Extracción y medida de peroxidasa en pulpa de arazá (Eugenia stipitata MC Vaugh)

A technique for both extraction and activity measurement of peroxidase extracted from arazá (Eugenia stipitata Mc Vaugh) is described. Peroxidase from arazá pulp fruit was extracted using a combination of protein precipitation with acetone and extraction with 50 mM sodium buffer phosphate (pH 6.0). Optimum activity using guaiacol as H-donor was obtained at pH from 5.0 to 6.5, temperature from 60 to 75 °C, H2O2 between 10 to 15 mM and guaiacol from 80 to 160 mM. Thermal inactivation showed a first-order inactivation kinetic. Reactivation was observed when extracts were heated at 80 °C and afterwards incubated at 25 °C.

Ni(II), Cu(II), AND Zn(II) COMPLEXES DERIVED FROM A NEW SCHIFF BASE 2-((Z)-(3-METHYLPYRIDIN-2- YLEIMINO)METHYL)PHENOL AND SYNTHESIS OF NANO SIZED METAL OXIDE PARTICLES FROM THESE COMPOUNDS

Synthesis, spectral identification, and magnetic properties of three complexes of Ni(II), Cu(II), and Zn(II) are described. All three compounds have the general formula [M(L)2(H2O)2], where L = deprotonated phenol in the Schiff base 2-((z)-(3-methylpyridin-2-yleimino)methyl)phenol. The three complexes were synthesized in a one-step synthesis and characterized by elemental analysis, Fourier transform infrared spectroscopy, electronic spectra, X-ray diffraction (XRD), and room temperature magnetic moments. The Cu(II) and Ni(II) complexes exhibited room temperature magnetic moments of 1.85 B.M. per copper atom and 2.96 B.M. per nickel atom. The X-band electron spin resonance spectra of a Cu(II) sample in dimethylformamide frozen at 77 K (liquid nitrogen temperature) showed a typical ΔMS = ± 1 transition. The complexes ([M(L)2(H2O)2]) were investigated by the cyclic voltammetry technique, which provided information regarding the electrochemical mechanism of redox behavior of the compounds. Thermal decomposition of the complexes at 750 ºC resulted in the formation of metal oxide nanoparticles. XRD analyses indicated that the nanoparticles had a high degree of crystallinity. The average sizes of the nanoparticles were found to be approximately 54.3, 30.1, and 44.4 nm for NiO, CuO, and ZnO, respectively.

DEVELOPMENT AND VALIDATION OF ANALYTICAL METHOD FOR PALM OIL DETERMINATION IN MICROCAPSULES PRODUCED BY COMPLEX COACERVATION

The microencapsulation of palm oil may be a mechanism for protecting and promoting the controlled release of its bioactive compounds. To optimize the microencapsulation process, it is necessary to accurately quantify the palm oil present both external and internal to the microcapsules. In this study, we developed and validated a spectrophotometric method to determine the microencapsulation efficiency of palm oil by complex coacervation. We used gelatin and gum arabic (1:1) as wall material in a 5% concentration (w/v) and palm oil in the same concentration. The coacervates were obtained at pH 4.0 ± 0.01, decanted for 24 h, frozen (−40 ºC), and lyophilized for 72 h. Morphological analyzes were then performed. We standardized the extraction of the external palm oil through five successive washes with an organic solvent. We then explored the best method for rupturing the microcapsules. After successive extractions with hexane, we determined the amount of palm oil contained in the microcapsules using a spectrophotometer. The proposed method was shown to be of low cost, fast, and easy to implement. In addition, in the validation step, we confirmed the method to be safe and reliable, as it proved to be specific, accurate, precise, and robust.

Biological and molecular characterization of an isolate of Tobacco streak virus obtained from soybeans in Brazil

A virus was isolated from soybean (Glycine max) plants with symptoms of dwarfing and bud blight in Wenceslau Braz County, Paraná, Brazil. The host range and properties resembled those of Tobacco streak virus (TSV). The purified virus showed three peaks in a frozen sucrose gradient. Antiserum was produced and the virus was serologically related to TSV. Electron microscopy detected 28 nm spherical particles. Coat protein (CP) had a Mr of 29.880 Da. A fragment of 1028 nt was amplified, cloned and sequenced. One open reading frame with 717 nt was identified and associated to the CP. The CP gene shared 83% identity with the sequence of TSV CP from white clover (Trifolium repens) (GenBank CAA25133). This is the first report of the biological and molecular characterization of TSV isolated from soybeans. It is proposed that this isolate be considered a strain of TSV named TSV-BR.

Conduta terapêutica atual no adenocarcinoma da cárdia e da junção esofagogástrica

Adenocarcinomas of the cardia and gastroesophageal junction are peculiar entities with three different origins, which differ somewhat from other adenocarcinomas of the stomach in their clinical presentation and pathogenesis, and have a poorer prognosis. In this article the authors reviewed definitions, incidence and epidemiology, etiologic factors, genetic implications, clinical presentation, diagnosis, staging and treatment, with emphasis on the surgical approach, discussing the current management of these cancers. The prognostic factors related specifically to the cardia cancers are: esophageal invasion greater than 3cm, microscopic residual tumor and wall penetration (>T2). Preoperative workup should include computed tomography, and endoscopic ultrasonography and laparoscopy when available. Preoperative recognition of T3/ T4/N2 lesions should indicate inclusion in neo-adjuvant protocols whenever possible. The authors present the results of 46 resected cases of adenocarcinomas of the cardia and GE junction of the Instituto Nacional do Câncer- Brazil (1981-1995). Cure was intended in 29 and palliation in 17 patients. The most common type of resection was total gastrectomy with abdominal esophagectomy (28 cases). Morbidity (major and minor) occurred in 50% of the patients. The main causes were of respiratory origin and fistulas (19.6% each). Death occurred in 44% of the patients with fistula. Postoperative death until the 30th day occurred in 17.24% of the curative cases and in 23.52% of the palliative ones. The median survival time was 68.5 months for stage I, 25 months for stage II, 31 months for stage III and 12.5 months for stage IV diseases. The median survival time was 8 months for palliation and 28.5 months for cure. No long-term survival was obtained with the palliative group, whereas 25% survived five years of more in the curative group. The authors conclude that the surgical approach should be the one the surgeon feels more comfortable with. Complete removal of the disease proved by frozen section, splenectomy and D2 lymphadenectomy should be the standard therapy with curative intent.

Adenoma viloso da ampola de Vater

The aim of this paper is to present a case of a 37-year-old female patient with a benign tumor of the Ampulla of Vater and a brief review of the literature. The patient presented with progressive obstructive jaundice and weigth loss due to the presence of two adenomas of the second portion of duodenum. Laboratory tests confirmed the presence of obstruction of the biliary tree. Ultrasound and CT scan of the abdomen revealed bile duct dilatation. ERCP showed a tumor at the site of the Ampulla of Vater. The biopsies revealed tubular adenoma. She was submitted to local resection of the tumors and sphincteroplasty, since the frozen biopsy at the time of surgery showed no malignancy. During the post-operative follow-up she presented recurrence of symptoms. An upper GI endoscopy revealed a tumor at the Ampulla of Vater. She was then submitted to Whipple procedure with an uneventful recovery.

Leiomiossarcoma de jejuno

We describe a case of leiomyosarcoma of the jejunum in which abdominal computed tomography showed an intestinal tumor with a "sui generis" metalic artefact inside, which made us think of a benign disease (foreign body granuloma), because the patient lived in the rural area and he had a manioc flour mill, which is one the basic foods of the majority of the population of the north in Brazil. Because of the aspect of the tumor, we decided on a large scale resection, considering the possibility of a malignant tumor since we don't have frozen sections. This assured us of an adequate treatment for the tumor. This type of pathology should be remembered even though it only accounts for 0,2-2% of the intestinal tract tumors.

Tumor do apêndice vermiforme

Appendiceal tumors are rare and usually presented as acute appendicitis. They are incidentally discovered at an emergency surgery, in which case the decision-making is very difficult. The purpose of this report is discuss to the most appropriate management for appendiceal tumors. A retrospective analysis of one case treated at HU-UEL was undertaken and the literature was reviewed. The conclusion is that frozen section should be done whenever the appendiceal findings are atypical. For appendiceal carcinoids greater than 2 cm and adenocarcinomas, a right hemicolectomy is recommended. Appendectomy is apropriate for patients whose carcinoid tumors are 1 cm in diameter or less and for carcinoid tumors between 1 cm and 2 cm without extensive lymphatic permeation and mesenteric invasion. Every case should be searched for synchronous tumors.

Epidemiological evaluation of hepatic trauma victims undergoing surgery

Objective : to evaluate the epidemiological variables and diagnostic and therapeutic modalities related to hepatic trauma patients undergoing laparotomy in a public referral hospital in the metropolitan region of Vitória-ES. Methods : we conducted a retrospective study, reviewing charts of trauma patients with liver injuries, whether isolated or in association with other organs, who underwent exploratory laparotomy, from January 2011 to December 2013. Results : We studied 392 patients, 107 of these with liver injury. The male: female ratio was 6.6 : 1 and the mean age was 30.12 years. Penetrating liver trauma occurred in 78.5% of patients, mostly with firearms. Associated injuries occurred in 86% of cases and intra-abdominal injuries were more common in penetrating trauma (p <0.01). The most commonly used operative technique was hepatorrhaphy and damage control surgery was applied in 6.5% of patients. The average amounts of blood products used were 6.07 units of packed red blood cells and 3.01 units of fresh frozen plasma. The incidence of postoperative complications was 29.9%, the most frequent being infectious, including pneumonia, peritonitis and intra-abdominal abscess. The survival rate of patients suffering from blunt trauma was 60%, and penetrating trauma, 87.5% (p <0.05). Conclusion : despite technological advances in diagnosis and treatment, mortality rates in liver trauma remain high, especially in patients suffering from blunt trauma in relation to penetrating one.

A análise seminal deve ser requisitada para homens com histórico de fertilidade prévia?

OBJETIVOS: determinar se a história prévia de fertilidade pode predizer o atual status de fertilidade de um paciente masculino examinado por infertilidade do casal. MÉTODOS: estudo retrospectivo envolvendo análises seminais de 183 pacientes consecutivos subférteis avaliados entre setembro de 2002 e março de 2004. Foram excluídos do estudo os pacientes que haviam se submetido a radioterapia, quimioterapia, orquiectomia ou vasectomia. Os valores médios de todas as análises foram usados em pacientes com múltiplas análises de sêmen. Pacientes com concentração espermática superior a 20x10(6) espermatozóides/mL, motilidade superior a 50% e espermatozóides com morfologia estrita superior a 14% foram considerados normais. Os pacientes foram divididos em dois grupos, segundo o status de fertilidade: infertilidade primária (118 pacientes) e infertilidade secundária (65 pacientes). Os dados foram analisados pelos testes estatísticos chi2 e teste t de Student. RESULTADOS: não houve diferença na idade média entre os pacientes com infertilidade primária, 37,3±6,3, e infertilidade secundária, 38,1±5,9; p=0,08. No grupo de pacientes com infertilidade primária, 51,9% (61 pacientes) tiveram uma concentração espermática normal, 70,3% (83 pacientes) tiveram a motilidade espermática normal e 26,37% (31 pacientes), por sua vez, morfologia normal. No grupo de pacientes com infertilidade secundária, 53,8% (35 pacientes) tiveram concentração espermática normal, 75,4% (49 pacientes) tiveram motilidade espermática normal e 32,3% (21 pacientes), morfologia normal. Nenhuma diferença significativa foi detectada na concentração espermática (21,3x10(6)/mL versus 23,1x10(6)/mL; p=0,07), motilidade (45,2 versus 48,1%; p=0,08) e morfologia (6,1 versus 6,4%; p=0,09) entre os grupos de pacientes com infertilidade primária e secundária. CONCLUSÕES: a análise seminal deve ser solicitada mesmo em casos de fertilidade masculina prévia. Os médicos não devem presumir que um paciente possui uma análise seminal normal, baseados no fato de este possuir história de estabelecimento de uma gravidez no passado.

Criopreservação de sêmen humano: comparação entre métodos de congelação e tipos de envase

OBJETIVOS: comparar duas diferentes técnicas de congelação e dois tipos de envase do sêmen humano durante processo de criopreservação. MÉTODOS: estudo experimental, no qual foi analisada a criopreservação de 18 amostras de sêmen de 18 voluntários. Após a adição de meio crioprotetor, "Test-yolk buffer" , as amostras de sêmen foram envasadas em palhetas com capacidade de 0,25 mL ou em criotubos de 2 mL e submetidas à criopreservação por dois métodos, um lento e outro rápido, totalizando quatro tratamentos distintos: RP (congelação pelo método rápido e envasado em palheta), RT (rápido-criotubo), LP (lento-palheta) e LT (lento-criotubo). As amostras, após 24 horas, foram descongeladas em temperatura ambiente e mantidas a 37ºC. Os dados coletados foram analisados através do teste t de Student, com p<0,05, utilizando o programa de computador SPSS for Windows® versão 11.0.0. RESULTADOS: houve redução da motilidade espermática após o processo de criopreservação. A taxa de motilidade inicial foi 58,1% e as motilidades após os diferentes métodos de criopreservação foram: 19,2% (RP), 27% (RT), 21,1% (LP) e 30,3% (LT). Houve redução significativa na morfologia normal. A taxa de morfologia normal inicial foi 14,2% e as morfologias após os diferentes métodos de criopreservação foram: 12,8% (RP), 12,6% (RT), 12,6% (LP) e 12,4% (LT). CONCLUSÕES: o método de criopreservação lento com envase em criotubo esteve associado à melhor motilidade espermática após o descongelamento. Não houve diferença entre os métodos quando avaliada a morfologia espermática.

Experimental poisoning by Baccharis megapotamica var. weirii in buffalo

Five male 6-8 month-old Murrah buffalo calves were orally dosed with the fresh aerial parts of Baccharis megapotamica var. weirii at doses of 1, 3, 4, 5 and 10g/kg body weight (bw) (~1-10mg macrocyclic trichothecenes/kg/bw). The B. megapotamica used for the experiment was harvested on a farm where a recent spontaneous outbreak of poisoning caused by such plant had occurred. Clinical signs appeared 4-20 hours and 4 buffaloes died 18-49 hours after the ingestion of the plant. Clinical signs were apathy, anorexia, and watery diarrhea, fever, colic, drooling, muscle tremors, restlessness, laborious breathing and ruminal atony, and dehydration. The most consistent gross findings were restricted to the gastrointestinal (GI) tract consisted of varying degrees of edema and reddening of the mucosa of the forestomach. Histopathological findings consisted of varying degrees of necrosis of the epithelial lining of the forestomach and of lymphocytes within lymphoid organs and aggregates. Fibrin thrombi were consistently found in sub-mucosal vessels of the forestomach and in the lumen of hepatic sinusoids. It is suggested that dehydration, septicemia and disseminated intravascular coagulation participate in the pathogenesis of the intoxication and play a role as a cause of death. A subsample of B. megapotamica var. weirii was frozen-dried and ground and analyzed using UHPLC (Ultra High Performance Liquid Chromatography) with high resolution Time of Flight mass spectrometry and tandem mass spectrometry, it was shown that the plant material contained at least 51 different macrocyclic trichothecenes at a total level of 1.1-1.2mg/g. About 15-20% of the total trichothecenes contents was found to be monosaccharide conjugates, with two thirds of these being glucose conjugates and one third constituted by six aldopentose conjugates (probably xylose), which has never been reported in the literature.

Comparison of two methods of tear sampling for protein quantification by Bradford method

The aim of this study was to compare two methods of tear sampling for protein quantification. Tear samples were collected from 29 healthy dogs (58 eyes) using Schirmer tear test (STT) strip and microcapillary tubes. The samples were frozen at -80ºC and analyzed by the Bradford method. Results were analyzed by Student's t test. The average protein concentration and standard deviation from tears collected with microcapillary tube were 4.45mg/mL ±0.35 and 4,52mg/mL ±0.29 for right and left eyes respectively. The average protein concentration and standard deviation from tears collected with Schirmer Tear Test (STT) strip were and 54.5mg/mL ±0.63 and 54.15mg/mL ±0.65 to right and left eyes respectively. Statistically significant differences (p<0.001) were found between the methods. In the conditions in which this study was conducted, the average protein concentration obtained with the Bradford test from tear samples obtained by Schirmer Tear Test (STT) strip showed values higher than those obtained with microcapillary tube. It is important that concentration of tear protein pattern values should be analyzed according the method used to collect tear samples.

Sperm morphological and morphometric evaluation in captive collared peccaries (Pecari tajacu)

The aim of this study was to compare different staining methods for the evaluation of sperm morphology by light microscopy and also to describe the morphometry of the entire sperm in collared peccaries (Pecari tajacu). Semen from 10 males was obtained by electroejaculation and evaluated for sperm motility, vigor, and concentration. Semen smears were prepared through three different staining methods: Bengal rose, brome-phenol blue, and eosin-nigrosin. Smears were evaluated under light microscopy and sperm morphologic alterations were determined in percentage. In addition, sperm morphometric analysis was conducted by light microscopy coupled to image analyzer software. The smears stained with Bengal Rose provide the best results for the visualization of the sperm tail, midpiece, and head. The use of eosin-nigrosin stain did not allow an adequate impregnation, and some sperm presented a few contrasts with the background. A higher incidence of bent coiled tails was verified in the use of brome-phenol blue staining (P<0.05). Through morphometric evaluation, it was observed that the tail occupies the greatest proportion (89%) of the sperm which presents a discretely elongated head. According to the results, the use of the Bengal Rose stain is recommended for the morphologic evaluation of the collared peccary sperm.

The seasonal and ovarian status effects on in vitro production of domestic cat embryos between Equator and Tropic of Capricorn

From the Tropic of Capricorn to Equator, the seasonality of domestic cat is known to be absent, i.e., these animals are considered non-seasonal breeders at these regions. We hypothesized that this particularity might have some influence on in vitro embryo production. The aim of this experiment was to determine the percentage of cleavage and morulae and blastocyst formation produced from oocytes recovered from queen ovaries of three distinct status - follicular, luteal or inactive - during two different reproductive seasons experienced by cats in southeast of Brazil (22°53'09" S and 48°26'42" W) - non breeding season (NBS), comprehending January to March; and breeding season (BS), August to October. Thirty queens were neutered. Ovaries were classified according to their status and were sliced in PBS for cumulus oocyte complex (COC) releasing. Grade I COC were washed three times in H-MEM supplemented with BSA, glutamine, sodium pyruvate, cysteine, streptomycin and penicillin. Oocytes were incubated in groups of 20-30 in 400µL of DMEM supplemented with FSH, LH, estradiol, IGF-I and basic fibroblast growth factor under mineral oil for 30 or 36 hours at 38°C in humidified environment of 5% de O2, 5% CO2 and 90% N2. COC were fertilized in Ham's F-10 medium supplemented with BSA, cysteine, pyruvate and streptomycin/penicillin (culture medium) with fresh semen selected through swim up technique. Eighteen hours later, the presumptive zygotes were denuded, the percentage of cleavage was determined and every 10 zygotes were transferred to 100mL drops of culture medium for culture during three days. After 72 hours of culture the percentage of morulae formation was evaluated and these structures were transferred to drops of the same culture medium. At the eighth day of culture blastocyst formation was analyzed. During NBS, from a total of 272 (inactive), 162 (luteal) and 134 (follicular) fertilized oocytes, the percentage of cleaved zygotes, morulae and blastocysts derived from inactive ovaries were 24.63, 16.54 and 8.09 respectively; for those derived from luteal ovaries, the percentage was 21.6, 12.96 and 8.64, and for those from follicular ovaries, they were 24.62, 16.41 and 8.21. Considering BS, from a total of 102 (inactive), 198 (luteal) and 86 (follicular) fertilized oocytes, the relative frequency (%) of cleaved zygotes, morulae and blastocysts derived from inactive ovaries were 64.7, 41.17 and 23.53 respectively; for those derived from luteal ovaries, the percentage was 64.14, 40.41 and 23.73, and for those from follicular ovaries, they were 63.95, 39.54 and 24.41. The results of this experiment demonstrate that no statistically significant difference (P<0.05) was verified in the frequency of cleaved embryos and morulae and blastocyst formation when comparing the three ovarian conditions in the same season. However the breeding season presented better results considering cleavage and morulae and blastocyst formation.