Cloning and phylogenetic relationship of red drum somatolactin cDNA and effects of light on pituitary somatolactin mRNA expression.

The nucleotide sequence for red drum somatolactin (SL) cDNA was determined and the expression of pituitary SL mRNA was examined in red drum kept under various light conditions. A full length of SL cDNA (1629 bp) was isolated and characterized from a red drum pituitary cDNA library. The SL cDNA has an open reading frame of 696 nucleotides which encodes a 24-amino-acid signal peptide and a 207-amino-acid mature peptide.

Structural and expression analyses of gonadotropin Ibeta subunit genes in goldfish (Carassius auratus).

Gonadotropin (GTH) is a pituitary glycoprotein hormone that regulates gonadal development in vertebrates. In teleosts, it is considered that two types of GTH, GTH I (follicle-stimulating hormone-like GTH) and GTH II (luteinizing hormone-like GTH), are produced in the pituitary, and their molecules are comprised of common alpha and distinct beta subunits. In this study, we describe the complete structure and 5'-flanking regulatory region of two distinct genes encoding GTH Ibeta in goldfish, Carassius auratus.

Isolation of a 65-kDa protein from white muscle of warm temperature-acclimated goldfish (Carassius auratus).

A 65-kDa protein expressed in association with warm temperature acclimation of goldfish (Carassius auratus) was purified from epaxial muscle by successive ion-exchange, gel filtration, and reversed-phase columns while monitoring immuno-reaction with a specific antibody. A total of 517 micrograms of the 65-kDa protein was obtained from 23.4 g of the muscle of 30 degrees C-acclimated fish.

[Basic and clinical studies on tazobactam/piperacillin in pediatric field].

A drug susceptibility test of the combination drug TAZ/PIPC, which consists of a newly developed beta-lactamase inhibitor, tazobactam (TAZ), and one of penicillin antibiotics, piperacillin (PIPC), with combination ratio of 1:4 in potency, was conducted with stock strains and clinical isolates. The clinical efficacy and safety of its injection was also evaluated in children with a variety of infectious diseases. The results were as follows: 1.

Ontogenic development of salmon GnRH and chicken GnRH-II systems in the brain of masu salmon (Oncorhynchus masou).

Ontogenic development of salmon gonadotropin-releasing hormone (GnRH) and chicken GnRH-II systems in masu salmon (Oncorhynchus masou) was examined. Salmon GnRH was first detected by radioimmunoassay in the embryo on day 36 after fertilization. Salmon GnRH-immunoreactive fibers were detected initially by immunocytochemistry in the vicinity of the olfactory placode of the embryo (day 36) and were distributed widely in the brain as well as in the pituitary gland of fish just after hatching (day 80).

A cross-sectional evaluation of spontaneous platelet aggregation in relation to complications in patients with type II diabetes mellitus.

To clarify the relationship between platelet function and diabetic complications, we investigated spontaneous platelet aggregation (SPA) and agonist-induced platelet aggregation by a particle counting method using light scattering (LS) and by a conventional light transmission method (LT) in 23 age- and sex-matched control subjects and 74 patients with type II diabetes mellitus. We also observed platelets using the FIC-2 (TOA Medical Electronics, Kobe, Japan) flow cytometer and imaging device. Observation by the FIC-2 device showed microaggregates of platelets in samples with increased SPA-LS.

Hyperglycaemic hormones inhibit protein and mRNA synthesis in in vitro-incubated ovarian fragments of the marine shrimp Penaeus semisulcatus.

The present work shows for the first time that peptides belonging to the Crustacean hyperglycaemic hormone family (CHH-family hormones) from Penaeus japonicus affect protein and mRNA synthesis in in vitro-incubated ovarian explant fragments removed from vitellogenic females of Penaeus semisulcatus. Reduced levels of protein synthesis, determined by TCA-precipitable 35S-labeled proteins, were found in the presence of crude sinus gland extracts from both P. semisulcatus and P.

Two differing salmon GnRH precursor mRNAs are co-expressed in the brain of sockeye salmon.

The localization of two salmon-type gonadotropin-releasing hormone (sGnRH) precursors, pro-sGnRH-I (short type) and pro-sGnRH-II (long type), was investigated by using in situ hybridization techniques in the brain of the landlocked sockeye salmon, Oncorhynchus nerka. We used 30-mer oligonucleotide probes complementary to pro-sGnRH-I and pro-sGnRH-II cDNA. No significant differences were observed in the localization of sGnRH neurons expressing pro-sGnRH-I and pro-sGnRH-II mRNAs; both were expressed in the olfactory nerve, the olfactory bulbs, the regions between the olfactory bulb and telencephalon, the ventral telencephalon, the preoptic area, and the hypothalamus.

[Abnormal hemoglobinopathy (HbE) diagnosed from microcytic hypochromic red blood cells in a 31 year-old Bangladeshian male].

Microcytic hypochromic red blood cells (RBC) were discovered in a 31 year-old Bangladeshi man. Additional laboratory data revealed only slight elevations of LDH and transaminase activities. The patient was clinically asymptomatic and showed no signs of anemia.

Purification and molecular cloning of a chitinase expressed in the hepatopancreas of the penaeid prawn Penaeus japonicus.

A protein with chitinase activity was purified from the hepatopancreas of the penaeid prawn Penaeus japonicus, and the amino acid sequences of several amino acid fragments were determined. A cDNA clone was isolated and sequenced which contained the coding sequence of the enzyme. The conceptually translated protein (named Pjchi-3) was a member of the chitinase family based on sequence similarities to chitinases of non-crustacean species, as was the case with Pjchi-1 and 2, two chitinase homologues previously isolated from P.

Mouse glycine N-methyltransferase is sexually dimorphic and regulated by growth hormone.

The secretion pattern of putuitary growth hormone (GH) in rodents is sexually dimorphic and regulates the expression of some hepatic genes. We report cloning of mouse glycine N-methyltransferase (GNMT) cDNA. The mouse GNMT mRNA is expressed at much lower levels in male than in female livers.

Molecular cloning of a molt-inhibiting hormone cDNA from the kuruma prawn Penaeus japonicus.

The crustacean molt-inhibiting hormone (MIH) is released from the X-organ sinus gland complex and suppresses ecdysteroid synthesis by the Y-organ. In the present study, we have isolated a cDNA which encodes a MIH (Pej-SGP-IV) of the kuruma prawn Penaeus japonicus in order to study its expression and characterize the structure of its precursor. A cDNA fragment was isolated using RT-PCR with two degenerate oligonucleotide primers that were designed based on the peptide sequence of Pej-SGP-IV, and this fragment was used as a probe to screen an eyestalk cDNA library.

Induction of male-type gonadotropin secretion by implantation of 11-ketotestosterone in female goldfish.

In goldfish, plasma gonadotropin levels increase during spawning in both males and females (GTH surge). A female-typical GTH surge induces ovulation (ovulatory surge), and a male-typical surge triggers milt production in response to sex pheromones released from ovulatory females. This study examined whether the male-typical GTH surge occurs in adult females that are implanted with 11-ketotestosterone (KT), which induces male-typical sexual behavior in adult female goldfish.

Steroid hormone-dependent overexpression of cytochromes P450 2A in liver tumors of TGF alpha transgenic male mice.

To clarify the mechanism underlying the male preference of liver tumor in transforming growth factor (TGF) alpha transgenic mice, we analyzed the sexually dimorphic expression of two P450s, i.e., female-specific mouse 15 alpha hydroxylase P450 (2A4) and coumarin 7-hydroxylase P450 (2A5).

Characterization, guanosine 5'-O-(3-thiotriphosphate) modulation, daily variation, and localization of melatonin-binding sites in the catfish (Silurus asotus) brain.

Characteristics, guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS) modulation, daily variation, and localization of melatonin-binding sites in the brain of a nocturnal teleost, the catfish Silurus asotus, were studied by radioreceptor assay using 2-[125I]iodomelatonin as the radioligand. The specific binding was rapid, stable, saturable, and reversible. The radioligand binds to a single class of receptor site with an affinity (Kd) of 30.

Photic and circadian regulations of melatonin rhythms in fishes.

Photic and circadian regulations of melatonin rhythms in the pineal organ and the retina of several teleosts were studied to investigate the regulatory mechanisms of melatonin rhythms in fishes. In the eyecup preparations of the goldfish, Carassius auratus, both time of day and lighting conditions affected melatonin production, with high melatonin production observed only in the dark-treated group incubated during the 'subjective' night. Thus, in the goldfish retina, local photoreceptors and an ocular circadian clock seem to regulate melatonin production, as in the zebrafish retina and in the pineal organ of a number of teleosts, including the goldfish.

Transcriptional regulation by HNF-4 of the steroid 15alpha-hydroxylase P450 (Cyp2a-4) gene in mouse liver.

The mouse P450 gene Cyp2a-4 encodes the hepatic steroid 15apha-hydroxylase. We have defined in the 5'-flanking sequence of Cyp2a-4 gene, a composite regulatory element (-61AGACCAAAGTCCGGCCTTC-42) which contains a potential CpG methylation site at position -50. Gel-shift assays indicate that this element consists of overlapped binding sites for a hepatocyte-enriched transcription factor HNF-4 and a Sp1-like protein.

[Plasma cell leukemia associated with monocytosis].

A 51-year-old man was admitted to our hospital in December 1993, because of fatigue. Peripheral blood tests showed a WBC of 49,400/microliter with 36% plasma cells and 35% monocytes, Hb 14.5 g/dl, and Plt 137,000/microliter.

[Predisposition of subclones of pancreatic carcinoma cells, AsPC-1, to changes in functional and histopathological features of xenograft tumors with response to extracellular matrix].

We cloned two characteristics subclones from a human pancreatic carcinoma cell line, AsPC-1, according to their distinctive cell shapes; one an epithelial morphology and designated as "Beto-1" and the other a fibroblastic morphology and designated as "Fib-1". Fib-1 grew faster than Beto-1, but the growth rate of the cells on plastics was as high as that of the cells on the extracellular matrix extracts, matrigel. The pancreatic tumor-marker proteins, alpha-amylase, insulin, CEA, POA, PP, and AFP, but not CA 19-9, were positive in both subclones.

Ocular melatonin rhythms in the goldfish, Carassius auratus.

Ocular melatonin rhythms in the goldfish were studied and compared to those in the pineal organ and plasma. Under light:dark (LD) of 12 h light:12 h dark, melatonin contents in the eye as well as the pineal organ and plasma exhibited clear day-night changes with higher levels at mid-dark than at mid-light. However, melatonin contents in the eye at mid-light and mid-dark were approximately 100 and 9 times greater than those in the pineal organ, respectively.

Reciprocal size-effect relationship of the key residues in determining regio- and stereospecificities of DHEA hydroxylase activity in P450 2a5.

Collectively, the P450 2a4/2a5 system hyrdoxylates DHEA in at least three positions (7alpha, 7beta, and 2alpha). An individual P450, however, exhibits high specificity to one of these products. Using site-directed mutagenesis of mP450 2a5 from the wild mouse Mus minutoides and bacterial expression, we have associated the function of residues 117, 209, and 481 with the respective specificity observed in each P450.

Mitochondrial DNA mutations in Japanese detected by polymerase chain reaction--restriction fragment--single strand conformation polymorphism analysis.

PCR-restriction fragment-SSCP (PCR-RF-SSCP) analysis of mitochondrial DNA by HhaI/HincII in Japanese revealed 46 polymorphic patterns. The determinations of nucleotide sequence of these 46 patterns revealed 56 mutations compared with the Cambridge Sequence.

Cloning and sequence analysis of a cDNA encoding a crustacean hyperglycemic hormone from the Kuruma prawn Penaeus japonicus.

The crustacean hyperglycemic hormones (CHHs) are released from the X-organ sinus gland complex and regulate the glucose level in the hemolymph. In the present study, we have isolated a complementary DNA that encodes a CHH (Pej-SGP-III) of the Kuruma prawn Penaeus japonicus to study its expression and the structure of its precursor. In a cDNA clone of 774 base pairs (bp), an open reading frame of 345 bp was found.