Allelic variants of Ly-5 in inbred and natural populations of mice

Allelic variants of Ly-5 in inbred commensal and other natural populations of mice were analyzed by patterns of restriction fragment length polymorphisms (RFLP) and Southern hybridization using an Ly-5 cDNA probe and by cell-surface staining with a panel of antibodies directed against polymorphic and nonpolymorphic Ly-5 determinants. New Ly-5 alleles were defined by RFLPs generated by both Eco RI and Bam HI restriction enzyme digests. The Mus musculus subspecies and other species within the genus Mus showed a strong correlation between allelic variants defined by restriction enzymes and serologic specificities. The data also suggest the conservation of the Ly-5 gene throughout the genus Mus.     

Involvement of the acyl chain of ceramide in carbohydrate recognition by an anti-glycolipid monoclonal antibody: the case of an anti-melanoma antibody,M2590, to GM3-ganglioside

The effect of the chain length of the fatty acid residue of the ceramide moiety of ganglioside G_M3 on the binding ability of monoclonal antibody M2590, which is specific for the carbohydrate structure of G_M3-ganglioside, was examined by means of a direct binding assay on thin layer chromatography plates (TLC immunostaining) and a quantitative enzyme-linked immunosorbent assay (ELISA). Derivatives of G_M3 with a long fatty acid chain reacted with the M2590 antibody, but those with a short fatty acid chain showed no reaction in either assay system. These results suggested that the acyl fatty acid moiety of the ganglioside played an important role in the formation or maintenance of the antigenic structure of the carbohydrate moiety of the ganglioside.     

The production of pheromone clouds by spraying in the melon fly,Dacus cucurbitae coquillett (Diptera: Tephritidae)

Pheromone clouds sprayed by melon fly males were visually detected by focusing a beam of light at them during dusk when the males were vibrating their wings. The clouds were sprayed to the front, rear and upper sides of the male. We found that special morphological structures are used for spraying the pheromone clouds. When a male melon fly engages in calling behavior, sex pheromone droplets are excreted from his anus. This excretion is wiped off with the tarsus of his hind leg, and then it is deposited on the sexually dimorphic cubital cell hairs on the wing. During wing vibration, the targal bristles on the 3rd abdominal segment, which are peculiar to males, are rubbed against the specialized hairs of the cubital cell. Calling males sprayed clouds of pheromone with these actions. This paper was presented at the 32nd Annual Meeting of the Japanese Society of Applied Entomology and Zoology (Kochi, April, 1988).     

Replication of ColE2 and ColE3 plasmids: Two ColE2 incompatibility functions

Summary We have identified and localized two incompatibility determinants (IncA and IncB) within a 1.3 kb segment of ColE2 sufficient for autonomous replication. The IncA determinant is localized in a region shorter than 250 bp and expresses incompatibility against both ColE2 and ColE3. The region which determines sensitivity to the IncA determinant seems to overlap with the region specifying the IncA determinant. The expression of the trans-acting factor(s) specifically required for replication of ColE2 interferes with expression of the IncA determinant against ColE2 but not against ColE3. The IncA determinant might be at least partly responsible for the copy number control of the plasmid. The IncB determinant is localized in a 50 bp region (origin) which is sufficient for initiation of replication in the presence of the trans-acting factor(s). The IncB determinant is specific for ColE2 and seems to be due to titration of the trans-acting essential replication factor(s) by binding.     

Biosynthesis of a novel transformation-sensitive heat-shock protein that binds to collagen. Regulation by mRNA levels and in vitro synthesis of a functional precursor

The synthesis of a major collagen-binding glycoprotein of molecular weight 47,000 was previously shown to be altered by malignant transformation as well as by heat shock in chick embryo fibroblasts (Nagata, K., and Yamada, K.M. (1986) J. Biol. Chem. 261, 7531-7536 and Nagata, K., Saga, S., and Yamada, K.M. (1986) J. Cell Biol. 103, 223-229). In this paper, we examined the synthesis of this heat shock protein (hsp47) in terms of possible functional precursors and its regulation after heat shock and transformation by Rous sarcoma virus. Actinomycin D inhibited the induction of hsp47 after heat shock. Messenger RNAs purified from chick embryo fibroblasts (CEF), heat-treated CEF, and transformed CEF were analyzed in an in vitro translation system. In vitro translated products readily bound to gelatin-Sepharose, and levels were increased after heat shock and decreased after transformation. The increase in mRNA after heat shock was shown more directly by Northern assay using a synthetic oligonucleotide probe. We identified two putative precursors of hsp47 using an in vitro translation/processing system and tunicamycin: one is a 42-kDa primary translation product and the second is a 41-kDa polypeptide lacking signal peptide and carbohydrate moieties. Both of these precursors are biologically active as determined by gelatin-binding activity, in contrast to the lack of binding activity of precursors in several other membrane-associated receptor systems.     

Microbial transformation of neutral fraction and upgraded neutral fraction of Polish tall oil

Summary Microbial transformations of neutral fraction (NF) and upgraded neutral fraction (UNF) of Polish tall oil byMycobacterium sp. MB 3683 were performed. Final metabolites and yields were compared to bioconversion of pure -sitosterol. Additionally, origin of a new metabolite —5-androsta-3,6,17-trione was proved by transformation of UNF in the presence of labeled -sitosterol.     

Association of Lps gene with natural resistance of mouse macrophages against Legionella pneumophila

We have cloned a 1.6-kb region of chromosomal DNA from Thermoplasma acidophilum into Escherichia coli using as a probe part of the Methanococcus vannielii fus-gene. The sequence of the clone was highly homologous to part of the corresponding Methanococcus vannielii gene. By chromosome walking, a 4.7-kb EcoRI fragment containing the complete gene was isolated. Nucleotide sequencing revealed an open reading frame of 2196 nucleotides. The deduced amino acid sequence contains the known peptide sequence around the ADP-ribosylation site of T. acidophilum elongation factor 2, which unequivocally confirms that the fus-gene has been cloned. The amino acid sequence was compared to that of hamster and E. coli, as well as to known archaebacterial EF-2 sequences.     

Improvement of first-service pregnancy rate in cows with gonadotropin-releasing hormone analog

The effect of an intramuscular injection of a new analog of gonadotropin-releasing hormone (GnRH), fertirelin, on the first-service pregnancy rate in cows was investigated by a double blind experiment. A total of 1,194 cows was injected intramuscularly either with 100 mug of GnRH or placebo (physiological saline solution) at the time of first insemination postpartum. Pregnancy rate (number of cows calved/ number of cows serviced) was 57.2 % in 605 cows treated with GnRH, while the performance was 49.7 % in 589 cows of the placebo group. The difference of pregnancy rates in both groups was significant (P<0.05). GnRH injected at insemination was effective, especially in cows at the first and third lactations, cows at 101 days postpartum or later, cows with daily milk yield of 26-30 kg, and also in cows from the area where a regional average fertility was relatively low.     

Hormonal induction of all stages of spermatogenesis in germ-somatic cell coculture from immature Japanese eel testis

In cultivated male eel, spermatogonia are the only germ cells present in testis. Our previous studies using an organ culture system have shown that gonadotropin and 11-ketotestosterone (11-KT, a potent androgen in teleost fishes) can induce all stages of spermatogenesis in vitro. for detailed investigation of the control mechanisms of spermatogenesis, especially of the interaction between germ cells and testicular somatic cells during 11-KT-induced spermatogenesis in vitro, we have established a new culture system in which germ cells and somatic cells are cocultured after they are aggregated into pellets by centrifugation. Germ cells (spermatogonia) and somatic cells (mainly Sertoli cells) were isolated from immature eel testis. Coculture of the isolated germ cells and somatic cells without forming aggregation did not induce spermatogenesis, even in the presence of 11-KT. In contrast, when isolated germ cells and somatic cells were formed into pellets by centrifugation and were then cultured with 11-KT for 30 days, the entire process of spermatogenesis from premitotic spermatogonia to spermatozoa was induced. However, in the absence of 11-KT in the culture medium spermatogenesis was not induced, even when germ cell and somatic cells were aggregated. These results demonstrate that physical contact of germ cells to Sertoli cells is required for inducing spermatogenesis in response to 11-KT.