Semen cryopreservation in golden‐headed lion tamarin,Leontopithecus chrysomelas

Wild animal genetic resource banking (GRB) represents a valuable tool in conservation breeding programs, particularly in cases involving endangered species such as the golden‐headed lion tamarin (Leontopithecus chrysomelas). Thus, we aimed to assess a sperm freezing protocol for golden‐headed lion tamarins using two different exenders: BotuBOV® (BB) and Test Yolk Buffer® (TYB). Ejaculates were collected by penile vibrostimulation from animals housed at São Paulo Zoological Park Foundation, São Paulo, Brazil, and after immediate analysis, two aliquots were diluted in BB and TYB. Postthawing samples were evaluated for total and progressive motility, plasma membrane and acrosome integrities, mitochondrial activity, susceptibility to oxidative stress, and sperm–egg‐binding. No differences between BB and TYB were found for most seminal parameters, except for acrosome integrity and susceptibility to oxidative stress (in both cases BB showed higher values). However, in spite of these differences and regardless of the extender used, postthaw sperm motility and viability with the described protocol were encouraging (on average >50% and >80%, respectively), indicating that sperm cryopreservation may be a short‐term measure for the conservation of golden‐headed lion tamarins.     

Inhibition of tyrosine autophosphorylation of the solubilized insulin receptor by an insulin-stimulating peptide derived from bovine serum albumin

A polypeptide from a tryptic digest of bovine serum albumin potentiates glucose oxidation stimulated by insulin in isolated rat adipocytes. We studied whether this effect is related to a modification of the insulin receptor kinase. In a solubilized rat adipocytes receptor system, the peptide caused dose-dependent inhibition of the stimulation by insulin of phosphorylation of the 95,000 dalton subunit of insulin receptor. The peptide also inhibited stimulation by vanadate of tyrosine autophosphorylation of the beta subunit of the receptor, though it enhanced vanadate-stimulated glucose oxidation. During the phosphorylation reaction, no phosphorylated forms of the peptide could be detected. The peptide had no effect on dephosphorylation of the phosphorylated beta subunit of the insulin receptor. These results strongly suggest that the inhibition of phosphorylation by the peptide is due not to either simple substrate competition or activation of phosphoprotein phosphatase, but to specific inhibition of tyrosine-specific protein kinase.     

Molecular cloning and sequence analysis of cDNA for human hepatocyte growth factor

Amino acid sequences of four peptide fragments of human hepatocyte growth factor purified from the plasma of patients with fulminant hepatic failure were determined. Based on the amino acid sequence of one of the fragments, two oligodeoxyribonucleotide mixtures were synthesized and used to screen a human placenta cDNA library. On the screening, two overlapping cDNA clones for human hepatocyte growth factor were isolated and the nucleotide sequence of the cDNA was determined. The entire primary structure of the protein was deduced from the sequence. The protein consists of 728 amino acid residues, including a possible signal peptide at the N-terminus. The sequence revealed that the heavy and light chains which comprise the protein are encoded by the same mRNA and are produced from a common translation product by proteolytic processing.     

Phoresy ofTelenomus sp. (Scelionidae: Hymenoptera), an egg parasitoid of the tussock mothEuproctis taiwana

The phoretic relationship between the egg parasitoidTelenomus sp. cf.euproctidis Wilcox and its host the tussock mothEuproctis taiwana was studied in Okinawa, Japan. One third of the female moths studied in the field carried female parasitoid adults. No male moths carried parasitoids. Parasitoids were observed only in the anal tuft of the moth. Laboratory observation revealed that most of the parasitoids left the body of the moth at the time of the first oviposition of their host and proceeded to lay eggs on the moth egg masses.     

Exposure of Leaves of Cucumis sativus L. to Low Temperatures in the Light Causes Uncoupling of Thylakoids II. Non-Destructive Measurements with Intact Leaves

To examine the effects of chilling of leaves of cucumber (Cucumissativus L.) in moderate light on the coupling state of thylakoidsin situ, changes in fluorescence, changes in light scatteringand flash-induced changes in absorbance at 518 nm were examinedin intact leaves. After chilling of leaves at 5?C in the lightfor 5 h, the non-photochemical quenching of fluorescence, ameasure of energisation of thylakoids, was largely suppressed.The treatment also caused a suppression of light-induced changesin the light scattering by leaves, which depends on the formationof a pH gradient across thylakoid membranes. When thylakoidswere prepared by very gentle methods from the leaves chilledin the light, through a step of preparation of intact chloro-plasts,the transport of electrons from H₂O to ferricyanide was uncoupled,being insensitive to an uncoupler, methylamine. These data provide consistent evidence that the thylakoids areuncoupled in situ by the chilling of leaves in the light and,as a consequence of the uncoupling, the energisation of themembranes is suppressed. However, the decay of the flash-inducedchange in absorbance at 518 nm in leaves was not markedly acceleratedby the treatment. The thylakoids isolated from leaves chilledin the light, which were in the uncoupled state, also did notshow a rapid decay, unless an efficient uncoupler such as gramicidinwas added. These results suggest that even a considerable uncouplingof thylakoids, brought about by chilling of leaves in the light,is not sufficient to cause a marked acceleration of the decayof the flash-induced change in absorbance at 518 nm. Therefore,analysis at 518 nm is not always a sensitive method for assessingthe coupling state of thylakoids. (Received July 1, 1991; Accepted October 4, 1991)     

An oxygen-evolving complex with a simple subunit structure - ‘a water-plastoquinone oxidoreductase” - from the thermophilic cyanobacterium Synechococcus sp.

An oxygen-evolving complex has been highly purified from the thermophilic cyanobacterium Synechococcus sp. The complex, which reproducibly showed 5 major polypeptide bands of 47, 40, 35, 30 and 9 kDa on SDS-polyacrylamide gel electrophoresis and contained 3.2 Mn per Q_A, had an oxygen-evolving activity of 300–400 μmol/mg chl per h in the presence of 5 mM MnCl₂; or CaCl₂. The complex most likely represents a minimum functional unit of the photosynthetic oxygen evolution.     

Studies on the biological activity of an insulin-stimulating peptide from a tryptic digest of bovine serum albumin

Summary A two-chain polypeptide, which corresponds to amino acid residues 115–143 and 144–184(185) of bovine serum albumin, connected to each other by a disulfide bridge, potentiated the effects of insulin on glucose transport and glucose metabolism in isolated rat adipocytes. Although the peptide alone had little activity, it shifted the concentration-response curves of insulin-stimulated D-[I-¹⁴C]glucose oxidation, 2-deoxyglucose transport, and lipid synthesis from D-[U-¹⁴C]glucose to lower insulin concentrations. It also increased the maximal responses of these parameters to insulin. However, it did not affect insulin binding to adipocytes. The peptide protected insulin considerably from degradation, but this effect alone cannot account for its effect in increasing the maximal responses to the hormone, and even when degradation of a submaximal concentration of insulin was suppressed by bacitracin, the peptide still had an enhancing effect. These results suggest not only that the peptide influences a step distal to receptor-mediated insulin binding but also that inhibition of insulin degradation alone cannot explain its total effect.The peptide lost its insulin-stimulating activity completely when it was further digested with V8 or lysinespecific endopeptidase, or when it was reduced and then carboxamidomethylated or oxidized with performic acid. Similar active tryptic fragments were obtained from human and rat albumins.Insulin-stimulating peptides should be useful in studies on the mechanisms of insulin action including both the sensitivities and responsiveness of target cells to the hormone.Abbreviations ISP insulin-stimulating peptide - HEPES N-(2-hydroxyethyl)piperazine-N-2-ethanesulfonic acid - HPLC high-performance liquid chromatography - SDS sodium dodecyl sulfate     

Alteration of opioid receptors in seizure-susceptible El mouse brain

The distribution density of opioid receptors in the brain of El mice (seizure-susceptible strain) was examined to determine the relation between seizures and the opioid system. Saturation curves and Scatchard plots of [³H]2-d-alamine-5-d-leucine enkephalin binding revealed that the opioid delta receptor density in adult El mice during interictal periods was significantly increased in the cerebral cortex, hippocampus, and septal area. It was further shown that the concentration of such receptors in 25-day-old El mice that had no seizures was also significantly increased in the hippocampus and septal area, with no changes in apparent affinities, as compared with in the corresponding regions in ddY mice (seizure-nonsusceptible strain; the mother strain of El). Such up-regulation of opioid receptors in the El mouse brain could result from deficits in endogenous opioid peptides, which could be associated with the pathogenesis of seizure diathesis in the El mouse.