RESUMO
The energy spacing between the spin-doublet bound state of _{Λ}^{4}He(1^{+},0^{+}) was determined to be 1406±2±2 keV, by measuring γ rays for the 1^{+}â0^{+} transition with a high efficiency germanium detector array in coincidence with the ^{4}He(K^{-},π^{-})_{Λ}^{4}He reaction at J-PARC. In comparison to the corresponding energy spacing in the mirror hypernucleus _{Λ}^{4}H, the present result clearly indicates the existence of charge symmetry breaking (CSB) in ΛN interaction. By combining the energy spacings with the known ground-state binding energies, it is also found that the CSB effect is large in the 0^{+} ground state but is vanishingly small in the 1^{+} excited state, demonstrating that the ΛN CSB interaction has spin dependence.
RESUMO
Evidence for the neutron-rich hypernucleus (Λ)(6)H is presented from the FINUDA experiment at DAΦNE, Frascati, studying (π+,π-) pairs in coincidence from the K(stop)(-) + (6)Li â(Λ)(6)H + π+ production reaction followed by (Λ)(6)H â (6)He + π- weak decay. The production rate of (Λ)(6) undergoing this two-body π- decay is determined to be (2.9 ± 2.0) × 10(-6)/K(stop)(-). Its binding energy, evaluated jointly from production and decay, is BΛ((Λ)(6)H) = (4.0 ± 1.1) MeV with respect to (5)H+Λ. A systematic difference of (0.98 ± 0.74) MeV between BΛ values derived separately from decay and from production is tentatively assigned to the (Λ)(6)H 0(g.s.)(+) â 1+ excitation.
RESUMO
The Θ(+) pentaquark baryon was searched for via the π(-)pâK(-)X reaction with a missing mass resolution of 1.4 MeV/c(2) (FWHM) at the Japan Proton Accelerator Research Complex (J-PARC). π(-) meson beams were incident on the liquid hydrogen target with a beam momentum of 1.92 GeV/c. No peak structure corresponding to the Θ(+) mass was observed. The upper limit of the production cross section averaged over the scattering angle of 2° to 15° in the laboratory frame is obtained to be 0.26 µb/sr in the mass region of 1.51-1.55 GeV/c(2). The upper limit of the Θ(+) decay width is obtained to be 0.72 and 3.1 MeV for J(Θ)(P)=1/2(+) and J(Θ)(P)=1/2(-), respectively, using the effective Lagrangian approach.
RESUMO
BACKGROUND: Children living at high altitude in San Antonio de los Cobres (SAC), Argentina, were shown to have lower high-density lipoprotein cholesterol (HDL-C) levels than Buenos Aires (BA) children. HDL antioxidant capacity is mainly attributed to paraoxonase1 (PON1). OBJECTIVE: To compare PON1 activity in indigenous SAC vs. BA children. METHODS: A cross-sectional study compared 158 SAC vs. 97 BA children (6-16 years). Anthropometric data and lipoprotein profile were measured. PON1 was evaluated employing paraoxon (PON) and phenylacetate (ARE) activity. RESULTS: The prevalence of overweight/obesity was lower in SAC than in BA children (18.3 vs. 30.9%). Triglycerides (1.34 vs. 0.90â mmol/l), apo B (0.84 vs.0.72â g/l), apo A-I (1.33 vs. 1.27â g/l), and ARE activity (100 vs. 90â µmol/ml/min) were higher, while HDL-C (1.16 vs. 1.32â mmol/l) and PON activity (170 vs. 203â nmol/ml/min) were lower in SAC than in BA. Separate multiple linear regression analyses showed that SAC children had significantly higher triglyceride (Beta -0.38), apo B (Beta -0.34), and ARE (Beta -0.36) plus lower HDL-C (Beta 0.33) and PON (Beta 0.25) compared with BA; adjusted for age, gender, and BMI. CONCLUSION: SAC showed an unfavorable lipoprotein profile, lower PON and higher ARE activities compared with BA children, suggesting the presence of altered HDL metabolism and antioxidant capacity.
Assuntos
Arildialquilfosfatase/sangue , Obesidade Infantil/enzimologia , Adolescente , Altitude , Apolipoproteína A-I/sangue , Argentina/epidemiologia , Argentina/etnologia , Arildialquilfosfatase/genética , Arildialquilfosfatase/metabolismo , Criança , HDL-Colesterol/sangue , Estudos Transversais , Feminino , Humanos , Modelos Lineares , Lipídeos/sangue , Masculino , Obesidade Infantil/epidemiologia , Fenilacetatos/metabolismo , Fatores de Risco , Triglicerídeos/sangueRESUMO
We report here the different ways in which four subunits of the basal transcription/repair factor TFIIH (XPB, XPD, p62 and p44) and the damage recognition XPC repair protein can enter the nucleus. We examined their nuclear localization by transiently expressing the gene products tagged with the enhanced green fluorescent protein (EGFP) in transfected 3T3 cells. In agreement with the identification of more than one putative nuclear localization signal (NLS) in their protein sequences, XPB, XPC, p62 and p44 chimeras were rapidly sorted to the nucleus. In contrast, the XPD-EGFP chimeras appeared mainly localized in the cytoplasm, with a minor fraction of transfectants showing the EGFP-based fluorescence also in the nucleus. The ability of the XPD chimeras to enter the nucleus was confirmed by western blotting on fractionated cell extracts and by functional complementation of the repair defect in the UV5 rodent cells, mutated in the XPD homologous gene. By deletion mutagenesis, we were unable to identify any sequence specific for nuclear localization. In particular, deletion of the putative NLS failed to affect subcellular localization and, conversely, the C-terminal part of XPD containing the putative NLS showed no specific nuclear accumulation. These findings suggest that the nuclear entry of XPD depends on its complexation with other proteins in the cytoplasm, possibly other components of the TFIIH complex.
Assuntos
Núcleo Celular/metabolismo , DNA Helicases , Reparo do DNA , Fatores de Transcrição TFII , Fatores de Transcrição/metabolismo , Células 3T3 , Animais , Transporte Biológico , Sobrevivência Celular/genética , Sobrevivência Celular/efeitos da radiação , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Proteínas de Fluorescência Verde , Células HeLa , Humanos , Proteínas Luminescentes/genética , Proteínas Luminescentes/metabolismo , Camundongos , Microscopia de Fluorescência , Subunidades Proteicas , Proteínas/genética , Proteínas/metabolismo , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Fator de Transcrição TFIIH , Fatores de Transcrição/genética , Transfecção , Proteína Grupo D do Xeroderma PigmentosoRESUMO
The drug-sensitive mutant UVS1, isolated from the Chinese hamster cell line CHO9, was previously found to complement the UV sensitivity of the excision repair-defective rodent mutants representative of groups 1 to 8 (Hata et al., Cancer Res., 51: 195-198, 1991; M. Numata et al., personal communication). Recently two new complementation groups of UV-sensitive CHO mutants, e.g., groups 9 and 10, have been identified (Stefanini et al., Cancer Res., 51: 3965-3971, 1991). In this paper we demonstrate that the repair defect in UVS1 cells is genetically different from those present in the mutants CHO7PV and CHO4PV, representing groups 9 and 10, respectively. Therefore, UVS1 represents a new complementation group of UV-sensitive rodent cell lines, the eleventh group.
Assuntos
Células CHO/efeitos da radiação , Reparo do DNA , DNA/efeitos da radiação , Raios Ultravioleta , Animais , Cricetinae , MutaçãoRESUMO
In this paper we demonstrate that the mutants CHO7PV and CHO4PV isolated by us from the CHO-K1 prol- cell line represent two new complementation groups of UV-sensitive excision repair-defective rodent mutants. We have classified the mutant CHO7PV as representative of Group 9 and CHO4PV as representative of Group 10. Cellular and biochemical characterization of these mutants indicates that they are moderately sensitive to a broad spectrum of mutagens (UV and mono- and bifunctional alkylating agents), partially unable to perform UV-induced DNA repair synthesis, and partially defective in the incision step of the DNA excision repair pathway and in the removal of the two main lesions caused by UV [cyclobutane pyrimidine dimers and (6-4) photo-products]. In terms of UV survival and incision, CHO4PV is apparently more defective than CHO7PV (40% and 50% of wild-type survival, respectively, and 55% and 75% of wild-type incision), whereas when repair DNA synthesis and lesion removal are compared, CHO7PV seems to be more severely affected (30% of wild-type unscheduled DNA synthesis in CHO7PV and 60% in CHO4PV). This suggests a subtlety in the relation between removal of these specific lesions and overall repair capacity and survival.
Assuntos
Linhagem Celular/fisiologia , Reparo do DNA/genética , DNA/efeitos da radiação , Animais , Linhagem Celular/efeitos da radiação , Cricetinae , Cricetulus , DNA/genética , Dano ao DNA/genética , Teste de Complementação Genética , Células Híbridas/fisiologia , Mutação/efeitos da radiação , Raios UltravioletaRESUMO
Xeroderma pigmentosum (XP)/Cockayne syndrome (CS) complex is a combination of clinical features of two rare genetic disorders in one individual. A sun-sensitive boy (XP20BE) who had severe symptoms of CS, with dwarfism, microcephaly, retinal degeneration, and mental impairment, had XP-type pigmentation and died at 6 y with marked cachexia (weight 14.5 lb) without skin cancers. We evaluated his cultured cells for characteristic CS or XP DNA-repair abnormalities. The level of ultraviolet (UV)-induced unscheduled DNA synthesis was less than 5% of normal, characteristic of the excision-repair defect of XP. Cell fusion studies indicated that his cells were in XP complementation group G. His cells were hypersensitive to killing by UV, and their post-UV recovery of RNA synthesis was abnormally low, features of both CS and XP. Post-UV survival of plasmid pSP189 in his cells was markedly reduced, and post-UV plasmid mutation frequency was higher than with normal cells, as in both CS and XP. Sequence analysis of the mutated plasmid marker gene showed normal frequency of plasmids with multiple base substitutions, as in CS, and an abnormally increased frequency of G:C-->A:T mutations, a feature of XP. Transfection of UV-treated pRSVcat with or without photoreactivation revealed that his cells, like XP cells, could not repair either cyclobutane pyrimidine dimers or non-dimer photoproducts. These results indicate that the DNA-repair features of the XP20BE (XP-G/CS) cells are phenotypically more like XP cells than CS cells, whereas clinically the CS phenotype is more prominent than XP.
Assuntos
Síndrome de Cockayne/complicações , Síndrome de Cockayne/genética , Reparo do DNA , Mutagênese , Raios Ultravioleta/efeitos adversos , Xeroderma Pigmentoso/complicações , Xeroderma Pigmentoso/genética , Sobrevivência Celular/efeitos da radiação , Criança , Síndrome de Cockayne/patologia , DNA/efeitos da radiação , Fibroblastos/efeitos da radiação , Teste de Complementação Genética , Humanos , Masculino , Plasmídeos/genética , RNA/biossíntese , Xeroderma Pigmentoso/patologiaRESUMO
A new mitomycin C (MMC)-sensitive rodent line, UV40, has been identified in the collection of ultraviolet light- (UV-) sensitive mutants of Chinese hamster ovary (CHO) cells isolated at the previous Facility for Automated Experiments in Cell Biology (FAECB). It was isolated from an UV mutant hunt using mutagenesis of AA8 cells with the DNA intercalating frameshift mutagen ICR170. It is complemented by CHO-UV-1, irsl, irs3, irslSF, MC5, V-C8 and V-H4 with respect to its MMC sensitivity based on cell survival. Despite having approx. 4 X normal UV sensitivity and increased sensitivity to UV inhibition of DNA replication, it has near-normal incision kinetics of UV irradiated DNA, and normal (6-4) photoproducts removal. It also is not hypermutable by UV, and shows near normal levels of UV inhibition of RNA synthesis. UV40 also has approx. 11 x .10 x .5 x and 2 x AA8 sensitivity to MMC, ethyl methanesulfonate (EMS), methyl methanesulfonate (MMS), and X-rays, respectively. Thus, its defect apparently does not involve nucleotide excision repair but rather another process, possibly in replicating past lesions. The spontaneous chromosomal aberration frequency is elevated to 20% in UV40, and the baseline frequency of sister chromatid exchange is also approximately 4-fold increased. The phenotype of UV40 appears to differ from all other rodent mutants that have so far been described.
Assuntos
Células CHO , Mitomicina/farmacologia , Mutagênicos/toxicidade , Adenina/análogos & derivados , Adenina/farmacologia , Aminoacridinas/farmacologia , Animais , Células CHO/efeitos dos fármacos , Adesão Celular , Linhagem Celular , Sobrevivência Celular , Aberrações Cromossômicas , Cricetinae , Replicação do DNA/efeitos da radiação , Teste de Complementação Genética , Mutagênese Sítio-Dirigida , Compostos de Mostarda Nitrogenada/farmacologia , Fenótipo , RNA/biossíntese , RNA/efeitos da radiação , Tolerância a Radiação , Troca de Cromátide Irmã , Raios UltravioletaRESUMO
Results of cellular and genetic characterization of UV sensitive clones (UVs) isolated from CHO-K1 cell line are reported. The cross-sensitivity to agents inducing a variety of DNA lesions, the induction of chromosome aberrations and of 6-thioguanine and ouabain resistant mutants, the occurrence of methotrexate resistant cells were analyzed in clones showing different degrees of UV sensitivity. Genetic analysis was performed by complementation analysis of hybrids obtained by fusion of our mutants with UVs cells belonging to the six complementation groups (c.g.) so far identified. Three clones were assigned to c.g. 2, one clone to c.g. 5. Two clones (CHO7PV and CHO4PV), were able to complement each other and showed complementation after fusion with any of the six c.g.; these clones were considered carriers of two new mutations in genes presumably involved in DNA repair.
Assuntos
Mutação , Tolerância a Radiação , Raios Ultravioleta , Animais , Linhagem Celular , Cricetinae , CricetulusRESUMO
The results obtained in 201 cases of haemorrhage of the higher tract of the digestive tube are reported. The role of early endoscopy (less than 24 h) and that of a "rigorous" selection of patients for surgery treatment are emphasized. The value of surgical resection in serious haemorrhage of chronic peptic ulcus is confirmed by the satisfactory results obtained.
Assuntos
Hemorragia Gastrointestinal/cirurgia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Gastrectomia , Hemorragia Gastrointestinal/diagnóstico , Hemorragia Gastrointestinal/etiologia , Gastroscopia , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Fatores de TempoRESUMO
BACKGROUND: The development of intestinal transplant (Tx) programs introduces thymoglobulin donor treatment as well as an almost complete warm dissection of the abdominal organs to allocate them to different recipients. Our aim is to assess the reproducibility and feasibility of the surgical technique of multi-organ procurement with the use of thymoglobulin donor pre-treatment and report the short- and long-term outcomes of every graft harvested as part of multi-organ procurement (MTOp), including the intestine. METHODS: Data were collected of all organs harvested from MTOp, including the intestines allocated to our center from March 2006 to July 2011. Data from 92 recipients and 116 organs procured from 29 MTOp were analyzed. Twelve hearts, 2 lungs, and 1 cardio-pulmonary block were transplanted; primary graft dysfunction developed in 4 of the 12 hearts and in the cardio-pulmonary block. RESULTS: The survival rate was 75% and 100% for hearts and lungs, respectively. Nineteen livers, 9 kidney-pancreas, 19 kidneys, and 29 intestines were transplanted. Delayed graft function (DGF) of the pancreas developed in 3 of 9 kidney-pancreas, and the other 3 exhibited DGF of the kidney; 4 of 19 Tx kidneys had DGF. The survival was 84%, 78%, 95%, and 65.5% for livers, kidney-pancreas, kidneys, and intestines, respectively. CONCLUSIONS: Organs procured during MTOp including the intestine can be safely used, increasing organ availability and transplant applicability without compromising allocation, quality, and long-term results of the non-intestinal-procured organs.
Assuntos
Transplante de Órgãos , Coleta de Tecidos e Órgãos/métodos , Obtenção de Tecidos e Órgãos , Adolescente , Adulto , Soro Antilinfocitário , Criança , Pré-Escolar , Estudos de Viabilidade , Feminino , Sobrevivência de Enxerto , Humanos , Lactente , Intestinos/transplante , Masculino , Reprodutibilidade dos Testes , Estudos Retrospectivos , Taxa de Sobrevida , Coleta de Tecidos e Órgãos/efeitos adversos , Coleta de Tecidos e Órgãos/mortalidade , Resultado do Tratamento , Adulto JovemAssuntos
Apoptose/efeitos da radiação , Dano ao DNA/genética , Reparo do DNA/genética , Queratinócitos/efeitos da radiação , Raios Ultravioleta/efeitos adversos , Apoptose/genética , Ciclo Celular/genética , Ciclo Celular/efeitos da radiação , Células Cultivadas , DNA/efeitos da radiação , Relação Dose-Resposta à Radiação , Fibroblastos/metabolismo , Fibroblastos/efeitos da radiação , Humanos , Queratinócitos/metabolismo , Dímeros de Pirimidina/metabolismo , Dímeros de Pirimidina/efeitos da radiação , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/efeitos da radiaçãoRESUMO
Trichothiodystrophy (TTD) is an autosomal recessive disorder with symptoms affecting several tissues and organs. The most relevant features are hair abnormalities, physical and mental retardation, ichthyosis, signs of premature aging and cutaneous photosensitivity. The clinical spectrum of TTD varies widely from patients with only brittle, fragile hair to patients with the most severe neuroectodermal symptoms. To date, four genes have been identified as responsible for TTD: XPD, XPB, p8/TTDA, and TTDN1. Whereas the function of TTDN1 is still unknown, the former three genes encode subunits of TFIIH, the multiprotein complex involved in basal and activated transcription and in nucleotide excision repair (NER). Ongoing investigations on TTD are elucidating not only the pathogenesis of the disease, which appears to be mainly related to transcriptional impairment, but also the modalities of NER and transcription in human cells and how TFIIH operates in these two fundamental cellular processes.