Radioimmunological Quantification of C21, C19 and C18 Steroids in Haemolymph of the Insect Locusta migratoria

Summary

Titers of pregnenolone, progesterone, testosterone, 5α-dihydrotestosterone, estrone and estradiol were measured by radioimmunoassay in purified haemolymph extracts of larval and adult male and female Locusta migratoria. They varied between following values (in pg/ml): pregnenolone: 467–757; progesterone: 37–119; testosterone: 11–54; 5α-dihydrotestosterone: 13–41; estrone: 25–1392; estradiol: 12–26. Titers of pregnenolone progesterone, testosterone, 5α-dihydrotestosterone and estradiol did not substantially fluctuate among the developmental stages we examined. Peaks of estrone were found in males and females in the middle of the fifth larval instar and in 3 week old adult males. The titers of most of the above six steroids are about 5 to 10 times lower than the concentrations found in purified extracts of several tissues of this insect.     

Clinical experience with heparinoid sodium pentosan polysulfate (SP54) in the treatment of cerebral ischemia

Fifty-five patients suffering from acute cerebral ischaemia were treated with pentosan polysulphate (SP54) intravenously and subsequently orally. The control group consisting of 35 patients received the infusion solution and vasodilator drugs. The statistically significant changes appearing following infusion of SP54 include the following parameters: haematocrit, blood viscosity and cholesterol concentrations. In individual cases the euglobulin lysis time was shortened within two hours after the beginning of the infusion. As demonstrated by the statistically reliable results in patients treated with SP54 the neurological symptoms improved significantly within the first two weeks after the onset of the disease compared to the control groups. Based on case reports, recommendations are given for the indication and contraindication of SP54 treatment in cerebral ischaemia.     

Lactobacillus harbinensis sp. nov., consisted of strains isolated from traditional fermented vegetables 'Suan cai' in Harbin, Northeastern China and Lactobacillus perolens DSM 12745

Taxonomical analysis of two genetically distinguished Lactobacillus strains isolated from traditional Chinese fermented vegetables ‘Suan cai’ was performed. They formed l-lactate from glucose, were facultatively heterofermentative, and had a DNA G+C content of 53–54 mol%. They fermented d- and l-arabinose. They produced lactate, ethanol and acetate from gluconate at a molar ratio of 1.1:0.4:0.7. Phylogenetic analysis of 16S rDNA revealed that the two strains were closely related to L. perolens. DNA–DNA hybridization analysis revealed that the two strains were different from L. perolens type strain DSM 12744 and formed a separate cluster with L. perolens DSM 12745. G+C molar content of DNA of the former is 51%, whereas those of the latter strains were in the range of 53–54%. Based on the results, we propose that the new species be named L. harbinensis sp. nov. and that L. perolens DSM 12745 be reclassified as L. harbinensis DSM 12745. The type strain of L. harbinensis DSM 16991^T (=AHU 1762^T=SBT 10908^T).     

The type species of the Ordovician agnostid trilobiteGeragnostus Howell, 1935

The type of the widespread Ordovician agnostid trilobiteGeragnostus, G. sidenbladhi (Linnarsson, 1869) from the upper Tremadocian Ceratopyge Limestone of Sweden, is redescribed and a lectotype is selected.Geragnostus Howell, 1935 bears a strong similarity toArthrorhachis Hawle & Corda, 1847. Obviously the two genera are closely allied, andGeragnostus may eventually be regarded as a junior subjective synonym ofArthrorhachis. It is suggested, however, that the distinction betweenGeragnostus andArthrorhachis might be upheld if the latter genus is taken to include metagnostids in which the posterior lobe (M3) of the pygidial rhachis is distinctly shorter than the length (sag.) of the anterior and middle lobes combined (M1 + M2).     

Impact of treating hyperlipidemia or hypertension to reduce the risk of death from coronary artery disease

OBJECTIVE: To compare the prevalence of modifiable risk factors for cardiovascular disease among hypertensive and nonhypertensive adults and to estimate the effect of treating hyperlipidemia or hypertension to reduce the risk of death from coronary artery disease. METHODS: The authors evaluated a sample of 7814 subjects aged 35-74 years free of clinical cardiovascular disease from the Canadian Heart Health Surveys to estimate the prevalence of cardiovascular risk factors. They identified hyperlipidemic subjects (ratio of total cholesterol to high-density lipoprotein cholesterol [total-C/HDL-C] 6.0 [corrected] or more for men and 5.0 [corrected] or more for women) and hypertensive subjects (systolic or diastolic blood pressure 160/90 mm Hg or greater, or receiving pharmacologic or nonpharmacologic treatment). A life expectancy model was used to estimate the rate of death from coronary artery disease following specific treatments. RESULTS: An elevated total-C/HDL-C ratio was significantly more common among hypertensive than nonhypertensive men aged 35-64 (rate ratio [RR] 1.56 for age 35-54, 1.28 for age 55-64) and among hypertensive than nonhypertensive women of all ages (RR 2.73 for age 35-54, 1.58 for age 55-64, 1.31 for age 65-74). Obesity and a sedentary lifestyle were also more common among hypertensive than among nonhypertensive subjects. According to the model, more deaths from coronary artery disease could be prevented among subjects with treated but uncontrolled hypertension by modifying lipids rather than by further reducing blood pressure for men aged 35-54 (reduction of 50 v. 29 deaths per 100,000) and 55-64 (reduction of 171 v. 104 deaths per 100,000) and for women aged 35-54 (reduction of 44 v. 39 deaths per 100,000). Starting antihypertensive therapy in subjects aged 35-74 with untreated hypertension would achieve a greater net reduction in deaths from coronary artery disease than would lipid lowering. Nonetheless, the benefits of lipid therapy were substantial: lipid intervention among hypertensive subjects aged 35-74 represented 36% of the total benefits of treating hyperlipidemia in the total hyperlipidemic population. INTERPRETATION: The clustering of hyperlipidemia and the potential benefits of treatment among hypertensive adults demonstrate the need for screening and treating other cardiovascular risk factors beyond simply controlling blood pressure.     

Molecular weight determination of methyl esters of mycolic acids using thermospray mass spectrometry.

Methyl esters of normal fatty acids, corynomycolate and corynomycolenate were used as model compounds for thermospray mass spectrometric procedures for molecular weight determination of the related nocardial mycolic acids. By using ammonium acetate at the positive ion generator, in both cases, a family of ions was produced. The following members were found and corresponded to the adducts: (1) M + H; M + NH4 and M + H + NH4 for methyl esters of normal fatty acids, whereas M + H, M + 2H and M + H + NH4 were the adducts most frequently observed with methyl corynomycolates. The methyl esters of C40-C48 mycolic acids from Rhodococcus rhodochrous exhibited prominent peaks corresponding to adducts M + H + NH4 whereas those corresponding to M + 2H showed slightly lower intensities. The structure M + H had no significant representatives with this subclass of mycolic acids. A similar pattern was observed with methyl esters of C50-C54 mycolic acids from Nocardia asteroides GUH-2. Ion peaks C50-C54 representing adducts M + 2H and M + H + NH4 prevailed in the mass spectrum. In this case, the intensities of peaks corresponding to M + 2H were slightly higher than those of the M + H + NH4. Essentially three main species of nocardomycolic acids were detected: (1) monounsaturated C50:1, C52:1 and C54:1; (2) diunsaturated C50:2, C52:2 and C54:2 and (3) triunsaturated C52:3 and C54:3 mycolic acids. The most abundant mycolic acid was C52:2 followed in decreasing abundance by C52:1, C54:2, C50:2, C52:3 and C54:3 mycolic acids.     

Straight-chain fatty alcohols in the hyperthermophilic archaeon Pyrococcus furiosus

Two straight-chain fatty alcohols (n-hexadecanol and n-octadecanol) were found in the neutral lipid fraction extracted from Pyrococcus furiosus cells. They were identified by thin-layer and gas-liquid chromatography, mass and infrared spectra, and chemical modification. The fatty alcohols accounted for 54% of the neutral lipid of the cell. Received: March 8, 2000 / Accepted: May 8, 2000     

Pathogenic Aeromonas hydrophila serogroup O:14 and O:81 strains with an S layer

Five autoagglutinating Aeromonas hydrophila isolates recovered from eels and humans were assigned to serogroups O:14 and O:81 of the Sakazaki and Shimada (National Institutes of Health) scheme. They had the following properties in common: positive precipitation after boiling, moderate surface hydrophobicity (salt-aggregation-test value around 1.2), pathogenicity for fish and mice (50% lethal dose, 10(4.61) to 10(7.11)), lipopolysaccharides that contained O-polysaccharide chains of homogeneous chain length, and an external S layer peripheral to the cell wall observed by electron microscopy. A strong cross-reactivity was detected by immunoblotting between the homogeneous O-polysaccharide fraction of O:14 and O:81 strains but not between them and the lipopolysaccharide of A. hydrophila TF7 (O:11 reference strain). Outer membrane fractions of these strains contained a predominant 53- to 54-kDa protein which was glycine extractable under low-pH (pH 2.8) conditions and was identified as the surface array protein. The S-layer proteins of the O:14 and O:81 A. hydrophila strains seemed to be primarily different from those previously purified from strains A. hydrophila TF7 and Aeromonas salmonicida A450 on the basis of colony hybridizations with both the structural genes vapA and ahsA. This is the first report of the presence of an S layer in mesophilic Aeromonas strains not belonging to serogroup O:11.     

THE ISOLATION AND PROPERTIES OF LARGE BASIC PEPTIDES FROM BOVINE SPINAL CORD

Abstract— Two basic peptides (B1 and B2) were derived from bovine spinal cord following in situ proteolysis at 37°C for 10–24 h. These peptides do not arise as degradation products from the A1 protein as shown by amino acid composition and end group analysis; rather they appear to originate from some larger basic protein in the spinal cord having similarities to the P2 protein, a basic protein found in peripheral nerve myelin. The peptides were purified following defatting, acid extraction, and ammonium sulphate fractionation, by chromatography on Amberlite IRC-50 resin using guanidinium chloride. The peptides, found generally in a 4:1 ratio of B1 to B2, appeared homogeneous on gel electrophoresis and immunodiffusion. Approximately 25–60 mg of peptides was obtained per 100 g wet spinal cord.
In contrast to the basic A1 protein from myelin, neither of these peptides nor their pepsin digests were encephalitogenic. They do not cross-react immunologically with the basic A1 protein, but cross-react with each other. These peptides further differ from the A1 protein in their tryptic peptide map, size (B1, 63 residues; B2, 54 residues), and composition particularly the high lysine: arginine ratio, and low histidine content. Like the A1 protein, however, they contain a tryptophan residue and a blocked NH₂-terminal amino acid; peptide Bl has COOH-terminal valine. It was concluded that the basic peptides represent a fragment of a hitherto unidentified protein(s) of the nervous system.     

Association of the 54-nucleotide repeat polymorphism of hPer3 with heroin dependence in Han Chinese population

Circadian clock genes have the function of producing circadian rhythm. They are also implicated in the origin or development of many diseases such as cancers and neuropsychiatric diseases. The purpose of this study is to determine whether the 54-nucleotide repeat polymorphism of hPer3 , one of the circadian clock genes, associates with heroin dependence. DNA samples were obtained from 209 Chinese heroin-dependent subjects and 249 Chinese healthy controls. The 54-nucleotide repeat polymorphism was detected by polymerase chain reaction and DNA agarose gel electrophoresis. The frequency of four-repeat allele was significantly higher ( χ ²= 10.64, P = 0.001; corrected for multiple tests, P = 0.003) in the mixed gender heroin-dependent subject group (four repeat: 0.89, five repeat: 0.11) than in the mixed gender control group (four repeat: 0.81, five repeat: 0.19); the frequency of four-repeat allele was also significantly higher ( χ ²= 10.00, P = 0.002; corrected for multiple tests, P = 0.006) in the male heroin-dependent subject group (four repeat: 0.89, five repeat: 0.11) than in the male control group (four repeat: 0.81, five repeat: 0.19); for females, no significant trend was observed with the 54-nucleotide repeat polymorphism between the heroin-dependent subject group and the control group. Our results suggest that the 54-nucleotide repeat polymorphism of hPer3 significantly associates with heroin dependence at the allele frequency level and may be a potential risk factor for the development of heroin dependence.     

Karyotype evolution in Curimatidae (Teleostei, Characiformes) from the Amazon region. II. Centric fissions in the genus Potamorhina

Using cytogenetic analysis following Giemsa staining, nucleolar organizer region (NOR) staining, and C-banding, three distinct karyotypes in three species of curimatids belonging to the fish genus Potamorhina were identified: 2n = 54/44 M + 10 SM (P. pristigaster), 2n = 56/52 M + 2 SM + 2 ST (P. latior), and 2n = 102/2 M + 2 SM + 98 A (P. altamazonica). A 2n = 54 was considered to be the ancestral diploid number and the different karyotypes were probably the result of centric fissions. Both the NOR pattern and constitutive heterochromatin pattern are species specific.     

The postweaning rise of tonic luteinizing hormone secretion in anestrous cows is not prevented by chronic milking or the physical presence of the calf

The objective of the following study was to examine the ability of frequent milking, the physical presence of the calf, and their combination to prevent a postweaning rise in tonic luteinizing hormone (LH) secretion, estrus, and ovulation. Thirty Hereford cows were allowed to suckle their calves ad libitum until 17-21 days post partum and confirmed as anestrus. They were then assigned alternately by order of calving to 1 of 5 treatment groups: (1) Suckled (S) ad libitum; (2) Nonsuckled (NS)--calf removed for 102 h; (3) Nonsuckled--calf present (NSC)--calf remained with cow, but muzzled to prevent suckling for 102 h; (4) Nonsuckled--milked 8 times a day (NSM)--calf removed for 102 h and cow hand-milked for 10 min every 2 h from 0700 to 2100 h; (5) Nonsuckled--calf present--milked 8 times a day (NSMC)--combination of 3 and 4. Luteinizing hormone secretion patterns, estrous activity, and ovulation were monitored throughout the experiment. Prior to treatment (Day 0), mean pulse frequency (pulses/6 h), mean concentrations (ng/ml), and median concentrations (ng/ml) of LH did not differ (p greater than 0.45) between groups, and were 0.7 +/- 0.15, 2.8 +/- 0.14, and 2.6 +/- 0.11, respectively. Marked rises (p less than 0.01--p less than 0.03) in LH pulse frequency were observed in all groups except S between 48 and 54 h after onset of treatment. Mean and median concentrations of LH were lower (p less than 0.02) in S cows than in all other groups at 48-54 h.(ABSTRACT TRUNCATED AT 250 WORDS)     

Formation of bulges associated with penicillin production in high-producing strains ofPenicillium chrysogenum

Bulges were formed in the hyphae of a high penicillin-producing strain ofPenicillium chrysogenum AS-P-78 when penicillin was accumulated in the broth. The mycelium of cultures grown in the presence of 50 mM lysine, which specifically inhibits penicillin formation, showed a greatly reduced number of bulges. The size and number of bulges increased during the fermentation in parallel with penicillin accumulation. A smaller number of bulges was formed in the mycelium of the low-producing strain Wis 54-1255 than in the high-producing mutant. Three mutants blocked in penicillin biosynthesis did not form these globose structures. Bulges were not osmotically sensitive, although some of them burst out. They may be formed by weakening of the cell wall of hyphae following accumulation of high concentrations of penicillin.     

Mechanism of interferon action. II. Induction and decay kinetics of the antiviral state and protein P54 in human amnion U cells treated with gamma interferon

The kinetics of induction and decay of the antiviral state and polypeptide p54 expression induced by recombinant human interferon gamma (rIFN-gamma) were examined in human amnion U cells. The kinetics of induction of the antiviral state, as measured by the single-cycle yield reduction of vesicular stomatitis virus, were first order over the period of about 6-12 h following a lag of about 2-4 h. The induction of p54 synthesis by rIFN-gamma slightly preceded the induction of the antiviral state. The kinetics of p54 induction were first order over a period of about 2-8 h after a lag of about 1 h. The rate of polypeptide p54 synthesis induced by rIFN-gamma decayed significantly within 1 day after the removal of IFN. However, polypeptide p54 was comparatively stable, displaying a half-life of about 3 days. The antiviral state likewise decayed significantly within 3-4 days following removal of IFN-gamma, and by 5-8 days, the virus yields were comparable to those of untreated control cell cultures. These results suggest that polypeptide p54 may play an important role in the antiviral action of rIFN-gamma in human amnion U cells.     

Differential ovarian expression of KiSS-1 and GPR-54 during the estrous cycle and photoperiod induced recrudescence in Siberian hamsters (Phodopus sungorus)

Kisspeptins, coded by the KiSS-1 gene, regulate aspects of the reproductive axis by stimulating GnRH release via the G protein coupled receptor, GPR54. Recent reports show that KiSS/GPR54 may be key mediators in photoperiod-controlled reproduction in seasonal breeders, and that KiSS-1/GPR54 are expressed in the hypothalamus, ovaries, placenta, and pancreas. This study examined the expression of KiSS-1/GPR54 mRNA and protein in ovaries of Siberian hamsters (Phodopus sungorus). Ovaries from cycling hamsters were collected during proestrus (P), estrus (E), diestrus I (DI), and diestrus II (DII). To examine KiSS-1/GPR54 during stimulated recrudescence, additional hamsters were maintained either in long day (LD 16L:8D, control) or short day (SD 8L:16D) for 14 weeks and then transferred to LD for 0-8 weeks. Staining of KiSS-1/GPR54 protein was detected by immunohistochemistry in steroidogenic cells of pre-antral and antral follicles, and corpora lutea. Immunostaining peaked in P and E, but decreased in the diestrus stages (P < 0.05). In recrudescing ovaries, KiSS-1/GPR54 immunostaining was low after 14 weeks of SD exposure (post-transfer [PT] week 0), and increased during the early weeks of recrudescence. Expression of KiSS-1/GPR54 mRNA was low with short day exposure, but increased during recrudescence and was higher at PT week 8 as compared to PT weeks 0 and 2 (P < 0.05). The elevated KiSS-1/GPR54 expression during P and E suggests a potential role in ovulation in Siberian hamsters. Transient increases in KiSS-1/GPR54 expression following LD stimulation are also suggestive of possible involvement in ovulation and/or restoration of ovarian function.     

Functional and structural organization of chlorophyll in the developing photosynthetic membranes of Euglena gracilis Z. V-separation and characterization of pigment-protein complexes of the differentiated thylakoids

Low temperature sodium dodecyl sulfate polyacrylamide gel electrophoresis following mild solubilization of Euglena thylakoid components allowed to resolve, in addition to the main CP1, CPa and LHCP chlorophyll-protein complexes, the additional CP1a and LHCP green bands. A carotenoid enriched band CPc can be separated from CPa using high acrylamide concentration. Pigment and polypeptide composition of these complexes were analyzed by absorption and fluorescence measurements and two dimensional gel electrophoresis. Spectral properties of CP1 and CP1a indicate an heterogenous organization of chlorophyll and the presence of significant amount of chlorophyll b in these complexes. They both contain a major 68 kilodalton polypeptide associated with three minor low molecular weight polypeptides in CP1a. CPa and CPc exhibit a characteristic fluorescence emission at 687 nm and they each contain one polypeptide of 54 and 41 Kda respectively. LHCP and LHCP are less abundant than in higher plant thylakoids and they contain a lower proportion of chl b (chl a: chl b=3). They include two polypeptides of 26 and 29 Kda.Abbreviations chl chlorophyll - SDS Sodium Dodecyl Sulfate - EDTA Ethylene Diamine Tetraacetic Acid - DTT Dithiothreitol     

The metastasis suppressor gene KiSS-1 encodes kisspeptins, the natural ligands of the orphan G protein-coupled receptor GPR54.

Natural peptides displaying agonist activity on the orphan G protein-coupled receptor GPR54 were isolated from human placenta. These 54-, 14,- and 13-amino acid peptides, with a common RF-amide C terminus, derive from the product of KiSS-1, a metastasis suppressor gene for melanoma cells, and were therefore designated kisspeptins. They bound with low nanomolar affinities to rat and human GPR54 expressed in Chinese hamster ovary K1 cells and stimulated PIP(2) hydrolysis, Ca(2+) mobilization, arachidonic acid release, ERK1/2 and p38 MAP kinase phosphorylation, and stress fiber formation but inhibited cell proliferation. Human GPR54 was highly expressed in placenta, pituitary, pancreas, and spinal cord, suggesting a role in the regulation of endocrine function. Stimulation of oxytocin secretion after kisspeptin administration to rats confirmed this hypothesis.     

Antioxidant properties of natural p-terphenyl derivatives from the mushroom Thelephora ganbajun

The antioxidant activity in vitro of three poly(phenylacetyloxy)-substituted 1,1':4',1"-terphenyl compounds from the edible mushroom Thelephora ganbajun were investigated. The IC50 values of compounds 1-3 for lipid peroxidation in rat liver homogenate were 400, 48, 54 microM, respectively. Compounds 1-3 increased superoxide dismutase (SOD) activity with EC50 values of 182, 74, 204 microM. They were also assessed on the DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity with EC50 values of 49, 1233, 55 microM.     

Fructose 2,6-bisphosphate, carbohydrate partitioning, and crassulacean Acid metabolism

To initiate structural studies of the ADPglucose pyrophosphorylase from spinach an improved purification procedure was devised. The modified purification scheme allowed the isolation of 20 to 30 milligrams pure enzyme from 10 kilogram of spinach leaves. Electrophoresis of the purified enzyme confirmed an earlier study which showed that the enzyme was putatively composed of two subunits (Copeland L, J Preiss 1981 Plant Physiol 68: 996-1001). The two subunits migrate as 51 and 54 kilodalton proteins upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Both proteins can be detected on Western blots of leaf homogenates prepared under denaturing conditions suggesting that both subunits exist in vivo. Anion-exchange chromatography in the presence of urea allowed resolution of the 51 and 54 kilodalton proteins. They possess different N-terminal amino acid sequences and tryptic peptide maps. Western blot analysis reveals that the 51 and 54 kilodalton proteins are antigenically dissimilar. The 51 but not the 54 kilodalton protein is immunologically related to the ADPglucose pyrophosphorylase from maize endosperm and potato tuber.     

Photoperiodic stimulation of pubertal development in male deer mice: involvement of the circadian system

Pubertal development in prairie deer mice (Peromyscus maniculatus bairdii) is accelerated by exposure of juveniles to a long-day photoperiod, and, conversely, retarded by exposure to short days. The purpose of the present study was to evaluate the possible involvement of the circadian system in the photoperiodic regulation of puberty. Weanling males, previously housed on a short-day light cycle of 6L:18D, were subjected to a "resonance" protocol in which they received one of the following light cycles: 6L:18D, 6L:30D, 6L:42D, 6L:54D, or 16L:8D. Post-weaning exposure to cycles of 16L:8D, 6L:30D, and 6L:54D stimulated reproductive organ growth as measured at 6 weeks of age. Exposure to cycles of 6L:18D and 6L:42D failed to stimulate reproductive development. These data support the hypothesis that young male deer mice use a circadian rhythm of responsiveness to light to measure photoperiodic time and, consequently, regulate pubertal development.