An investigation into the ecological niches and seasonal nature of Clostridium estertheticum and Clostridium gasigenes in the Irish beef farm environment.

Blown pack spoilage (BPS) of vacuum packaged beef is caused by psychrotolerant and psychrophilic Clostridium species, primarily Clostridium estertheticum and Clostridium gasigenes. The aim of this study was to investigate the environmental niches and impact of season on these BPS Clostridium spp. on Irish beef farms. On each of five different beef farms, faecal (10), soil (5), silage (5), air (5), bedding straw (5), drinking water (5) and puddle/ditch water (5) samples were collected during Spring, Summer, Autumn and Winter and tested for C. estertheticum and C. gasigenes using culture (direct plating and enrichment) and molecular (conventional PCR and quantitative PCR (qPCR)) based techniques. C. estertheticum and C. gasigenes were detected in all sample types, with qPCR detection rates ranging from 4% to 50% and at concentrations of up to 1·5 log10 CFU per g and 3·5 log10 CFU per g, respectively. The impact of season was not clear as the results were mixed depending on the detection method used. It was concluded that BPS-causing C. estertheticum and C. gasigenes are widely distributed in the beef farm environment.

Distinct beneficial effects of continuous vs accumulated exercise training on cardiovascular risk factors in Wistar rats.

We compared the effects of continuous exercise (CE) vs accumulated exercise (AE) training on CVD risk factors and heart of young male Wistar rats. The exercise training (ET) was performed in a swimming pool for 30-60 min/day, 5 days/week over 15 weeks. CE group performed the ET in a single long daily session (30-60 min), while AE group performed the ET at the same frequency, intensity, and duration of CE rats, but in three short bouts over the course of a day (10-20 min in three daily sessions). AE training was more efficient than CE in attenuating body and fat weight gain and inhibiting visceral adipocyte hypertrophy at the same food intake level. CE training was more efficient in improving systolic blood pressure, LDL/HDL cholesterol, and serum triglyceride. Both ET protocols increased heart function, decreased lipid peroxidation, and increased intracellular Hsp72 content in the heart. This work shows distinct beneficial effects of CE vs AE training suggesting that the prescription of one or other may be preferred to prevent the increase of a specific CVD risk factor.

Caloric restriction-induced weight loss with a high-fat diet does not fully recover visceral adipose tissue inflammation in previously obese C57BL/6 mice.

Caloric restriction (CR) reduces body weight and systemic inflammation, but the effects on adipose tissue under dietary lipid overload are controversial. We evaluated the effects of CR-induced weight loss with a high-fat diet on adipose tissue inflammation of obese mice. Male mice were assigned into low-fat diet (LF) and high-fat diet (HF) groups. After 8 weeks, the mice in the HF group were reassigned for another 7 weeks into the following 3 conditions: (i) kept in the HF condition; (ii) changed to low-fat diet ad libitum (LFAL); and (iii) changed to high-fat calorie-restricted (RHF) diet to reach LFAL body weight. Serum markers, adipocytokines, morphology, and inflammatory infiltrates in retroperitoneal adipose tissue (RAT) were accessed. The body weights of the LFAL and RHF groups were reduced, equaling the body weights of the LF group. The LFAL mice had restored almost all inflammatory markers as the LF mice, except tumor necrosis factor-alpha (TNF-α), monocyte chemoattractant protein-1 (MCP-1), and adiponectin. Compared with the HF group, the RHF group had lowered visceral adiposity, retroperitoneal adipocyte sizes, and RAT inflammatory cell infiltration, as well as TNF-α, interleukin-6, and hepatic and serum C-reactive protein, which were higher than that of the LFAL group; adiponectin and MCP-1 did not change. CR with high-fat diet reduced body weight and attenuated visceral adiposity but did not fully recover visceral tissue inflammation. Novelty Caloric restriction in a high-fat diet ameliorated visceral adiposity. Caloric restriction in a high-fat diet did not recover visceral adipose tissue inflammation.

Propagation of a Brazilian isolate of Anaplasma marginale with appendage in a tick cell line (BME26) derived from Rhipicephalus (Boophilus) microplus.

Anaplasma marginale is a tick-borne pathogen of cattle responsible for the disease anaplasmosis. Data suggest that Rhipicephalus (Boophilus) microplus and R. annulatus may be the major tick vectors of A. marginale in tropical and subtropical regions of the world. In this work we demonstrated the first infection and propagation of a Brazilian isolate of A. marginale (UFMG1) in the BME26 cell line derived originally from embryos of R. (Boophilus) microplus. The establishment of A. marginale infection in a cell line derived from R. (Boophilus) microplus is relevant for studying the A. marginale/tick interface.

Low temperature affects cattle tick reproduction but does not lead to transovarial transmission of Anaplasma marginale.

Anaplasma marginale is an obligate intracellular pathogen that infects the erythrocytes of calves, causing bovine anaplasmosis. This rickettsia is biologically transmitted by several species of ticks. In tropical and subtropical regions of the world, Rhipicephalus microplus is the main vector. Due to their mobility and longevity, the adult males play an important role in the transmission of A. marginale to calves. Some studies have demonstrated that A. marginale can be intrastadially and interstadially transmitted in R. microplus, but the transovarial transmission has not been demonstrated so far. In the present study, we investigated the effects of low temperature on both the A. marginale migration from infected females to their offspring and reproductive parameters of the tick R. microplus. The larvae of R. microplus fed on a calf infected with the strain Jaboticabal of A. marginale. At the end of the parasitic phase, fully engorged females were incubated at either 18°C or 28°C for oviposition. Although A. marginale was detected in the salivary glands of the females, demonstrating that the ticks were successfully infected, the presence of rickettsia was not detected in the offspring. However, the preoviposition period of the non-infected females maintained at 18°C was longer than that of those maintained at 28°C. In addition, the average weight of the mass of eggs as well as the egg production efficiency (ratio of the egg mass weight to the female weight) of the females maintained at 18°C were significantly lower than those of the females incubated at 28°C. There was no larval hatching from the eggs maintained exclusively at 18°C, even at 65 days after female detachment. Hatching occurred only when the eggs maintained at 18°C were transferred to 28°C at 20 days after female detachment (18°C/28°C). We also verified a significantly higher larvae conversion efficiency (ratio of the larvae mass weight to the egg mass weight) in the group of females maintained exclusively at 28°C compared to those from the 18°C/28°C group. Collectively, our results reinforce that low temperature exerts negative effects on female fertility and egg development in R. microplus, although it has no influence on A. marginale transmission to the progeny. In the field, the detrimental effects of temperatures on tick reproductive fitness lead to a reduction of tick population, which may cause a decrease in the incidence of bovine anaplasmosis.

Crossed testicular ectopia.

Crossed testicular ectopia (CTE) is a rare anomaly, characterized by migration of one testis towards the opposite inguinal canal. Presented here is a case of crossed ectopia of the right testis, treated by extraperitoneal transposition of the gonad and right orchiopexy. Embriology and surgical findings suggest that CTE is a common consequence of many unclear ethiologic factors, specially mechanical ones, and can be associated with Muller duct persistence. Review of literature suggests a classification of CTE into 3 types: I--associated with inguinal hernia alone; II--associated with persistent mullerian remnants; III--associated with other anomalies without mullerian remnants. Treatment includes transeptal orchiopexy or extraperitoneal transposition of the testis, research for mullerian remnants and other anomalies, and long term postoperative follow-up, due to the risk of becoming malignant.

[Hepatic mesenchimal hamartoma in neonates]. / Hamartoma mesenquimal hepático no recém-nascido.

OBJECTIVES: To approach the diagnosis and management of two different features of hepatic mesenchymal hamartoma (HMH) in the neonate. METHODS: Report of two cases of HMH in newborns and review of the literature (MEDLINE and LILACS). RESULTS: Two cases of newborns with huge abdominal masses are presented, one of them complicated by rupture and hemorrhage during ultrasound manipulation, leading to hypovolemic shock. Urgent operation showed a ruptured giant hepatic cyst containing hemorrhagic yellowish fluid. The cyst was removed by edge resection at the level of hepatic segment 6 and the postoperative period was uneventful. The pathological analysis of the cyst including immunohistochemical tests revealed HMH. In case 2, with prenatal diagnosis of hepatic tumor, the mass progressively enlarged after birth, and a left hepatectomy was performed. The macroscopic appearance suggested hemangioma but the histopathological aspect indicated mesenchymal hamartoma with endothelial predominance. Nowadays both children are healthy and asymptomatic. CONCLUSIONS: The management of HMH is surgical treatment, because it does not disappear spontaneously. Some cases evolve to giant forms even in the neonatal period leading to fatal complications. Preoperative differential diagnosis from hemangioma is quite difficult for cystic types, and immunohistochemical analysis is important for pathological confirmation.

[WinDiet. Support decision system for the assessment of nutritional status and diet prescription]. / WinDiet. Sistema de apoio à decisão para avaliação do estado nutricional e prescrição de dietas.

A "Sistema de Apoio à Decisão para Avaliação do Estado Nutricional e Prescrição de Dietas" called "WinDiet" was developed. The system was developed on a computer compatible to IBM-PC standard. For system processing procedures through the computer, that is, interpretation, compilation, table generation en others, it was selected the Delphi version 1.0 software, from Borland International, for its using facilities, data basis accomplishment, communication environment and mainly an ample availability of mathematic, impression and programming resources besides allowing for all the programming operations to be accomplished on Windows environment. Knowing the procedures for nutritional evaluation and diet prescription was possible by consulting to text books, periodicals and the area experts. This system allows for individual nutritional assessment applying the anthropometric methods such as weight, height, skinfold thickness measurings, the lab methods such as albumin, transferrin, height/creatinin index and others, the dietary methods by food consumption recall, and the clinical methods that give emphasis to pathological antecedents and indicative sings of nutritional status. The software also turns possible to prescribe and calculate diets from recommendation besides having an actual data basis which may be changed or modified. Yet it maintains the individual description from all evaluation and menus, emitting its complete reports. It was verified that it reduce significantly the working time and increase the precision of estimate and nutritional diagnosis. Thus, the WinDiet may be used in nutritionist's offices, ambulatories, hospitals as well as on nutritional researches and education.

Energy expenditure: components and evaluation methods.

INTRODUCTION: The determination of energy expenditure, considering the physical activity level and health status, is very important to adjust the individuals' nutritional supply. Energy expenditure can be determined by using indirect calorimetry, bioelectrical impedance, doubly labeled water, predictive equations, among others. All these methods have been used in clinical and research areas. However, considering the inconsistence in several research results, there is no consensus yet about the applicability of many of these methods. OBJECTIVES: The aim of this review is to describe the components of energy expenditure and the methods for its determination and estimation, summarizing their main advantages and limitations. RESULTS AND DISCUSSION: Indirect calorimetry and doubly labeled water are considered more accurate methods, but expensive. On the other hand, even though other methods present limitations, they are convenient and less expensive, and can be used with some caution.

Induction of a gloverin-like antimicrobial polypeptide in the sugarcane borer Diatraea saccharalis challenged by septic injury.

Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) is an important pest for Brazilian sugarcane. In the present study, we detected two distinct spots in hemolymph from septic injured larvae (HDs1 and HDs2), which are separated by 2DE gel electrophoresis. Both spots were subjected to in-gel tryptic digestion and MALDI-TOF/TOF analysis, which revealed the sequence VFGTLGSDDSGLFGK present in both HDs1 and HDs2. This sequence had homology and 80% identity with specific Lepidoptera antimicrobial peptides called gloverins. Analyses using the ImageMaster 2D software showed pI 8.94 of the HDs1 spot, which is similar to that described to Hyalophora gloveri gloverin (pI 8.5). Moreover, the 14-kDa molecular mass of the spot HDs1 is compatible to that of gloverins isolated from the hemolymph of Trichoplusia ni, Helicoverpa armigera and H. gloveri. Antimicrobial assays with partially purified fractions containing the HDs1 and HDs2 polypeptides demonstrated activity against Escherichia coli. This is the first report of antimicrobial polypeptides in D. saccharalis, and the identification of these peptides may help in the generation of new strategies to control this pest.

Antimicrobial activity in the tick Rhipicephalus (Boophilus) microplus eggs: Cellular localization and temporal expression of microplusin during oogenesis and embryogenesis.

Arthropods display different mechanisms to protect themselves against infections, among which antimicrobial peptides (AMPs) play an important role, acting directly against invader pathogens. We have detected several factors with inhibitory activity against Candida albicans and Micrococcus luteus on the surface and in homogenate of eggs of the tick Rhipicephalus (Boophilus) microplus. One of the anti-M. luteus factors of the egg homogenate was isolated to homogeneity. Analysis by electrospray mass spectrometry (ESI-MS) revealed that it corresponds to microplusin, an AMP previously isolated from the cell-free hemolymph of R. (B.) microplus. Reverse transcription (RT) quantitative polymerase chain reactions (qPCR) showed that the levels of microplusin mRNA gradually increase along ovary development, reaching an impressive highest value three days after the adult females have dropped from the calf and start oviposition. Interestingly, the level of microplusin mRNA is very low in recently laid eggs. An enhance of microplusin gene expression in eggs is observed only nine days after the onset of oviposition, achieving the highest level just before the larva hatching, when the level of expression decreases once again. Fluorescence microscopy analysis using an anti-microplusin serum revealed that microplusin is present among yolk granules of oocytes as well as in the connecting tube of ovaries. These results, together to our previous data, suggest that microplusin may be involved not only in protection of adult female hemocele, but also in protection of the female reproductive tract and embryos, what points this AMP as a considerable target for development of new methods to control R. (B.) microplus as well as the vector-borne pathogens.

Zinc and iron bioavailability of genetically modified soybeans in rats.

The aim of this work was to evaluate zinc and iron bioavailability of UFV-116, a new variety without 2 lipoxygenases, with better taste and flavor than a commercial variety OCEPAR 19, containing all 3 isozymes. To evaluate zinc absorption using 65Zn whole body retention and femur 65Zn uptake, rats were given 3 g of a 65ZnCl2 labeled test meal (0.25 microCi). The 2 varieties were tested at the level of 9 and 30 ppm of zinc as defatted soy flour. Two other groups (control) received egg white as source of protein and ZnS04.H20 as the zinc source. To evaluate iron absorption, using 59Fe whole body retention, animals were given a 3 g 59FeCl3 labeled test meal (0.2 microCi). The 2 varieties were tested at 12 and 25 ppm iron as defatted soy flour. Whole fat soy flour of variety 1 (UFV-116) was higher (P < 0.05) in Ca, K, Mg, phytic acid, and oxalate than variety 2 (OCEPAR-19). No difference was observed among the soybean varieties (P > 0.05) for femur 65Zn retention, at different levels of zinc. However, whole body retention was lower (P < 0.05) for UFV-116 than for OCEPAR-19. Femur 65Zn uptake was correlated with the whole body retention; however, whole body retention was more sensitive. Whole body 59Fe retention from UFV-116 was lower (P < 0.05) than from OCEPAR-19. Zinc and iron bioavailability was lower for UFV-116, possibly due to its higher content of antinutrient factors, especially phytate.

Induction of a gloverin-like antimicrobial polypeptide in the sugarcane borer Diatraea saccharalis challenged by septic injury

Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) is an important pest for Brazilian sugarcane. In the present study, we detected two distinct spots in hemolymph from septic injured larvae (HDs1 and HDs2), which are separated by 2DE gel electrophoresis. Both spots were subjected to in-gel tryptic digestion and MALDI-TOF/TOF analysis, which revealed the sequence VFGTLGSDDSGLFGK present in both HDs1 and HDs2. This sequence had homology and 80 percent identity with specific Lepidoptera antimicrobial peptides called gloverins. Analyses using the ImageMaster 2D software showed pI 8.94 of the HDs1 spot, which is similar to that described to Hyalophora gloveri gloverin (pI 8.5). Moreover, the 14-kDa molecular mass of the spot HDs1 is compatible to that of gloverins isolated from the hemolymph of Trichoplusia ni, Helicoverpa armigera and H. gloveri. Antimicrobial assays with partially purified fractions containing the HDs1 and HDs2 polypeptides demonstrated activity against Escherichia coli. This is the first report of antimicrobial polypeptides in D. saccharalis, and the identification of these peptides may help in the generation of new strategies to control this pest.

Increased natural killer cell activity in uterine cervix cancer patients undergoing radiation therapy.

Natural killer (NK) cell activity in peripheral blood mononuclear cells (PBMCs) from women with carcinoma of the uterine cervix was studied using a 4-hour 51Cr release assay and K562 cells as the sensitive target. PBMCs were obtained from 21 previously untreated patients at different stages of disease according to the International Federation of Gynecology and Obstetrics classification. PBMCs were also obtained from 36 patients treated with radiation therapy at different disease stages. Seventeen healthy age-matched volunteer women were used as controls. Mean cytotoxicity for the normal donors was 25.1 +/- 6.56% at a 100:1 effector-target cell ratio, 33.8 +/- 7.96% for the previously untreated cancer patients and 52 +/- 18.4% for the treated cancer patients. Most of the treated patients (86%) showed a complete response to radiation therapy and the mean cytotoxicity of the whole group (36 treated patients) was significantly increased compared to controls (p < 0.05). It is suggested that radiation therapy may produce cell alterations leading to an increase in NK cell activity in patients treated for uterine cervical cancer. The significance of this increase is discussed.

Natural killer cell activity in Hodgkin's disease patients undergoing radiation therapy or chemotherapy and radiation therapy.

Natural killer (NK) cell activity in peripheral blood mononuclear cells (PBMCs) from patients with Hodgkin's disease was studied using 4 h 51Cr release assay and K562 cells as sensitive targets. PBMCs were obtained from 15 previously untreated patients at different stages of their disease. PBMCs were also obtained from 46 patients treated by radiation therapy or combined chemotherapy and radiation therapy. Twenty healthy age-matched volunteer donors were used as controls to the treated patients. For these normal donors the mean cytotoxicity was 24.8 +/- 5.67% at a 100:1 effector-target cell ratio; and 43.7 +/- 12.1% for the treated cancer patients. Fifteen healthy age-matched volunteer donors were used as controls to the untreated patients. The mean cytotoxicity for these normal donors was 20.8 +/- 3.61% at a 100:1 effector-target cell ratio; and 37.6 +/- 6.65% for the previously untreated cancer patients. The mean cytotoxicity for all 35 normal donors was 23.1 +/- 5.22% at a 100:1 effector-target cell ratio. Most treated patients (93.5%) had a complete response to therapy and a significant difference was found between the mean cytotoxicity of the whole group (46 treated patients), compared with controls (P < 0.001). A significant difference (P < 0.05) was also observed when the same 11 patients were studied before and after treatment.

[Applications of videolaparoscopic surgery in children] / Aplicações da cirurgia videolaparoscópica em crianças.

OBJECTIVES: To present the videolaparoscopies performed by the Childrens Surgery Service, and study the main indications for pediatric laparoscopic surgeries, considering advantages and disadvantages over conventional open procedures. METHODS: Retrospective analysis of 612 videolaparoscopies in children aged between 8 days and 17 years treated from November/95 to 2000. Basic principles of videolaparoscopy and the postoperative management of several pediatric diseases are described. The results, advantages, and complications were analyzed after a 5-year follow-up period. RESULTS: Laparoscopic surgery allowed for a wide series of abdominal procedures conventionally carried out through open surgery, mainly for the treatment of gastroesophageal reflux disease, inflammatory acute abdomen, adhesive intestinal obstruction, biliary lithiasis, tumors, cryptorchidism, ovarian diseases, splenectomies, aganglionosis, trauma and others. Morbidity was low (1%), and mortality due to laparoscopy was nonexistent. Conversion to laparotomy occurred in only 14 cases (2.3%), mainly because of trauma. The principal advantages included minimal surgical trauma, pain and reflex ileum, short hospital stay, almost no incisional hernias and better cosmetic scars. CONCLUSIONS: Videolaparoscopy seems to be a great advance in modern pediatric surgery, allowing safer and less invasive treatment of a wide series of abdominal diseases at all pediatric ages.

Clinical implications of natural killer cytotoxicity in patients with squamous cell carcinoma of the penis.

Impairment of natural cytotoxicity mediated by natural killer (NK) cells may play a role in the pathogenesis of penile carcinoma. The aim of this study was to examine the NK activity profile and its prognostic significance in patients with squamous cell carcinoma of the penis. The NK activity was measured in peripheral blood mononuclear cells (PBMCs) from 39 patients diagnosed histologically as having invasive squamous cell penile carcinoma and 4 patients with verrucous carcinoma of the penis. Of 39 patients with invasive squamous cell carcinoma, 4 had undergone previous penile amputation. According to the prognosis, the patients with invasive squamous cell carcinoma were divided into two groups: with metastasis and without metastasis. The patients were evaluated in relation to clinicopathologic variables using univariate analyses. NK cell activity was significantly decreased in all patients with penile carcinoma when compared with the control groups (p < 0.0001). There was no statistically significant difference between the groups with and without metastasis. We conclude that there is a decrease in NK activity in PBMCs from patients with penile carcinoma and that the presence of advanced disease or metastatic involvement is not responsible for this reduction.

Aspiración esofágica contínua en atresia de esófago: utilización del principio de Venturi / Constant esophageal suctioning in esophageal atrsia: usage of the Venturi principle

En los neonatos con atresia de esófago uno de los cuidados preoperatorios fundamentales es la aspiración de saliva del esófago proximal para evitar las complicaciones broncopulmonares,importante causa de morbimortalidad.La aspiración continua es más eficiente y segura que la aspiración intermitente y generalmente se realiza por medio de vacío.Se presenta un sistema de aspiración continua utilizando el principio de venturien Y o en T,sin uso de energía eléctrica o bombas de vació,de bajo costo,fácil manejo por enfermería y que brinda seguridad para los recién nacidos con atresia de esófago.El sistema de aspiración en Y fue aplicado en 24 pacientes,evitando la acumulación de saliva en la orofaringe y sin complicaciones broncopulmonares,El sistema se asoció a posición semisentada,antibioticoterapia y eventual gastrotomía.Dos niños con grandes fístulas traqueosesofágicas presentaron broncoaspiración por reflujo gástrico.Una niña murió por cardiopatía al tercer día de vida.Seis niños ya presentaban neumonía o atelectasis en la admisión y de éstos,3 mejoraron durante el uso del sistema Venturi.Los otros 18 pacientes no presentaron ninguna complicación por broncoaspiración.La aspiración del sistema Venturi es eficiente,económico,seguro,de fácil manejo y evita las complicaciones pulmonares en niños con atresia de esófago

Aspiración esofágica contínua en atresia de esófago: utilización del principio de Venturi / Constant esophageal suctioning in esophageal atrsia: usage of the Venturi principle

En los neonatos con atresia de esófago uno de los cuidados preoperatorios fundamentales es la aspiración de saliva del esófago proximal para evitar las complicaciones broncopulmonares,importante causa de morbimortalidad.La aspiración continua es más eficiente y segura que la aspiración intermitente y generalmente se realiza por medio de vacío.Se presenta un sistema de aspiración continua utilizando el principio de venturien Y o en T,sin uso de energía eléctrica o bombas de vació,de bajo costo,fácil manejo por enfermería y que brinda seguridad para los recién nacidos con atresia de esófago.El sistema de aspiración en Y fue aplicado en 24 pacientes,evitando la acumulación de saliva en la orofaringe y sin complicaciones broncopulmonares,El sistema se asoció a posición semisentada,antibioticoterapia y eventual gastrotomía.Dos niños con grandes fístulas traqueosesofágicas presentaron broncoaspiración por reflujo gástrico.Una niña murió por cardiopatía al tercer día de vida.Seis niños ya presentaban neumonía o atelectasis en la admisión y de éstos,3 mejoraron durante el uso del sistema Venturi.Los otros 18 pacientes no presentaron ninguna complicación por broncoaspiración.La aspiración del sistema Venturi es eficiente,económico,seguro,de fácil manejo y evita las complicaciones pulmonares en niños con atresia de esófago