In vitro phagocytic study of blood leucocytes and peritoneal macrophages of walking catfish Clarias batrachus ( Ham.) against Aeromonas hydrophila and Escherichia coli.

In observation of in vitro phagocytic activity against Aeromonas hydrophila isolate 34k (a virulent form) and Escherichia coli (an avirulent bacteria) of neutrophil- and monocyte-like cells of walking catfish Clarias batrachus showed phagocytosis. N eutrophils and monocytes phagocytized the avirulent form of bacterial isolate more than the virulent one. Other blood leucocytes did not show phagocytosis. Peritoneal macrophage of the fish were separated by glycogen elicitation and the macrophages were being adhered on plastic cover slips for studying their in vitro phagocytic activity. Most of the cells were alive after adherence and showed phagocytosis against the virulent and avirulent bacteria. The percent phagocytosis and phagocytic index were higher against the avirulent E. coli than the virulent A. hydrophila.

Energy substrate requirement for in vitro maturation of oocytes from unstimulated adult rhesus monkeys

The energy substrates lactate, pyruvate, and glucose were evaluated for supporting in vitro cytoplasmic maturation of rhesus monkey oocytes. A total of 321 cumulus-oocyte complexes (COCs) aspirated from greater than or equal to 1000 mum diameter follicles

Cryopreservation of rhesus macaque (Macaca mulatta) spermatozoa and their functional assessment by in vitro fertilization

Although spermatozoa from several species of nonhuman primates have been cryopreserved, there has been no report of success with rhesus macaque spermatozoa as judged by functional assays. Two Tris-egg yolk freezing media. TEST and TTE. which have: been successfully used for cynomolgus macaque (Macaca fascicularis) spermatozoa, were compared for cryopreservation of spermatozoa From four rhesus macaques (Macaca mulatta). The postthaw motility (percentage and duration) of spermatozoa cryopreserved in TTE was much higher than that for spermatozoa cryopreserved in TEST. The function of sperm cryopreserved in TTE was evaluated by in vitro fertilization or oocytes collected from gonadotropin-stimulated prepubertal rhesus macaques. Of the inseminated oocytes. 82 +/- 13% were fertilized and 63 +/- 22 and 39 +/- 21% of the resulting zygotes developed into morulae and blastocysts. respectively. These results indicate that rhesus macaque spermatozoa can be effectively cryopreserved in TTE medium. This finding will facilitate the application of in vivo and in vitro assisted reproductive technologies in this species. (C) 2001 Academic Press.

17 beta-Estradiol and progesterone improve in-vitro cytoplasmic maturation of oocytes from unstimulated prepubertal and adult rhesus monkeys

BACKGROUND: Effects of 17beta-estradiol and progesterone on rhesus monkey oocyte maturation in vitro were evaluated by embryo development subsequent to IVF. METHODS AND RESULTS: In experiment 1, immature cumulus-oocyte complexes collected from unstimulated adult females during the non-breeding season were matured in modified medium CMRL-1066 containing various combinations of gonadotrophins (FSH + LH), estradiol and/or progesterone. Formation of morulae and blastocysts was greatest in oocytes matured in medium containing estradiol and/or progesterone, with or without gonadotrophins (morula 38-46%, blastocyst 14-20%) than in control oocytes matured without estradiol or progesterone (morula 14%, blastocyst 0%). In experiment 2, cumulus-oocyte complexes from unstimulated prepubertal female monkeys were matured in medium with gonadotrophins, estradiol or progesterone. The best development to the morula stage was obtained with oocytes matured with gonadotrophins and estradiol or gonadotrophins and progesterone (43 and 25 morulae, respectively), while control oocytes matured with gonadotrophins but without steroid hormones gave the poorest morula developmental response (12%). However, there was no difference in blastocyst development across all groups (0-3%). CONCLUSIONS: These results demonstrate that during rhesus monkey oocyte maturation in vitro: (i) estradiol or progesterone can improve oocyte developmental competence; (ii) immature oocytes from prepubertal versus adult females have differential responses to challenge with estradiol or progesterone.

In vitro construction of a recombinant human embryonic brain-derived neurotrophin-4 gene and pEGFP-N1 vector

BACKGROUND: Neurotrophin-4 (NT-4) can promote neuronal growth, development, differentiation, maturation, and survival. NT-4 can also improve recovery and regeneration of injured neurons, but cannot pass through the blood-brain barrier, which limits its ac

IN-VITRO IMMUNOTOXICITY AND CYTOTOXICITY OF TRICHOSANTHIN AGAINST HUMAN NORMAL IMMUNOCYTES AND LEUKEMIA-LYMPHOMA CELLS

Trichosanthin (TCS) is a ribosome-inactivating protein from root tubers of Trichosanthes kirilowii Maxim. In this paper, the effects of TCS on the viability of human peripheral blood immunocytess, on the proliferation of lymphocytes, and its cytotoxicity to twelve cell lines of lymphoma or leukemia had been observed. TCS at high concentration (>12.5 mu g/ml) affected the viability of human B lymphocytes, but not that of human peripheral blood mononuclear cells (PBMCs), T lymphocytes and granulocytes. Human peripheral blood-derived monocytes/macrophages were highly sensitive to TCS (ID50 at 1.70 mu g/ml). TCS suppressed lymphocyte proliferation stimulated by Concanavalin A (Con A) or lipopolysaccharide (LPS). Human T cell lines and macrophage cell lines were more sensitive (ID50 < 0.9 mu g/ml) to TCS than B cell lines and myeloid lines. These results suggest that selective cytotoxicity of TCS to human macrophages/monocytes may be implicated in anti-HIV activity, and that selectively killing some leukemia-lymphoma cells by TCS merit further evaluation in treatment of some lymphoma and leukemia.