RESUMO Blastocistos de suínos foram submetidos ao MALDI-TOF para se identificarem os principais fosfolipídios (PL). Depois, parte destes embriões (D6) foram vitrificados (n=52), ou permaneceram frescos (grupo controle, n=42). Após o aquecimento, os blastocistos foram cultivados in vitro para se avaliar a reexpansão e a eclosão (BE) às 24 e 48 horas. Finalmente, às 48 horas, os BE foram submetidos ao RT-qPCR em busca dos genes BCL2A1, BAK, BAX e CASP3. No MALDI-TOF, a intensidade do íon foi expressa em unidades arbitrárias. O desenvolvimento embrionário foi comparado por qui-quadrado (P<0,05). Entre os PL mais representativos estavam as fosfatidilcolinas [PC (32: 0) + H] +; [PC (34: 1) + H] + e [PC (36: 4) + H] +. Além do PL, o MALDI revelou alguns triglicerídeos (TG), incluindo PPL (50: 2) + Na +, PPO (50: 1) + Na +, PLO (52: 3) + Na + e POO (52: 2) + Na. A reexpansão não diferiu (P>0,05) entre blastocistos frescos ou vitrificados às 24 (33,3%, 32,7%) e 48 horas (2,4%, 13,5%). As taxas de eclosão foram maiores (P<0,05) para o grupo fresco comparado ao vitrificado às 24 (66,7% x 15,4%) e 48 horas (97,6% x 36,0%). O BAX estava mais expresso (P<0,05) após a vitrificação. Concluindo, os blastocistos Piau podem ser criopreservados por Cryotop. Este estudo também demonstrou que a via apoptótica pode ser responsável pela baixa eficiência da criopreservação de embriões suínos.

ABSTRACT Piau porcine blastocysts were submitted to MALDI-TOF to identify the main phospholipids (PL). After that, in vivo blastocysts (D6) were vitrified (n=52), non-vitrified were used as control (n=42). After warming, blastocysts were in vitro cultured to assess re-expansion and hatching at 24 and 48 hours. Finally, at 48 hours, hatched blastocysts were submitted to RT-qPCR searching for BCL2A1, BAK, BAX and CASP3 genes. For MALDI-TOF, the ion intensity was expressed in arbitrary units. Blastocyst development was compared by Qui-square (P< 0.05). Among the most representative PL was the phosphatidylcholine [PC (32:0) + H]+; [PC (34:1) + H]+ and [PC (36:4) + H]+. Beyond the PL, MALDI revealed some triglycerides (TG), including PPL (50:2) + Na+, PPO (50:1) + Na+, PLO (52:3) + Na+ and POO (52:2) + Na. Re-expansion did not differ (P> 0.05) between fresh or vitrified blastocysts at 24 (33.3%; 32.7%) or 48 hours (2.4%; 13.5%). Hatching rates were higher (P< 0.05) for fresh compared to vitrified at 24 (66.7%; 15.4%) and 48 hours (97.6%; 36.0%). BAX was overexpressed (P< 0.05) after vitrification. In conclusion, Piau blastocysts can be cryopreserved by Cryotop. This study also demonstrated that the apoptotic pathway may be responsible for the low efficiency of porcine embryo cryopreservation.

ABSTRACT Eugenia uniflora L., Myrtaceae, popularly known as “pitanga”, is used in traditional medicine due the properties attributed to its chemical content, these being mainly hydrolysable tannins and flavonoids. This study provides a qualitative and quantitative evaluation of chemical profile from leaves of E. uniflora. The HPLC analysis was carried out on a C18 column (4.6 mm × 250 mm, 5 µm) by gradient elution with methanol and water (acidified with trifluoracetic acid); and silica gel Plates 60-F 254 with 10–12 µm and 5–6 µm particles, respectively for TLC and High HPTLC analysis. The chromatographic data obtained from HPLC, TLC and HPTLC presented bands and peaks related to flavonoids (myricitrin and derivatives) and tannins (gallic and ellagic acids), which were observed from different samples. The chromatographic similarities enabled the building of a typical fingerprint for the herbal material. The similarity analysis of the sample data by Pearson correlation showed R values >0.9 among peaks (HPLC) and bands (HPTLC). In addition, the analytical methodology developed by HPLC enabled the satisfactory quantification of marker substances [ellagic acid = 0.22% and 0.20% (m/m); gallic acid = 0.20% and 0.43%; myricitrin = 0.42 and 1.74% (m/m) in herbal drug and crude extract, respectively]. The procedure was also validated in accordance with the assays required by Brazilian legislation. Thus, the HPTLC and HPLC methods developed in this study provide helpful and simple tools for the quality evaluation both qualitatively and quantitatively of raw materials and extractives from leaves of E. uniflora.

RESUMO Populações de animais selvagens são reguladas por diversos fatores bióticos e abióticos, e parasitas são um fator biótico que afetam a dinâmica e a densidade de populações. De 2002 até 2014, 62 animais silvestres provenientes dos biomas Pantanal, Amazônia e Cerrado, que vieram a óbito no atendimento do hospital veterinário ou foram encontrados atropelados em rodovias, foram submetidos à necropsia parasitológica. Ao todo 36 espécies de parasitas foram identificadas em 24 espécies de hospedeiros. Entre os parasitas, a ordem mais prevalente foi Oxyurida (29,1%), seguida por Strongylida (20,9%), Spirurida (19,4%), Ascaridida (16,2%), Pentastomida (3,2%), Echinostomida (3,2%), Gygantorhynchia (3,2%), Rhabditida (1,6%), Plagiorchiida (1,6%) e Monilimorfida (1,6%), destacando-se os nematódeos, por seu maior potencial biótico e facilidade de adaptação ao meio do que as demais classes. A maior ocorrência de endoparasitas foi observada em animais endotérmicos que ectotérmicos, e este estudo registra onze novas ocorrências de hospedeiros para endoparasitas de animais selvagens. O estudo contribui para o conhecimento da biodiversidade de parasitas em animais silvestres dos três biomas do Centro-Oeste do Brasil.

ABSTRACT The population of wild animals is regulated by many biotic and abiotic factors, and parasites are a biotic factor that affects the dynamic and density of host populations. From 2002 to 2014, 62 wild animals from the biomes Pantanal, Amazon, and "Cerrado" (or Savanna), which died in attendance in the veterinary hospital or have been road-killed, underwent necropsy for parasitological examination. Overall, 36 species of parasites were identified from 24 host species. Among the parasites, the most prevalent order was Oxyurida (29.1%), followed by Strongylida (20.9%), Spirurida (19.4%), Ascaridida (16.2%), Pentastomida (3.2%), Echinostomida (3.2%), Gygantorhynchia (3.2%), Rhabditida (1.6%), Plagiorchiida (1.6%), and Monilimorfida (1.6%), especially nematodes, which have more biotic potential and is more easily adapted to the environment than other classes. The occurrence of endoparasites was observed more frequently in endothermic than ectothermic animals, and herein is reported eleven new host occurrences for endoparasites in wild animals. The study has contributed to the knowledge on the biodiversity of parasites in wild animals from three biomes in central-western Brazil.

AbstractThe aim of this study was to evaluate the influences of variables of preparation on total flavonoids content from extractive solution of Lippia sidoides Cham., Verbenaceae. Thus a 23 factorial design was used to study the importance of plant proportion, the extraction method and solvent on the extraction of flavonoid. The methodology of determination of chemicals in factorial design was validated according to the parameters required by Brazilian Health Agency. The extraction solution was selected through a full factorial design where the best conditions to achieve the highest content of flavonoids were: 7.5% (w/v) of plant with ethanol 50% (v/v) as solvent. The polyphenols content was determined by LC method and its relationship with the antioxidant and free radical scavenging activities was evaluated. The free radical scavenging activities and antioxidant potentials were determined for different concentrations using various in vitro models. Our results indicate that extracts exhibited a significant dose-dependent antioxidant effect as evaluated by TRAP/TAR assays. Besides, we observed an antioxidant activity against hydroxyl radicals and nitric oxide, and protection against lipid peroxidation in vitro. Our results suggest that the extract presents significant in vitro antioxidant potential indicating promising perspectives for its use as pharmaceutical/or food additive.

Rearranjos cromossômicos de célula única (SCCR) são achados incidentais em culturas de células. Não obstante, alguns autores os têm relacionado com o abortamento habitual. Noventa indivíduos classificados em 4 grupos foram estudados prospectivamente: A) indivíduos com abortamentos espontâneos, filhos malformados ou gravidezes molares (N = 36); B) homens azoospérmicos (N = 14); C) indivíduos com um ou mais filhos com anormalidades cromosômicas de novo ou herdadas (N = 26) e D) indivíduos sem problemas reprodutivos e com filhos saudáveis (grupo controle, N = 14). As preparações de cromossomos de linfócitos foram coradas para bandamento GTG e 50-100 metáfases foram avaliadas por indivíduo. Os rearranjos observados foram translocações, deleções, duplicações, isocromossomos, anéis, fragmentos e marcadores. Trinta e quatro indivíduos (21 homens e 13 mulheres) tiveram de 1 a 5 SCCR. Quatro tiveram rearranjos apenas dos cromossomos 7 e 14. A freqüência de SCCR (com exceção dos rearranjos envolvendo os cromossomos 7 e 14) foi de 0,0063, enquanto que para os rearranjos entre os cromossomos 7 e 14 foi 0,0010. Comparações estatísticas intergrupo (<FONT FACE="Symbol">c</font>2 com correção de Yates) não mostraram diferenças significativas. Assim, a ocorrência de SCCR em nossa amostra mostrou-se independente do modo de averiguação e do sexo.

Single cell chromosome rearrangements (SCCR) are incidental findings in cell cultures. Nevertheless, some authors have implicated them in habitual abortion. Ninety individuals classified in four groups were prospectively studied: A) individuals with spontaneous abortions, malformed children or molar pregnancies (N = 36); B) azoospermic males (N = 14); C) individuals with one or more children with either de novo or inherited, constitutional, chromosome abnormalities (N = 26), and D) individuals with healthy children and no reproductive failure (control group, N = 14). Lymphocyte chromosome preparations were stained for GTG bands, and 50-100 metaphases were scored per individual. The rearrangements observed were translocations, deletions, duplications, isochromosomes, rings, fragments and markers. Thirty-four individuals (21 males, 13 females) had a range of 1-5 SCCR. Four had rearrangements only of chromosomes 7 and 14. SCCR frequency (with the exception of rearrangements involving chromosomes 7 and 14) was 0.0063, while that of rearrangements between chromosomes 7 and 14 was 0.0010. Statistical intergroup comparisons (<FONT FACE="Symbol">c</font>2 with Yates correction) did not show significant differences. Hence, the occurrence of SCCR in our sample was found to be independent of ascertainment mode and sex.