Fluoride exposure inhibits protein expression and enzyme activity in the lung-stage larvae of Ascaris suum.

Sodium fluoride (NaF) is an anion that has been previously shown to block the moulting process of Ascaris suum larvae. This study describes moulting and development-specific protein expression profiles of A. suum lung-stage L3 (AsLL3) following NaF exposure. AsLL3s cultured in the presence or absence of NaF were prepared for protein analysis using two-dimensional (2D) electrophoresis. NaF exposure inhibited at least 22 proteins in AsLL3 compared with moulted larvae (i.e. AsLL4). A further comparison of AsLL4 with those of pre-cultured AsLL3 and NaF-exposed AsLL3 revealed 8 stage-specifically and 4 over-expressed proteins. Immunoblot analysis revealed an inhibition by NaF of 19 immunoreactive proteins. Enzyme assay and immunochemical data showed an inhibition of the moulting-specific inorganic pyrophosphatase activity by 41% and a decreased expression in NaF-treated larvae, indicating its significance in the moulting process. A protein spot associated with NaF inhibition was isolated and identified by peptide mass spectrometry and bioinformatics approaches to be a member of 3-hydroxyacyl-CoA dehydrogenase/short-chain dehydrogenase enzyme families. These results have implications for the identification of proteins specific to the moulting process as potential chemotherapeutic targets.

Enhanced phagocytic activity of neutrophils caused by administration of egg white derivatives (EWD) in cats injected with cyclophosphamide (CPA).

To examine in vivo effects of egg white derivatives (EWD), the numbers of peripheral blood cells and neutrophil phagocytosis were evaluated in cats injected intramuscularly with cyclophosphamide (CPA). There were no changes in the number of red blood cells (RBC) or packed cell volume (PCV) values regardless of oral administration of EWD or injection of CPA, but the numbers of platelets, white blood cells (WBC) and neutrophils in cats administered EWD significantly increased (p<0.05 to 0.01) when compared with those in control cats which received saline solution. In addition, the administration of EWD resulted in a significant enhancement in the phagocytic activity of neutrophils (p<0.01) when compared to control cats, suggesting that EWD has a stimulating effect on leukocyte progenitors. The numbers of platelets, WBC and neutrophils, and the phagocytic activity of neutrophils in cats injected with CPA alone were significantly lower (p<0.05 to 0.01) than those in control cats. However, co-administration of EWD to cats injected with CPA resulted in a significant increase in the numbers of platelets, WBC and neutrophils (p<0.05 to 0.01), and in the phagocytic response of neutrophils (p<0.01) when compared to cats injected with CPA alone. Therefore, these results suggest that co-administration of EWD may be effective in reducing some possible side effects in animals treated with immunosuppressive or antitumor agents.

Effects of the lipopolysaccharide-protein complex and crude capsular antigens of Pasteurella multocida serotype A on antibody responses and delayed type hypersensitivity responses in the chicken.

The effects of the lipopolysaccharide-protein complex (LPS) and crude capsular antigen (CCA) prepared from Pasteurella multocida serotype A isolated from a duck in the Philippines, on antibody responses to sheep red blood cells (SRBC) and Brucella abortus (BA) and delayed type hypersensitivity (DTH) responses to bovine serum albumin (BSA) in the chickens were studied. Chickens injected subcutaneously with LPS and CCA at 1 and 2 weeks of age and immunized intravenously with the mixed antigens of SRBC and BA, at 3 and 4 weeks of age showed significantly increased antibody responses against both SRBC and BA, when evaluated at 7 days after each immunization. In addition, these chickens sensitized intramuscularly with the emulsion of BSA in complete Freund's adjuvant at 5 weeks of age, and then injected into the wattle with BSA at 7 weeks of age also showed significantly increased DTH responses against BSA, when evaluated at 24 and 48 hr after challenge. These results indicate that LPS and CCA of P. multocida serotype A have a property enhancing humoral and cell-mediated immune responses.

Effects of four antigenic fractions of Pasteurella multocida serotype A on phagocytosis of chicken peripheral blood leukocytes.

The effects of four antigenic fractions of Pasteurella multocida serotype A isolated from a duck in the Philippines on the phagocytic activity of chicken peripheral blood leukocytes were studied by a flow cytometer. These fractions were the lipopolysaccharide-protein complex (LPS), crude capsular antigen (CCA), ribosomal fraction (RS) and outer cell layer (OCL). Among these four antigens, only CCA but not LPS RS and OCL, significantly increased the phagocytic activities of mononuclear cells (MNC) and polymorphonuclear cells (PMN). This result indicates that CCA has an immunological property enhancing the phagocytic activities of MNC and PMN.

[Pulsatile release of gonadotropins in athletic women].

In athletic women menstrual disorders such as luteal insufficiency, oligomenorrhea and amenorrhea are often seen. It has been suggested that such disorders may be related to hypothalamic-pituitary axis insufficiency caused by physical activity. To investigate the mechanism by which the disorder is promoted, episodic gonadotropin secretion was studied in 10 athletic women (normal ovulatory, 5; short luteal, 5) and 6 non-athletic controls. In the early follicular phase of the menstrual cycle, blood samples were obtained through an indwelling venous catheter at 15-minute intervals for 4 hours. The concentrations of luteinizing hormone (LH) and follicle-stimulating hormones (FSH) were measured by radioimmunoassay. The mean LH and FSH concentrations in the athletic groups were lower than those in the control group. LH pulse frequencies in the short luteal group were 2.8 +/- 0.2 times/four hours (mean +/- standard error) and decreased compared with that of the control group (3.7 +/- 0.2 times/four hours; p less than 0.001). The LH pulse amplitudes of all three groups were similar. Pulsatile FSH secretion was also observed in all the women. Dynamism of FSH was the same as that of LH except for low pulse amplitude in the athletic groups. These data suggest that hypothalamic-pituitary axis insufficiency, especially of hypothalamic origin may be caused by athletic activity.

Effect of naloxone on hormonal changes during exercise.

It is well established that prolactin release during exercise is one of the important factors in exercise-induced menstrual dysfunction. The purpose of this study is to clarify the mechanisms of prolactin release during exercise. Ten female athletes measured their BBT every morning. They performed incremental exercise on a cycle ergometer, with or without naloxone, on the 5th to 8th days of the follicular phase. Three minutes before the exercise, 0.4mg of naloxone was injected intravenously and a further 1.6mg/hr of naloxone was continuously infused during exercise. Blood samples were collected after 60 minutes bed rest (Rest), at the time when the heart rates reached 150 bpm (Submax), the point of exhaustion (Max) during exercise and after 60 minutes bed rest following exercise (After 1hr). The levels of prolactin in serum, dopamine, beta-endorphin. VIP and ACTH in the plasma were measured. Whereas prolactin increased significantly at Submax (p less than 0.05) and Max (p less than 0.001), the increase in prolactin was suppressed by the administration of naloxone (p less than 0.05). Dopamine showed no remarkable change during exercise, with or without naloxone. There were significant increases in beta-endorphin at Max (p less than 0.001), VIP at Submax and Max (p less than 0.001), but these increases were suppressed by the administration of naloxone (p less than 0.001). ACTH which had markedly increased at Submax (p less than 0.025) and Max (p less than 0.001) showed a slight tendency to decrease following the administration of naloxone, but there were no significant differences in both groups.(ABSTRACT TRUNCATED AT 250 WORDS)