or media. Symptomatic mice received 10(6) or 2.5x10(6) cells from 13 weeks of age. A third, pre-symptomatic, group received 10(6) cells from 9 weeks of age. Control groups were media-injected G93A and mice carrying the normal hSOD1 gene. Motor function tests and various assays determined cell effects. Administered cell distribution, motor neuron counts, and glial cell densities were analyzed in mouse spinal cords. Results showed that mice receiving 10(6) cells pre-symptomatically or 2.5x10(6) cells symptomatically significantly delayed functional deterioration, increased lifespan and had higher motor neuron counts than media mice. Astrocytes and microglia were significantly reduced in all cell-treated groups. Conclusions/Significance: These results demonstrate that multiple injections of MNC hUCB cells, even beginning at the symptomatic disease stage, could benefit disease outcomes by protecting motor neurons from inflammatory effectors. This multiple cell infusion approach may promote future clinical studies.

Impaired blood-brain/spinal cord barrier in ALS patients

Vascular pathology, including blood-brain/spinal cord barrier (BBB/BSCB) alterations, has recently been recognized as a key factor possibly aggravating motor neuron damage, identifying a neurovascular disease signature for ALS. However, BBB/BSCB competence in sporadic ALS (SALS) is still undetermined. In this study, BBB/BSCB integrity in postmortem gray and white matter of medulla and spinal cord tissue from SALS patients and controls was investigated. Major findings include (1) endothelial cell damage and pericyte degeneration, (2) severe intra- and extracellular edema, (3) reduced CD31 and CD105 expressions in endothelium, (4) significant accumulation of perivascular collagen IV, and fibrin deposits (5) significantly increased microvascular density in lumbar spinal cord, (6) IgG microvascular leakage, (7) reduced tight junction and adhesion protein expressions. Microvascular barrier abnormalities determined in gray and white matter of the medulla, cervical, and lumbar spinal cord of SALS patients are novel findings. Pervasive barrier damage discovered in ALS may have implications for disease pathogenesis and progression, as well as for uncovering novel therapeutic targets. (C) 2012 Elsevier B.V. All rights reserved.

Dynamic Viscosity of Implantable Autologous Materials Into the Vocal Fold

Objective. To compare the dynamic viscosity (DV) of superficial layer of temporalis fascia (SLTF) with that of other biological tissues traditionally used for vocal fold implants to treat vocal fold rigidity. Study Design. Experimental. Method. Measurement of DV of samples of SLTF, deep layer of temporalis fascia (DLTF), and abdominal fat of 12 cadavers. Results. DV values of the different samples were presented in the following increasing order: SLTF, DLTF, and abdominal fat. There was statistical difference between the samples. Conclusion. DV of SLTF is lower than of other tissues tested.

Morphological characterization of the progenitor blood cells in canine and feline umbilical cord

The umbilical cord blood (UCB) is an important source of hematopoietic stem cells with great deal of interest in regenerative medicine. The UCB cells have been extensively studied as an alternative to the bone marrow transplants. The challenge is to define specific methods to purify and characterize these cells in different animal species. This study is aimed at morphological characterization of progenitor cells derived from UCB highlighting relevant differences with peripheral blood of adult in dog and cats. Therefore, blood was collected from 18 dogs and 5 cats' umbilical cords from fetus in various developmental stages. The mononuclear cells were separated using the gradient of density Histopaque-1077. Characterization of CD34+ cells was performed by flow cytometric analysis and transmission electron microscopy. Granulocytes (ancestry of the basophiles, eosinophiles, and neutrophiles) and agranulocytes (represented by immature lymphocytes) were identified. We showed for the first time the ultrastructural features of cat UCB cells. Microsc. Res. Tech. 75:766770, 2012. (C) 2011 Wiley Periodicals, Inc.