Pantropic retroviral vector integration, expression, and germline transmission in medaka (Oryzias latipes)

The study focuses on adult children (n = 81) having the main responsibility for parents with dementia (study group). They were compared with children (n = 102) of non-demented parents (reference group). The children were interviewed about burden experienced. The interviews also secured information about the children's co-operation with the informal and formal network and their willingness to remain as caregivers during the progression of the disease or ageing process. The results showed that the daughters reported feeling more affection in their caregiving role than the sons. About one third of the participants in the study could not get relief from someone else. Eighty-nine per cent of the children in the study group and 76% of the children in the reference group were not willing to care for their parents in the family home during the progression of the disease, even if they were employed as caregivers.     

Inhibitory guanine-nucleotide-binding-regulatory protein alpha subunits in medaka (Oryzias latipes) oocytes--cDNA cloning and decreased expression of proteins during oocyte maturation

We have previously shown that pertussis-toxin-sensitive inhibitory guanine-nucleotide-binding-regulatory proteins (G proteins) are involved in the signal transduction of steroidal maturation-inducing hormone (MIH) of rainbow trout (Oncorhynchus mykiss) oocytes, 17alpha,20beta-dihydroxy-4-pregnen-3-one (17alpha,20beta-DP) [Yoshikuni, M. & Nagahama, Y. (1994) Dev. Biol. 166, 615-622]. In this study, we obtained five different cDNA fragments of G protein alpha subunits from medaka (Oryzias latipes) intact ovarian follicles (three subtypes of G(i alpha), G(i alpha a), G(i alpha b) and G(i alpha c); two subtypes of G(s alpha), G(s alpha d), and G(s alpha e)). Using a newly developed extraction method for medaka oocyte RNA, we demonstrated that oocytes expressed both G(i alpha a) and G(i alpha c), but not G(i alpha b). Full-length cDNA clones for G(i alpha a) and G(i alpha c) were then isolated from a medaka ovarian follicle cDNA library. The predicted amino acid sequences of G(i alpha a) and G(i alpha c) exhibited significant similarity with G(i alpha1) and G(i alpha2) of other species, respectively. Both G(i alpha a) and G(i alpha c) possessed a specific Cys residue in the C-terminal region that was the site for ADP-ribosylation by pertussis toxin. G(o alpha), another G protein that is ADP-ribosylated by pertussis toxin, was not detected in oocytes, although it was expressed in brain tissue. Western blot analyses using a specific antibody against G(i alpha1) and G(i alpha2) subunit proteins revealed that in both medaka and rainbow trout G(i alpha) subunit protein (40 kDa) contents were abundant in plasma membranes of postvitellogenic immature oocytes, decreased in mature oocytes, and were absent in ovulated eggs. Furthermore, specific 17alpha,20beta-DP binding to plasma membranes was higher in postvitellogenic immature oocytes than in ovulated eggs. Taken together, these results suggest that G(i alpha a) and/or G(i alpha c) may be involved in the transduction of the signal from 17alpha,20beta-DP receptors during oocyte maturation of fish oocytes.     

Gravity influences the position of the dorsoventral axis in medaka fish embryos (Oryzias latipes)

OBJECTIVE: To evaluate the use of a selective dopamine-1 agonist (fenoldopam) to provide selective splanchnic vasodilatation during sustained hypotensive endotoxaemia in sheep. DESIGN: Randomised, controlled, experimental study. SETTING: Animal research laboratory. SUBJECTS: 12 adult instrumented, midazolam-sedated sheep. INTERVENTIONS: The animals were randomised to receive a 20-min continuous infusion of dopamine (10 microg x kg(-1) x min(-1), fenoldopam (10 microg x kg(-1) x min(-1) and noradrenaline (1 microg x kg(-1) x min(-1)) under control conditions and 12 h after endotoxaemia was induced by a continuous infusion of Escherichia coli endotoxin producing a stable hyperdynamic state simulating human septic shock. This drug dosage was selected to produce a 25-30% increase in cardiac output by all three drugs during control conditions. MEASUREMENTS AND RESULTS: Systemic and splanchnic haemodynamic data were continuously obtained and systemic and splanchnic oxygen delivery (DO2) and consumption (VO2) were calculated. Hyperdynamic hypotensive endotoxaemia did not modify the splanchnic and renal reduction in DO2 and the vasoconstrictive reactivity to noradrenaline observed during control conditions. In contrast, endotoxaemia abolished the fenoldopam and dopamine-induced increase in splanchnic DO2 (especially in the coeliac trunk) observed during control conditions. CONCLUSIONS: During sustained hyperdynamic endotoxaemia, the dopaminergic-induced selective increase in coeliac trunk blood flow is abolished, most probably because of an already maximally vasodilated splanchnic circulation which prevented dopamine or fenoldopam to vasodilate this area further. Contrary to common belief, selective dopamine-1 agonist administration under these conditions may therefore not be beneficial to the splanchnic organs, though it improves whole body DO2 and VO2.     

A highly repetitive sequence isolated from genomic DNA of the medaka (Oryzias latipes)

In order to investigate the relationship between various temporomandibular joint (TMJ) pain levels and the detection of high signal intensity (joint effusion) on T2 weighted magnetic resonance imaging (MRI), 19 consecutive patients who complained of unilateral painful TMJ hypomobility (closed locking) were involved in this study. All patients were clinically examined in a routine manner, and all patients rated their pain levels by a visual analogue scale and eight pain questionnaire prior MRI study. T1 and T2 weighted MRI was taken in sagittal section at unilateral affected joint side. The presence or absence of a high signal intensity spot within the TMJ compartment were judged by three examiners. The high signal intensity was detected in 10 joints, but not in 9 joints. In between these two groups, the pain ratio was calculated and compared. The data showed that there was no significant statistical correlation between pain levels and the presence of high signals. This study disclosed that the MRI detection of high signal intensity in the closed locking TMJ did not directly relate to the presence of TMJ pain nor the increased pain level. These indicate the need of further larger studies.     

Organophosphate-induced acetylcholinesterase inhibition and embryonic retinal cell necrosis in vivo in the teleost (Oryzias latipes)

Recent monitoring of the Sacramento-San Joaquin River system (CA) indicates that levels of the organophosphate pesticide, diazinon, exceed National Academy of Science guidelines and these levels result in toxicity in USEPA acute toxicity tests with Ceriodaphnia dubia. Since organophosphates (OPs) inhibit acetylcholinesterase (AChE), the present study examined the effects of diazinon on the embryonic nervous system of a model teleost, medaka, Oryzias latipes. Preliminary histological screens revealed limited retinal cell necrosis in control embryos with apparent increased necrosis in diazinon-exposed embryos. Subsequently, embryos were exposed to 1.8 x 10(-5), 4.4 x 10(-5), or to 8.8 x 10(-5) M diazinon and replicates were frozen for biochemical analysis or were fixed for histopathological analysis at days 3, 5, and 7 of development. Diazinon exposure significantly inhibited AChE activity within whole embryos and in homogenates of retinas from treated animals. Histological examination of embryos indicated that as the retina underwent differentiation into distinct cell layers, between days 5 and 7, small foci of necrotic cells became apparent within the inner nuclear layer and isolated individual pyknotic cells were observed in the ganglion layer. Quantification of foci of necrotic cells revealed that 8.8 x 10(-5) M diazinon increased number and area of these lesions. Enzyme histochemistry localized AChE activity to regions equivalent to sites of necrosis. Separate exposures of embryos to the OP, diisopropylphosphorofluoridate, produced large foci of necrotic cells at sites equivalent to those seen following diazinon exposure.     

Identification of the sex chromosomes of the medaka, Oryzias latipes, by fluorescence in situ hybridization

In the medaka, Oryzias latipes, which does not have cytologically recognizable sex chromosomes, the sex is genetically determined and the mechanism of sex determination (XX/XY) can be revealed by genetic crosses using a particular pigment gene. In a previous study, we isolated a sex-linked DNA marker (SL1) using the genomic differences between inbred strains of medaka. In the present paper, we further isolated another sex-linked clone (pHO5.110). The pHO5. 110-related sequences were tightly linked to sex in O. latipes. We designated the locus of the pHO5.110-related sequence on sex chromosomes of medaka as Sex-Linked 2 (SL2). Southern blot analyses suggested that the pHO5.110-related sequence was tandemly repetitive in the medaka genome. Using the clone as a probe for FISH analysis, strong hybridization signals were obtained in a couple of chromosomes that formed one of two large submetacentric chromosome pairs. The pHO5.110-related sequences were repetitive in the genomes of other species of Oryzias (O. curvinotus, O. luzonensis and O. mekongensis) that are karyologically related to O. latipes (all are members of the so-called biarmed group). By contrast, the sequences were not detected as repetitive in other Oryzias species. Hence, it is thought that pHO5.110-related sequences were amplified in the genome of a common ancestor of the biarmed group.     

Photorepair and excision repair removal of UV-induced pyrimidine dimers and (6-4) photoproducts in the tail fin of the medaka, Oryzias latipes

Induction and repair of UV-B induced DNa damage in the tail fin of the Medaka, were examined immunohistochemicaly and by the enzyme-linked immunosorbent assay (ELISA). UV-induced DNA damage was detected only in the outermost layer of epithelial cells and did not differ in fishes having different degree of melanization. Both pyrimidine dimers and (6-4) photoproducts in the fin cells were removed by excision repair in the dark, the excision of (6-4) photoproducts being about twice as efficient as that of pyrimidine dimers. The rate of excision repair of UV-induced lesions in fin tissue was three to four times that in cultured Medaka cells, OL32. In the fin cells, reductions in the numbers of pyrimidine dimers and (6-4) photoproducts were seen after treatment with fluorescent light, whereas less reductions of pyrimidine dimers and no reductions of (6-4) photoproducts were observed in OL32 cells.     

Efficiency of cell culture derivation from blastula embryos and of chimera formation in the medaka (Oryzias latipes) depends on donor genotype and passage number

Bacteriocin typing, plasmid profiling and ribotyping were used to type 34 food and patient Clostridium perfringens isolates from 10 food poisoning cases, respectively, outbreaks. In nine cases/outbreaks bacteriocin patterns showed identical main groups. Subgroups differed within all cases/outbreaks. Plasmid profiles were identical for all isolates within each of three outbreaks. In eight food poisoning cases and outbreaks, all the ribotypes of each food and stool isolate were found to be identical. All three typing methods give valuable results for the characterization of C. perfringens beyond the species level. Bacteriocin typing represents a suitable addition to plasmid typing, particularly since the results do not show any correlation between losses of plasmids and changes in bacteriocin sensitivity patterns. Ribotyping was found to be a suitable tool to determine the genetic relationship of C. perfringens isolates in the context of food-borne poisoning.     

Ultraviolet radiation-absorbing mycosporine-like amino acids (MAAs) are acquired from their diet by medaka fish (Oryzias latipes) but not by SKH-1 hairless mice

The growth-factor-like phospholipid lysophosphatidic acid (LPA) mediates a wide variety of biological functions. We recently reported the cloning of the first G-protein-coupled receptor for LPA, called ventricular zone gene-1 (vzg-1/lpA1/edg-2) because its embryonic central nervous system (CNS) expression is restricted to the neocortical ventricular zone (Hecht et al. [1996] J. Cell Biol. 135:1071-1083). Vzg-1 neural expression diminishes at the end of the cortical neurogenetic period, just before birth. Here, we have investigated the subsequent reappearance of vzg-1 expression in the postnatal murine brain, by using in situ hybridization and northern blot analyses. Vzg-1 expression was undetectable by in situ hybridization at birth, but reappeared in the hindbrain during the 1st postnatal week. Subsequently, expression expanded from caudal to rostral, with peak expression observed around postnatal day 18. At all postnatal ages, vzg-1 expression was concentrated in and around developing white matter tracts, and its expansion, peak, and subsequent downregulation closely paralleled the progress of myelination. Double-label in situ hybridization studies demonstrated that vzg-1-expressing cells co-expressed mRNA encoding proteolipid protein (PLP), a mature oligodendrocyte marker, but not glial fibrillary acidic protein (GFAP), an astrocyte marker. Consistent with this, vzg-1 mRNA expression was reduced by 40% in the brains of jimpy mice, which exhibit aberrant oligodendrocyte differentiation and cell death. Together with our characterization of vzg-1 during cortical neurogenesis, these data suggest distinct pre- and postnatal roles for LPA in the development of neurons and oligodendrocytes and implicate lysophospholipid signaling as a potential regulator of myelination.     

Diagnostic criteria for degenerative, inflammatory, proliferative nonneoplastic and neoplastic liver lesions in medaka (Oryzias latipes): consensus of a National Toxicology Program Pathology Working Group

Women residing in villages in three districts of Pakistan were recruited, trained to deliver primary care and mobilize their communities for health, assigned to limited catchment areas, provided with supervisory and managerial support, and remunerated. Their comprehensive activities substantially reduced infant, child and maternal mortality within a year and generated positive perceptions of family planning in the communities. The programme was cost-effective and appeared suitable as a model for reforming the organization and provision of health care services.     

Phenotypic rescue of the albino mutation in the medakafish (Oryzias latipes) by a mouse tyrosinase transgene

Mutations of the tyrosinase gene are one common cause of a similar phenotype in all vertebrates, known as albinism. In an attempt to contribute to an understanding of the genetic hierarchy governing the development of pigmentation, we have used a mouse tyrosinase minigene under the control of its 5.2 kb upstream promoter region to rescue two different albino mutations in the medakafish, Oryzias latipes. Around hatching stages an almost perfect phenocopy of the wildtype pigmentation was obtained. Subsequent ectopic melanin overproduction indicated a possible incompatibility of the heterologous mouse promoter for stable expression during the entire ontogenesis. Like in some tyrosinase transgenic mouse lines a strong variegation effect was observed. The transgene-mediated pigmentation phenotype was obtained up to the eighth offspring generation. The phenotypic effects of the tyrosinase transgene in different albino mutant strains places the i3-locus upstream and the b-locus downstream of the tyrosinase locus i1 in the genetic hierarchy leading to wildtype pigmentation.     

Lysyl oxidase (ras recision gene) expression in human amnion: ontogeny and cellular localization

Certain human heritable forms of colon cancer have characteristically high frequencies of microsatellite alterations. These microsatellite changes are markers of genomic instability and the direct consequence of mutations in genes involved with DNA mismatch repair processes, which are in part responsible for maintaining the sequence integrity of the genome. Given that the B6C3F1 mouse is genetically predisposed to develop liver tumors we were interested in determining whether tumors derived in this strain of mouse may contain alterations in microsatellite sequences. The analysis of 48 tumors at 24 different microsatellite loci revealed that microsatellite alterations were detected in 12 of 48 tumors (25%). Although this frequency is relatively high, 11 of the 12 tumors exhibited only a single alteration and in 10 of those tumors this change was at the same microsatellite locus. Microsatellite alterations were also detected in the DNA isolated from 6 of 22 (27%) normal liver tissues with 4 of the 6 occurring at the same locus where the majority of changes were observed in the tumors. Based on these results, we conclude that the microsatellite alterations present in the mouse liver tumor tissue are most likely the result of spontaneous mutational events. Consequently, the genomic instability operational in a particular type of hereditary human colon cancer does not appear to be operational in the genetically predisposed B6C3F1 mouse liver. In addition, we demonstrated that the activation of the H-ras gene, which causes some forms of genetic instability in vitro, does not contribute to genetic instability within liver tumors as measured by microsatellite alterations.     

Ol-Prx 3, a member of an additional class of homeobox genes, is unimodally expressed in several domains of the developing and adult central nervous system of the medaka (Oryzias latipes)

Large-scale genetic screens for mutations affecting early neurogenesis of vertebrates have recently been performed with an aquarium fish, the zebrafish. Later stages of neural morphogenesis have attracted less attention in small fish species, partly because of the lack of molecular markers of developing structures that may facilitate the detection of discrete structural alterations. In this context, we report the characterization of Ol-Prx 3 (Oryzias latipes-Prx 3). This gene was isolated in the course of a large-scale screen for brain cDNAs containing a highly conserved DNA binding region, the homeobox helix-three. Sequence analysis revealed that this gene belongs to another class of homeobox genes, together with a previously isolated mouse ortholog, called OG-12 [Rovescalli, A. C., Asoh, S. & Nirenberg, M. (1996) Proc. Natl. Acad. Sci. USA 93, 10691-10696] and with the human SHOX gene [Rao, E., Weiss, B., Fukami, M., Rump, A., Niesler, B., et al. (1997) Nat. Genet. 16, 54-62], thought to be involved in the short-stature phenotype of Turner syndrome patients. These three genes exhibit a moderate level of identity in the homeobox with the other genes of the paired-related (PRX) gene family. Ol-Prx 3, as well as the PRX genes, are expressed in various cartilaginous structures of head and limbs. These genes might thus be involved in common regulatory pathways during the morphogenesis of these structures. Moreover, this paper reports a complex and monophasic pattern of Ol-Prx 3 expression in the central nervous system, which differs markedly from the patterns reported for the PRX genes, Prx 3 excluded: this gene begins to be expressed in a variety of central nervous system territories at late neurula stage. Strikingly, it remains turned on in some of the derivatives of each territory during the entire life of the fish. We hope this work will thus help identify common features for the PRX 3 family of homeobox genes.     

Structure of the mouse osteoclastogenesis inhibitory factor (OCIF) gene and its expression in embryogenesis

A new phenylpropanoid glycoside, alpha-(3,4-dihydroxyphenyl)ethyl-(2'-O-alpha-L-rhamno-pyranosyl- 3'-O-beta-D-apiofuranosyl-4'-O-E-caffeoyl)-beta-D-glucopyran oside, verbascoside, and two new flavone glucosides, nevadensin 7-O-beta-D-glucoside and nevadensin 7-O-[alpha-L-rhamnosyl(1-->6)]-beta-D-glucoside, have been isolated from the aerial parts of Lysionotus pauciflorus. The structures have been elucidated on the basis of spectroscopic data and chemical correlation.     

Human aldehyde dehydrogenase: chromosomal assignment of the gene for the isozyme that metabolizes gamma-aminobutyraldehyde

A cDNA clone of the E3 isozyme of human liver aldehyde dehydrogenase consisting of a 1320-base pair (bp) coding region and a 180-bp non-coding region at the 3' end was used for chromosomal localization of the E3 gene. Using a panel of human/hamster somatic cell hybrids we have localized, the gene coding for the E3 isozyme to human chromosome 1.     

The ontogeny of the expression of K+ channel-like gene (CHIF) in the rat kidney papilla

A whole genome scan was undertaken in a granddaughter design comprising 1158 progeny-tested bulls in order to map QTL influencing milk yield and composition. In this paper we report the identification of a locus on the centromeric end of bovine Chromosome (Chr) 14, with major effect on fat and protein percentage as well as milk yield. The genuine nature of this QTL was verified using the grand2-daughter design, that is, by tracing the segregating QTL alleles from heterozygous grandsires to their maternal grandsons and confirming the predicted QTL allele substitution effect.     

First case of missense mutation (LDH-H:R171P) in exon 4 of the lactate dehydrogenase gene detected in a Japanese patient

BACKGROUND: The quality of life, psychologic sequelae, and functional status of liver transplant recipients with recurrent hepatitis C virus (HCV) hepatitis have not been well defined. METHODS: Perceived quality of life, psychologic distress, depression, adaptive coping, and functional status were prospectively assessed in 59 liver transplant recipients at baseline (before transplantation) and 6 and 12 months after transplantation; comparisons were made between patients with recurrent HCV hepatitis and all other patients. RESULTS: Recurrent HCV hepatitis developed in 41% (14/34) of the patients with HCV. At 6 months, the patients with recurrent HCV hepatitis had significantly lower functional status (P=0.013) and experienced less gain in physical functioning from baseline than other patients (P=0.02). Quality of life, depression, and psychologic distress were not different at 6 months for patients with recurrent HCV hepatitis compared with all other patients. At 12 months, the patients with recurrent HCV hepatitis had significantly lower quality of life (P=0.003), greater depression (P=0.045), higher psychologic distress (P=0.05), and lower physical functioning (P=0.008) than all other patients. CONCLUSION: Recurrent HCV hepatitis in liver transplant recipients is associated with impairment in quality of life, functional status, and greater depression compared with patients who did not have HCV and those without HCV recurrence.     

Differential gene expression in multilocus isozyme systmes of the developing green sunfish

The patterns of expression of eight multilocous isozyme systems were investigated in the differentiated adult tissues and the early embryonic stages (0-210 hours after fertilization) of the green sunfish, Lepomis cyanellus. Enzymes encoded by approximately 23 gene loci were resolved by starch-gel electrophoresis and detected by specific histochemical staining. The developmental patterns of these isozyme systems appear to be the result of the diffential expression of the multiple gene loci. Isozymic forms of glucoseophosphate isomerase (GPI-A2), malate dehydrogenase (MDH-A2), and creatine kinase (CK-C2) were present in most differentiated tissues, in the unfertilized eggs, and in all stages of embryonic development. Closely homologous forms of these isozymes (GPI-B2, MDH-B2, and CK-A2) were expressed predominantly in skeletal muscle and were first detected at around the time of hatching (38-42 hours). The similar temporal and spatial patterns of gene expressions for the GPI, LDH, MDH, and CK loci suggest that the duplicates loci encoding enzymes, diverged in their regulation to patterns of differential gene expression which are similar for each enzyme system.