Immunohistochemical expression of HER - 2 and Ki - 67 in granulosa cell tumor in bitches

Granulosa cell tumour, an ovarian neoplasm of stromal origin, is an important tumour related to oestrogenic dominance syndrome and cystic endometrial hyperplasia–pyometra complex. In order to analyse ovarian tumour´s malignant potential, immunohistochemical markers can be used, such as anti-HER2 and anti-Ki-67. The aim of this study was to evaluate the expression of immunohistochemical markers HER-2 and Ki-67 in granulosa cell tumour from bitches´ ovaries. In HER-2 immunomarker analysis using the HercepTest^® method, most tumours were classified as 2+ (moderate labelling). Concerning Ki-67 immunomarker, only one case was described as having a high proliferative index. An association was found between immunostained cell percentage by anti-HER-2 antibodies and high pleomorphism, represented by the pattern of follicular/trabecular tumour arrangement. There was no correlation between anti-Ki-67 and anti-HER-2 antibody immunostaining intensities, probably due to only one case with a high Ki-67 index. With an effective protocol for HER-2 and Ki-67 immunohistochemical identification in granulosa cell tumours in bitches, it was possible to characterize this neoplasm proliferation profile.     

Risk factors for L. monocytogenes contamination of dairy products in Switzerland, 1990-1999

Our purpose was to identify the main hazards associated with the spread of Listeria monocytogenes in dairy products in Switzerland and to determine the changes in predominant serotypes of the isolates, using databases on dairy-processing and environments from the Swiss Dairy Research Station during the years 1990-1999. Overall, of 76,271 samples collected, 3722 (4.9%) were positive for the presence of L. monocytogenes. Cheese-ripening facilities had the highest proportion of positive samples (7.6%), followed by small-scale local dairies (4.4%). By sample type, the highest proportion of positive samples (9.5%) was observed in water samples used for cheese-washing, followed by cheese-surface swabs (5.0%). During the 10-year period, no positive samples were obtained from cream, ice cream, milk powder, yogurt, or fresh cheese. Of 3722 L. monocytogenes isolates, 1328 (35.7%) were serologically typeable. Serotypes 1/2a, 1/2b, and 4b accounted for 92.7% of the 1328 isolates. Until 1995, the most-prevalent serotype was 1/2b (annual proportional prevalence 39.3-72.2%)--whereas since 1996, 1/2a was the most prevalent (34.7-54.7%). During 1996-1999, serotype 1/2a increased by 88%, compared to the average of 1990-1995. In the final random-effect multivariable logistic model, the strongest predictor of a positive culture was samples from cheese-ripening plant (OR=1.54; 95% CI: 1.14, 2.08) and the second-strongest predictor was samples collected by someone who was employed by the plant (OR=1.48; 1.29, 1.71). Hard and semi-hard cheeses were more likely to be associated with serotype 1/2b and soft cheeses with serotype 1/2a.     

Die bodenbiologische Bedeutung von Collembolen und ihre Eignung als Testorganismen für die Ökotoxikologie

It is the purpose of this study to determine the suitability of arthropods living on the ground for ecotoxicological tests, so that subacute toxicity tests may be carried out in them. Among these, also Collembola are tested for the suitability. They belong to the group of primarily wingless insects (Apterygota) having a length of 0.5 to 5 mm. They are slightly sclerotized and mainly living on the ground. As test organisms they meet the following requirements:

1. Presence in all types of soil (Collembola live, very often in great numbers, on any soil in any climate).
2. Importance for soil biology (due to their great number and their omnivorous way of nutrition they play a considerable role in the treatment and transformation of wastes of vegetal origin).
3. Easy breeding (so that they are constantly available for tests; many species of Collembola are easy to breed, may be bred in large quantities and at relatively low expenditure on time and equipment).
4. Short period of development and high reproduction rate (primarily, the smaller species of Collembola have short periods of development and high reproduction rates when adequate temperatures are maintained).
5. Controllability (which is guaranted by appropriate selection and conservation methods).

The advantages mentioned above indicate that Collembola are very well suited for ecotoxicological tests of any kind. Practical testing at a larger scale, however, is still to be carried out.     

In vivo bioinsecticidal activity toward Ceratitis capitata (fruit fly) and Callosobruchus maculatus (cowpea weevil) and in vitro bioinsecticidal activity toward different orders of insect pests of a trypsin inhibitor purified from tamarind tree (Tamarindus indica) seeds

A proteinaceous inhibitor with high activity against trypsin-like serine proteinases was purified from seeds of the tamarind tree (Tamarindus indica) by gel filtration on Shephacryl S-200 followed by a reverse-phase HPLC Vidac C18 TP. The inhibitor, called the tamarind trypsin inhibitor (TTI), showed a Mr of 21.42 kDa by mass spectrometry analysis. TTI was a noncompetitive inhibitor with a Ki value of 1.7 x 10(-9) M. In vitro bioinsecticidal activity against insect digestive enzymes from different orders showed that TTI had remarkable activity against enzymes from coleopteran, Anthonomus grandis (29.6%), Zabrotes subfasciatus (51.6%), Callosobruchus maculatus (86.7%), Rhyzopertha dominica(88.2%), and lepidopteron, Plodia interpuncptella (26.7%), Alabama argillacea (53.8%), and Spodoptera frugiperda (75.5%). Also, digestive enzymes from Diptera, Ceratitis capitata (fruit fly), were inhibited (52.9%). In vivo bioinsecticidal assays toward C. capitata and C. maculatus larvae were developed. The concentration of TTI (w/w) in the artificial seed necessary to cause 50% mortality (LD50) of larvae was 3.6%, and that to reduce mass larvae by 50.0% (ED50) was 3.2%. Furthermore, the mass C. capitata larvae were affected at 53.2% and produced approximately 34% mortality at a level of 4.0% (w/w) of TTI incorporated in artificial diets.     

Genetic association between different criteria to define sexual precocious heifers with growth,carcass, reproductive and feed efficiency indicator traits in Nellore cattle using genomic information

The aim of this study was to estimate genetic parameters for different precocious calving criteria and their relationship with reproductive, growth, carcass and feed efficiency in Nellore cattle using the single‐step genomic BLUP. The reproductive traits used were probability of precocious calving (PPC) at 24 (PPC24), 26 (PPC26), 28 (PPC28) and 30 (PPC30) months of age, stayability (STAY) and scrotal circumference at 455 days of age (SC455). Growth traits such as weights at 240 (W240) and 455 (W455) days of age and adult weight (AW) were used. Rib eye area (REA), subcutaneous fat thickness (SFT), rump fat thickness (RFT) and residual feed intake (RFI) were included in the analyses. The estimation of genetic parameters was performed using a bi‐trait threshold model including genomic information in a single‐step approach. Heritability for PPC traits was moderate to high (0.29–0.56) with highest estimates for PPC24 (0.56) and PPC26 (0.50). Genetic correlation estimates between PPC and STAY weakened as a function of calving age. Correlation with SC455, growth and carcass traits were low (0.25–0.31; ?0.22 to 0.04; ?0.09 to 0.18, respectively), the same occurs with RFI (?0.09 to 0.08), this suggests independence between female sexual precocity and feed efficiency traits. The results of this study encourage the use of PPC traits in Nellore cattle because the selection for such trait would not have a negative impact on reproductive, growth, carcass and feed efficiency indicator traits. Stayability for sexual precocious heifers (PPC24 and PPC26) must be redefined to avoid incorrectly phenotype assignment.     

Histopathological findings in the early diagnosis of granulosa cell tumour in bitches

Granulosa cell tumour (GCT) is a majorly observed ovarian tumour in female dogs. It is essential to diagnose GCT in its initial phase before any symptoms occur, as histological and physiological differences may be observed based on the evolution of this neoplasia. This study aimed to analyse the anatomic histopathology of GCT in its initial stage, with findings of ovaries not yet with the suspicion of neoplasms in the Canis familiaris. A sample including 55 ovaries presented GCT in 40 female dogs. The histopathological analysis was performed considering the intensity of pleomorphism, vascularization and inflammatory infiltrate. Furthermore, we evaluated the mitoses count in 10 fields using 40× magnification. Out of the 40 animals evaluated, 62.5% (25/40) presented the tumour in only one ovary. The Call‐Exner corpuscle was present in 65% (26/40) of the cases. The follicular histological pattern was present in 52.5% (21/40) of the animals. The presence of the Call‐Exner bodies and the degree of tumour cell pleomorphism (p = 0.033) were associated. Moreover, the degree of vascularization and the intensity of the inflammatory infiltrate were also related (p = 0.001). In addition, there was a positive relationship between the increase in pleomorphism and the mean age of the animals (p = 0.044). This study confirmed that the appearance of this tumour may precede any clinical symptomatology. In this study, the most frequent histopathological pattern was the follicular. The characteristics of the granulosa cell tumour diagnosed early were poorly pleomorphic cells, low mitotic index and presence of Call‐Exner body.     

Are active organic matter fractions suitable indices of management effects on soil carbon? A meta-analysis of data from the Pampas

Active fractions of organic matter have been proposed as early indices of changes in soil organic carbon (organic-C) induced by management. We performed a meta-analysis of published results from 31 field experiments conducted in the Pampas in which tillage and rotation effects on organic-C, microbial biomass carbon (microbial-C), light fraction carbon (light-C), particulate carbon (particulate-C) and basal respiration (mineralized-C) were assessed. We compared the changes of organic-C and the four active fractions between management treatments sampled at the same date and depth within each experiment. Pooling all the experiments, active fractions-C varied on average 1.2- to 3.0-fold more than organic-C, depending on the fraction considered, but these average changes were significantly greater than organic-C changes only for particulate-C. This later fraction showed to be more sensitive to agricultural practices than organic-C. In experiments in which organic-C changes were lower than 15–25%, the four labile fractions may show opposite trends than organic-C. Above this threshold, changes of active fractions generally copied organic-C changes and were greater. Consequently, the active fractions may be used as indicators of changes in organic-C only when this latter variable has already suffered a huge change. In these cases, it will be easier to simply measure organic-C.     

In vitro antiviral activity of dehydroepiandrosterone and its synthetic derivatives against vesicular stomatitis virus

In this work the antiviral activity of 20 dehydroepiandrosterone (DHEA) analogs with different substituents at positions C-3, C-15, C-16 and C-17 were evaluated against vesicular stomatitis virus (VSV) in Vero cell cultures. The selectivity indexes (SI) obtained with DHEA and epiandrosterone (EA) were 50 and 72.6, respectively. The work showed that the compounds 21-norpregna-5,17(20)-dien-3β,16α-diyl-diacetate, 17,17-ethylendioxyandrostan-5,15-dien-3β-ol and 3β-hydroxypregn-17(20)-en-16-one had higher SI values than ribavirin, which was used as a reference drug. The antiviral mode of action of DHEA was also investigated against VSV replication in Vero cells, and time of addition experiments showed that DHEA mainly affected a late event in the virus growth cycle. Analysis of RNA and protein synthesis indicated that DHEA adversely affected positive strand RNA synthesis and viral mature particle formation.     

Photoperiod and Soil Munition Constituent Effects on Phytoaccumulation and Rhizosphere Interactions in Boreal Vegetation

Permafrost thaw is expected to alter biogeochemistry and hydrology, potentially increasing the mobility of soil constituents. Northern latitude boreal forests where permafrost thaw is occurring also experience extreme changes in day length during the growing season. As the effects of photoperiod on plant uptake of soil constituents or interactions with the rhizosphere are unknown, our objective was to determine these interactions with three plant species from different functional groups. A tree, forb, and grass common to military training ranges in this region were grown in soil spiked with or without lead, antimony, or 2,4-dinitrotoluene and grown under 16, 20, or 24 h of light. Plant biomass, soil constituent uptake, and rhizosphere bacterial communities were compared between treatments. Photoperiod had no effect on plant uptake of any soil constituent or on rhizosphere community, indicating that plants and their associated microbial communities adapted to this environment are resilient to extremes in photoperiod. Lead uptake was not significant in any plant species and had no effect on the rhizosphere. Antimony increased the percentage composition of Saprospirales in the rhizospheres of two of the three plants, indicating an interaction between this bacterial order and antimony. Antimony uptake by white spruce (Picea glauca) was considerable, with a mean concentration of 1731 mg kg^?1 in roots, while mean shoot concentration was only 155 mg kg^?1, indicating its potential to phytostabilize this heavy metal. Although antimony had the strongest impact on the rhizosphere bacterial community, it was also readily accumulated by the grass and tree.