RESUMEN
We constructed turkey herpesvirus (HVT) vector vaccines in which the VP2 gene of infectious bursal disease virus (IBDV) was inserted into the HVT genome in the following regions: UL3-4, UL22-23, UL45-46, and US10-SORF3. We then evaluated the relationship between the gene insertion site and the capacity of the virus to elicit antibodies. rHVT/IBD (US10) showed good growth activity in vitro, with growth comparable to that of the parent HVT. On the other hand, rHVT/IBD (UL3-4), rHVT/IBD (UL22-23), and rHVT/IBD (UL45-46) exhibited decreased growth activity in chicken embryo fibroblast (CEF) cells compared to the parent HVT. However, the rHVT/IBD (US10) elicited lower levels of virus-neutralizing (VN) antibodies compared to the other constructs. rHVT/IBD (UL3-4) and rHVT/IBD (UL45-46) appeared to be similar in their ability to elicit VN antibodies. Based on the results of in vitro and in vivo assays, rHVT/IBD (UL3-4) was selected for further testing. In a challenge assay, rHVT/IBD (UL3-4) protected chickens from challenge with virulent Marek's disease virus serotype 1 and IBDV. In conclusion, the site of gene insertion may have a strong effect on the growth of the vector virus in vitro and its antibody-eliciting capacity. Insertions in the UL3-4 region permitted a balance between growth activity and VN-antibody-eliciting capacity, and this region might therefore be an appropriate insertion site for IBDV VP2.
Asunto(s)
Anticuerpos Antivirales/inmunología , Herpesvirus Meleágrido 1/inmunología , Enfermedad de Marek/prevención & control , Enfermedades de las Aves de Corral/prevención & control , Proteínas Virales/inmunología , Vacunas Virales/inmunología , Animales , Pollos , Herpesvirus Meleágrido 1/química , Herpesvirus Meleágrido 1/genética , Herpesvirus Meleágrido 1/crecimiento & desarrollo , Herpesvirus Gallináceo 2/inmunología , Herpesvirus Gallináceo 2/fisiología , Enfermedad de Marek/inmunología , Enfermedad de Marek/virología , Mutagénesis Insercional , Enfermedades de las Aves de Corral/inmunología , Enfermedades de las Aves de Corral/virología , Pavos , Vacunación , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/química , Vacunas Sintéticas/genética , Vacunas Sintéticas/inmunología , Proteínas Virales/administración & dosificación , Proteínas Virales/química , Proteínas Virales/genética , Vacunas Virales/administración & dosificación , Vacunas Virales/química , Vacunas Virales/genéticaRESUMEN
BACKGROUND: Because psychiatric disorders are risk factors for suicide, psychiatric consultation should be an essential element of suicide prevention among individuals with a high risk of suicide. The aim of the present study was to compare the characteristics of individuals who had or had not received psychiatric consultation before they attempted suicide in Japan. METHODS: Clinical records were used to identify 300 consecutive persons who were admitted to the hospital for attempting suicide between April 2006 and March 2013. We divided the patients into two groups. One group consisted of patients who consulted a psychiatrist before their suicidal behaviours (the consultation group), and the other group consisted of patients who had not consulted a psychiatrist before their suicidal behaviours (the non-consultation group). Group differences were analysed with respect to gender, age, method of suicide attempts, psychiatric diagnosis (ICD-10), and duration of hospitalisation in the emergency unit. RESULTS: Females tended to be over-represented in the consultation group (73.0%), and males tended to be over-represented in the non-consultation group (59.8%). Poisoning by prescription drugs was used more frequently as a method of suicide in the consultation group than in the non-consultation group. Neuroticism and related disorders were higher in the non-consultation group (33.7%) than in the consultation group (18.9%). Mood disorders (32.6%) were nearly as common as neuroticism in the non-consultation group, and together they accounted for almost two-thirds of all diagnoses. Mood disorders were comparable between the consultation group (30.9%) and the non-consultation group (32.6%). Adult personality disorders (13.3%) and schizophrenia and related disorders (26.0%) were higher in the consultation group than in the non-consultation group. CONCLUSIONS: Measures have to be taken to encourage people with these diverse characteristics to consult psychiatrists, and psychiatrists have to regularly evaluate patients for suicide risk. Furthermore, we need further research on the relationship between psychiatric consultation and poisoning by prescribed drugs.
Asunto(s)
Trastornos Mentales/epidemiología , Derivación y Consulta/estadística & datos numéricos , Ideación Suicida , Intento de Suicidio/psicología , Adulto , Anciano , Anciano de 80 o más Años , Estudios Transversales , Servicio de Urgencia en Hospital , Femenino , Hospitalización , Humanos , Japón/epidemiología , Masculino , Persona de Mediana Edad , Trastornos del Humor/epidemiología , Factores de Riesgo , Adulto JovenRESUMEN
The prospect of space travel continues to capture the imagination. Several competing companies are now promising flights for the general population. Previously, it was recognized that many of the physiological changes that occur with spaceflight are similar to those seen with normal ageing. This led to the notion that spaceflight can be used as a model of accelerated ageing and raised concerns about the safety of individuals engaging in space travel. Paradoxically, however, space travel has been recently shown to be beneficial to some aspects of muscle health in the tiny worm Caenorhabditis elegans. C. elegans is a commonly used laboratory animal for studying ageing. C. elegans displays age-related decline of some biological processes observed in ageing humans, and about 35% of C. elegans' genes have human homologs. Space flown worms were found to have decreased expression of a number of genes that increase lifespan when expressed at lower levels. These changes were accompanied by decreased accumulation of toxic protein aggregates in ageing worms' muscles. Thus, in addition to spaceflight producing physiological changes that are similar to accelerated ageing, it also appears to produce some changes similar to delayed ageing. Here, we put forward the hypothesis that in addition to the previously well-appreciated mechanotransduction changes, neural and endocrine signals are altered in response to spaceflight and that these may have both negative (e.g. less muscle protein) and some positive consequences (e.g. healthier muscles), at least for invertebrates, with respect to health in space. Given that changes in circulating hormones are well documented with age and in astronauts, our view is that further research into the relationship between metabolic control, ageing, and adaptation to the environment should be productive in advancing our understanding of the physiology of both spaceflight and ageing.
Asunto(s)
Envejecimiento/fisiología , Caenorhabditis elegans/fisiología , Vuelo Espacial , Adaptación Fisiológica , Envejecimiento/genética , Animales , Caenorhabditis elegans/genética , Metabolismo Energético , Expresión Génica , Genes de Helminto , Humanos , Longevidad/fisiología , Modelos Animales , Modelos Biológicos , Músculos/fisiología , Ingravidez/efectos adversosRESUMEN
BACKGROUND: Senescence-accelerated mice (SAM) are a series of mouse strains originally derived from unexpected crosses between AKR/J and unknown mice, from which phenotypically distinct senescence-prone (SAMP) and -resistant (SAMR) inbred strains were subsequently established. Although SAMP strains have been widely used for aging research focusing on their short life spans and various age-related phenotypes, such as immune dysfunction, osteoporosis, and brain atrophy, the responsible gene mutations have not yet been fully elucidated. RESULTS: To identify mutations specific to SAMP strains, we performed whole exome sequencing of 6 SAMP and 3 SAMR strains. This analysis revealed 32,019 to 38,925 single-nucleotide variants in the coding region of each SAM strain. We detected Ogg1 p.R304W and Mbd4 p.D129N deleterious mutations in all 6 of the SAMP strains but not in the SAMR or AKR/J strains. Moreover, we extracted 31 SAMP-specific novel deleterious mutations. In all SAMP strains except SAMP8, we detected a p.R473W missense mutation in the Ldb3 gene, which has been associated with myofibrillar myopathy. In 3 SAMP strains (SAMP3, SAMP10, and SAMP11), we identified a p.R167C missense mutation in the Prx gene, in which mutations causing hereditary motor and sensory neuropathy (Dejerine-Sottas syndrome) have been identified. In SAMP6 we detected a p.S540fs frame-shift mutation in the Il4ra gene, a mutation potentially causative of ulcerative colitis and osteoporosis. CONCLUSIONS: Our data indicate that different combinations of mutations in disease-causing genes may be responsible for the various phenotypes of SAMP strains.
Asunto(s)
Envejecimiento/genética , Enfermedad/genética , Exoma/genética , Genómica , Mutación/genética , Análisis de Secuencia , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Exones/genética , Humanos , Proteínas de la Membrana/química , Proteínas de la Membrana/genética , Ratones , Datos de Secuencia Molecular , Fenotipo , Especificidad de la EspecieRESUMEN
11ß-hydroxysteroid dehydrogenase (HSD11B) catalyzes the interconversion between active and inactive glucocorticoid, and is known to exist as two distinct isozymes: HSD11B1 and HSD11B2. A third HSD11B isozyme, HSD11B1L (SCDR10b), has recently been identified. Human HSD11B1L, which was characterized as a unidirectional NADP(+)-dependent cortisol dehydrogenase, appears to be specifically expressed in the brain. We previously reported that HSD11B1 and abundant HSD11B2 isozymes are expressed in neonatal pig testis and the Km for cortisol of NADP(+)-dependent dehydrogenase activity of testicular microsomes obviously differs from the same activity catalyzed by HSD11B1 from pig liver microsomes. Therefore, we hypothesized that the neonatal pig testis also expresses the third type of HSD11B isozyme, and we herein examined further evidence regarding the expression of HSD11B1L. (1) The inhibitory effects of gossypol and glycyrrhetinic acid on pig testicular microsomal NADP(+)-dependent cortisol dehydrogenase activity was clearly different from that of pig liver microsomes. (2) A highly conserved human HSD11B1L sequence was observed by RT-PCR in a pig testicular cDNA library. (3) mRNA, which contains the amplified sequence, was evaluated by real-time PCR and was most strongly expressed in pig brain, and at almost the same levels in the kidney as in the testis, but at lower levels in the liver. Based on these results, neonatal pig testis appears to express glycyrrhetinic acid-resistant HSD11B1L as a third HSD11B isozyme, and it may play a physiologically important role in cooperation with the abundantly expressed HSD11B2 isozyme in order to prevent Leydig cell apoptosis or GC-mediated suppression of testosterone production induced by high concentrations of activated GC in neonatal pig testis.
Asunto(s)
11-beta-Hidroxiesteroide Deshidrogenasas/metabolismo , Sus scrofa/metabolismo , Testículo/enzimología , 11-beta-Hidroxiesteroide Deshidrogenasas/antagonistas & inhibidores , 11-beta-Hidroxiesteroide Deshidrogenasas/química , 11-beta-Hidroxiesteroide Deshidrogenasas/genética , Secuencia de Aminoácidos , Animales , Animales Recién Nacidos , Secuencia de Bases , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Perfilación de la Expresión Génica , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Humanos , Masculino , Microsomas Hepáticos/efectos de los fármacos , Microsomas Hepáticos/enzimología , Datos de Secuencia Molecular , NADP/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Testículo/efectos de los fármacosRESUMEN
INTRODUCTION: Agonists of peroxisome proliferator-activated receptor gamma (PPARγ) have been examined as chemopreventive and chemotherapeutic agents. The aim was to investigate the cytotoxicity and action mechanisms of 15-deoxy-Δ(12,14)-prostaglandin J(2) (15d-PGJ(2)), one of endogenous ligands for PPARγ, in terms of PPARγ-dependency and the mitogen-activated protein kinase (MAPK) and Akt pathway in three human renal cell carcinoma (RCC)-derived cell lines. METHODS: 786-O, Caki-2 and ACHN cells were used as human RCC-derived cell lines. Cell viability and caspase-3 activity was detected by fluorescent reagents, and chromatin-condensation was observed with a brightfield fluorescent microscope after staining cells with Hoechst33342. The expression levels of proteins were detected by Western blot analysis. RESULTS: 15d-PGJ(2) showed cytotoxicity in dose-dependent manner. 15d-PGJ(2) induced chromatin-condensation and elevated caspase-3 activity, and the cell viability was restored by co-treatment with a pan-caspase inhibitor, Z-VAD-FMK, indicating the involvement of caspase-dependent apoptosis. The cytotoxicity was not impaired by a PPARγ inhibitor, GW9662, suggesting that 15d-PGJ(2) exerted the cytotoxicity in a PPARγ-independent manner. Some antioxidants rescued cells from cell death induced by 15d-PGJ(2), but some did not, suggesting that reactive oxygen species (ROS) did not contribute to the apoptosis. 15d-PGJ(2) also increased the expression levels of phospho-c-Jun N terminal kinase (JNK) in Caki-2 cells, and decreased those of phospho-Akt in 786-O cells, indicating that the JNK MAPK and the Akt pathways participated in the anticancer effects of 15d-PGJ(2) in some cell lines. CONCLUSION: 15d-PGJ(2) exerted cytotoxic effects accompanying caspase-dependent apoptosis, and this effect was elicited in a PPARγ-independent manner in three cell lines. In addition, the JNK MAPK and Akt pathway was involved in the cytotoxicity of 15d-PGJ(2) to some extent in some cell line. Therefore, our study showed the 15d-PGJ(2) to potentially be an interesting approach for RCC treatment.
Asunto(s)
Carcinoma de Células Renales/metabolismo , Neoplasias Renales/metabolismo , MAP Quinasa Quinasa 4/metabolismo , PPAR gamma/fisiología , Prostaglandina D2/análogos & derivados , Proteínas Proto-Oncogénicas c-akt/metabolismo , Antioxidantes/farmacología , Western Blotting , Carcinoma de Células Renales/enzimología , Carcinoma de Células Renales/patología , Línea Celular Tumoral , Fluorometría , Humanos , Neoplasias Renales/enzimología , Neoplasias Renales/patología , L-Lactato Deshidrogenasa/metabolismo , Microscopía Fluorescente , Prostaglandina D2/toxicidad , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Backgrounds: Hikikomori, pathological social withdrawal, is becoming a crucial mental health issue in Japan and worldwide. We have developed a 3-day family intervention program for hikikomori sufferers based on Mental Health First Aid (MHFA) and Community Reinforcement and Family Training (CRAFT). This study aims to confirm the effectiveness of the 3-day program by a randomized controlled trial. Methods: This study was registered on the UMIN Clinical Trials Registry (UMIN000037289). Fifteen parents were assigned to the treat as usual (TAU) group (TAU only; Age Mean, 65.6; SD, 7.8), and 14 to the Program group (program + TAU; Age Mean, 67.9; SD, 8.6). This study was discontinued due to the COVID-19 pandemic; the recruitment rate was 36.3% of our target sample size of 80. Results: Perceived skills improved temporally and stigma temporally worsened in the TAU group. Confidence decreased and attitude showed no change in both groups. Aggressive behaviors of hikikomori sufferers were significantly worsened in the Program group; however, no serious domestic violence was reported. In the TAU group, Avoidance and irregular life patterns were improved. Activity levels were worsened in both groups. Two participants (16.7%) in the Program group and one participant (7.7%) in the TAU group reported actual behavioral changes (e.g., utilizing support). Conclusion: We could not draw general conclusions on the effectiveness of the program due to the study discontinuation. Nevertheless, this study indicates the necessity for revision of the program to improve family members' confidence in engaging with hikikomori sufferers, with safer approaching by families.
RESUMEN
Pig 11beta-hydroxysteroid dehydrogenase (11beta-HSD) type 1 cDNA was cloned from neonatal pig testis, and 15 nucleotides were found to differ from the sequence in GenBank (Accession No. NM_214248). It was an exclusive clone obtained as pig 11beta-HSD type 1, and the sequence of 11beta-HSD type 1 cDNA cloned from pig liver was identical to that from testis. Amino acid sequence, deduced from cloned cDNA, also had a conserved triad of catalytically important Ser, Tyr and Lys residues for the short-chain dehydrogenase/reductase family, a membrane-spanning domain consisting of hydrophobic amino acid and a glycine motif in the cofactor binding region. The protein translated from this clone on expression in mammalian HEK293 cells exhibited oxo-reduction activity of cortisone and oxidation activity of cortisol. Furthermore, this oxo-reduction activity of cortisone was stimulated by co-expression of human H6PDH, while oxidation activity of cortisol was suppressed by H6PDH co-expression in HEK293 cells. Based on these results, the sequence of newly cloned cDNA is considered to correspond to an active enzyme form of pig 11beta-HSD type 1.
Asunto(s)
11-beta-Hidroxiesteroide Deshidrogenasa de Tipo 1 , ADN Complementario , Isoenzimas , Testículo/enzimología , 11-beta-Hidroxiesteroide Deshidrogenasa de Tipo 1/genética , 11-beta-Hidroxiesteroide Deshidrogenasa de Tipo 1/metabolismo , Secuencia de Aminoácidos , Animales , Animales Recién Nacidos , Secuencia de Bases , Células Cultivadas , ADN Complementario/genética , ADN Complementario/metabolismo , Humanos , Hidrocortisona/metabolismo , Isoenzimas/genética , Isoenzimas/metabolismo , Masculino , Datos de Secuencia Molecular , Alineación de Secuencia , Homología de Secuencia de Aminoácido , PorcinosRESUMEN
BACKGROUNDS: Hikikomori, a severe form of social withdrawal, is increasingly a serious mental health issue worldwide. Hikikomori is comorbid with various psychiatric conditions including depression, social anxiety and suicidal behaviors. Family support is encouraged as a vital first step, however evidence-based programs have yet to be established. Mental Health First Aid (MHFA) is one of the most well-validated educational programs encouraging lay people such as family members, to support close persons suffering from various psychiatric conditions such as depression, anxiety and suicidal behaviors. METHODS: We newly developed an educational program for family members of hikikomori sufferers mainly based on MHFA and 'Community Reinforcement and Family Training (CRAFT)' with role-play and homework. As a single-arm trial, 21 parents (7 fathers and 14 mothers) living with hikikomori sufferers participated in our program with five once-a-week sessions (2 h per session) and six monthly follow-ups, and its effectiveness was evaluated using various self-rated questionnaires. RESULTS: Perceived skills toward a depressed hikikomori case vignette, stigma held by participants, and subscales of two problematic and one adaptive behaviors of hikikomori sufferers were improved throughout the sessions and follow-ups. In addition, positive behavioral changes of hikikomori sufferers such as improved social participation were reported by participants. LIMITATIONS: Single-arm design and evaluation using self-rated questionnaires are the main limitations of the present study. CONCLUSIONS: Our newly developed program has positive effects on family members in their contact and support of hikikomori sufferers. Future trials with control groups are required to validate the effectiveness of this program.
RESUMEN
In Caenorhabditis elegans, the downregulation of insulin-like signaling induces lifespan extension (Age) and the constitutive formation of dauer larvae (Daf-c). This also causes resistance to oxidative stress (Oxr) and other stress stimuli and enhances the expression of many stress-defense-related enzymes such as Mn superoxide dismutase (SOD) that functions to remove reactive oxygen species in mitochondria. To elucidate the roles of the two isoforms of MnSOD, SOD-2 and SOD-3, in the Age, Daf-c and Oxr phenotypes, we investigated the effects of a gene knockout of MnSODs on them in the daf-2 (insulin-like receptor) mutants that lower insulin-like signaling. In our current report, we demonstrate that double deletions of two MnSOD genes induce oxidative-stress sensitivity and thus ablate Oxr, but do not abolish Age in the daf-2 mutant background. This indicates that Oxr is not the underlying cause of Age and that oxidative stress is not necessarily a limiting factor for longevity. Interestingly, deletions in the sod-2 and sod-3 genes suppressed and stimulated, respectively, both Age and Daf-c. In addition, the sod-2/sod-3 double deletions stimulated these phenotypes in a similar manner to the sod-3 deletion, suggesting that the regulatory pathway consists of two MnSOD isoforms. Furthermore, hyperoxic and hypoxic conditions affected Daf-c in the MnSOD-deleted daf-2 mutants. We thus conclude that the MnSOD systems in C. elegans fine-tune the insulin-like-signaling based regulation of both longevity and dauer formation by acting not as antioxidants but as physiological-redox-signaling modulators.
Asunto(s)
Proteínas de Caenorhabditis elegans/metabolismo , Caenorhabditis elegans/metabolismo , Regulación del Desarrollo de la Expresión Génica , Longevidad/fisiología , Animales , Secuencia de Bases , Caenorhabditis elegans/genética , Proteínas de Caenorhabditis elegans/genética , Eliminación de Gen , Insulina/metabolismo , Factor I del Crecimiento Similar a la Insulina/metabolismo , Longevidad/genética , Metamorfosis Biológica/genética , Datos de Secuencia Molecular , Mutación , Estrés Oxidativo , Parasitología/métodos , Interferencia de ARN , Transducción de Señal/genética , Superóxido Dismutasa/genética , Superóxido Dismutasa-1RESUMEN
The first International Caenorhabditis elegans Experiment (ICE-First) was carried out using a Russian Soyuz spacecraft from April 19-30, 2004. This experiment was a part of the program of the DELTA (Dutch Expedition for Life science Technology and Atmospheric research) mission, and the space agencies that participate in the International Space Station (ISS) program formed international research teams. A Japanese research team that conducted by Japan aerospace Exploration Agency (JAXA) investigated the following aspects of the organism: (1) whether meiotic chromosomal dynamics and apoptosis in the germ cells were normal under microgravity conditions, (2) the effect of the space flight on muscle cell development, and (3) the effect of the space flight on protein aggregation. In this article, we summarize the results of these biochemical and molecular biological analyses.
RESUMEN
17beta-Hydroxysteroid dehydrogenase (17beta-HSD) Type3 is an NADPH-dependent membrane-bound enzyme that is specifically expressed in testis and catalyzes the conversion of androstenedione to testosterone. To date, the sequence of Type3 enzymes has been clarified in humans, mice and rats; however, the sequence of the pig enzyme remains unknown. In this study, we determined the cDNA sequence of pig testicular 17beta-HSD Type3. PCR primers for partial pig testicular 17beta-HSD Type3 were designed from rat and human enzyme consensus sequences. Full-length cDNA was obtained by 3'- and 5'-RACE based on partial PCR products. The cDNA coding region was 933 bp in length, which is the same as the human enzyme, and shared 84.7% sequence identity with the human cDNA coding region. The monomer was estimated to have a molecular weight of 34,855 and to contain 310 amino acid residues. The predicted pig amino acid sequence showed 81.9, 75.5 and 72.9% sequence identity with the human, rat and mouse sequences, respectively. To elucidate 17beta-HSD Type3 activity, the expression vector pCMV/pig17beta-HSD3 was established and transfected into human embryo kidney 293 cells. Subsequently, 17beta-HSD activity (androstenedione conversion to testosterone) was strongly detected in cell lysates.
Asunto(s)
17-Hidroxiesteroide Deshidrogenasas/genética , 17-Hidroxiesteroide Deshidrogenasas/metabolismo , Porcinos/metabolismo , Testículo/enzimología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Línea Celular , ADN Complementario , Expresión Génica , Humanos , Masculino , Datos de Secuencia Molecular , ARN Mensajero/metabolismo , TransfecciónRESUMEN
The insulin/IGF-1 signalling (IIS) pathway plays an important role in the regulation of larval diapause, the long-lived growth arrest state called dauer arrest, in Caenorhabditis elegans. In this nematode, 40 insulin-like peptides (ILPs) have been identified as putative ligands of the IIS pathway; however, it remains unknown how ILPs modulate larval diapause. Here we show that the secretory polarity of INS-35 and INS-7, which suppress larval diapause, is changed in the intestinal epithelial cells at larval diapause. These ILPs are secreted from the intestine into the body cavity during larval stages. In contrast, they are secreted into the intestinal lumen and degraded during dauer arrest, only to be secreted into the body cavity again when the worms return to developmental growth. The process that determines the secretory polarity of INS-35 and INS-7, thus, has an important role in the modulation of larval diapause.
Asunto(s)
Proteínas de Caenorhabditis elegans/genética , Caenorhabditis elegans/genética , Polaridad Celular , Células Epiteliales/metabolismo , Crecimiento y Desarrollo/genética , Mucosa Intestinal/metabolismo , Somatomedinas/genética , Animales , Western Blotting , Proteínas de Caenorhabditis elegans/metabolismo , Hidrolasas de Éster Carboxílico/genética , Hidrolasas de Éster Carboxílico/metabolismo , Insulinas , Mucosa Intestinal/citología , Larva , Estadios del Ciclo de Vida , Microscopía Fluorescente , Hormonas Peptídicas/genética , Hormonas Peptídicas/metabolismo , Receptor de Insulina/genética , Receptor de Insulina/metabolismo , Somatomedinas/metabolismoRESUMEN
In the mouse genome, we found one processed Itpa gene-like sequence and two processed Itpa pseudogenes as well as the Itpa gene itself with introns, located on chromosome 2F3, which was isolated by a retro-recombination method. We also identified three types (A, B, C) of Itpa transcripts in mouse tissues. The processed Itpa gene-like sequence located on chromosome 2E1 has a complete open reading frame for exactly the same polypeptide as ITPA encoded by the type A transcript, with a polyadenylation signal. However, no transcribed sequence derived from the Itpa gene-like sequence was detectable in any of the mouse tissues examined, thus naming the sequence as Itpa processed pseudogene alpha. The type A Itpa mRNA, which was expressed in all mouse tissues examined, only encodes mouse ITPA polypeptide consisting of 198 amino acid residues with a capacity to hydrolyze dITP into dIMP. Itpa mRNA was detected in all tissues examined, and its expression is especially high in the testis, brain, and thymus. ITPA protein was mostly detected in the cytoplasm, to a lesser extent in the nuclei of neurons in the brain, and also those of hepatocytes, epithelial cells lining the bile duct, and endothelial cells lining the portal vein in the liver.
Asunto(s)
Genoma , Pirofosfatasas/genética , Empalme Alternativo , Secuencia de Aminoácidos , Animales , Células 3T3 BALB , Secuencia de Bases , Cerebelo/metabolismo , Hipocampo/metabolismo , Intrones , Hígado/metabolismo , Ratones , Datos de Secuencia Molecular , Especificidad de Órganos/genética , Seudogenes , Pirofosfatasas/metabolismo , ARN Mensajero/metabolismo , Alineación de Secuencia , Homología de Secuencia , Inosina TrifosfatasaRESUMEN
Royal jelly (RJ) produced by honeybees has been reported to possess diverse health-beneficial properties and has been implicated to have a function in longevity across diverse species as well as honeybees. 10-Hydroxy-2-decenoic acid (10-HDA), the major lipid component of RJ produced by honeybees, was previously shown to increase the lifespan of Caenorhabditis elegans. The objective of this study is to elucidate signaling pathways that are involved in the lifespan extension by 10-HDA. 10-HDA further extended the lifespan of the daf-2 mutants, which exhibit long lifespan through reducing insulin-like signaling (ILS), indicating that 10-HDA extended lifespan independently of ILS. On the other hand, 10-HDA did not extend the lifespan of the eat-2 mutants, which show long lifespan through dietary restriction caused by a food-intake defect. This finding indicates that 10-HDA extends lifespan through dietary restriction signaling. We further found that 10-HDA did not extend the lifespan of the long-lived mutants in daf-15, which encodes Raptor, a target of rapamycin (TOR) components, indicating that 10-HDA shared some longevity control mechanisms with TOR signaling. Additionally, 10-HDA was found to confer tolerance against thermal and oxidative stress. 10-HDA increases longevity not through ILS but through dietary restriction and TOR signaling in C. elegans.
RESUMEN
Although ghrelin acts as a modulator of feeding behavior and energy metabolism in the central nervous system, recent studies have implicated the peripheral actions of ghrelin in reproductive tissues. Here, we investigated the expression of ghrelin and its receptor (GHS-R) in mouse oocyte and preimplantation embryos, and we examined the role of ghrelin in the regulation of early embryo development. Both ghrelin and GHS-R mRNAs were detected in morula or more advanced embryo stages. As for the origin of ghrelin, both ghrelin mRNA and protein were identified in the uterine endometrium. The levels of ghrelin in uterine fluid as well as plasma were significantly increased in fasting mice compared with animals with free access to foods. Addition of ghrelin to culture media inhibited the development of two-cell embryos to the hatched blastocysts, and the inhibitory effects of ghrelin were abolished by an antagonist for the GHS-R. In addition, ghrelin significantly decreased the number of total cells, inner cell mass, and trophectoderm cells in blastocysts. These observations suggest that ghrelin could inhibit the development of preimplantation embryos during fasting. Thus, ghrelin may act as a peripheral factor to avoid the excess metabolic demands imposed by pregnancy during malnutritional states.
Asunto(s)
Blastocisto/fisiología , Regulación del Desarrollo de la Expresión Génica , Hormonas Peptídicas/genética , Hormonas Peptídicas/metabolismo , Receptores Acoplados a Proteínas G , Animales , Blastocisto/citología , Líquidos Corporales/química , Recuento de Células , Endometrio/química , Endometrio/fisiología , Ayuno/fisiología , Femenino , Ghrelina , Técnicas In Vitro , Ratones , Microscopía Fluorescente , Trastornos Nutricionales/fisiopatología , Oocitos/fisiología , Hormonas Peptídicas/análisis , Embarazo , ARN Mensajero/análisis , Receptores de Superficie Celular/genética , Receptores de GhrelinaRESUMEN
We investigated the effects of grapefruit juice (GFJ) and orange juice (OJ) on drug transport by MDR1 P-glycoprotein (P-gp) and multidrug resistance protein 2 (MRP2), which are efflux transporters expressed in human small intestine. We examined the transcellular transport and uptake of [(3)H]vinblastine (VBL) and [(14)C]saquinavir in a human colon carcinoma cell line (Caco-2) and in porcine kidney epithelial cell lines transfected with human MDR1 cDNA and human MRP2 cDNA, LLC-GA5-COL150, and LLC-MRP2, respectively. In Caco-2 cells, the basal-to-apical transports of [(3)H]VBL and [(14)C]saquinavir were greater than those in the opposite direction. The ratio of basal-to-apical transport to apical-to-basal transport of [(3)H]VBL and [(14)C]saquinavir by Caco-2 cells was reduced in the presence of MK571 (MRPs inhibitor), verapamil (P-gp inhibitor), cyclosporin A (inhibitor of both), 50% ethyl acetate extracts of GFJ and OJ, or their components (6',7'-dihydroxybergamottin, bergamottin, tangeretin, hepatomethoxyflavone, and nobiletin). Studies of transport and uptake of [(3)H]VBL and [(14)C]saquinavir with MDR1 and MRP2 transfectants showed that VBL and saquinavir are transported by both P-gp and MRP2. GFJ and OJ components inhibited the transport by MRP2 as well as P-gp. However, their inhibitory potencies for P-gp or MRP2 were substrate-dependent. The present study has revealed that GFJ and OJ interact with not only P-gp but also MRP2, both of which are expressed at apical membranes and limit the apical-to-basal transport of VBL and saquinavir in Caco-2 cells.
Asunto(s)
Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/metabolismo , Bebidas , Citrus/química , Proteínas de Transporte de Membrana/metabolismo , Proteínas Asociadas a Resistencia a Múltiples Medicamentos/metabolismo , Algoritmos , Antineoplásicos Fitogénicos/farmacocinética , Transporte Biológico Activo/efectos de los fármacos , Células CACO-2 , Bloqueadores de los Canales de Calcio/farmacología , Ciclosporina/farmacología , Interacciones Alimento-Droga , Inhibidores de la Proteasa del VIH/farmacocinética , Humanos , Inmunosupresores/farmacología , Proteínas de Transporte de Membrana/genética , Proteína 2 Asociada a Resistencia a Múltiples Medicamentos , Proteínas Asociadas a Resistencia a Múltiples Medicamentos/genética , Extractos Vegetales/química , Propionatos/farmacología , Quinolinas/farmacología , Saquinavir/farmacocinética , Transfección , Verapamilo/farmacología , Vinblastina/farmacocinéticaRESUMEN
The free radical theory of aging proposes that oxidative stress is one of the determinants of an organism's life span. In Caenorhabditis elegans, genetic or environmental changes have been shown to modulate life span. Here we discuss whether changes in the generation and destruction of free radicals are implicated in these life span modulations. Changes in culture oxygen concentrations that are considered to reflect free radical generation perturb the life span. The life spans under high and low oxygen concentrations were shorter and longer, respectively, than those under normoxic conditions. Short-term exposure to high oxygen concentration lengthens the life span. This is considered to be the result of an increase in antioxidant defense induced by short-term oxidative stress. Mutations in genes such as age-1 and daf-2 that compose the insulin-like signaling network conferred oxidative stress resistance and an increase in Mn-SOD gene expression as well as life span extension.
Asunto(s)
Proteínas de Caenorhabditis elegans , Caenorhabditis elegans/fisiología , Longevidad , Estrés Oxidativo , Animales , Caenorhabditis elegans/genética , Proteínas del Helminto/metabolismo , Insulina/metabolismo , Esperanza de Vida , Modelos Animales , Mutación , Oxígeno/metabolismo , Fenotipo , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal/fisiología , Superóxido Dismutasa/metabolismoRESUMEN
In order to simply and safely treat radioactive iodine waste, a study of the removal of iodide ion from radioactive waste using electrodialysis with an anion exchange paper membrane, in which trimethylhydroxylpropylammonium groups were homogeneously dispersed with high density. In Na125I and Na36Cl concentration-cell system, electric ion and water conductances, phenomenological coefficients, have been experimentally determined on basis of nonequilibrium thermodynamics. Prepared paper membrane had higher permselectivity of 125I ion than 36Cl ions by approximately 21%. On the other hand, water flux that was accompanied by an ionic transference in prepared paper membrane was greatly larger than that in typical synthesized membrane. It is suggested that a depression of water mobility is important to practice an ideal radioactive iodide waste electrodialysis system with a novel anion exchange paper membrane.
Asunto(s)
Cromatografía por Intercambio Iónico/métodos , Diálisis/métodos , Electroquímica/métodos , Residuos Industriales/prevención & control , Radioisótopos de Yodo/aislamiento & purificación , Modelos Químicos , Contaminantes Radiactivos del Agua/aislamiento & purificación , Aniones , Simulación por Computador , Residuos Industriales/análisis , Membranas Artificiales , Papel , Radiofármacos/aislamiento & purificación , Purificación del Agua/métodosRESUMEN
How microgravitational space environments affect aging is not well understood. We observed that, in Caenorhabditis elegans, spaceflight suppressed the formation of transgenically expressed polyglutamine aggregates, which normally accumulate with increasing age. Moreover, the inactivation of each of seven genes that were down-regulated in space extended lifespan on the ground. These genes encode proteins that are likely related to neuronal or endocrine signaling: acetylcholine receptor, acetylcholine transporter, choline acetyltransferase, rhodopsin-like receptor, glutamate-gated chloride channel, shaker family of potassium channel, and insulin-like peptide. Most of them mediated lifespan control through the key longevity-regulating transcription factors DAF-16 or SKN-1 or through dietary-restriction signaling, singly or in combination. These results suggest that aging in C. elegans is slowed through neuronal and endocrine response to space environmental cues.