RESUMEN
Bioglass 45S5, a silica-based glass, has pioneered a new field of biomaterials. Bioglass 45S5 promotes mineralization through calcium ion release and is widely used in the dental field, including toothpaste formulations. However, the use of Bioglass 45S5 for bone grafting is limited owing to the induction of inflammation, as well as reduced degradation and ion release. Phosphate-based glasses exhibit higher solubility and ion release than silica-based glass. Given that these glasses can be synthesized at low temperatures (approximately 1,000°C), they can easily be doped with various metal oxides to confer therapeutic properties. Herein, we fabricated zinc- and fluoride-doped phosphate-based glass (multicomponent phosphate [MP] bioactive glass) and further doped aluminum oxide into the MP glass (4% Al-MP glass) to overcome the striking solubility of phosphate-based glass. Increased amounts of zinc and fluoride ions were detected in water containing the MP glass. Doping of aluminum oxide into the MP glass suppressed the striking dissolution in water, with 4% Al-MP glass exhibiting the highest stability in water. Compared with Bioglass 45S5, 4% Al-MP glass in water had a notably reduced particle size, supporting the abundant ion release of 4% Al-MP glass. Compared with Bioglass 45S5, 4% Al-MP glass enhanced the osteogenesis of mouse bone marrow-derived mesenchymal stem cells. Mouse macrophages cultured with 4% Al-MP glass displayed enhanced induction of anti-inflammatory M2 macrophages and reduced proinflammatory M1 macrophages, indicating M2 polarization. Upon implanting 4% Al-MP glass or Bioglass 45S5 in a mouse calvarial defect, 4% Al-MP glass promoted significant bone regeneration when compared with Bioglass 45S5. Hence, we successfully fabricated zinc- and fluoride-releasing bioactive glasses with improved osteogenic and anti-inflammatory properties, which could serve as a promising biomaterial for bone regeneration.
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Sustitutos de Huesos , Cerámica , Fluoruros , Vidrio , Zinc , Fluoruros/química , Animales , Ratones , Cerámica/química , Sustitutos de Huesos/química , Vidrio/química , Osteogénesis/efectos de los fármacos , Materiales Biocompatibles/química , Ensayo de MaterialesRESUMEN
Aldehyde dehydrogenase 2 (ALDH2) detoxifies toxic aldehydes and has a key role in protecting the liver. An elevated gamma-glutamyl transferase (GGT) level is related to oxidative stress and nonalcoholic fatty liver disease (NAFLD). We herein investigated the association between inactive ALDH2*2 allele (rs671) and the risk of NAFLD, including the relationship to the GGT level. A retrospective follow-up study (mean 5.4±1.1 years) was conducted among 341 Japanese health screening program participants. The receiver operating characteristic curve indicated that the GGT level predicted the development of NAFLD (area under the curve: 0.65, P<0.05) with a cutoff value of 25.5 IUl(-1). The longitudinal risk of NAFLD was higher in the ALDH2*2 allele carriers than in the noncarriers (odds ratio (OR): 2.30, 95% confidence interval (CI): 1.21-4.40), and the risk was further increased among the *2 allele carriers with GGT values ⩾25.5 IUl(-1) (OR: 4.28, 95% CI: 1.80-10.19). On the other hand, there were no significant changes in the subjects' body weight and body mass index during observation period. The ALDH2*2 allele, in relation to the GGT level, may potentially be a novel risk factor for NAFLD.
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Aldehído Deshidrogenasa/genética , Alelos , Enfermedad del Hígado Graso no Alcohólico/genética , Aldehído Deshidrogenasa Mitocondrial , Índice de Masa Corporal , Femenino , Estudios de Seguimiento , Humanos , Estudios Longitudinales , Masculino , Estrés Oxidativo/genética , Curva ROC , Estudios Retrospectivos , Factores de Riesgo , gamma-Glutamiltransferasa/genética , gamma-Glutamiltransferasa/metabolismoRESUMEN
AIM: The common variants p.I27L (rs1169288), p.A98V (rs1800574) and p.S487N (rs2464196) of the hepatocyte nuclear factor 1-α (HNF1A) gene have been inconsistently associated with impaired glucose tolerance and/or an increased risk of type 2 diabetes mellitus (T2DM). The present study aimed to investigate whether these associations are affected by weight. METHODS: A cross-sectional analysis was conducted among 861 Japanese subjects (males: 65.5%; 61.8±12.3years) attending a health-screening programme. Interactive effects between HNF1A variants and weight status on risk of T2DM or dysglycaemic status were determined. RESULTS: The 27L variant carriers were at higher risk of T2DM and dysglycaemic status than non-carriers, but only in normal-weight subjects [odds ratio (OR): 2.04, P=0.03 and OR: 2.56, P=0.01, respectively]. An interactive effect of the p.I27L (rs1169288) variant and weight status on the risk of dysglycaemic status was found (P=0.04). Age, but not body mass index (BMI), was a risk factor for dysglycaemic status in the 27L carriers (OR: 1.05, P=0.0003), whereas BMI was a risk factor in non-carriers (OR: 1.23, P=0.008). No carriers of 98V were identified, and 487N was not associated with either T2DM or dysglycaemic status in our study population. CONCLUSION: These findings suggest that the HNF1A p.I27L (rs1169288) variant may be a significant risk factor of T2DM in normal-weight subjects and that earlier inconsistent results may have been due, in part, to subjects' weight status. Further investigations in larger cohorts are needed to verify these findings.
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Peso Corporal/fisiología , Diabetes Mellitus Tipo 2/epidemiología , Diabetes Mellitus Tipo 2/genética , Factor Nuclear 1-alfa del Hepatocito/genética , Adulto , Anciano , Estudios Transversales , Diabetes Mellitus Tipo 2/fisiopatología , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
BACKGROUND: It has recently been recognized that there is a close relationship between spinal cord tethering (SCT) and congenital anorectal malformation (ARM). PATIENTS AND METHODS: We evaluated spinal MRI examinations of 28 patients with ARM (14 boys and 14 girls) aged 5 months to 9 years. All patients diagnosed with SCT subsequently underwent operation. Patients were divided into high and low type ARM groups. We reviewed the relationship between SCT and ARM, and evaluated the untethering surgery. RESULTS: We evaluated 14 boys (high, 9; low, 5) and 14 girls (high, 4; low, 10). Of these 28 patients, 13 had SCT on MRI. Five out of 13 patients with high type ARM and 8 out of 15 patients with low type ARM had SCT. Seven out of 10 girls with low type ARM had SCT. Ten of these 13 patients with SCT experienced bowel/urological/orthopedic symptoms. SCT symptoms progressed prior to operation in the 2 patients who underwent untethering surgery a few years after their initial MRI examination. Postoperatively, orthopedic symptoms disappeared completely in all patients, but other symptoms did not. CONCLUSIONS: Based on the results of this study, we recommend routine MRI examination of patients with ARM and early untethering surgery in cases with SCT.
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Anomalías Múltiples , Canal Anal/anomalías , Anomalías del Sistema Digestivo/diagnóstico , Defectos del Tubo Neural/cirugía , Procedimientos Neuroquirúrgicos/métodos , Recto/anomalías , Niño , Preescolar , Diagnóstico Diferencial , Femenino , Estudios de Seguimiento , Humanos , Lactante , Masculino , Defectos del Tubo Neural/diagnóstico , Estudios Retrospectivos , Factores de Tiempo , Resultado del TratamientoRESUMEN
The inflammatory reaction is known to be controlled in neonates. We clarified the characteristics of cytokine profile in neonatal patients and assess its clinical significance. Serum levels of interleukin (IL)-6 and IL-1 receptor antagonist (ra) were determined in 152 pediatric patients and 33 each of maternal and cord bloods. Supernatant IL-1ra levels of cultured monocytes and granulocytes stimulated with IL-1beta or LPS, and IL-1ra mRNA expression of granulocytes were assayed in 15 each of cord and healthy adult bloods. Although surgical stress in neonates was heavier than that in infants, there was no difference in the occurrence of postoperative morbidity and mortality. In neonates, the perioperative serum level of IL-1ra was significantly raised, and the postoperative IL-6 response was well controlled. The serum concentration of IL-1ra in cord blood was not different from that in maternal blood, whereas, the serum concentration of IL-6 in cord blood was significantly reduced than that in maternal blood. In granulocytes, significantly more IL-1ra was produced from cord than from adult blood. An IL-1ra predominant immune status in neonates may be a naturally acquired adaptation system and play a crucial role in attenuating acute inflammatory reaction in a vulnerable host defense.
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Citocinas/metabolismo , Estrés Fisiológico/metabolismo , Procedimientos Quirúrgicos Operativos , Temperatura Corporal , Proteína C-Reactiva/análisis , Niño , Femenino , Sangre Fetal/química , Granulocitos/metabolismo , Humanos , Lactante , Recién Nacido , Proteína Antagonista del Receptor de Interleucina 1/metabolismo , Interleucina-6/metabolismo , Monocitos/metabolismo , Embarazo , Estrés Fisiológico/etiologíaRESUMEN
The aim of our study was to evaluate the diagnostic sensitivity of the new thyrotropin receptor antibody (TRAb) assays (Cosmic TRAb CT, ELISA and Yamasa DYNOtest TRAb). TRAb was positive in 43 of 44 (97.7%) untreated patients with Graves' disease by both TRAb CT and/or ELISA and NYNOtest TRAb. Thus the new TRAb assays were clearly more sensitive than the conventional assay (positivity: 85%). There was a strong positive correlation between the data obtained in TRAb CT and/or ELISA and those obtained in DYNOtest TRAb (r = 0.942, p < 0.0001). There was a significant correlation between the new TRAb and TSAb (r = 0.696, p < 0.0001). Although there was a significant correlation between the new TRAb and thyroid stimulation-blocking antibody (TSBAb), the correlation coefficient was low (r = 0.605, p < 0.0001). The increased sensitivity of the new TRAb assays for Graves' disease provides an advantage over conventional assay.
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Autoanticuerpos/sangre , Enfermedad de Graves/diagnóstico , Inmunoglobulinas Estimulantes de la Tiroides/sangre , Receptores de Tirotropina/inmunología , Autoanticuerpos/inmunología , Enfermedad de Graves/inmunología , Humanos , Inmunoensayo , Inmunoglobulinas Estimulantes de la Tiroides/inmunología , Sensibilidad y EspecificidadRESUMEN
The interstitial nucleus of the posterior limb of the anterior commissure (IPAC) receives inputs from several autonomic/limbic regions in the forebrain, including the agranular insular cortex, bed nucleus of the stria terminalis, the amygdaloid complex, and the lateral hypothalamic area. We sought to identify the distribution of afferent sources to the IPAC and to determine whether these IPAC projection fibers issue collaterals to the nucleus of the solitary tract (NTS), the principal relay of primary visceral afferents. Two fluorescent tracers, FluoroGold and FluoroRed, were centered stereotaxically on the IPAC and the NTS on chloral hydrate-anesthetized Sprague-Dawley rats. Although the majority of IPAC and NTS afferents were spatially segregated, small but substantial numbers of dually labeled neurons (three to four cells/section) were observed in the dorsal bank of the posterior agranular insular cortex, exclusively in layer V. Collateral projection neurons were also found in the posterior part of the lateral hypothalamic area (two to six cells/section). The branching projections identified here may represent a potential link between affective or motivated behavior and viscerosensory processing.
Asunto(s)
Amígdala del Cerebelo/citología , Cuerpo Estriado/citología , Vías Nerviosas/citología , Núcleos Septales/citología , Núcleo Solitario/citología , Estilbamidinas , Amígdala del Cerebelo/fisiología , Animales , Vías Autónomas/citología , Vías Autónomas/fisiología , Benzoatos , Corteza Cerebral/citología , Corteza Cerebral/fisiología , Cuerpo Estriado/fisiología , Emociones/fisiología , Colorantes Fluorescentes , Área Hipotalámica Lateral/citología , Área Hipotalámica Lateral/fisiología , Masculino , Vías Nerviosas/fisiología , Neuronas/citología , Neuronas/fisiología , Ratas , Ratas Sprague-Dawley , Núcleos Septales/fisiología , Núcleo Solitario/fisiología , Aferentes Viscerales/citología , Aferentes Viscerales/fisiologíaRESUMEN
Aquaporin-2, a water-channel protein, is known to increase water permeability due to vasopressin binding to V2 receptors at the renal collecting duct and is excreted into the urine. It is still unclear whether a hyponatremic state is caused by vasopressin-dependent aquaporin-2 in patients clinically diagnosed with the syndrome of inappropriate secretion of antidiuretic hormone. To determine this, we measured urinary aquaporin-2 and vasopressin by radioimmunoassay in normonatremic or hyponatremic patients after cerebral infarction and in healthy controls. In the normonatremia group, urinary aquaporin-2 and plasma AVP levels were higher than in controls. In the hyponatremia group, plasma AVP was relatively high despite low plasma osmolality in each patient. However, urinary aquaporin-2 in hyponatremia was significantly increased when compared with the other two groups. In conclusion, AQP-2 increment does not directly reflect non-osmotic AVP secretion in a hyponatremic state. This result indicates that the urinary excretion of AQP-2 is not only AVP-dependent in hyponatremic states.
Asunto(s)
Acuaporinas/orina , Infarto Cerebral/orina , Hiponatremia/orina , Síndrome de Secreción Inadecuada de ADH/orina , Anciano , Anciano de 80 o más Años , Acuaporina 2 , Acuaporina 6 , Circulación Cerebrovascular/fisiología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios Prospectivos , RadioinmunoensayoRESUMEN
BACKGROUND: A study was undertaken in patients with undiagnosed sleep apnoea/hypopnoea syndrome (OSAS) to document the use of prescribed medications, especially those used in cardiovascular diseases, in the year before the OSAS diagnosis was confirmed. METHODS: A total of 549 patients with OSAS (401 men of mean age 47.2 years, mean body mass index (BMI) 35.5 kg/m(2), mean apnoea/hypopnoea index (AHI) 47.2 and148 women of mean age 50.2 years, BMI 39.6 kg/m(2), AHI 32.6) were each matched to one general population control by age, sex, geographical location, and family physician. Medication use was evaluated for patients and controls using a database containing information about all prescriptions completed in the province of Manitoba, Canada. RESULTS: In the year before OSAS was diagnosed, prescribed medication costs were $155.91 (Canadian dollars) (95% CI $91.34 to $220.49) greater for cases than for controls. Cases were dispensed 3.3 (95% CI 1.5 to 5.2) more prescriptions, were on 1.2 (95% CI 0.8 to 1.6) more medications, and were supplied with 157.4 (95% CI 95.9 to 218.8) more daily doses of medication. The odds ratio of OSAS cases being on a prescribed medication was 1.88 relative to controls (95% CI 1.38 to 2.54, p<0.0001). In the same year 36.6% of cases and 19.7% of controls were using medications for cardiovascular disease (OR 2.82, 95% CI 2.05 to 3.89, p<0.0001), consuming 79.4 (95% CI 48.9 to 109.8) more daily doses of medication, having been dispensed 1.7 (95% CI 1.0 to 2.4) more prescriptions, and at a $75.26 (95% CI $44.03 to $106.50) greater cost. The odds ratio of patients with OSAS being on medications indicated for the treatment of systemic hypertension was 2.71 (95% CI 1.96 to 3.77) relative to controls; however, such medications might also be prescribed for other indications such as angina pectoris and congestive heart failure, and for the secondary prevention of myocardial infarction. The use of medications indicated for the treatment of systemic hypertension was predicted significantly by age (odds ratio (OR) 1.10 per year), BMI (OR 1.05 per unit), and AHI (OR 1.01 per unit). CONCLUSIONS: In the year before OSAS was diagnosed, patients with OSAS were heavy users of medications, particularly those used to treat cardiovascular diseases.
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Fármacos Cardiovasculares/uso terapéutico , Síndromes de la Apnea del Sueño/tratamiento farmacológico , Índice de Masa Corporal , Fármacos Cardiovasculares/economía , Costos de los Medicamentos , Electrocardiografía/métodos , Electromiografía/métodos , Femenino , Humanos , Masculino , Persona de Mediana Edad , Oportunidad Relativa , Polisomnografía/métodos , Síndromes de la Apnea del Sueño/economíaRESUMEN
Macrophage colony-stimulating factor (M-CSF) is one of several hematologic growth factors capable of regulating the survival, proliferation, and differentiation of macrophages, but its role in modulation of the accumulation and function of alveolar macrophages (AMs) in vivo is not well defined. Osteopetrotic (Op/Op) mice have no detectable M-CSF and show variable tissue-specific reductions in macrophage numbers. It was hypothesized that AMs would be decreased in number and have altered function in Op/Op mice because of the absence of M-CSF. Lung macrophages identified by Mac-3 staining in lung sections were decreased in number in 20-day-old Op/Op mice (P <.001) but not Op/Op mice older than 4 months (P =.68) compared with findings in age-matched littermate controls. The numbers of AMs recovered by bronchoalveolar lavage (BAL) were also reduced in young but not adult Op/Op mice compared with controls. Expression of interleukin-3 (IL-3) was increased in the lungs of Op/Op mice compared with controls as determined by quantification of IL-3 cytokine levels (P =.04), bioactivity (P =.02), and messenger RNA transcript levels. AMs of Op/Op mice spontaneously released higher levels of matrix metalloproteinases (MMPs) than AMs of controls as determined by immunohistochemical staining of AMs and zymographic assessment of BAL fluid and AM lysates. Consistent with an increased release of MMP, Op/Op mice had abnormal elastin deposition and spontaneously developed emphysema in the absence of molecular or cellular evidence of lung inflammation. These data show that the AM deficiency observed in young Op/Op mice is spontaneously corrected with age and is associated with increased lung levels of IL-3, spontaneous MMP expression by AMs, and destruction of lung tissue.
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Factor Estimulante de Colonias de Macrófagos/fisiología , Macrófagos Alveolares/enzimología , Macrófagos Alveolares/patología , Metaloproteinasas de la Matriz/metabolismo , Osteopetrosis/patología , Factores de Edad , Animales , Enfisema/enzimología , Enfisema/etiología , Enfisema/patología , Interleucina-3/metabolismo , Pulmón/enzimología , Pulmón/metabolismo , Pulmón/patología , Factor Estimulante de Colonias de Macrófagos/deficiencia , Factor Estimulante de Colonias de Macrófagos/farmacología , Macrófagos Alveolares/efectos de los fármacos , Metaloproteinasas de la Matriz/efectos adversos , Ratones , Ratones Mutantes , Osteopetrosis/genéticaRESUMEN
STUDY DESIGN: The functional recovery of rats that underwent spinal cord transection in infancy was evaluated by multimodal examination (functional tests, electrophysiologic evaluation, tract-tracing) to determine the basis for the recovery. OBJECTIVES: To determine whether the hind limb function in rats that underwent spinal cord transection in infancy is regained completely, which descending tracts regenerate after the transection, and whether the functional recovery is correlated with axonal reconnection. SUMMARY OF BACKGROUND DATA: It is widely accepted that a newborn rat recovers its hind limb function after spinal cord transection even without any specific treatments. This functional recovery might be attributed to possible regeneration of some descending pathways, although there is a counterargument that well-trained spinal cord reflexes may bring about functional compensation. METHODS: The thoracic spinal cord of infant rats was completely transected at Th10 when they were 2 weeks of age. Multimodal functional tests and electrophysiologic studies were performed 5 weeks later. Some recovered rats (i.e., those able to walk after the transection) underwent spinal cord retransection, with subsequent reevaluation of locomotion and muscle-evoked potentials. At 6 weeks after the initial transection, tract-tracing studies were performed in some animals. RESULTS: A motor performance score detected the functional differences between the control and the recovered rats. Muscle-evoked potentials of hind limbs after electrical stimulation to the brain were recorded in some of the recovered rats, but never in the unrecovered rats. Moreover, the muscle-evoked potentials of the recovered rats disappeared after spinal cord retransection that resulted in loss of voluntary movement. Morphologic studies in two rats provided evidence that reconnection of rubrospinal, vestibulospinal, and reticulospinal tracts had occurred, whereas corticospinal regeneration was not detected. CONCLUSIONS: It can be concluded that the hind limb function of rats that underwent spinal cord transection in infancy was partially regained; that axonal regeneration of the rubrospinal, vestibulospinal, or reticulospinal tracts was demonstrated, whereas the reconnection of the corticospinal tract was not observed; and that the axonal regeneration of these tracts is involved in the functional recovery.
Asunto(s)
Animales Recién Nacidos/fisiología , Regeneración Nerviosa , Traumatismos de la Médula Espinal/fisiopatología , Médula Espinal/fisiopatología , Animales , Animales Recién Nacidos/crecimiento & desarrollo , Vías Eferentes/fisiopatología , Electrofisiología , Potenciales Evocados Motores , Miembro Posterior/fisiopatología , Actividad Motora/fisiología , Tractos Piramidales/fisiopatología , Ratas , Ratas Wistar , Recuperación de la Función , Núcleo Rojo/fisiopatología , Análisis de Supervivencia , Transmisión SinápticaRESUMEN
Four of the 708 snorers (0.56%), referred to our sleep breathing disorders clinic for the past 2 years were diagnosed as having narcolepsy-cataplexy. Detecting HLA DRB1*1501/DQB1*0602 positive was informative for differentiating genuine narcolepsy from non-sleep apnea syndrome (non-SAS) hypersomnia in our clinic. A non-SAS obese boy, diagnosed as having essential hypersomnia syndrome, was found to be HLA DRB1*1502/DQB1*0601 positive. His hypocretin concentration was 206 pg/mL in the cerebrospinal fluid.
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Trastornos de Somnolencia Excesiva/complicaciones , Trastornos de Somnolencia Excesiva/epidemiología , Péptidos y Proteínas de Señalización Intracelular , Narcolepsia/complicaciones , Narcolepsia/epidemiología , Síndromes de la Apnea del Sueño/complicaciones , Adolescente , Adulto , Anciano , Instituciones de Atención Ambulatoria , Proteínas Portadoras/líquido cefalorraquídeo , Trastornos de Somnolencia Excesiva/inmunología , Femenino , Antígenos HLA-DQ/inmunología , Cadenas beta de HLA-DQ , Antígenos HLA-DR/inmunología , Cadenas HLA-DRB1 , Humanos , Masculino , Persona de Mediana Edad , Narcolepsia/inmunología , Neuropéptidos/líquido cefalorraquídeo , OrexinasRESUMEN
In six patients with chronic bradydysrhythmias, polysomnographies were performed before cardiac pacemaker implantation and over the week following implantation. A patient with third-degree atrioventricular block (AVB) and two patients with sinus node dysfunction (SND) were associated with sleep-disordered breathing (SDB). Their cardiac pacemaker therapies, with the increase in the average heart rate, led to a reduction of apnea-hypopnea index and/or an improvement of Cheyne-Stokes breathing. It seems that chronic bradydysrhythmia is one of the causative factors leading to SDB.
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Bradicardia/complicaciones , Marcapaso Artificial , Síndromes de la Apnea del Sueño/complicaciones , Síndromes de la Apnea del Sueño/diagnóstico , Anciano , Anciano de 80 o más Años , Respiración de Cheyne-Stokes , Enfermedad Crónica , Femenino , Bloqueo Cardíaco/complicaciones , Bloqueo Cardíaco/terapia , Frecuencia Cardíaca/fisiología , Humanos , Masculino , Persona de Mediana Edad , Polisomnografía , Periodo Posoperatorio , Cuidados Preoperatorios , Índice de Severidad de la Enfermedad , Resultado del TratamientoRESUMEN
Although acute infection of feline foamy virus (FeFV) is normally highly cytopathogenic in Crandell feline kidney (CRFK) cells, a noncytopathic persistent infection was established in the cells after cocultivation of the initially infected cells with uninfected cells four times. To investigate reactivation of persistent infection, CRFK cells chronically infected with FeFV were treated with trichostatin A (TA), a histone deacetylase inhibitor. TA induced higher FeFV production from the Coleman strain carrier culture and also induced marked syncytium formation. In contrast, human foamy virus, which contains less homologous long terminal repeat (LTR) and putative internal promoter (IP) sequences, persistently infecting baby hamster kidney cells was not reactivated by TA. The Sammy-1 strain of FeFV, from which a part of the U3 region in the LTR is naturally deleted, showed less reactivation. The Coleman LTR promoter-based beta-Gal-expressing plasmid was activated in the persistently Coleman-infected cells in the presence of TA, whereas the Sammy-1 LTR was not activated. Furthermore, the amounts of Gag protein expressed did not change in the presence or absence of TA. Because the putative IP region was very similar between the two strains, the initiation by TA is relatively specific for LTR sequences, and, therefore, histone deacetylation is at least in part responsible for reactivation of FeFV from carrier cell culture.
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Ácidos Hidroxámicos/farmacología , Spumavirus/fisiología , Activación Viral/efectos de los fármacos , Animales , Secuencia de Bases , Gatos , Línea Celular , Productos del Gen gag/genética , Genoma Viral , Riñón , Datos de Secuencia Molecular , Spumavirus/efectos de los fármacos , Secuencias Repetidas Terminales/genéticaRESUMEN
OBJECTIVE: To develop radioimmunoassay for somatostatin receptor type 2 (SSTR2) and search for its presence in certain rat tissues. METHODS: Anti-SSTR2 antiserum has been raised in New Zealand white rabbits immunized with a conjugate of synthetic SSTR2 with bovine serum albumin. Radioiodination of SSTR2 was performed by chloramin T method followed by purification of radioiodinated material on Sephadex G-25 column. RESULTS: The obtained antibody did not crossreact with SSTR1, SSTR3, SSTR4, SSTR5, hypothalamic hormones, pituitary hormones, neuropeptides or gut hormones. The assay was performed with a double antibody system. SSTR2 was extracted from the tissues with acid acetone. The dilution curve of acid acetone-extracts of rat hypothalamus in the radioimmunoassay system was parallel to the standard curve. The recovery of tissue SSTR2 was about 89 %, and the intra-assay and inter-assay variations were 4.9 % and 7.8 %, respectively. SSTR2 was found in the hypothalamus, cerebrum, cerebellum, pituitary, stomach and testis. CONCLUSIONS: These data suggest that this assay system is suitable for the estimation of SSTR2 in the tissues.
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Receptores de Somatostatina/análisis , Acetona , Animales , Especificidad de Anticuerpos , Química Encefálica , Cerebelo/química , Hipotálamo/química , Sueros Inmunes , Radioisótopos de Yodo , Marcaje Isotópico , Masculino , Hipófisis/química , Control de Calidad , Conejos , Ratas , Ratas Wistar , Estómago/química , Telencéfalo/química , Testículo/químicaRESUMEN
The term rapidly adapting pulmonary stretch receptor (RAR) refers to one of the major pulmonary sensory receptors that responds to inflation and deflation of the lungs as well as to irritant stimuli with rapidly adapting irregular discharges. The functional role and central pathways are largely unknown. The aim of this study was to elucidate morphological characteristics of second-order neurons (RAR cells) activated by vagal afferent fibers originating from RARs. A mixture of horseradish peroxidase (HRP) and Neurobiotin was injected intracellularly into physiologically identified RAR cells in Nembutal-anesthetized, immobilized, and artificially ventilated Wister rats. Direct visualization of individual RAR cells (n = 12), including their somata, dendritic arborizations, and fine axonal branches with terminal boutons, was possible for the first time. Their somata were located in the commissural or medial subdivision of the nucleus of the solitary tract, caudal to the level of the area postrema. The RAR cells had, in addition to dendrites extending into the NTS area, one or two long dendrites extending laterally and/or ventrolaterally into the medullary reticular formation. The stem axons issuing from the RAR cells first coursed ventrolaterally toward the reticular formation in the vicinity of the ambiguus nucleus and then bifurcated into ascending and descending axons: three RAR cells possessed only ascending axons. Some of the ascending axons could be traced as far as the level of the facial nucleus and some of the descending axons beyond the spinomedullary junction. These ascending and/or descending axons gave off extensive axon collaterals distributing boutons within and in the vicinity of the ambiguus nucleus. These results, showing an anatomical substrate for the network implicated in RAR-evoked reflexes, provide useful clues for study of the RAR system.
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Biotina/análogos & derivados , Pulmón/inervación , Neuronas/citología , Receptores de Estiramiento Pulmonares/fisiología , Ratas Wistar/anatomía & histología , Núcleo Solitario/citología , Vías Aferentes , Animales , Axones/fisiología , Biotina/análisis , Vías Eferentes , Electrofisiología , Peroxidasa de Rábano Silvestre , Neuronas/química , Neuronas/fisiología , Puente/citología , Ratas , Médula Espinal/citologíaRESUMEN
We examined regional and intracellular distribution of Ca(2+)/calmodulin-dependent protein kinase kinase beta (CaM-KK beta), which activated Ca(2+)/calmodulin-dependent protein kinase I and IV (CaM-K I and IV) immunohistochemically in the central nervous system of the rat by light and electron microscopy. Although most neurons in the brain and spinal cord exhibited the immunoreactivity, no labeled neurons were observed in the globus pallidus or entopeduncular nucleus, and only a small number of neurons showed weak immunoreactivity in the substantia nigra pars reticulata. In general, the immunoreactivity was observed both in the cytoplasm and cellular nucleus, although the immunoreactivity was not found in the cellular nucleus in some large neurons such as in the mesencephalic trigeminal nucleus, lateral vestibular nucleus or gigant cellular reticular formation. As to motoneurons of the cranial nerve nuclei and the anterior horn of the spinal cord, they revealed the immunoreactivity both in the cytoplasm and nucleus. The reaction product appeared as fine granules in the cytoplasm and nucleus under light microscopy. Electron microscopic observations confirmed that the reaction product was localized mainly on the Golgi apparatus or on the nuclear chromatin. Immunolabeling for antibody against CaM-KK beta was discussed with the distribution of CaM-K I, IV and another CaM-KK, CaM-KK alpha, in the central nervous system.
Asunto(s)
Encéfalo/enzimología , Neuronas/enzimología , Proteínas Serina-Treonina Quinasas/metabolismo , Animales , Encéfalo/citología , Quinasa de la Proteína Quinasa Dependiente de Calcio-Calmodulina , Proteína Quinasa Tipo 1 Dependiente de Calcio Calmodulina , Proteína Quinasa Tipo 4 Dependiente de Calcio Calmodulina , Proteínas Quinasas Dependientes de Calcio-Calmodulina/análisis , Cerebelo/enzimología , Diencéfalo/enzimología , Inmunohistoquímica , Mesencéfalo/enzimología , Proteínas del Tejido Nervioso/análisis , Proteínas del Tejido Nervioso/metabolismo , Neuronas/citología , Neuronas/ultraestructura , Puente/enzimología , Proteínas Serina-Treonina Quinasas/análisis , Ratas , Telencéfalo/enzimologíaRESUMEN
Cry4A is a dipteran-specific delta-endotoxin produced by Bacillus thuringiensis, and toxic to Culex pipiens (mosquito) larvae. The immunohistochemical staining of the midgut sections of C. pipiens larvae revealed that Cry4A bound in vitro and in vivo to the microvilli of the epithelial cells of posterior midgut and gastric caecae. The binding of digoxigenin-labeled Cry4A (DIG-Cry4A) to the apical microvilli was almost abolished in the presence of excess unlabeled Cry4A, suggesting that the binding of Cry4A to the microvilli was specific. Several Cry4A-specific binding proteins were detected using the ligand blotting technique with DIG-Cry4A. Moreover, an insertion assay was done, where the binding of DIG-Cry4A to the BBMVs was completely irreversible and did not compete with excess unlabeled Cry4A. On the basis of these results, we propose a schematic interpretation for the binding process of Cry4A.
Asunto(s)
Proteínas Bacterianas/metabolismo , Toxinas Bacterianas , Culex/metabolismo , Sistema Digestivo/metabolismo , Endotoxinas/metabolismo , Animales , Bacillus thuringiensis , Toxinas de Bacillus thuringiensis , Sitios de Unión , Células Epiteliales/metabolismo , Proteínas Hemolisinas , Concentración de Iones de Hidrógeno , Técnicas In Vitro , Larva/metabolismo , Microvellosidades/metabolismoRESUMEN
We tried to investigate the present problems, concerning the reference individual and interval in thyroid function tests and find the solutions for them. We are now using healthy adults for the reference individual and interval. Recently, we found the sex-difference and age-related changes for reference individual and interval in free T3 measurement. We raised the questions on whether there are any sex-differences and/or age-related changes or not. Surprisingly, there were almost no detailed data about them especially in Japan. Therefore, we examined the Europe data, and found out some kind of sex-differences and age-related changes. We propose the following examinations using many Japanese population in order to provide a precise and proper reference individual and interval: 1. Whether there are any sex-difference in thyroid function tests? 2. Whether there are age-related change in thyroid function tests, for instance, simply dividing population into the immature, adult and the aged?