Regulation of T cell homeostasis during fetal and early postnatal life

Before parturition the fetal lamb develops a large pool of long-lived recirculating T cells which provides a large population of naive T cells with a diverse TcR repertoire. After birth and concomitant with exposure to environment antigens, fetal T cells are rapidly replaced by short-lived cells formed postnatally. The majority of thymic emigrants homing to spleen in postnatal lambs are short-lived, in contrast to emigrants targeting lymph nodes where a population appears to be long-lived. The lifespan of thymic emigrants in the fetus is unknown as in the relative importance of antigen-driven processes versus developmental programming in regulating T cell homeostasis in early postnatal life.     

The detection of CD2+, CD4+, CD8+, and WC1+ T lymphocytes, B cells and macrophages in fixed and paraffin embedded bovine tissue using a range of antigen recovery and signal amplification techniques

In order to develop procedures to label the main bovine leucocyte populations in paraffin embedded sections, the immunoreactivity of 25 monoclonal antibodies (mAbs) to different leucocyte antigens was assessed with formal dichromate (FD5) and 10% formalin fixation, a battery of antigen retrieval (AR) methods, and the biotin-tyramide amplification system. All the leucocyte populations investigated (CD2+, CD4+, CD8+, WC1+ T lymphocytes, B cells and macrophages) were strongly and specifically detectable under an appropriate combination of mAb, AR method and signal amplification system. CD4 and CD8 required the most stringent conditions and could only be demonstrated in FD5 fixed sections. For detection of CD2, WC1+ T lymphocytes, B cells and macrophages, all the mAbs produced immunoreactivity in FD5 or formalin fixed tissues. The need to check a range of different AR methods is stressed, as the method of choice varied for each individual mAb. The incorporation of the signal amplification system was necessary to observe a strong signal and the complete distribution of CD4, CD8 and B cells. Fixation by FD5 proved to be better than formalin for the preservation of surface antigens but it was inferior for the detection of markers which were found to show cytoplasmic immunoreactivity, such as the macrophage marker MAC387 or the B cell markers BAQ155 or IL-A59.     

Functional and structural comparison of cytokines in different species

As the number of recombinant cytokines increases, so does our knowledge of their structure and function in different species. The biological cross-reactivity of cytokines from one species on cells from a different species has been reported on in the literature but this information is scattered over many publications and some of it has not yet been published. Comparing sequence information combined with three-dimensional and receptor-binding information (i.e. biological cross-reactivity) in different species provides insight into the underlying rules governing cross-reactivity and conservation. It was observed that there is quite a strict threshold of 60% amino acid identity above which cytokines tend to cross-react. Below this threshold few cytokines cross-react on cells from a different species. When comparing frequencies of reported species cross-reactivities between cytokines belonging to different cytokine-folding families it is obvious that not all cytokines within these folding families are equally cross-reactive. The underlying reason for these differences may lay in the ability of certain folding families to accumulate more mutations and still produce a protein, which is able to fold in the desired tridimensional structure. For example, cytokines belonging to the short 4-alpha-helix bundle can accumulate mutations in the 4-alpha-helices and large loops connecting the 4-alpha-helices and are the least cross-reactive. In contrast, cytokines belonging to the beta-sheet based folds (beta-trefoil and beta-sandwich) are the most cross-reactive and also the most conserved cytokines amongst the different species studied.     

Global increase in quaternary explosive volcanism

The worldwide distribution of volcanic ash has been determinmined from 320 deep-sea sections drilled during the Deep Sea Drilling Project. The ash distribution in the deep-sea sections, which span the last 20 million years, indicates that there has been a much higher rate of explosive volcanism from both island arc and hot spot volcanoes during the last 2 million years. This episode, and perhaps another in the Middle Miocene cprrelates with previously reported widespread synchronism in increased volcanicity in certain oceanic islands. Increased Quaternary volcanism coincides approximately with that episode of the Cenozoic marked by major and rapidly fluctuating climatic change.     

MUSCLE: VOLUME CHANGES IN ISOLATED SINGLE FIBERS

Volumetric experiments on single fibers isolated from semiten-dinosus muscles of frogs, some performed in correlation with measurements of membrane potential, confirm the data obtained on whole muscles, but only for the specific range of conditions in which most of the latter experiments have been done. These conditions are restricted to media in which the anion ( Cl usually) is permanent and the K is 10 to 12.5 meqlliter, or four to five times above the normal level in Ringer's solution. When other ionic conditions are employed, phenomena are disclosed which have not previously been described. The findings throw doubt upon the validity of some generally accepted views regarding the permeability properties of the membrane of frog muscle fibers and regarding the nature of the mechanisms which regulate their volume.     

Herd management practices associated with paratuberculosis seroprevalence in Dutch dairy herds

We describe the paratuberculosis management practices applied in dairy herds in the Netherlands. The findings from paratuberculosis seronegative and seropositive herds were compared to discover possible risk factors. In total, 370 randomly selected herds with > or =20 dairy cows were surveyed. A questionnaire was used to collect data on current and previous paratuberculosis management practices. All cattle aged > or =3 years were serologically tested for paratuberculosis using an enzyme-linked immunosorbent assay. Herds with >33 tested cattle, of which only one was seropositive, were excluded to reduce the risk of including false-positive herds in the analysis. A comparison of the management data of the seronegative herds (n = 166) and the seropositive herds (n = 143) showed that in both groups important management measures for the prevention of paratuberculosis, such as calving in a cleaned calving area, removing the calf immediately after birth, and feeding paratuberculosis non-suspect roughage to calves, were used only rarely. However, such measures should be regarded as the critical first step to control the disease and/or reduce its prevalence. Using univariable analysis, four factors were statistically different between seronegative and seropositive herds: herd size, cows with clinical signs of paratuberculosis, prompt selling of clinically diseased cattle and feeding milk replacer. Using a multivariable logistic regression model, only herd size was a significantly different factor. These results indicate that most of the paratuberculosis preventive management measures were executed on these Dutch dairy farms only to a limited extent.     

Cellular and humoral local immune responses in sheep experimentally infected with Oestrus ovis (Diptera: Oestridae)

Cellular and humoral local responses were investigated following repetitive artificial Oestrus ovis infections in lambs. The presence of larvae induced a huge local recruitment of either leucocytes (T and B lymphocytes, macrophages) or granulocytes (eosinophils, mast cells and globule leucocytes). This cellular response was more pronounced in the ethmoid and sinus (development sites of second and third instar larvae) than in the septum or turbinates where first instar larvae migrate. Infected lambs produced Oestrus ovis specific IgG and IgA antibodies in their mucus. This local humoral response was mainly directed against larval salivary gland antigens and not against larval digestive tract antigens. Compared to the control animals, the sinusal mucosa of infected animals was extremely thickened and the epithelium exhibited hyperplasia, metaplasia and eosinophilic exocytosis. The possible roles of these local immune responses in the regulation of O. ovis larvae populations in sheep are discussed.