Tentative identification of a vasopressin–neurophysin and an oxytocin–neurophysin in the rat

1. Rat neurohypophysial extracts have been examined by polyacrylamide-gel electrophoresis. 2. Three of the proteins were tentatively identified as neurophysins by their acidic nature and their disappearance after dehydration of the animals. 3. These proteins were radioactive 24h after intracisternal injection of [(35)S]cysteine. 4. Two of the proteins were present in much greater quantities than the third, and these two were present in the gland in the same ratio as the hormones vasopressin and oxytocin. 5. One of these proteins was absent from glands of rats homozygous for diabetes insipidus but present in heterozygous animals. 6. It is suggested that these two proteins are the vasopressin-neurophysin and oxytocin-neurophysin of the rat.     

Endogenous opioid peptides: do they mediate the acute antihypertensive action of clonidine in humans?

The involvement of endogenous opioid peptides in the antihypertensive action of acutely administered clonidine, a centrally acting adrenergic agonist, was studied in humans. Eight hypertensive subjects received clonidine 0.2 mg orally, naloxone 8 mg i.v. followed by a 0.13 mg/min infusion, and both drugs together on separate days. Clonidine resulted in a significant decrease in mean blood pressure, which was not affected by concomitant treatment with naloxone. Naloxone alone or with clonidine caused significant elevations in plasma aldosterone, not mediated by increased plasma renin activity. Plasma beta-endorphin was not increased after clonidine administration. In humans, the antihypertensive effects of acute clonidine administration do not appear to be mediated by the release or action of endogenous opioids.     

Do inhalation general anesthetic drugs induce the neuronal release of endogenous opioid peptides?

The antagonism of some effects of inhalation general anesthetic agents by naloxone suggests that there may be an opioid component to anesthetic action. There is evidence that this opioid action component is due to neuronal release of endogenous opioid peptides. The strongest evidence is provided by studies that monitor changes in the concentration of opioid peptides in the perfused brain following inhalation of the anesthetic. Indirect or circumstantial evidence also comes from studies of anesthetic effects on regional brain levels of opioid peptides, antagonism of selected anesthetic effects by antisera to opioid peptides and anesthetic-induced changes radioligand binding to opioid receptors. It is likely that some inhalation general anesthetics (e.g., nitrous oxide) can induce neuronal release of opioid peptides and that this may contribute to certain components of general anesthesia (e.g., analgesia). More definitive studies utilizing in vivo microdialysis or autoradiography in selected areas of the brain during induction and successive states of general anesthesia have yet to be conducted.     

Do endogenous opioid peptides mediate the effects of photoperiod on release of luteinizing hormone and prolactin in ovariectomized ewes?

Three experiments were done to determine if endogenous opioid peptides (EOPs) mediate the effects of photoperiod on release of luteinizing hormone (LH) and prolactin (Prl) in ovariectomized (OVX) ewes. Intravenous infusions of 0.5 naloxone X h-1 X kg body weight-1 for 3.5 h increased (P less than 0.01) mean plasma concentrations of LH and decreased (P less than 0.025) mean interpulse interval (period) of LH pulses in OVX ewes exposed to long day lengths (16L:8D). Infusions of either 1.0 or 2.5 mg morphine-SO4 X h-1 X kg-1 for 3 h increased (P less than 0.005) the period of LH pulses and increased (P less than 0.005) concentrations of Prl in OVX ewes during the breeding season. In OVX ewes exposed to long (16L:8D) or short (8L:16D) day lengths infusions of naloxone increased (P less than 0.05) mean concentrations of LH, whereas morphine decreased (P less than 0.01) mean concentrations of LH. These effects were attributed to changes in period of LH pulses (P less than 0.001). The drug X photoperiod interactions were not significant for LH parameters. Naloxone did not affect Prl release in either long- or short-day groups, but morphine increased (P less than 0.001) Prl release during long and short day lengths. The effect of morphine on Prl release was more pronounced in ewes exposed to long day lengths than in those exposed to short day lengths. In conclusion, EOPs inhibit the LH pulse generator in OVX ewes. However, it is doubtful that the EOPs mediate the steroid-independent effects of photoperiod on LH release. The results also suggest that photoperiod may influence Prl release via opiate neurons.     

Caffeine and reduction of fetal ossification in the rat: fact or artifact?

This study was done to determine the gestational period during which the rat fetus is susceptible to reduction of skeletal ossification by caffeine. Caffeine, 100 mg/kg/day by gavage, caused the greatest reduction in ossification, as assessed by staining with alizarin red S, in fetuses exposed between day 16 to 19 of gestation, less in those treated between day 7 to 19, and markedly less in those receiving it between day 7 to 16; a single dose on day 19 had very little effect. This indicates that the fetus is most susceptible late in pregnancy. Bones in early stages of mineralization on day 20 showed a greater reduction in staining than those in later stages. Thus, caffeine appears to lower the rate of ossification rather than reduce its final extent. In the day 7 to 19 caffeine treatment group, but not in the day 16 to 19 group, maternal and fetal body weights were reduced, and 1.6% of the fetuses had aphalangia. The day 7 to 16 caffeine treatment reduced fetal body weight. This argues against an association between reduction in fetal weight and ossification. None of the treatments affected rates of resorption or litter size. A novel and important observation made is that the different caffeine treatments affected the staining by alizarin of both claws and bones in a qualitatively and quantitatively similar manner. Since claws are devoid of osteoid, this observation questions the specificity of alizarin for the assessment of the state of fetal ossification and raises doubt as to the significance of the observed action of caffeine on ossification.     

Are 5-HT receptors involved in the sprouting of serotoninergic terminals following neonatal 5,7-dihydroxytryptamine treatment in the rat?

The neonatal administration of 5,7-dihydroxytryptamine to rats (100 mg kg^?1 s.c. on the 1st and 2nd day after birth) resulted in marked reductions in serotoninergic presynaptic markers ([³H]-5-HT synaptosomal uptake, tryptophan hydroxylase activity and endogenous 5-HT content) in various forebrain areas, particularly the cerebral cortex and the hippocampus. In contrast, this treatment produced an increased outgrowth of serotoninergic terminals in the brain stem as judged by the significant increments of these presynaptic markers in this region. Both in the hippocampus and the brain stem, these 5,7-dihydroxytryptamine-induced changes in serotoninergic innervation were associated with a transient increase in 5-HT-sensitive adenylate cyclase activity. No significant alteration of the specific high affinity binding of [³H]-5-HT to synaptosomal membranes from various brain regions was detected in 5,7-dihydroxytryptamine-treated rats for at least the first postnatal month.The chronic blockade of 5-HT receptors by metergoline (5 mg kg^?1 day^?1 from day 3 to day 22 after birth) altered neither the changes in presynaptic markers nor the evolution of [³H]-5-HT high affinity binding in 5,7-dihydroxytryptamine-treated rats.These findings further illustrate that the high affinity binding sites for [³H]-5-HT do not correspond to postsynaptic 5-HT receptors coupled to adenylate cyclase in the rat brain. Apparently, 5-HT receptors play no role in the increased outgrowth of serotoninergic systems in the brain stem following neonatal 5,7-dihydroxy-tryptamine treatment.     

Association of plasma Aß peptides with blood pressure in the elderly

Background

Aß peptides are often considered as catabolic by-products of the amyloid ß protein precursor (APP), with unknown physiological functions. However, several biological properties have been tentatively attributed to these peptides, including a role in vasomotion.We assess whether plasma Aß peptide levels might be associated with systolic and diastolic blood pressure values (SBP and DBP, respectively).

Methodology/Principal Findings

Plasma Aß_1-40 and Aß_1-42 levels were measured using an xMAP-based assay in 1,972 individuals (none of whom were taking antihypertensive drugs) from 3 independent studies: the French population-based 3C and MONA-LISA (Lille) studies (n = 627 and n = 769, respectively) and the Australian, longitudinal AIBL study (n = 576). In the combined sample, the Aß_1-42/ Aß_1-40 ratio was significantly and inversely associated with SBP (p = 0.03) and a similar trend was observed for DBP (p = 0.06). Using the median age (69) as a cut-off, the Aß_1-42/Aß_1-40 ratio was strongly associated with both SBP and DBP in elderly individuals (p = 0.002 and p = 0.03, respectively). Consistently, a high Aß_1-42/ Aß_1-40 ratio was associated with a lower risk of hypertension in both the combined whole sample (odds ratio [OR], 0.71; 95% confidence interval [CI], 0.56-0.90) and (to an even greater extent) in the elderly subjects (OR, 0.53; 95% CI, 0.37–0.75). Lastly, all these associations appeared to be primarily driven by the level of plasma Aß_1-40.

Conclusion

The plasma Aß_1-42/Aß_1-40 ratio is inversely associated with SBP, DBP and the risk of hypertension in elderly subjects, suggesting that Aß peptides affect blood pressure in vivo. These results may be particularly relevant in Alzheimer''s disease, in which a high Aß_1-42/Aß_1-40 plasma ratio is reportedly associated with a decreased risk of incident disease.     

Are monophyly and synapomorphy the same or different? Revisiting the role of morphology in phylogenetics

Species are groups of organisms, marked out by reproductive (replicative) properties. Monophyletic taxa are groups of species, marked out by synapomorphies. In Nelson’s analysis, monophyly and synapomorphy are identical relations. Monophyly and synapomorphy, however, are not equivalent relations. Monophyly is epistemically not accessible, whereas synapomorphy is epistemically accessible through character analysis. Monophyly originates with speciation, the two sister‐species that come into being through the splitting of the ancestral species lineage forming a monophyletic taxon at the lowest level of inclusiveness. Synapomorphy provides the empirical evidence for monophyly, inferred from character analysis in the context of a three‐taxon statement. If synapomorphy and monophyly were equivalent, phylogenetic systematists should find a single tree, instead of multiple equally parsimonious trees. Understanding synapomorphy as the relevant evidence for phylogenetic inference reveals a category mistake in contemporary phylogenetics: the treatment of morphological characters mapped onto molecular trees as synapomorphies and homoplasies. The mapping of morphological characters onto nodes of a molecular tree results in an empirically empty procedure for synapomorphy discovery. Morphological synapomorphies and homoplasies can only be discovered by morphological and combined analyses. The use of morphology in phylogenetic inference in general is defended by examples from Laurales and Squamata in particular. To make empirical evidence scientifically relevant in order to search for concordance, or dis‐concordance, of phylogenetic signal, is certainly more fruitful for phylogenetics than the uncritical mapping of morphological traits on a molecular scaffold. © The Willi Hennig Society 2010.     

Mapping antibody binding sites on cytochrome c with synthetic peptides: Are results representative of the antigenic structure of proteins?

Crystallographic work on antigen-antibody complexes has revealed that extensive surface areas of proteins may interact with antibodies. On the other hand, most experimental approaches to locate and define antigenic determinants of protein antigens rely on the linear sequence of the polypeptide chain. Hence the question arises whether mapping of antibody binding sites by analysis of the reactivity of anti-protein antibodies with synthetic peptides can provide a representative picture of the antigenic structure of a protein antigen. We have addressed this question using yeast iso-1 cytochrome c as a protein antigen against which antisera were raised in rabbits. The reaction of the antisera with 103 synthetic hexapeptides covering the entire sequence of cytochrome c was tested by the pepscan procedure in which peptides are coupled to polyethylene rods and tested by ELISA. For the assay, anti-cytochrome c antibodies were fractionated by affinity chromatography on native yeast iso-1 cytochrome c and on apo-cytochrome c; the latter is a random coil. It was found that only antibodies retained by the apo-cytochrome c affinity column react with synthetic peptides. These antibodies comprise a small fraction, probably less than 2%, of all cytochrome c-specific antibodies. The majority of antigenic determinants, which seem to consist of strongly conformation-dependent topographic epitopes, could not be uncovered by the peptide approach. Epitope mapping with short peptides seems of limited usefulness in the case of small, globular, and conformationally stable proteins like cytochrome c.     

Interaction of acetochlor with estrogen receptor in the rat uterus. Acetochlor--possible endocrine modulator?

The aim of the present study was to investigate the ability of the chloroacetanilide herbicide acetochlor to interact with the endocrine system. The modulation of the binding of [3H]estradiol-17beta to protamine sulphate-precipitated uterine nuclear and cytoplasmic estrogen receptors was analysed for this purpose. Two different stages of reproductive life cycle of female rats, virgin and uniparous, were used. Our results demonstrate that acetochlor interacts in a specific manner with high-affinity limited-capacity uterine estrogen receptors. A significant difference (p < 0.001) in estrogen receptor density was observed between two control groups: uniparous rats (Bmax = 43.634 +/- 9.516) and virgin rats (Bmax = 154.375 +/- 21.462), suggesting an intrinsic difference in the nuclear estrogen receptor levels between female rats in different reproductive life cycle stages. Consequently, a different response to acetochlor treatment was noted. Exposure to acetochlor significantly (p < 0.001) increased estrogen receptor binding activity in the nuclear fraction of uniparous female rats (Bmax = 123.324 +/- 5.666) in comparison to control (Bmax = 43.634 +/- 9.516). In exposed virgin female rats, no significant difference was detected when compared to the corresponding control group. These results should prompt us to more thoroughly evaluate potential hazards of acetochlor to human and wildlife endocrine systems.     

Australian Entomology: isolated, or in touch with the rest of the world?

Abstract   Australia is isolated from the rest of the world geographically but not scientifically: Australian entomology has a long and distinguished record of collaboration with a diversity of other countries. Much of the focus relates to keeping new pests out of Australia (biosecurity and quarantine) and managing those that do get in (biological control and insect pest management). Many projects have cooperated with neighbouring countries, to assist them to control pests and thereby reduce threats to Australia. Biological control projects have also had two-way benefits, and examples are given of various collaborations in these fields.     

Are research efforts on Animalia in the South Pacific associated with the conservation status or population trends?

Analyses of knowledge gaps can highlight imbalances in research, encouraging greater proportionality in the distribution of research efforts. In this research we used generalized linear mixed models (GLMM) with the aim to determine if research efforts for the period 2005–2015 for terrestrial vertebrates of Amphibia, Aves, Mammalia and Reptilia in the South Pacific region were correlated with conservation status (critically endangered (CR), endangered (EN), vulnerable (VU), least concern (LC) and near threatened (NT)) or population trends (increasing, stable, decreasing and unknown) through the International Union for Conservation of Nature (IUCN) database. Our results showed that research distribution was uneven across different classes. Out of 633623 investigated papers, the average number of publications per species was 43.7, 306.7, 717.6 and 115.3 for Amphibia (284 species), Aves (1306 species), Mammalia (243 species) and Reptilia (400 species), respectively. Consistently, the lower publication effort on Amphibia compared to other taxonomic classes was revealed as significant by GLMM analysis. There was no significant differences in research effort among levels of conservation status. However, we found significantly different publication efforts among population trends of all examined species in that species with “unknown” population trends gained significantly lower researchers’ attention compared to species with “decreasing” trend. Results also indicated that, although it was not significant, the highest attention is given to species with “increasing” population trend over all taxonomic classes. Using the Information Theoretic approach we also generated a set of competing models to identify most important factors influencing research efforts, revealing that the highest ranked model included taxonomic class and population trend.     

Mutant vasopressin precursor in the endoplasmic reticulum of the Brattleboro rat. Ultrastructural evidence from individual “vasopressin” cells localized with the light microscope by use of a new gold/silver method for immunostain enhancement

Summary This ultrastructural study demonstrates that the vasopressin immunoreactivity found in the occasional, densely stained cells in the hypothalamus of the homozygous Brattleboro rat is localized in the rough endoplasmic reticulum. 50-m Vibratome sections were stained with anti-vasopressin serum by use of a peroxidase method with 3,3-diaminobenzidine as chromogen. The diaminobenzidine end-product has a specific capability to bind gold particles from a chloroauric acid solution and the bound gold was used to precipitate silver grains from a silver developer. The stained sections were flat embedded in resin and ultrathin sections were cut of areas containing the immuno-identified occasional cells. In these densely stained, vasopressin-immunoreactive cells of homozygous Brattleboro rats the rough endoplasmic reticulum was dilated. The lumen of the reticulum contained both end-products of diaminobenzidine and gold/silver grains, but some parts of the reticulum appeared unstained. No other cell organelles were immunostained and no secretory granules were found. In control rats, gold/silver deposits were found throughout the cytoplasm of vasopressin-immunoreactive cells. In these immunostained cells secretory granules were seen.     

Are the early holocene cattle in the eastern sahara domestic or wild?

Questions relating to the antiquity of domestic cattle in the Sahara are among the most controversial in North African prehistory. It is generally believed that cattle were first domesticated in southwest Asia, particularly Anatolia, or in southeast Europe, where their remains have been found in several sites dated between 9,000 and 8,000 years ago.1 The discovery, in several small sites in the Western Desert of Egypt, of large bovid bones identified as domestic cattle and having radiocarbon dates ranging between 9,500 and 8,000 B.P. has raised the possibility that there was a separate, independent center for cattle domestication in northeast Africa (Fig. 1).^2–4 However, it has not been universally accepted that these bones are from cattle or, if so, that the cattle were domestic.     

Are we missing half of the viruses in the ocean?

Viruses are abundant in the ocean and a major driving force in plankton ecology and evolution. It has been assumed that most of the viruses in seawater contain DNA and infect bacteria, but RNA-containing viruses in the ocean, which almost exclusively infect eukaryotes, have never been quantified. We compared the total mass of RNA and DNA in the viral fraction harvested from seawater and using data on the mass of nucleic acid per RNA- or DNA-containing virion, estimated the abundances of each. Our data suggest that the abundance of RNA viruses rivaled or exceeded that of DNA viruses in samples of coastal seawater. The dominant RNA viruses in the samples were marine picorna-like viruses, which have small genomes and are at or below the detection limit of common fluorescence-based counting methods. If our results are typical, this means that counts of viruses and the rate measurements that depend on them, such as viral production, are significantly underestimated by current practices. As these RNA viruses infect eukaryotes, our data imply that protists contribute more to marine viral dynamics than one might expect based on their relatively low abundance. This conclusion is a departure from the prevailing view of viruses in the ocean, but is consistent with earlier theoretical predictions.