Evaluation of homologous, heterologous, and affinity conjugates for the serodiagnosis of Toxoplasma gondii and Neospora caninum in maned wolves (Chrysocyon brachyurus)

Use of serological tests in the diagnosis of infectious diseases in wild animals has several limitations, primarily the difficulty of obtaining species-specific reagents. Wild canids, such as maned wolves (Chrysocyon brachyurus), are highly predisposed to infection by Toxoplasma gondii and, to a lesser extent, to Neospora caninum. The aim of the present study was to evaluate homologous, heterologous, and affinity conjugates in enzyme-linked immunosorbent assays (ELISAs) and indirect fluorescent antibody tests (IFATs) for detecting immunoglobulin (Ig) G antibodies against T. gondii and N. caninum in maned wolves. Serum samples were obtained from 59 captive animals in Brazil and tested by ELISA for T. gondii serology and IFAT for N. caninum serology using 3 different enzymatic and fluorescent conjugates: homologous (guinea pig anti-maned wolf IgG-peroxidase and -fluorescein isothiocyanate [FITC]), heterologous (rabbit anti-dog IgG-peroxidase and -FITC), and affinity (protein A-peroxidase and -FITC). Seropositivity to T. gondii was comparable among the homologous (69.5%), heterologous (74.6%), and affinity (71.2%) enzymatic conjugates. A significant positive correlation was found between the antibody levels determined by the 3 enzymatic conjugates. The highest mean antibody levels (ELISA index = 4.5) were observed with the protein A-peroxidase conjugate. The same seropositivity to N. caninum (8.5%) was found with the homologous and heterologous fluorescent conjugates, but protein A-FITC was not able to detect or confirm any positive samples with homologous or heterologous conjugates. Our results demonstrate that homologous, heterologous, and affinity conjugates might be used in ELISA for serological assays of T. gondii in wild canids, whereas for N. caninum infection, only the homologous or heterologous fluorescent conjugates have been shown to be useful. © American Society of Parasitologists 2005.

An overview of functional and stereological evaluation of spermatogenesis and germ cell transplantation in fish

Although there are almost thirty-thousand species of fish living in a great variety of habitats and utilizing vast reproductive strategies, our knowledge of morphofunctional and quantitative aspects of testis structure and spermatogenesis is still incipient for this group of vertebrates. In this review, we discuss aspects that are important to better understanding of testis structure and function, and of the development of germ cells (GC) during spermatogenesis. To achieve this, we have recently completed a number of studies presenting morphometric and functional data related to the numbers of GC and Sertoli cells (SC) per each type of spermatogenic cyst, the number of spermatogonial generations, the SC efficiency, and the magnitude of GC loss that normally occurs during spermatogenesis. We also investigated SC proliferation and the relationship of this important event to early spermatogenic cysts. The available data strongly suggest that SC proliferation in sexually mature tilapia is the primary factor responsible for the increase in testis size and for determination of the magnitude of sperm production. The influence of temperature on the duration of spermatogenesis in tilapia was also evaluated and we have used this knowledge to deplete endogenous spermatogenesis in this teleost, in order to develop an experimental system for GC transplantation. This exciting technique results in new possibilities for investigation of spermatogenesis and spermatogonial stem cell biology, creating also an entirely new and promising scenario in biotechnology - transgenic animal production and the preservation of the genetic stocks of valuable animals or endangered species. © Springer Science+Business Media B.V. 2008.

Bone augmentation and autogenous transplantation of premolar to the site of the fissure in a cleft palate patient

This article presents a case report of autogenous tooth transplantation to the site of the fissure, in addition to bone augmentation with graft of autogenous bone harvested from the iliac crest, performed in a cleft palate patient, who had insufficient bone volume. A non-syndromic 10-year-old girl, with a unilateral cleft lip and palate, incisal transforamen fissures, agenesis of the maxillary left central incisor and both maxillary lateral incisors, was treated with autogenous bone graft in the cleft area. The orthodontic treatment plan was to replace the missing lateral incisors with the maxillary canines and to extract the mandibular first premolars. One of the mandibular premolars was extracted from its site with 2/3 of its root formation completed and transplanted to the maxillary left central incisor area. After orthodontic treatment, the anatomic crowns were characterized with composite resin. Autogenous tooth transplantation can be performed in the area of the fissure in young cleft palate patients, by performing bone graft augmentation before transplantation of the tooth, to gain sufficient recipient alveolar bone volume. A multidisciplinary approach is mandatory for the success of this clinical procedure, especially in cleft palate patients. © 2012 John Wiley & Sons A/S.

Aspectos clínicos do enxerto conjuntival 360° e do implante da membrana amniótica criopreservada no tratamento de úlceras de córnea em cães

To evaluate the clinical application of implant of the canine cryopreserved amniotic membrane (DMEM plus DMSO 1:1) and 360° conjunctival flap in the treatment of progressive corneal ulceration. 10 dogs of the different breeds, males and females, aging four months to four years old with deep corneal ulceration and different clinical progression were divided in two groups: G1=360° conjunctival graft (n=5) and G2=implant of amniotic membrane, sutured at the edge of the ulcer with epithelial side facing up, associated with the third eyelid flap (n=5). The comparative analysis between groups was: complications, blepharospasm, ocular secretion, corneal vascularization, epithelial defect and corneal opacification in six moments (first emergency care, surgery and 3, 7, 15 and 30 days of postoperative). Without epithelial defect was evaluated quality of the scar. It was used score scale for subjective to qualify of the ocular signs. In G1, it was observed the non-adherence of the conjunctival graft to the ulcer (n=2), dehiscence of the suture (n=2), anterior synechia (n=2) and intense chemosis (n=1). In G2, it was not observed these complications. It was not significant difference between the groups to others ocular parameters, but it was different among the start and end moments of the same groups (ocular secretion, corneal vascularization, epithelial defect). The corneal opacity was more intense in G1. According to the clinical results, the cryopreserved amniotic membrane implant proved to be as effective in the corneal ulceration in comparison to the 360° conjunctival flap, because probably, the membrane promoted a trophic support for epithelialization, anti-inflamatory effect associated with important to the end result phenotype.

Healing of fresh frozen bone allograft with or without platelet-rich plasma: A histologic and histometric study in rats

Purpose: This study histomorphometrically analyzed the effect of autogenous platelet-rich plasma (PRP) on healing of fresh frozen bone allograft (FFBA) in bony defects in rat calvaria. Materials and Methods: A 5mm-diameter defect was created in the calvarium of 30 rats. Animals were divided into three groups: C (defect was filled by blood clot only), FFBA (defect was filled with 0.01mL of FFBA), and FFBA/PRP (defect was filled with 0.01mL of FFBA combined with 100μL of PRP). All animals were euthanized at 30 days postoperatively. Histomorphometry and histology analyses were performed. Data were statistically analyzed (analysis of variance, Tukey, p<.05). Results: FFBA had a statistically smaller new bone area than groups FFBA/PRP and C. No statistically significant differences were observed between groups FFBA and FFBA/PRP with regard to remaining bone graft particle area. Conclusion: It can be concluded that (1) PRP improved the incorporation of FFBA, increasing the amount of new bone formed; (2) PRP has not influenced the resorption of nonviable particles of the FFBA; and (3) presence of remaining FFBA particles might have accounted for the smaller amount of new bone observed in group FFBA when compared with control group. © 2011 Wiley Periodicals, Inc.

Quasi-homologous spherically symmetric branes and their symmetry breaking

We revisit the dynamical system-based approach of spherically symmetric vacuum braneworlds, pointing out and studying the existence of a transcritical bifurcation as the dark pressure parameter changes its sign, we analyze some consequences of not discard the brane cosmological constant. For instance, it is noteworthy that the existence of an isothermal state equation between the dark fluid parameters cannot be obtained via the requirement of a quasi-homologous symmetry of the vacuum. © 2013 Springer-Verlag Berlin Heidelberg and Società Italiana di Fisica.

Effect of the bone marrow cell transplantation on elevated plus-maze performance in hippocampal-injured mice

Several reports have shown that the hippocampus plays an important role in different aspects of the emotional control. There is evidence that lesions in this structure cause behavioral disinhibition, with reduction of reactions expressing fear and anxiety. Thus, to portray the aptitude of cell therapy to abrogate injuries of hippocampal tissue, we examined the behavioral effects of bone marrow mononuclear cells (BMMCs) transplantation on C57BL/6 mice that had the hippocampus damaged by electrolytic lesion. For this purpose, mice received, seven days after bilateral electrolytic lesion in the dorsal hippocampus, culture medium or BMMCs expressing the enhanced green fluorescent protein (EGFP) transgene. One week after transplantation, animals were tested in the elevated plus-maze (EPM). On the whole, three assessment sessions in the EPM were carried out, with seven days separating each trial. Thirty-five days after the induction of injury, mice were sacrificed and their brains removed for immunohistochemistry. The behavioral evaluation showed that the hippocampal lesion caused disinhibition, an effect which was slightly lessened, from the second EPM test, in transplanted subjects. On the other hand, immunohistochemical data revealed an insignificant presence of EGFP+ cells inside the brains of injured mice. In view of such scenario, we hypothesized that the subtle rehabilitation of the altered behavior might be a result from a paracrine effect from the transplanted cells. This might have been caused by the release of bioactive factors capable of boosting endogenous recuperative mechanisms for a partial regaining of the hippocampal functions. © 2013 Elsevier B.V.

Padrão de cicatrização de defeitos de deiscência periodontal tratados com enxerto de tecido conjuntivo subepitelial ou matriz dérmica acelular: estudo histológico e histométrico em cães

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Cicatrização de enxertos de osso alógeno fresco congelado (OAFC) associados ou não ao plasma rico em plaquetas (PRP): estudo histológico e histométrico em mandíbulas de cães

Pós-graduação em Odontologia - FOA

Evaluation of nutritional indicators and body composition in patients with advanced liver disease enrolled for liver transplantation

PURPOSE: Malnutrition is prevalent in patients with advanced liver disease (LD) related to multifactorial causes. Fluid retention can underestimate the nutritional status based on anthropometric measures. We evaluated nutritional indicators and body composition (BC) in patients with liver cirrhosis and correlated them with LD severity. METHODS: Forty three patients with LD enrolled for liver transplantation were evaluated by Anthropometric measures, subjective evaluation (Global Assessment of Nutritional Status - SGA) and biochemical indicators. Single-frequency electrical bioimpedance (SFE-BIA) was used to evaluate body composition (BC). It measured resistance (R), reactance (Xc) and the phase angle (PA). LD severity was estimated by Child-Pugh and Meld criteria (Model for End-Stage Liver Disease). RESULTS: Child-Pugh index between patients was 7.11±1.70 and Meld was 12.23±4.22. Arm Circumference, Arm Muscle Circumference and Arm Muscle Area, SGA, hemoglobin, hematocrit and albumin showed better correlation with disease severity. Xc and PA showed correlation both with Meld and Child-Pugh score when BC were evaluated. PA was depleted in 55.8% of the patients. CONCLUSIONS: Diagnosis of malnutrition varied according to the method. Global assessment of nutritional status showed better correlation with disease severity than with objective methods. Single-frequency electrical bioimpedance for body composition analysis in cirrhotic patients must be cautiously used; however, primary vectors seems to be valid and promising in clinical practice.

Grafts of porcine small intestinal submucosa seeded with cultured homologous smooth muscle cells for bladder repair in dogs

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

Effect of platelet-rich plasma on bone healing of fresh frozen bone allograft in mandibular defects: a histomorphometric study in dogs

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Cardiomyopathy and cell therapy: ejection fraction improvement and cardiac muscle mass increasing, after a year of bone marrow stem cells transplantation, by magnetic resonance image

The idiopathic dilated cardiomyopathy (IDC) is one of the major public health problems in the western world. Patients with IDC in functional class IV (New York Health Association - NYHA), even after therapeutic optimization, have high mortality. Stem cell therapy has emerged as a potential therapeutic option for cell death-related heart diseases and several positive effects were assigned to cell therapy in cardiomyopathy. The aim of this study was identify short-term result of cell transplantation in idiopathic dilated cardiomyopathy patients (IDC) who were treated by transplantation of autologous bone marrow mononuclear cells (BMMC). Intracoronary injections of autologous BMMC were performed in eight patients with severe ventricle dysfunction (mean of left ventricle ejection fraction – LEVF=20.03%), cardiac mass muscle around 156.2 g and NYHA between III and IV grades, other 8 IDC patients received placebo. The IDCs were followed - up for one and two years, by magnetic resonance imaging (MRI). The results after one year showed significant improvement in LVEF (mean=181.4) and muscle mass increasing (mean=181.4 g), after two years the LVEF continued improving, reaching a mean of 32.69% and the cardiac muscle mass kept stable (mean=179.4 g). Excepted for one patient, all the other had improvement in the NYHA functional class. The placebo group did not show any improvement. We believe that BMMC implant may be a beneficial therapeutic option for IDC patients.