RESUMEN
The p.Asp211Gly homozygous HYLS1 mutation is so far known to cause only hydrolethalus syndrome, a lethal malformation syndrome. We report living sibling patients with a homozygous no-stop mutation in exon 4 of HYLS1, NM_145014.2:c.900A>C (p.Ter300TyrextTer11) in the second decade of life. The proband has Joubert syndrome (JS). The younger brother also has JS and an enlarged posterior fossa that was initially diagnosed as Dandy-Walker malformation. The present mutation is unique as it affects the stop codon. The product protein HYLS1 plays an essential role in the formation of the primary cilium. This report provides insight into the spectrum of disorders involving midline brain defects closely related to cilium dysfunction or ciliopathy.
Asunto(s)
Anomalías Múltiples/genética , Cerebelo/anomalías , Anomalías del Ojo , Predisposición Genética a la Enfermedad/genética , Mutación , Proteínas/genética , Retina/anomalías , Anomalías Múltiples/diagnóstico por imagen , Adolescente , Secuencia de Bases , Cerebelo/diagnóstico por imagen , Niño , Consanguinidad , Exoma/genética , Anomalías del Ojo/diagnóstico por imagen , Anomalías del Ojo/genética , Femenino , Homocigoto , Humanos , Enfermedades Renales Quísticas/diagnóstico por imagen , Enfermedades Renales Quísticas/genética , Masculino , Linaje , Retina/diagnóstico por imagen , Análisis de Secuencia de ADN , HermanosRESUMEN
The effects of reduced dimensions and the interfaces on antiferromagnetic quantum criticality are studied in epitaxial Kondo superlattices, with alternating n layers of heavy-fermion antiferromagnet CeRhIn_{5} and seven layers of normal metal YbRhIn_{5}. As n is reduced, the Kondo coherence temperature is suppressed due to the reduction of effective Kondo screening. The Néel temperature is gradually suppressed as n decreases and the quasiparticle mass is strongly enhanced, implying dimensional control toward a quantum critical point. Magnetotransport measurements reveal that a quantum critical point is reached for the n=3 superlattice by applying small magnetic fields. Remarkably, the anisotropy of the quantum critical field is opposite to the expectations from the magnetic susceptibility in bulk CeRhIn_{5}, suggesting that the Rashba spin-orbit interaction arising from the inversion symmetry breaking at the interface plays a key role for tuning the quantum criticality in the two-dimensional Kondo lattice.
RESUMEN
Although exercise training improves hippocampus-related cognition, the optimum exercise intensity is still disputed. Based on the lactate threshold (LT, approximately 20 m/min on treadmill) of rats, we have shown that 2 weeks of training with stress-free mild exercise (ME,
Asunto(s)
Hipocampo/fisiología , Memoria/fisiología , Condicionamiento Físico Animal/fisiología , Animales , Cognición/fisiología , Corticosterona/sangre , Ácido Láctico/sangre , Masculino , Aprendizaje por Laberinto/fisiología , Músculo Esquelético/fisiología , Ratas Wistar , Estrés PsicológicoAsunto(s)
Canal Anal/fisiopatología , Laparoscopía/métodos , Neoplasias del Recto/cirugía , Vagina/cirugía , Adulto , Anciano , Canal Anal/cirugía , Quimioradioterapia Adyuvante , Femenino , Humanos , Escisión del Ganglio Linfático , Persona de Mediana Edad , Terapia Neoadyuvante , Estadificación de Neoplasias , Tratamientos Conservadores del Órgano , Neoplasias del Recto/tratamiento farmacológico , Neoplasias del Recto/patología , Resultado del TratamientoAsunto(s)
Colectomía/métodos , Colon Transverso/cirugía , Laparoscopía/métodos , Páncreas/cirugía , HumanosRESUMEN
Background: There is a lack of large studies focusing on the prognostic significance of lateral lymph node (LLN) metastasis following LLN dissection (LLND) in rectal cancer. The aim of this study was to evaluate the prognostic impact of LLN metastases on survival of patients with advanced low rectal cancer. Methods: Consecutive patients with locally advanced, but not metastatic, extraperitoneal rectal cancer treated with neoadjuvant (chemo)radiotherapy plus total mesorectal excision between 2004 and 2015 were included in the study. LLND was performed when pretreatment imaging documented enlarged LLNs (7 mm or greater in size). Localization of nodal metastases and long-term outcomes were analysed. Kaplan-Meier analysis was used to compare the survival of patients with ypN0 disease with that of patients with mesorectal ypN+/LLN- status and patients with positive LLNs. The Cox proportional hazards model was used to evaluate predictors of disease-free survival (DFS) and local recurrence. Results: A total of 613 patients were included in the study; LLND was performed in 212 patients (34·6 per cent) and 57 (9·3 per cent) had LLN metastasis. Patients with LLN metastasis had improved DFS and local recurrence cumulative incidence rates compared with patients with mesorectal ypN2+/LLN- disease (DFS: P = 0·014; local recurrence: P = 0·006). Although the DFS rate of patients with LLN metastasis was worse than that of patients with ypN0 disease (P < 0·001), the cumulative incidence of local recurrence was similar (P = 0·491). In multivariable analysis, residual LLN metastasis was not an independent predictor of worse DFS or local recurrence. Conclusion: LLN metastasis is not an independent predictor of local recurrence or survival. Survival of patients presenting with LLN metastasis after (chemo)radiotherapy was intermediate between that of patients with ypN0 status and those with mesorectal ypN2 positivity.
Antecedentes: No existen en la literatura grandes estudios dirigidos a investigar la importancia pronóstica de las metástasis en los ganglios linfáticos laterales (lateral lymph nodes, LLN) después de la disección de los mismos (LLN dissection, LLND) en pacientes con cáncer de recto. El objetivo de este estudio fue evaluar el impacto pronóstico de las metástasis en los LLN sobre la supervivencia de los pacientes con cáncer de recto. Métodos: Se analizaron 613 pacientes consecutivos con cáncer de recto localmente avanzado extraperitoneal y no metastásico tratados con (quimio)radioterapia neoadyuvante seguida de resección total del mesorrecto (total mesorectal excision, TME) entre 2004 y 2015. Se realizó una LLND cuando el estudio mediante pruebas de imagen previo el tratamiento mostró LLN aumentados de tamaño ≥ 7 mm. Se analizó la localización de las metástasis ganglionares y los resultados a largo plazo. El análisis de supervivencia se realizó mediante el método de KaplanMeier para comparar las supervivencias de los pacientes ypN0 frente a los pacientes ypN con positividad mesorrectal/LLN negativos y frente a los pacientes LLN positivos. Se utilizó el modelo de riesgo proporcional de Cox para evaluar los factores predictivos de supervivencia libre de enfermedad y de recidiva local. Resultados: Se realizó una LLND en 212 (34,6%) pacientes, y 57 (9,3%) pacientes presentaban metástasis en los LLN. Los pacientes con metástasis en los LLN presentaron mejores curvas de incidencia acumulada de recidiva local y de supervivencia libre de enfermedad en comparación con los pacientes con ganglios mesorrectales ypN2 positivos/LLN negativos (respectivamente, P = 0,0135 y P = 0,0060). Aunque la curva de la supervivencia libre de enfermedad de los pacientes con metástasis en los LLN fue peor que la de los pacientes ypN0 (P < 0,0001), la incidencia acumulada de recidiva local fue similar (P = 0,4905). En el análisis multivariable, la metástasis residual en los LLN no fue un factor predictivo independiente de peor supervivencia libre de enfermedad ni de recidiva local. Conclusión: Las metástasis en los LLN no es un factor predictivo independiente de recidiva local o supervivencia. Los pacientes que presentaron metástasis en los LLN después de (quimio)radioterapia mostraron características de supervivencia intermedias entre ypN0 y pacientes con ganglios mesorrectales ypN2 positivos.
Asunto(s)
Metástasis Linfática/terapia , Terapia Neoadyuvante/métodos , Recurrencia Local de Neoplasia/diagnóstico , Proctectomía , Neoplasias del Recto/terapia , Anciano , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Quimioradioterapia Adyuvante/métodos , Supervivencia sin Enfermedad , Femenino , Fluorouracilo/uso terapéutico , Humanos , Incidencia , Estimación de Kaplan-Meier , Leucovorina/uso terapéutico , Escisión del Ganglio Linfático , Ganglios Linfáticos/patología , Ganglios Linfáticos/cirugía , Metástasis Linfática/patología , Masculino , Persona de Mediana Edad , Recurrencia Local de Neoplasia/epidemiología , Recurrencia Local de Neoplasia/patología , Recurrencia Local de Neoplasia/prevención & control , Estadificación de Neoplasias , Neoplasia Residual , Compuestos Organoplatinos/uso terapéutico , Pronóstico , Neoplasias del Recto/mortalidad , Neoplasias del Recto/patología , Recto/patología , Recto/cirugía , Estudios RetrospectivosRESUMEN
Previously, we obtained a protein that has considerable amino acid sequence homology with secretory phospholipase A(2) (PLA(2)) from a bullfrog pituitary fraction obtained during the purification of thyrotropin (TSH). Subsequently, partial amino acid sequence (N-terminal 45 amino acid residues) analysis revealed this protein to be identical to the N-terminal amino acid sequence of otoconin-22, the major protein of aragonitic otoconia in the Xenopus saccule. In this study we developed an antibody against the N-terminal peptide of the bullfrog protein and applied it for immunocytochemical study of the pituitary and its surrounding tissue. Western blotting analysis showed that this antibody recognizes a 20.4-kD protein that has a molecular mass close to that of otoconin-22. Immunohistochemical reaction with the antibody was not found in any anterior pituitary cells but was intense in the monolayer epithelial cells of the endolymphatic sac surrounding the pituitary gland, which is a major storage site of calcium carbonate in amphibians. An electron microscopic study revealed that the cuboidal cells in the endolymphatic sac contained large, polymorphic secretory granules in their apical cytoplasm. Immunogold particles indicating the presence of a PLA(2)-like protein were observed predominately in these secretory granules. These findings support the view that this PLA(2)-like protein obtained during purification of TSH was derived from the endolymphatic sac adhering to the pituitary and that this protein is a bullfrog otoconin. (J Histochem Cytochem 49:631-637, 2001)
Asunto(s)
Saco Endolinfático/metabolismo , Glicoproteínas/metabolismo , Fosfolipasas A/química , Hipófisis/metabolismo , Animales , Western Blotting , Proteínas de Unión al Calcio , Saco Endolinfático/ultraestructura , Femenino , Glicoproteínas/química , Inmunohistoquímica , Masculino , Microscopía Electrónica , Peso Molecular , Hipófisis/ultraestructura , Rana catesbeiana , Proteínas de XenopusRESUMEN
An insoluble steroid derivative was prepared by the coupling of desoxycorticosterone with aminoethylated agarose beads. When the naked full-grown Xenopus oocytes were incubated in contact with the steroid-bound agarose beads, the dissociation of oocyte nuclei or the initial step in the maturation of amphibian oocyte was induced in 30-100% of total oocytes. The induction did not occur in the oocytes covered with follicle cells and when the naked oocytes were placed apart from the steroid-bound agarose beads in the medium. The above findings confirmed that the oocyte surface was a primary site for this particular steroid action, eliminating the possibility of participation of free steroid artifactually released from the agarose. The 105,000g supernatant fraction of oocytes showed no sign of the presence of steroid receptor. This was not inconsistent with the assumption mentioned above.
Asunto(s)
Desoxicorticosterona/farmacología , Oocitos/fisiología , Óvulo/fisiología , Animales , Femenino , Oocitos/efectos de los fármacos , Progesterona/metabolismo , Receptores de Superficie Celular/aislamiento & purificación , Receptores de Superficie Celular/metabolismo , Sefarosa , XenopusRESUMEN
Four gonadotropin components were purified from the basic protein fraction of bullfrog pituitary glands. All the components had high potencies not only in the ovulation assay using Xenopus laevis ovaries but also in the competitive binding assay for rat follicle-stimulating hormone (FSH) using Xenopus laevis testis. The isoelectric points of the four components determined by isoelectric focusing were at pH 8.8, 9.0, 9.1, and 9.3, respectively. No appreciable difference in the physical and chemical properties other than isoelectric points was found among the four components. Their Stokes radius estimated by gel filtration was 2.36 nm. This value was appreciably lower than that reported for mammalian luteinizing hormone (LH). On sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis, reduced preparations of the four components each showed two bands, indicating a subunit structure similar to that of mammalian gonadotropins. Their mobilities were greater than those of mammalian LH. The molecular weights of subunits estimated by high-speed gel filtration in 6 M guanidine hydrochloride were lower than those of mammalian LH. The amino acid composition of the four basic components of bullfrog gonadotropin was appreciably different from that of mammalian LH.
Asunto(s)
Gonadotropinas Hipofisarias/aislamiento & purificación , Hipófisis/análisis , Aminoácidos/análisis , Animales , Fenómenos Químicos , Química , Hormona Folículo Estimulante/aislamiento & purificación , Focalización Isoeléctrica , Hormona Luteinizante/aislamiento & purificación , Rana catesbeianaRESUMEN
We examined the electrophysiological effect of pituitary adenylate cyclase activating polypeptide (PACAP) in isolated Xenopus laevis oocytes in vitro. In conventional two-electrode voltage clamp experiments, PACAP (1-10 microM) activated an inward rectifier current at membrane potentials more negative than -60 mV without causing any significant change in currents at potentials more positive than -60 mV both in the follicle-enclosed oocyte and in the defolliculated oocyte. This current reversed at -22.5 mV, close to the theoretical value of Cl- equilibrium potential and the reversal potential of this current was shifted positively by reducing [Cl-]o. This current was blocked by Cl- channel blocker SITS and Ba2+. Furthermore, VIP and adenylate cyclase activator forskolin did not elicit the currents. In conclusion, PACAP elicited the hyperpolarization-activated Cl- current in Xenopus laevis oocytes. This current may modulate the membrane potential of the oocyte, thereby affecting the oocyte physiology.
Asunto(s)
Canales de Cloruro/efectos de los fármacos , Canales de Cloruro/fisiología , Neuropéptidos/farmacología , Oocitos/efectos de los fármacos , Oocitos/fisiología , Animales , Canales de Cloruro/metabolismo , Femenino , Potenciales de la Membrana/efectos de los fármacos , Oocitos/metabolismo , Folículo Ovárico/efectos de los fármacos , Folículo Ovárico/fisiología , Técnicas de Placa-Clamp , Polipéptido Hipofisario Activador de la Adenilato-Ciclasa , Receptores de Somatotropina/efectos de los fármacos , Receptores de Péptido Intestinal Vasoactivo/efectos de los fármacos , Péptido Intestinal Vasoactivo/farmacología , Proteínas de Xenopus , Xenopus laevisRESUMEN
We investigated therapeutic effects of 4-methylcatechol (4-MC), a non-amine catechol compound, on streptozotocin (STZ)-induced diabetic neuropathy in rats. 4-MC is one of the potent stimulators of endogenous nerve growth factor (NGF) synthesis both in vitro and in vivo. Diabetic rats showed a statistically significant reduction in motor nerve conduction velocity (MNCV), mean myelinated axon diameter, and NGF content in the sciatic nerve during the experimental period of 8 weeks. The 4-MC treatment started 4 weeks after the STZ injection resulted in significantly greater NGF content, faster MNCV, and larger mean myelinated nerve fiber diameter and axon diameter than in untreated diabetic rats. These findings suggest that a decreased NGF level in the diabetic sciatic nerves may be involved in the pathogenesis of diabetic neuropathy and that 4-MC treatment could be useful for diabetic neuropathy.
Asunto(s)
Antioxidantes/uso terapéutico , Catecoles/uso terapéutico , Neuropatías Diabéticas/tratamiento farmacológico , Factores de Crecimiento Nervioso/biosíntesis , Animales , Axones/ultraestructura , Diabetes Mellitus Experimental/fisiopatología , Neuropatías Diabéticas/patología , Neuropatías Diabéticas/fisiopatología , Electrofisiología , Masculino , Neuronas Motoras/fisiología , Conducción Nerviosa/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Nervio Ciático/efectos de los fármacos , Nervio Ciático/patología , Nervio Ciático/fisiopatologíaRESUMEN
Acrylamide monomer (ACR) causes central-peripheral distal axonopathy. We induced neuropathy in rats by means of ACR injection as an experimental model of the dying-back type of peripheral neuropathy to assess the potential efficacy of 4-methylcatechol (4-MC), a potent stimulator of endogenous nerve growth factor (NGF) synthesis, as a therapeutic agent for the axonal nerve lesion. ACR-induced neuropathy in rats resulted in a dying- back type of axonal degeneration, and a statistically significant reduction in motor nerve conduction velocity (MNCV), and density of large myelinated fibers. We administered 4-MC and ACR together to rats intraperitoneally and found improved clinical signs, and significantly more NGF content in sciatic nerves, faster MNCV, and greater myelinated fiber density than in rats given ACR alone. These findings suggest that 4-MC can prevent the progression of ACR-induced neuropathy and decreased NGF levels may be involved in the pathogenesis of ACR neuropathy.
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Acrilamidas/efectos adversos , Antioxidantes/farmacología , Catecoles/farmacología , Factores de Crecimiento Nervioso/biosíntesis , Enfermedades del Sistema Nervioso/inducido químicamente , Conducción Nerviosa/efectos de los fármacos , Animales , Masculino , Enfermedades del Sistema Nervioso/fisiopatología , Conducción Nerviosa/fisiología , Ratas , Ratas Sprague-Dawley , Nervio Ciático/ultraestructura , Factores de TiempoRESUMEN
UNLABELLED: We experienced an autopsy case, small testes and tall stature, which suggested Klinefelter's syndrome. DNA analysis was performed to confirm the genetic abnormality. CASE HISTORY: A 28-year-old man who was single and lived with his parents. He suddenly lost his consciousness in a sitting room and died. Autopsy findings: He was 176 cm in height and 57 kg in weight. The post-mortem hypostasis was red-purple on his back, and rigor mortis was strong in each joint of the whole body. The heart weighted 340 g, in which dark red fluidal blood (300 ml) without coagulation was contained. The testes were smaller than normal adult male (left and right testes with epididymides weighted 8.1 g and 6.0 g, respectively). As a results of pathological examination, clumped Leydig cells, sclerotic and hyalined tubules were observed. Some germ cells with spermatozoid were also present. DNA Analysis: Generally, Klinefelter's syndrome is determined by karyotype analysis and/or the detection of sex chromatin. However, in this case, karyotype analysis and the detection of sex chromatin could not be demonstrated, because the blood which was collected in the autopsy became too old. Therefore, we tried sex determination and STR analysis (HPRT, HUMARA and DXS 1470) using DNA extracted from stored blood materials. Consequently, in the sex determination, no different situation was found in the X- and Y-specific bands from normal male's and as results of STR analyses, we could not corroborate the Klinefelter's syndrome.
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Autopsia/métodos , Análisis Mutacional de ADN/métodos , Síndrome de Klinefelter/genética , Repeticiones de Minisatélite/genética , Análisis para Determinación del Sexo/métodos , Adulto , Electroforesis en Gel de Poliacrilamida/métodos , Humanos , Cariotipificación , Masculino , Reacción en Cadena de la Polimerasa/métodosRESUMEN
The gene encoding extracellular poly(3-hydroxybutyrate) depolymerase from a thermophilic poly(3-hydroxybutyrate)-degrading bacterium, strain HS, was cloned and intracellularly expressed in Escherichia coli. The gene was found to consist of 1485-bp nucleotide sequence coding for a 22-amino-acid signal peptide and a 473-amino-acid mature protein. Phylogenetic analysis and domain structure showed that the enzyme was clustered with type II PHB depolymerases. The gene was expressed in E. coli under the control of the tac promoter. A 46-kDa protein was detected in the cell extract by SDS-PAGE. The N-terminal sequence of the protein agreed with that of the original enzyme. The crude enzyme was able to degrade PHB particles at 70 degrees C.
RESUMEN
A 1200-bp cDNA encoding Xenopus laevis deoxyribonuclease I (X. laevis DNase I) was constructed from the total RNA of a X. laevis pancreas using a rapid amplification of cDNA ends method. When the cDNA was transiently transfected into COS-7 cells, the recombinant polypeptide exhibited similar enzymological properties to those of the native pancreatic DNase I. The recombinant enzyme was considerably more labile than most other vertebrate DNase I enzymes. The X. laevis DNase I polypeptide was larger than any other known vertebrate DNase I, containing a unique Cys-rich stretch of 68 or 70 amino acid residues at the carboxyl terminus, and it had less well conserved binding sites for the Ca2+, G-actin and DNA, and two DNase I signature motifs. These alterations might account for its heat instability.
Asunto(s)
Desoxirribonucleasa I/genética , Xenopus laevis/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Células COS , Chlorocebus aethiops , Clonación Molecular , ADN Complementario , Desoxirribonucleasa I/química , Desoxirribonucleasa I/metabolismo , Humanos , Cinética , Datos de Secuencia Molecular , Páncreas/enzimología , Filogenia , Proteínas Recombinantes/metabolismo , Transfección , VertebradosRESUMEN
Sex determination from blood and teeth by PCR amplification of the alphoid satellite family according to the method of Witt and Erickson was investigated. Amplification of the X-(131 bp) and Y-(172 bp) specific sequences in males and that of the X-specific sequence in females was satisfactory in almost all samples. However, of the 22 samples (blood: 5, dental pulp: 12, tooth hard tissue: 5), a false positive fragment was detected at 172 bp among the female samples (blood: 17, dental pulp: 30, tooth hard tissue: 23) as a result of silver staining. These bands were not amplified by the dual PCR method and were different from the Y-specific band. Subsequent restriction enzyme digestion (DdeI, HinfI, and NciI) of these bands resulted in different patterns from the typical Y-specific band. Furthermore, in five male samples of hard tooth tissue specimens (n = 14), a Y-specific band was detected as a weak band, and in two of these five samples, this weak specific band was not amplified by dual PCR in spite of the male origin of the samples. These results showed that this method is useful for sex determination, but that dual PCR and restriction analysis should be used together in practical applications.
Asunto(s)
Manchas de Sangre , ADN Satélite/análisis , Odontología Forense , Análisis para Determinación del Sexo/métodos , Secuencia de Bases , Enzimas de Restricción del ADN/análisis , Femenino , Humanos , Masculino , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Cromosoma X/genética , Cromosoma Y/genéticaRESUMEN
VNTR polymorphism at the D4S43 locus was investigated, and allele frequencies in the Japanese population sample were determined by the polymerase chain reaction. Eleven different alleles and 16 genotypes were observed in 131 unrelated Japanese. The most common allele was one repeat unit (60.3%), the heterozygosity index of the present study was 58.7%, and the polymorphic information content was calculated at 0.55. In addition, we found four new size variations in the one-repeat-unit allele, which was the most common allele among the Japanese population. These variations were believed to be different from variations by repeats of the basic 14-bp unit. The one-repeat-unit allele at the D4S43 locus, which comes in four sizes, was detected in all of the samples, including degraded DNA samples obtained from hard tooth tissue. Therefore, this new variation is useful in personal identification by DNA analysis using forensic scientific degraded DNA samples.
Asunto(s)
ADN/metabolismo , Odontología Forense/métodos , Repeticiones de Minisatélite/genética , Diente/metabolismo , Alelos , ADN/genética , Dermatoglifia del ADN , Cartilla de ADN/química , Frecuencia de los Genes , Humanos , Japón , Reacción en Cadena de la PolimerasaRESUMEN
Direct digital X-ray technology was applied to dental identification of victims in two murder cases using the Compuray system. In both cases the digital radiography proved to be simple to use, quick and effective, allowing superimposition, enlargement and transportability to a mortuary. These are the first reported uses of the technology in Japan and further development promises the transmitability of data and images electronically to remote locations, further enhancing its usefulness. Comparing the skull with the dental ante-mortem X-ray films and records of a specific person who was reported "missing", we found many identical points between the two, especially in regard to the X-ray findings with the Compuray. In both cases we obtained a large number of X-ray images in a remarkably short time and this was very useful for identification by means of the teeth.