Identification of Closely Related Hydrobaenus Species (Diptera: Chironomidae) Using the Second Internal Transcribed Spacer (ITS2) Region of Ribosomal DNA

Three species of Japanese Hydrobaenus Fries (H. Kondoi, H. biwaquartus and H. kisosecundus) can be separated by variation in the sequence of the second internal transcribed spacer (ITS2) regions of ribosomal DNA. Suggested synonymy of H. kondoi with H. biwaquartus is refuted, and previously suggested diagnostic morphology (virga length, antennal, leg and venarum ratios, shape of tergite IX and gonostylus length) distinguishes the taxa. No difference in larval morphology was found. H. kisosecundus is readily distinguished on adult and larval morphology from H. kondoi and H. biwaquartus, and is more distant by molecular similarity measures. ITS2 regions apparently provide useful information for distinguishing closely related species in Chironomidae.     

An increase in in vivo release of LHRH and precocious puberty by posterior hypothalamic lesions in female rhesus monkeys (Macaca mulatta)

We have previously shown that a decrease in gamma-aminobutyric acid (GABA) tone and a subsequent increase in glutamatergic tone occur in association with the pubertal increase in luteinizing hormone releasing hormone (LHRH) release in primates. To further determine the causal relationship between developmental changes in GABA and glutamate levels and the pubertal increase in LHRH release, we examined monkeys with precocious puberty induced by lesions in the posterior hypothalamus (PH). Six prepubertal female rhesus monkeys (17.4 +/- 0.1 mo of age) received lesions in the PH, three prepubertal females (17.5 +/- 0.1 mo) received sham lesions, and two females received no treatments. LHRH, GABA, and glutamate levels in the stalk-median eminence before and after lesions were assessed over two 6-h periods (0600-1200 and 1800-2400) using push-pull perfusion. Monkeys with PH lesions exhibited external signs of precocious puberty, including significantly earlier menarche in PH lesion animals (18.8 +/- 0.2 mo) than in sham/controls (25.5 +/- 0.9 mo, P<0.001). Moreover, PH lesion animals had elevated LHRH levels and higher evening glutamate levels after lesions, whereas LHRH changes did not occur in sham/controls until later. Changes in GABA release were not discernible, since evening GABA levels already deceased at 18-20 mo of age in both groups and morning levels remained at the prepubertal levels. The age of first ovulation in both groups did not differ. Collectively, PH lesions may not be a good tool to investigate the mechanism of puberty, and, taking into account the recent findings on the role of kisspeptins, the mechanism of the puberty onset in primates is more complex than we initially anticipated.     

False killer whales (Pseudorca crassidens) from Japan and South Africa: Differences in growth and reproduction

Age and reproductive information for 65 false killer whales stranded in South Africa in 1981 are compared with similar material from 156 animals examined from drive fisheries in Japan in 1979 and 1980. Sizes at birth, sexual maturation, and physical maturity all indicated that both sexes were 10%–20% larger in Japan than South Africa. Females reached sexual maturation at similar ages (8–10.5 yr) in both populations, and although sample sizes were too small to establish male ages at puberty precisely the ranges in Japan (10.5–18.5 yr) and South Africa (5.25–17.5 yr) were not inconsistent. The initial ovulation rate for females from South Africa was 65% lower (and the apparent pregnancy rate 82% lower) than those from Japan and there were fewer animals ≤2 yr old within the school, but the magnitude of these differences suggests that the stranded school's reproductive performance was probably impaired. Collectively these comparisons and the literature indicate substantive size differences between false killer whales in different populations, although the patterns of growth appear similar. Firm conclusions about any geographical differences in reproduction require additional data.     

Phylogenetic placement of Geastrum melanocephalum and polyphyly of Geastrum triplex

Geastrum melanocephalum, originally described as Trichaster melanocephalus, is characterized by large basidiomata and an evanescent endoperidium. Although Trichaster was recently treated as a synonym of Geastrum and the specific name G. melanocephalum has often been used, it is still controversial whether Trichaster is an independent genus. Although a close affinity of G. melanocephalum and G. triplex has been suggested based on some morphological similarities, it is highly likely that G. triplex is polyphyletic because of its high morphological variability. To clarify the phylogenetic position of G. melanocephalum, it is therefore critical to evaluate the monophyly of G. triplex. This study sampled ITS, LSU, and atp6 genes from 144 specimens of Geastrales including G. melanocephalum and G. triplex from several continents. Results of phylogenetic analyses demonstrated G. melanocephalum is nested within Geastrum and is most closely related to the European and North American group of G. triplex. Morphological similarities of G. melanocephalum and European and North American G. triplex are also suggested. Based on phylogenetic and morphological evidence, we confirm Trichaster is a synonym of Geastrum, and the scientific name Geastrum melanocephalum should be accepted. Moreover, the present study revealed that taxa tentatively identified as “G. triplex” are highly polyphyletic, and a taxonomic revision of “G. triplex” is therefore needed.     

The effect of water loading on the urinary ratio of cortisone to cortisol in healthy subjects and a new approach to the evaluation of the ratio as an index for in vivo human 11β-hydroxysteroid dehydrogenase 2 activity

Factors that give rise to a large variation in the urinary ratio of free cortisone to cortisol (UFE/UFF) were investigated to accurately estimate 11β-hydroxysteroid dehydrogenase 2 (11β-HSD2) activity in humans in vivo. A water loading test was first carried out in two healthy subjects to examine the effect of water intake or urine volume on the urinary ratio of free cortisone to cortisol (UFE/UFF). The ratio was found to increase by water loading. We also examined urinary concentrations and amounts of cortisol, cortisone, creatinine, Na(+), K(+), and Cl(-), and urine volume, as possible factors affecting the urinary ratio (UFE/UFF), in 60 urine samples obtained from 15 healthy volunteers. Among these factors tested, the urinary concentration of cortisol was most highly correlated with the UFE/UFF ratio (r=-0.858), indicating that the in vivo activity of 11β-HSD2 (UFE/UFF) should fluctuate with the changes of the urinary concentration of cortisol. Based on the findings, we proposed a new estimation method of in vivo activity of 11β-HSD2 in humans, using the UFE/UFF ratio correlated with the urinary concentration of cortisol (UFE/UFF-cortisol concentration). Taking into consideration the intra-individual variabilities in the urinary concentration of cortisol, there were no significant within-day variations in 11β-HSD2 activity. The findings indicate that 11β-HSD2 activities can be accurately evaluated by simply measuring free cortisol and cortisone concentrations in spot urine samples. Furthermore, administrations of glycyrrhetinic acid in three healthy volunteers were performed to confirm the usefulness of the present assessment for the activity of 11β-HSD2.     

Improved synthesis and evaluation of 17-substituted aminoalkylgeldanamycin derivatives applicable to drug delivery systems

The 17-methoxy group of geldanamycin was substituted with 1,3-diaminopropane and 1,3-diamino-2-hydroxypropane to introduce a primary amino group useful for conjugation with targeting moieties and drug carriers. We have developed a procedure that has provided improved yield and reproducibility of the syntheses. Both geldanamycin derivatives demonstrated antiproliferative activity towards the human ovarian carcinoma cell line, A2780.     

Evolutionally conserved intermediates between ubiquitin and NEDD8

The investigation of common structural motifs provides additional information on why proteins conserve similar topologies yet may have non-conserved amino acid sequences. Proteins containing the ubiquitin superfold have similar topologies, although the sequence conservation is rather poor. Here, we present novel similarities and differences between the proteins ubiquitin and NEDD8. They have 57% identical sequence, almost identical backbone topology and similar functional strategy, although their physiological functions are mutually different. Using variable pressure NMR spectroscopy, we found that the two proteins have similar conformational fluctuation in the evolutionary conserved enzyme-binding region and contain a structurally similar locally disordered conformer (I) in equilibrium with the basic folded conformer (N). A notable difference between the two proteins is that the equilibrium population of I is far greater for NEDD8 (DeltaG(0)(NI)<5 kJ/mol) than for ubiquitin (DeltaG(0)(NI)=15.2(+/-1.0) kJ/mol), and that the tendency for overall unfolding (U) is also far higher for NEDD8 (DeltaG(0)(NU)=11.0(+/-1.5) kJ/mol) than for ubiquitin (DeltaG(0)(NU)=31.3(+/-4.7) kJ/mol). These results suggest that the marked differences in thermodynamic stabilities of the locally disordered conformer (I) and the overall unfolding species (U) are a key to determine the functional differences of the two structurally similar proteins in physiology.