Purpose: To establish a low-cost method to prepare platelet-rich plasma (PRP) and evaluates the potential of platelet derived factors to enhance wound healing in the surgical wounds in equine. Methods: To obtain a PRP gel, calcium gluconate and autologous thrombin were added to platelet-rich plasma. For the tests six saddle horses were used and two surgical incisions were made in each animal. Wounds were treated with PRP gel or untreated. Sequential wound biopsies collected at Treatment 1: at days 5 and 30 and Treatment 2: at days 15 and 45 post wounding permitted comparison of differentiation markers and wound repair. Results: The optimal platelets enrichment over 4.0 time's baseline values was obtained using 300 g for 10 min on the first centrifugation and 640 g for 10 min on the second centrifugation. Conclusion: Wounds treated with PRP gel exhibit more rapid epithelial differentiation and enhanced organization of dermal collagen compared to controls in equine Key words: Platelet-Rich Plasma. Wound Healing. Skin. Horses. RESUMOObjetivo: Estabelecer um método econômico na preparação de plasma rico em plaquetas (PRP) e avaliar se os fatores derivados destas plaquetas aceleram a cicatrização de feridas cirúrgicas em cavalos. Métodos: Gluconato de cálcio e trombina autógena foram adicionados ao PRP para a obtenção do gel de PRP. Foram usados seis cavalos de sela, cada um dos quais sofreu duas incisões cirúrgicas. Uma destas incisões foi tratada com gel de PRP e a outra suturada de maneira tradicional (controle). A biópsia das feridas foi coletada de maneira seqüencial; Tratamento 1. nos dias 5 e 30 e Tratamento 2. nos dias 15 e 45 do período pós-operatório permitindo uma comparação na diferenciação epitelial e no reparo das feridas. Resultados: O enriquecimento das plaquetas obtido através de uma primeira centrifugação usando 300 g por 10 minutos e uma segunda 640 g por 10 minutos acelerou quatro vezes a reparação tecidual em relação ao controle. Conclusão: As feridas tratadas com gel de PRP apresentaram uma mais rápida diferenciação epitelial e acelerou a organização do colágeno da derme comparado ao grupo controle em cavalos. Descritores: Plasma Rico em Plaquetas. Cicatrização de Feridas. Pele. Cavalos.

BackgroundVisceral leishmaniasis (VL) is characterized by parasite-specific immunosuppression besides an intense pro-inflammatory response. Lipopolisaccharide (LPS) has been implicated in the immune activation of T-cell deficient diseases such as HIV/AIDS and idiopathic lymphocytopenia. The source of LPS is gram-negative bacteria that enter the circulation because of immunological mucosal barrier breakdown. As gut parasitization also occurs in VL, it was hypothesized that LPS may be elevated in leishmaniasis, contributing to cell activation.Methodology/Principal FindingsFlow cytometry analysis and immunoassays (ELISA and luminex micro-beads system) were used to quantify T-cells and soluble factors. Higher LPS and soluble CD14 levels were observed in active VL in comparison to healthy subjects, indicating that LPS was bioactive; there was a positive correlation between these molecules (r = 0.61;p<0.05). Interestingly, LPS was negatively correlated with CD4+ (r = −0.71;p<0.01) and CD8+ T-cells (r = −0.65;p<0.05). Moreover, higher levels of activation-associated molecules (HLA-DR, CD38, CD25) were seen on T lymphocytes, which were positively associated with LPS levels. Pro-inflammatory cytokines and macrophage migration inhibitory factor (MIF) were also augmented in VL patients. Consistent with the higher immune activation status, LPS levels were positively correlated with the inflammatory cytokines IL-6 (r = 0.63;p<0.05), IL-8 (r = 0.89;p<0.05), and MIF (r = 0.64;p<0.05). Also, higher plasma intestinal fatty acid binding protein (IFABP) levels were observed in VL patients, which correlated with LPS levels (r = 0.57;p<0.05).Conclusions/SignificanceElevated levels of LPS in VL, in correlation with T-cell activation and elevated pro-inflammatory cytokines and MIF indicate that this bacterial product may contribute to the impairment in immune effector function. The cytokine storm and chronic immune hyperactivation status may contribute to the observed T-cell depletion. LPS probably originates from microbial translocation as suggested by IFABP levels and, along with Leishmania antigen-mediated immune suppression, may play a role in the immunopathogenesis of VL. These findings point to possible benefits of antimicrobial prophylaxis in conjunction with anti-Leishmania therapy.

RESUMO ABSTRACTThis study, realized from October 2000 to January 2003 describes the clinical epidemiological characteristics of visceral leishmaniasis in Três Lagoas, Mato Grosso do Sul State, Brazil. A total of 149 cases were confirmed, with a predominance of the male gender (71.1%). The principal age group was aged 0 to 4 years old (42%). The clinical picture included fever (97.3%), esplenomegaly (85.9%) and anemia (75.8%). Associated infections were seen in 32 patients (21.5%), pneumonia being most common. Changes registered in the laboratory included a median hemoglobin level of 8mg/dl and 3,100 leucocytes/mm 3 . Bone marrow smears were positive in 90.6% of patients. Of the 97.9% patients treated, 78.2% used pentavalent antimony. Mortalities occurred in 8% of cases, half of these with associated infection. Taken together, these data suggest changes in the physiographical occurrence of visceral leishmaniasis in this locality, with expansion and urbanization of the disease, requiring greater attention for early diagnosis and treatment.

A rapid indirect enzyme-linked immunosorbent assay (ELISA) was developed for measuring antibodies against Anaplasma marginale using a partially soluble antigen prepared from semi-purified initial bodies from erythrocytes with 80.0% of rickettsiaemia. This technique utilized alkaline phosphatase and p-nitrophenyl phosphate as reaction indicators. The high sensitivity (100.0%) was confirmed with sera from 100 calves experimentally-infected with A. marginale. All of these animals showed seroconversion before or at the same time of the first rickettsiaemia or even when it was not detected. Also the elevated specificity (94.0%) was confirmed by the low percentage of cross-reactions with sera from animals experimentally-infected with Babesia bigemina and Babesia bovis (1.4 and 6.6%, respectively). Performances of ELISA and indirect fluorescent antibody test (IFAT) with 324 sera from enzootically stable area did not show statistical difference (P>0.05), since the former showed 96.9% and the latter 97.2% of positive reactions. The advantage of this ELISA is a shorter execution time than others developed until now, allowing more samples to be analyzed.

The use of eosin methylene blue according to Giemsa as photosensitizer is presented for the first time in this paper. The present study evaluated the potential application of chlorophyllin sodium copper salt (CuChlNa) and eosin methylene blue according to Giemsa (EMB) as antimicrobial photosensitizers (aPS) for photodynamic inactivation (PDI) of Staphylococcus aureus (gram-positive) and Escherichia coli (gram-negative) bacteria. The experiments were performed using S. aureus stain ATCC 25923 and E. coli ATCC 25922 in which five aPS concentrations (0.0, 1.0, 2.5, 5.0, 10.0, and 20.0 μM for S. aureus and 0.0, 5.0, 10.0, 20.0, 40.0, and 50.0 μM for E. coli) were prepared and added in 2 mL of a saline solution containing the bacterial inoculum. After aPS incubation, the samples were divided into two groups, one kept in the dark and another submitted to the illumination. Then, the bacterial inactivation was determined 18 h after the incubation at 37 °C by counting the colony-forming units (CFU). The results revealed that both EMB and CuChlNa can be used as aPS for the photoinactivation of S. aureus, while only EMB was able to photoinactivate E. coli. Nevertheless, a more complex experimental setup was needed for photoinactivation of E. coli. The data showed that EMB and CuChlNa presented similar photoinactivation effects on S. aureus, in which bacterial growth was completely inhibited at photosensitizer (PS) concentrations over 5 μM, when samples were previously incubated for 30 min and irradiated by a light dose of 30 J cm as a result of an illumination of 1 h at 8.3 mW cm by using a red light at 625 nm with a 1 cm beam diameter and output power of 6.5 mW. In the case of E. coli, bacterial growth was completely inhibited only when combining a PS incubation period of 120 min with concentrations over 20 μM.

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RESUMO.-Objetivou-se avaliar a atividade antibacteriana in vitro da própolis marrom, por meio da determinação da Concentração Inibitória Mínima (CIM). O extrato alcoólico de própolis foi obtido de 35 g de própolis bruta macerada em 65 mL de álcool de cereais. As concentrações do extrato de própolis usadas foram: 75mg/mL; 56,4mg/mL; 37,5mg/mL; 18,9mg/mL; 9,3mg/mL; 4,5mg/mL e 2,25mg/mL. INTRODUÇÃOA própolis é uma substância resinosa que as abelhas coletam de diversas partes das plantas, como brotos, botões florais, cascas e exsudatos resinosos (Park et al. 1998, Pereira et al. 2002. As abelhas utilizam a própolis para fechar pequenos orifícios, mumificar insetos que morrem no interior da colmeia, defender contra invasão de microrganismos, fortalecer os favos e proteger a entrada das colmeias The aim of this study was to evaluate the in vitro antibacterial activity of brown propolis, by determining the minimum inhibitory concentration (MIC). The alcoholic extract of propolis was obtained from 35g of crude propolis macerated in 65mL of cereals alcohol. Dilutions of propolis extracts used were: 75mg/mL; 56.4mg/mL; 37.5mg/mL; 18.9mg/mL; 9.3mg/mL; 4.5mg/mL, and 2.25mg/mL. Thirty-two Gram-positive bacteria isolates were used: Rhodococcus equi, Staphylococcus aureus, Staphylococcus hyicus, Staphylococcus spp. and Streptococcus spp.; and the thirty-two Gram-negative bacteria isolated were: Enterobacter agglomerans, Escherichia coli, Klebsiella pneumoniae, Klebsiella sp., Pseudomonas aeruginosa, Pseudomonas sp., Salmonella sp., and Serratia rubidaea, obtained from infectious clinical processes of domestic animals, and stored in the Laboratory of Bacteriology of FAMEZ/ UFMS. The brown propolis extract showed antibacterial activity with MIC ranging from 2.25 to 18.9mg/ml for Gram-positive bacteria and 4.5 to 18.9mg/ml for Gram-negative bacteria, and bacteria from cattle and dogs were more resistant. It was concluded that brown propolis has antibacterial action, but the effect depends on the species of the bacterium and its origin.INDEX TERMS: Biological activity, brown propolis, bacteria, bactericidal, minimum inhibitory concentration.

The Brazilian city of Três Lagoas, Mato Grosso do Sul State, has experienced an urban outbreak of visceral leishmaniasis since 2000. In 2002, due to the increase in the number of cases, 46 families with cases of visceral leishmaniasis were studied to verify the prevalence of asymptomatic infection in household contacts. Indirect immunofluorescence and ELISA showed a 36.4% positive infection rate. There were no cases of symptomatic disease among these contacts. There was no statistically significant difference in gender or age. Median age was 21 years, and the 10-19-year age bracket was the most heavily affected (23%). As for family characteristics, no differences were observed in schooling or family income; most families (58.7%) owned their homes, which were built of masonry (97.8%) and had adequate infrastructure. All the families reported what were probably phlebotomine sand flies in the peridomicile. In conclusion, asymptomatic visceral leishmaniasis infection is frequent and occurs in both males and females, regardless of age.