Reproductive morphology of the male Tuatara,Sphenodon punctatus

Over the past decade, studies on reproductive morphology in the Squamata (snakes and lizards) have expanded tremendously. With the accumulation of these studies and revisions of the terminology based on structural similarities and differences, it is imperative to review the work on tuataras to determine whether the structural organization fits the revised terminology of vertebrates. We investigated the morphology of the male reproductive system in the Tuatara, Sphenodon punctatus (Rhynchocephalia), the sister taxon to the Squamata. Previous studies on the Tuatara used a nomenclature for the testicular ducts different from the current terminology for amniotes. The reproductive system in the Tuatara is consistent with reports in the Squamata. Two rete testis tubules exit the testis within a connective tissue sheath similar to that shown in other squamate species and the protherian Echidna. Each rete testis divides into multiple ductuli efferentes that fuse with the epididymis. The epididymis transitions into the ductus deferens where the sperm become more concentrated into spherical bundles. The ductus deferens enters the cloacal urodeum separately from the ureter. An ampulla ureter or ampulla urogenital papilla was not observed, which differs from previous studies of lepidosaurians. Furthermore, a sexual segment of the kidney (SSK) was not observed, consistent with previous studies on the Tuatara.     

Differential induction of activation markers in macrophage cell lines by interferon-gamma

Macrophage cell lines were used in these studies as a model system to dissect the biochemical and functional mosaic of the macrophage activation process. In particular, the requirements for the induction of tumoricidal and bactericidal activity in the RAW 264.7 and WEHI-3 cell lines by interferon-gamma (IFN-gamma) and bacterial lipopolysaccharide (LPS) were determined. Changes in expression of a series of macrophage markers traditionally associated with macrophage activation were monitored during stimulation of the cells in order to determine whether a detectable pattern of activation-associated changes is associated with the development of a particular functional activity. These markers included changes in the cell surface expression of major histocompatibility complex-encoded Class I and Class II antigens and antigens in the Mac-1/LFA-1 family, alterations in the levels of membrane enzymes (5' nucleotidase and alkaline phosphodiesterase), and production of secretory products including hydrogen peroxide and the monokines interleukin-1, interferons-alpha/beta, and tumor necrosis factor-alpha. Our results demonstrate that a given homogeneous macrophage population expresses a distinct subset of functional activities in response to single, defined activating signals such as IFN-gamma and LPS. The display of a variety of macrophage surface antigens, enzymes, and secreted products is activated simultaneously by such treatment; however, the particular pattern of such activation-associated markers cannot reproducibly be used to predict the ability of an activated cell to perform a particular function. The results also suggest that macrophage cell lines expressing differential response patterns following IFN-gamma stimulation provide a valuable system for dissection of the molecular and cell biology of macrophage activation.     

The diagnosis of antileptospiral antibodies in human subjects by solid-phase immunoenzyme analysis

The possibility of using the enzyme-linked immunosorbent assay (ELISA) for the diagnosis of leptospirosis has been shown. This method has proved to be more simple and sensitive than the leptospiral microagglutination and lysis test. The data on obtaining genus-specific leptospiral antigens are presented. As revealed in this study, the antigens obtained by the complex treatment of microbial cells with ultrasound and detergents show the maximum activity in ELISA. The optimum parameters of the ELISA system for the diagnosis of leptospirosis have been established.     

The role of fibronectin in the streptococcal adhesion process

The work shows that fibronectin obtained from human plasma is capable of binding with streptococci of different groups with almost equal effectiveness. Fibronectin bound to bacterial cells inhibits the adhesion of group A streptococci onto vaginal cells, but it produces no effect on the adhesion of group B streptococci. The binding constant of fibronectin 125I is equal to 10(6) -M-1, which indicates that the level of the specificity of interaction is not sufficiently high.     

The possibility of using equine serum albumin in place of bovine serum albumin and ovalbumin in radioimmunological and immunoenzyme analyses and in virological practice

Horse serum albumin has been shown to meet the requirements to protein preparations for microanalysis and thus to be suitable for use in kits of reagents for the radioimmunological determination of insulin and myoglobin, for the determination of tick-borne encephalitis virus antigen by the method of the enzyme immunoassay and for the stabilization of proteins in the hemagglutination test and the hemagglutination inhibition test.     

Identification of the protein HC receptor

In the present study, we demonstrate for the first time the presence of a specific receptor for protein HC on the surface of human cells using the human histiocytic lymphoma cell line U937. Cells treated for 4 days with the maturation inducer phorbol 12-myristate 13-acetate, were found to increase both the number of cells binding protein HC (76% higher than for untreated cells) and the expression of protein HC receptors. Protein HC bound to these cells in a specific and saturable manner. Scatchard analysis at 4 degrees C, using radioiodinated protein HC, indicated a single class of low-affinity receptor (Ka = 2-5 x 10(7) M-1) and 20,000-30,000 receptors per cell. Monoclonal antibodies against protein HC abrogated specific binding of this protein to U937. In contrast, monoclonal antibodies that did not react with protein HC (anti-LFA-1 alpha, anti-MO1 alpha) were without effect on the binding reaction.     

Blood genetic marker studies of a sheep-goat hybrid and its back-cross offspring

Summary. Blood samples from a female sheep-goat hybrid and its back-cross male offspring were tested for electrophoretic variants of plasma albumin, transferrin and esterase, and of red cell carbonic anhydrase, nucleoside phosphorylase, NADH-diaphorase, 'X'-protein, superoxide dismutase, malic enzyme and haemoglobin. Red cells were also tested for blood group antigens. Both animals showed variants that could not be attributed to either sheep or goat alone, thus confirming previous chromosomal data that the female was a genuine sheep-goat hybrid.     

The nitrogen cycle in shallow water sediment systems of rice fields Part III: The influence of N-application on the yield of rice

Fertilizer application to rice-fields in the river-deltas in the Mediterranean area is a potential menace for wildlife protection, through eutrophication.Fertilizer use shows a trend of increasing rates of N application. A rate for N of 200 kg ha^–1 has become normal and a rate of 400 kg ha^–1 has already been recorded.Denitrification causes large losses of N with the result that more fertilizer is applied. This is especially true for the Camargue (S-France), where N is applied long before the rice (Aryza sativa) can take it up.Therefore we have tried to develop techniques which need the application of smaller amounts of N which are used more efficiently. In order to do this we tried to establish a N budget for rice-fields.Experiments were therefore set up in the field (plots of 550 m²) and in pots (2–3 l). Our results suggest that a late application of N (e.g. when the rice shows signs of N-deficiency by becoming yellowish), but at lower concentrations (70 kg ha^–1) can produce the same ultimate yield. The introduction of carp without any further input of N produced the same final yield.The N budget shows that 15±1.5 g m^–2 of N is needed for a normal crop. N losses due to denitrification may be as high as 12.2–13.6 g m^–2 of N. The input by irrigation water may provide up to about 20% of the input; N fixation is negligible. We estimate that 25–50% of the N missing in the budget comes from minderalization of the organic N pool in the soil. Denitrification may render part of this pool bio-available by oxidation. In sum, this work has revealed some surprising effects with potentially important consequences for farming practice and, in consequence, for conservation.     

Tissue distribution and kininogen gene expression after acute-phase inflammation

A kinin-directed monoclonal antibody to kininogens has been developed by the fusion of murine myeloma cells with mouse splenocytes immunized with bradykinin-conjugated hemocyanin. The hybrid cells were screened by an enzyme-linked immunosorbent assay (ELISA) and a radioimmunoassay (RIA) for the secretion of antibodies to bradykinin. Ascitic fluids were produced and purified by a bradykinin-agarose affinity column. The monoclonal antibody (IgG1) bound to bradykinin, Lys-bradykinin, Met-Lys-bradykinin, and kininogens in ELISA. Further, this target-directed monoclonal antibody recognized purified low and high molecular weight bovine, human, or rat kininogens and T-kininogen in Western blotting. After turpentine-induced acute inflammation, rat kininogen levels increased dramatically in liver and serum as well as in the perfused pituitary, heart, lung, kidney, thymus, and other tissues, as identified by the kinin-directed kininogen antibody in Western blot analyses. The results were confirmed by measuring kinin equivalents of kininogens with a kinin RIA. During an induced inflammatory response, rat kininogens were localized immunohistochemically with the kinin-directed monoclonal antibody in parenchymal cells of liver, in acinar cells and some granular convoluted tubules of submandibular gland, and in the collecting tubules of kidney. Northern and cytoplasmic dot blot analyses using a kinin oligonucleotide probe showed that kininogen mRNA levels in liver but not in other tissues increase after turpentine-induced inflammation. The results indicated that rat kininogens are distributed in various tissues in addition to liver and only liver kininogen is induced by acute inflammation. The target-directed kininogen monoclonal antibody is a useful reagent for studying the structure, localization, and function of kininogens or any protein molecule containing the kinin moiety.