ABSTRACT The Aptian-Albian paleoichthyofauna in Brazil and Africa is quite diverse, yet their geographic patterns still require accounts. This work consists of a panbiogeographical analysis of paleoichthyofauna from Aptian-Albian through track analysis. Geographic coordinates were obtained and converted into paleocoordinates. Individual Tracks (ITs) and Generalized Tracks (GTs) were built with the aid of ArcView GIS v3.2 program using Trazos2004 extension. As a result, four GTs and tree Biogeographic Nodes (A, B and C) had been identified. GTs 1 and 2 shows two different faunas. GT3 indicates similarity between Brazil and Africa paleoichthyofauna suggesting the break-up of Gondwana as the event that separated it. GT4 represents a probable local seaway produced by transgressions. In the Node A a unique fauna was found indicating the presence of a third biota with its own characteristics. Node B is associated with the break-up of the supercontinent Gondwana. Node C was related to fauna exchange promoted by marine ingression and epicontinental seas that occurred during Aptian-Albian. We concluded that there is a relationship between the distributional patterns of biota and events such as the break-up of Gondwana, the formation of Atlantic Ocean and eustatic movements that affected South America and Africa.

ABSTRACT The effectiveness of bacteriophage P100, nisin and sodium lactate, individually and in combination, in inhibiting Listeria monocytogenes in ready-to-eat pork ham slices was assessed. The antimicrobials were applied to the surfaces of ready-to-eat pork ham slices, which were inoculated with a mixture of L. monocytogenes. Among the individual antimicrobial treatments, bacteriophage P100 was the most effective, decreasing L. monocytogenes to undetectable levels at zero and 72 h post-infection. Sodium lactate was the least effective treatment. Treatment with nisin at zero h significantly reduced initial cell density (p < 0.05). However, this pattern was not observed at 72 h of storage. A significant difference (p < 0.05) existed between the results of separate bacteriophage and nisin treatments after refrigerated storage, but not immediately upon inoculation of the bacteria. The results showed that the use of bacteriophage P100 is the method of choice for the control of bacteria.

RESUMO O presente trabalho teve como objetivo avaliar se contatos extradurais durante hemilaminectomia em cães com extrusão de disco intervertebral causariam piora neurológica no pós-operatório imadiato e/ou tardio. Dezenove cães com extrusão toracolombar de disco intervertebral foram submetidos à hemilaminectomia para descompressão medular e remoção do material extruso. Durante o procedimento cirúrgico, os contatos meningomedulares foram quantificados. Antes da cirurgia, 11/19 cães apresentavam paraplegia (com nocicepção) e 8/19 cães, paraparesia. Ao fim do estudo, apenas dois cães (2/19) mostravam paraplegia com dor profunda e um (1/19), paraparesia. Observou-se maior quantidade de contatos extradurais quando o material discal extruso encontrava-se em posição ventrolateral. Os contatos extradurais não mostraram influência estatística na evolução neurológica dos animais, bem como no tempo de recuperação das funções motora. Vinte e quatro e 48 horas após a cirurgia, 13/19 cães apresentavam o mesmo grau neurológico de antes da cirurgia. Após sete e 90 dias de pós-operatório, 13/19 e 17/19 demonstraram melhora neurológica em comparação com o pré-operatório, respectivamente. A quantidade de contatos extradurais não influenciou na recuperação neurológica dos cães. Esses achados indicam que uma inspeção minuciosa do canal vertebral pode ser recomendada, a fim de remover o máximo de material discal extruso, evitando-se piora neurológica por compressão medular.

ABSTRACT The objective of the present study was to evaluate if extradural contact during hemilaminectomy would cause neurological deterioration in the early and/or late postoperative period in dogs with intervertebral disc extrusion. Nineteen dogs with thoracolumbar intervertebral disc extrusion underwent hemilaminectomy for spinal cord decompression and removal of extruded disc material. Meningeal contacts during surgery were quantified. Paraplegia (with nociception) and paraparesis were observed in 11/19 and 8/19 of dogs, respectively, before surgery. At the end of our study, only two (2/19) had paraplegia and one (1/19), paraparesis. There were more extradural contacts when extruded intervertebral disc material was at a ventrolateral position. Extradural contacts during surgery had no influence on neurological progression nor on time to recovery of motor function. Immediately (24 and 48 hours) after surgery, 13/19 dogs had the same neurological stage before surgery. At 7 and 90 days, 13/19 and 17/19 dogs, respectively, showed neurological improvement, compared with their preoperative stage. There was no influence of the number of extradural contacts on neurological recovery. These findings indicate that a careful inspection of the vertebral canal for removal of as much extruded disc material as possible does not cause neurologic deterioration.

Objetivou-se avaliar as implicações e os resultados obtidos frente aos tratamentos conservativo e cirúrgico em 37 cães sem distinção de sexo, raça ou idade, que apresentaram fraturas e luxações vertebrais (FLV) toracolombares. Cada paciente passou por anamnese, seguida por exames clínico, neurológico e radiográficos para se obter o diagnóstico neuroanatômico e a severidade da lesão. Os cães foram submetidos ao tratamento conservativo ou cirúrgico de acordo com as características de cada caso. Dos 37 animais deste estudo, 37,7% tinham percepção de dor profunda (PDP) intacta. Vinte animais foram submetidos ao tratamento conservativo, 14 ao tratamento cirúrgico e três animais faleceram antes do tratamento. Dos animais que mantiveram a PDP intacta, a taxa de recuperação foi de 100%, com tempo de recuperação total variando de sete a 75 dias, ao passo que, dos animais que perderam a PDP, nenhum recuperou a deambulação voluntária. Dos 14 animais operados, 42,8% apresentaram alterações vertebrais não visibilizadas ao exame radiográfico convencional, sendo necessária mudança na técnica cirúrgica planejada em 14,2% desses casos. Nos animais que mantiveram a PDP, não houve diferença significante nas taxas nem no tempo de recuperação da deambulação voluntária, independentemente do tipo de tratamento. Entretanto, houve diferença altamente significante entre os graus que mantiveram a PDP intacta com os animais que perderam a percepção de dor profunda em relação à taxa de recuperação. A taxa de eutanásia foi menor que em outros trabalhos descritos na literatura. Dos animais sem PDP submetidos ao tratamento conservativo ou cirúrgico, 31,25% readquiriram a capacidade de caminhar sem recuperar a PDP, sendo essa deambulação involuntária atribuída ao caminhar espinal.

Our aim of this paper was to study the implications and results associated with conservative and surgical treatment of dogs with thoracolumbar Vertebral fractures and luxations (VFL) so that the information obtained can be used to obtain guidelines that lead to greater recovery rates. For such, 37 dogs presenting VFL were examined, without distinction of sex, breed, or age. Each patient had detailed history obtained, followed by clinical, neurologic and radiographic exams for obtaining neuroanatomic location and severity of the nervous and vertebral lesions. The dogs were submitted to conservative or surgical treatment according to each case. Deep pain perception (DPP) was intact in 37.7% of the 37 dogs in this study. Twenty dogs received conservative treatment, 14 underwent surgery and three dogs died before treatment. Of the dogs with intact DPP, 100% recovered, with a recovery time varying between seven and 75 days, while none of the dogs that lost DPP recovered voluntary motion. Of the 14 dogs submitted to surgery, 42.8% had vertebral lesions that weren't detected using conventional radiography, which demonstrates the low sensitivity of this exam, and led to a need for a change in the planned surgical technique in 14.2% of these cases. In dogs with intact DPP there was no significant difference in time or rate of recovery of voluntary motion, regardless of the treatment method. However, there was a highly significant difference between the dogs which retained and lost DPP regarding recovery rates. The percentage of euthanasia was less than in other reports, probably due to the greater awareness and protectiveness of owners nowadays. Of the dogs without DPP treated either surgically or conservatively, 31.25% recovered ambulation without recovery of DPP, characterizing this ambulation as involuntary, attributed to spinal walk.

Apolipoprotein E (APOE=gene, apoE=protein) is a known factor regulating the inflammatory response that may have regenerative effects during tissue recovery from injury. We investigated whether apoE deficiency reduces the healing effect of alanyl-glutamine (Ala-Gln) treatment, a recognized gut-trophic nutrient, during tissue recovery after 5-FU-induced intestinal mucositis. APOE-knockout (APOE-/-) and wild-type (APOE+/+) C57BL6J male and female mice (N=86) were given either Ala-Gln (100 mM) or phosphate buffered saline (PBS) by gavage 3 days before and 5 days after a 5-fluorouracil (5-FU) challenge (450 mg/kg, via intraperitoneal injection). Mouse body weight was monitored daily. The 5-FU cytotoxic effect was evaluated by leukometry. Intestinal villus height, villus/crypt ratio, and villin expression were monitored to assess recovery of the intestinal absorptive surface area. Crypt length, mitotic, apoptotic, and necrotic crypt indexes, and quantitative real-time PCR for insulin-like growth factor-1 (IGF-1) and B-cell lymphoma 2 (Bcl-2) intestinal mRNA transcripts were used to evaluate intestinal epithelial cell turnover. 5-FU challenge caused significant weight loss and leukopenia (P<0.001) in both mouse strains, which was not improved by Ala-Gln. Villus blunting, crypt hyperplasia, and reduced villus/crypt ratio (P<0.05) were found in all 5-FU-challenged mice but not in PBS controls. Ala-Gln improved villus/crypt ratio, crypt length and mitotic index in all challenged mice, compared with PBS controls. Ala-Gln improved villus height only in APOE-/- mice. Crypt cell apoptosis and necrotic scores were increased in all mice challenged by 5-FU, compared with untreated controls. Those scores were significantly lower in Ala-Gln-treated APOE+/+ mice than in controls. Bcl-2 and IGF-1 mRNA transcripts were reduced only in the APOE-/--challenged mice. Altogether our findings suggest APOE-independent Ala-Gln regenerative effects after 5-FU challenge.

Este estudo descreve o efeito da esfingosina 1-fosfato (S1P) no desenvolvimento de folículos pré-antrais, portanto da ativação e viabilidade de folículos caprinos cultivados in vitro. Fragmentos de ovários foram cultivados por um ou sete dias em meio essencial mínimo com diferentes concentrações de S1P (0, 1, 10, 50, 100 ou 200ng/mL). Os fragmentos de ovário foram processados para análise histológica em microscopia óptica, microscopia eletrônica e microscopia de fluorescência. O tratamento usando 1ng/mL de S1P foi capaz de manter a porcentagem de folículos normais durante o período de cultivo de sete dias. Ao final do período de cultivo, houve uma redução significativa (p<0,05) na porcentagem de folículos primordiais em todos os grupos tratados com S1P, comparados com os grupos controle (FC e CC), seguida por um aumento do número de folículos ativados (intermediários, primários e secundários). Adicionalmente, na cultura por sete dias com meio suplementado com S1P (1ng/mL), houve preservação da ultraestrutura das organelas e manteve-se a viabilidade dos folículos pré-antrais avaliados por microscopia de fluorescência. Em conclusão, após sete dias de cultura, o meio suplementado com 1ng/mL de S1P ativa o desenvolvimento de folículos pré-antrais de caprino, cultivados in situ e mantém as viabilidades oocitária e folicular.

This study describes the effect of sphingosine 1-phosphate (S1P) for development of preantral follicle, therefore the activation and follicular viability of caprine follicles cultured in vitro. Ovarian fragments were cultured for 1 or 7 days in Minimum Essential Medium with different S1P concentrations (0, 1, 10, 50, 100 or 200ng/mL). All ovarian fragments were processed for histological analysis in optical microscopy, transmission electron microscopy and fluorescence analysis. The treatment using 1ng/mL of S1P was able to maintain the percentage of normal follicles with the progression of the culture from day 1 to 7. At end of the 7-day culture period there was a significant reduction (P<0.05) in the percentage of primordial follicles in all groups treated with S1P, compared with fresh control (FC) and Control Culture (CC), which was followed by an increase of activated follicles (intermediary, primary and secondary). In addition, the culture for 7 days with media supplemented with S1P with 1ng/mL preserved the ultrastructure of organelles and kept the preantral follicular viability when evaluated by fluorescence microscopy. In conclusion, after 7 days of culture, the 1ng/mL of S1P activates the development of preantral caprine follicles, cultured in situ and maintains the oocitary and follicular viability.

O objetivo desse estudo foi investigar se o TGF-β afeta a sobrevivência, ativação e crescimento de folículos primordiais caprinos inclusos no córtex ovariano após o cultivo in vitro. Ovários de cabras foram coletados em abatedouro e fragmentos de tecido ovariano foram cultivados por um e sete dias em meio essencial mínimo alfa (α-MEM+) sozinho ou suplementado com TGF-β (1, 5, 10 ou 50ng/mL). Fragmentos ovarianos não cultivados e cultivados foram processados para análise histológica e ultraestrutural. Os resultados mostraram que, comparado ao controle fresco, houve diminuição no percentual de folículos morfologicamente normais em todos os tratamentos somente após sete dias de cultivo. O TGF-β não afetou a ativação folicular independente da concentração testada, contudo, o diâmetro folicular foi superior (P<0.05) no tratamento com 10ng/mL de TGF-β quando comparado ao controle fresco e aos demais tratamentos. Além disso, essa mesma concentração manteve a ultraestrutura normal dos folículos após sete dias de cultivo. Em conclusão, o TGF-β apresentou efeito adicional no crescimento folicular e na manutenção da integridade ultraestrutural de folículos pré-antrais caprinos inclusos no tecido ovariano quando utilizado na concentração de 10ng/mL durante sete dias de cultivo.

The objectives of this study were to investigate whether TGF-β affect the survival, activation and further growth of goat primordial follicles enclosed in ovarian cortex after in vitro culture. Goat ovaries were collected from an abattoir and pieces of ovarian tissues were cultured for one or seven days in a supplemented alpha Minimum Essential Medium, alone or containing TGF-β (1, 5, 10 or 50ng/mL). Ovarian tissues from the fresh control as well as those cultured were processed for histological and ultrastructural studies. The results showed that when compared with fresh control, there was decrease in the percentages of histologically normal follicles in all treatments only after seven days culture. TGF-β did not affect the activation of preantral follicles regardless of its concentration, however, larger follicles diameter (P<0.05) was observed using 10ng/mL TGF-β than in the fresh control and other treatments. Moreover, this concentration maintained the normal ultrastructure after seven days of culture. In conclusion, TGF-β showed additional effect on the follicle growth and the maintenance of ultrastructural integrity of goat preantral follicles enclosed in ovarian tissue when used at 10ng/mL during seven days of culture.

The objective of this study was to determine the effects of GDF-9, IGF-I, and GH alone or combined on preantral follicle survival, activation and development after 1 and 7 days of in vitro culture. Either fresh (non-cultured) or cultured ovarian tissue was processed for histological and fluorescence analysis. For all media tested, the percent of normal follicles was greater when compared to minimum essential medium supplemented (MEM+) alone, except when ovarian tissue was cultured with GDF-9/IGF-I or GDF-9/GH (P < 0.05). Fluorescence analysis showed that the percent of viable follicles after 7 days of culture was similar for non-cultured tissue and for all treatments tested. The percent of primordial follicles was reduced (P < 0.05) and there was a significant and concomitant increase in the percent of intermediate and primary follicles in all treatments tested after 7 days of culture when compared to non-cultured tissue. After 7 days of culture, the highest percent of intermediate follicles was observed with IGF-I/GH (61.3%), and the highest percent of primary follicles was achieved with IGF-I (57.7%). After 7 days of culture in MEM+ containing GDF-9, IGF-I and GH alone or in all associations, a significant increase in follicular diameter was observed when compared to MEM+ alone and non-cultured tissue. In conclusion, GDF-9, IGF-I and GH alone or in combination maintain preantral follicle survival and promote primordial follicle activation. Nevertheless, the data showed that IGF-I/GH and IGF-I alone are efficient in promoting the transition from primordial to intermediate follicles and from intermediate to primary follicles, respectively.

Porcine pituitary follicle stimulating hormone (pFSH) is known to regulate the production of growth factors that have an essential role in early foliculogenesis. However, the effects of different preparations of pFSH on the survival and development of caprine follicles are not yet known. The aim of this study was to evaluate the effects of different pFSH (Stimufol® and Folltropin®) on the in vitro survival and growth of caprine preantral follicles. Pieces of caprine ovarian tissues were cultured for either one or seven days in a supplemented Minimum Essential Medium, alone or containing either Stimufol® (50 ng/mL) or Folltropin® (10, 50, 100 and 1000 ng/mL). Fresh control ovarian tissues as well as cultured tissued were processed for histological and ultrastructural studies. The results showed that after seven days, only Stimufol® maintained follicular morphology similar to control. Moreover, follicular degeneration was higher in medium alone or with Folltropin® at 50, 100 and 1000 ng/mL. However, at day seven, the percentage of growing follicles was higher in 100 ng/mL of Folltropin® than Stimufol®. In conclusion, FSH preparations affect differently the performance of in vitro culture of caprine preantral follicles. Stimufol® was better to preserve follicular morphology while Folltropin® was more efficient to promote follicular growth.

The alpha2ß1 integrin is a major collagen receptor that plays an essential role in the adhesion of normal and tumor cells to the extracellular matrix. Alternagin-C (ALT-C), a disintegrin-like protein purified from the venom of the Brazilian snake Bothrops alternatus, competitively interacts with the alpha2ß1 integrin, thereby inhibiting collagen binding. When immobilized in plate wells, ALT-C supports the adhesion of fibroblasts as well as of human vein endothelial cells (HUVEC) and does not detach cells previously bound to collagen I. ALT-C is a strong inducer of HUVEC proliferation in vitro. Gene expression analysis was done using an Affimetrix HU-95A probe array with probe sets of ~10,000 human genes. In human fibroblasts growing on collagen-coated plates, ALT-C up-regulates the expression of several growth factors including vascular endothelial growth factor, as well as some cell cycle control genes. Up-regulation of the vascular endothelial growth factor gene and other growth factors could explain the positive effect on HUVEC proliferation. ALT-C also strongly activates protein kinase B phosphorylation, a signaling event involved in endothelial cell survival and angiogenesis. In human neutrophils, ALT-C has a potent chemotactic effect modulated by the intracellular signaling cascade characteristic of integrin-activated pathways. Thus, ALT-C acts as a survival factor, promoting adhesion, migration and endothelial cell proliferation after binding to alpha2ß1 integrin on the cell surface. The biological activities of ALT-C may be helpful as a therapeutic strategy in tissue regeneration as well as in the design of new therapeutic agents targeting alpha2ß1 integrin.

The pathogenic fungus Sporothrix schenckii is the causative agent of sporotrichosis. This subcutaneous mycosis may disseminate in immunocompromised individuals and also affect several internal organs and tissues, most commonly the bone, joints and lung. Since adhesion is the first step involved with the dissemination of pathogens in the host, we have studied the interaction between S. schenckii and several extracellular matrix (ECM) proteins. The binding of two morphological phases of S. schenckii, yeast cells and conidia, to immobilized type II collagen, laminin, fibronectin, fibrinogen and thrombospondin was investigated. Poly (2-hydroxyethyl methacrylate) (poly-HEMA) was used as the negative control. Cell adhesion was assessed by ELISA with a rabbit anti-S. schenckii antiserum. The results indicate that both morphological phases of this fungus can bind significantly to type II collagen, fibronectin and laminin in comparison to the binding observed with BSA (used as blocking agent). The adhesion rate observed with the ECM proteins (type II collagen, fibronectin and laminin) was statistically significant (P<0.05) when compared to the adhesion obtained with BSA. No significant binding of conidia was observed to either fibrinogen or thrombospondin, but yeast cells did bind to the fibrinogen. Our results indicate that S. schenckii can bind to fibronectin, laminin and type II collagen and also show differences in binding capacity according to the morphological form of the fungus.