RESUMEN
Emergency granulopoiesis, also known as demand-adapted granulopoiesis, is defined as the response of an organism to systemic bacterial infections, and it results in neutrophil mobilization from reservoir pools and increased myelopoiesis in the bone marrow. Indirect and direct initiating mechanisms of emergency granulopoiesis have been hypothesized. However, the detailed mechanism of hyperactive myelopoiesis in the bone marrow, which leads to granulocyte left shift, remains unknown. In this study, we report that TLR4 is expressed on granulo-monocytic progenitors, as well as mobilized human peripheral blood CD34+ cells, which account for 0.2% of monocytes in peripheral blood, and â¼ 10% in bone marrow. LPS, a component of Gram-negative bacteria that results in a systemic bacterial infection, induces the differentiation of peripheral blood CD34+ cells into myelocytes and monocytes in vitro via the TLR4 signaling pathway. Moreover, CD34+ cells directly responded to LPS stimulation by activating the MAPK and NF-κB signaling pathways, and they produced IL-6 that promotes emergency granulopoiesis by phosphorylating C/EBPα and C/EBPß, and this effect was suppressed by the action of an IL-6 receptor inhibitor. This work supports the finding that TLR is expressed on human hematopoietic stem and progenitor cells, and it provides evidence that human hematopoietic stem and progenitor cells can directly sense pathogens and produce cytokines exerting autocrine and/or paracrine effects, thereby promoting differentiation.
Asunto(s)
Granulocitos/metabolismo , Células Madre Hematopoyéticas/metabolismo , Interleucina-6/metabolismo , Transducción de Señal/fisiología , Células Madre/metabolismo , Receptor Toll-Like 4/metabolismo , Adaptación Fisiológica/fisiología , Antígenos CD34/metabolismo , Médula Ósea/metabolismo , Proteína beta Potenciadora de Unión a CCAAT/metabolismo , Diferenciación Celular/fisiología , Citocinas/metabolismo , Regulación de la Expresión Génica/fisiología , Células Precursoras de Granulocitos/metabolismo , Trasplante de Células Madre Hematopoyéticas/métodos , Humanos , Monocitos/metabolismo , Mielopoyesis/fisiologíaRESUMEN
BACKGROUND: Malocclusion is a multifactorial condition associated with genetic and environmental factors. The purpose of this study was to investigate the prevalence of occlusal traits, oral habits, and nose and throat conditions by age and to assess the association between malocclusion and its environmental factors in Japanese preschool children. METHODS: A total of 503 Japanese children (258 boys and 245 girls aged 3-6 years) were recruited. Occlusal traits were assessed visually to record sagittal, vertical, and transverse malocclusion, and space discrepancies. Lip seal was recorded by an examiner, and oral habits (finger sucking, lip sucking or lip biting, nail biting, chin resting on a hand) and nose and throat conditions (tendency for nasal obstruction, allergic rhinitis, palatine tonsil hypertrophy) were assessed by a questionnaire completed by the parents. The prevalence of each item was calculated, and binary logistic regression was used to examine the factors related to malocclusion. RESULTS: 62.0% of preschool children in the present study exhibited malocclusion, and 27.8% exhibited incompetent lip seal. Nail biting was the most frequent oral habit with a prevalence of 18.9%. Nasal obstruction was recorded in 30.4% of children. The results of binary logistic regression showed that incompetent lip seal was significantly related to malocclusion, and that nail biting was significantly negatively related. CONCLUSIONS: Incompetent lip seal is significantly associated with malocclusion, but nail biting may not necessarily be a deleterious habit for the occlusion in Japanese preschool children.
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Succión del Dedo , Maloclusión , Hábito de Comerse las Uñas , Obstrucción Nasal , Preescolar , Femenino , Humanos , Masculino , Pueblos del Este de Asia , Succión del Dedo/efectos adversos , Hábitos , Labio , Maloclusión/epidemiología , Maloclusión/etiología , Hábito de Comerse las Uñas/efectos adversos , Obstrucción Nasal/complicaciones , Factores de Riesgo , NiñoRESUMEN
BACKGROUND: Identification of carbapenemase-producing Enterobacteriaceae (CPE) in faecal specimens is challenging. This fact is particularly critical because low-level carbapenem-resistant organisms such as IMP-producing CPE are most prevalent in Japan. We developed a modified selective medium more suitable for IMP-type CPE. METHODS: Fifteen reference CPE strains producing different types of ß-lactamases were used to evaluate the commercially available CHROMagar KPC and chromID CARBA as well as the newly prepared MC-ECC medium (CHROMagar ECC supplemented with meropenem, cloxacillin, and ZnSO4) and M-ECC medium (CHROMagar ECC supplemented with meropenem and ZnSO4). A total of 1035 clinical samples were then examined to detect CPE using chromID CARBA and M-ECC medium. RESULTS: All tested strains producing NDM-, KPC-, and OXA-48-carbapenemases were successfully cultured in the media employed. Although most of the IMP-positive strains did not grow in CHROMagar KPC, chromID CARBA, or MC-ECC, all tested strains grew on M-ECC. When faecal samples were applied to the media, M-ECC medium allowed the best growth of IMP-type CPE with a significantly higher sensitivity (99.3%) than that of chromID CARBA (13.9%). CONCLUSIONS: M-ECC medium was determined as the most favourable selective medium for the detection of IMP-type CPE as well as other types of CPE.
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Proteínas Bacterianas , Técnicas de Tipificación Bacteriana/métodos , Medios de Cultivo , Infecciones por Enterobacteriaceae , Enterobacteriaceae , Inosina Monofosfato/metabolismo , beta-Lactamasas , Enterobacteriaceae/enzimología , Enterobacteriaceae/aislamiento & purificación , Enterobacteriaceae/metabolismo , Infecciones por Enterobacteriaceae/diagnóstico , Infecciones por Enterobacteriaceae/microbiología , Heces/microbiología , HumanosRESUMEN
Trisomy 8 (+8), one of the most common chromosomal abnormalities found in patients with myelodysplastic syndromes (MDS), is occasionally seen in patients with otherwise typical aplastic anemia (AA). Although some studies have indicated that the presence of +8 is associated with the immune pathophysiology of bone marrow (BM) failure, its pathophysiology may be heterogeneous. We studied 53 patients (22 with AA and 31 with low-risk MDS) with +8 for the presence of increased glycosylphosphatidylinositol-anchored protein-deficient (GPI-AP(-) ) cells, their response to immunosuppressive therapy (IST), and their prognosis. A significant increase in the percentage of GPI-AP(-) cells was found in 14 (26%) of the 53 patients. Of the 26 patients who received IST, including nine with increased GPI-AP(-) cells and 17 without increased GPI-AP(-) cells, 14 (88% with increased GPI-AP(-) cells and 41% without increased GPI-AP(-) cells) improved. The overall and event-free survival rates of the +8 patients with and without increased GPI-AP(-) cells at 5 yr were 100% and 100% and 59% and 57%, respectively. Examining the peripheral blood for the presence of increased GPI-AP(-) cells may thus be helpful for choosing the optimal treatment for +8 patients with AA or low-risk MDS.
Asunto(s)
Proteínas Ligadas a GPI/deficiencia , Granulocitos/metabolismo , Hemoglobinuria Paroxística/genética , Hemoglobinuria Paroxística/metabolismo , Trisomía , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Anemia Aplásica , Médula Ósea/metabolismo , Médula Ósea/patología , Enfermedades de la Médula Ósea , Trastornos de Fallo de la Médula Ósea , Cromosomas Humanos Par 8 , Femenino , Proteínas Ligadas a GPI/metabolismo , Hemoglobinuria Paroxística/diagnóstico , Hemoglobinuria Paroxística/tratamiento farmacológico , Hemoglobinuria Paroxística/epidemiología , Hemoglobinuria Paroxística/mortalidad , Humanos , Inmunosupresores/uso terapéutico , Incidencia , Masculino , Persona de Mediana Edad , Prevalencia , Estudios Retrospectivos , Análisis de Supervivencia , Resultado del Tratamiento , Adulto JovenRESUMEN
Inherited Protein S (PS) deficiency is an autosomal dominant thrombotic disorder. We encountered a case of inherited type I PS deficiency following a close examination for recurrent pregnancy loss and identified the mutation responsible; a novel splice donor site mutation in intron 13 of the PROS1 gene appeared to have caused a frameshift with premature termination at amino acid +551. These results will contribute to the creation of an accurate database and define the molecular basis for PS deficiency.
RESUMEN
Idiopathic aplastic anemia (AA) is a common cause of acquired BM failure. Although autoimmunity to hematopoietic progenitors is thought to be responsible for its pathogenesis, little is known about the molecular basis of this autoimmunity. Here we show that a substantial proportion of AA patients harbor clonal hematopoiesis characterized by the presence of acquired copy number-neutral loss of heterozygosity (CNN-LOH) of the 6p arms (6pLOH). The 6pLOH commonly involved the HLA locus, leading to loss of one HLA haplotype. Loss of HLA-A expression from multiple lineages of leukocytes was confirmed by flow cytometry in all 6pLOH(+) cases. Surprisingly, the missing HLA-alleles in 6pLOH(+) clones were conspicuously biased to particular alleles, including HLA-A*02:01, A*02:06, A*31:01, and B*40:02. A large-scale epidemiologic study on the HLA alleles of patients with various hematologic diseases revealed that the 4 HLA alleles were over-represented in the germline of AA patients. These findings indicate that the 6pLOH(+) hematopoiesis found in AA represents "escapes" hematopoiesis from the autoimmunity, which is mediated by cytotoxic T cells that target the relevant auto-antigens presented on hematopoietic progenitors through these class I HLAs. Our results provide a novel insight into the genetic basis of the pathogenesis of AA.
Asunto(s)
Anemia Aplásica/genética , Cromosomas Humanos Par 6/genética , Variaciones en el Número de Copia de ADN , Antígenos HLA/genética , Antígenos HLA-B/genética , Pérdida de Heterocigocidad , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Alelos , Anemia Aplásica/inmunología , Anemia Aplásica/terapia , Niño , Preescolar , Femenino , Frecuencia de los Genes , Haplotipos , Células Madre Hematopoyéticas/inmunología , Células Madre Hematopoyéticas/metabolismo , Humanos , Lactante , Masculino , Persona de Mediana Edad , Polimorfismo de Nucleótido Simple , Adulto JovenRESUMEN
Although myelodysplastic syndromes are heterogeneous disorders comprising a benign subset of bone marrow failure similar to aplastic anemia, no laboratory test has been established to distinguish it from bone marrow failures that can evolve into acute myeloid leukemia. Plasma thrombopoietin levels were measured in 120 patients who had myelodysplastic syndrome with thrombocytopenia (< 100 × 10(9)/L) to determine any correlation to markers associated with immune pathophysiology and outcome. Thrombopoietin levels were consistently low for patients with refractory anemia with excess of blasts, while patients with other myelodysplatic syndrome subsets had more variable results. Patients with thrombopoietin levels of 320 pg/mL and over had increased glycosylphosphatidylinositol-anchored protein-deficient blood cells (49.1% vs. 0%), were more likely to have a low International Prognostic Scoring System (IPSS) score (≤1.0, 100% vs. 65.5%), a higher response rate to immunosuppressive therapy (84.2% vs. 14.3%), and a better 5-year progression-free survival rate (94.1% vs. 63.6% for refractory cytopenia with unilineage dysplasia; 100.0% vs. 44.4% for refractory cytopenia with multilineage dysplasia). In conclusion, increased plasma thrombopoietin levels were associated with a favorable prognosis of bone marrow failure and could, therefore, represent a reliable marker for a benign subset of myelodysplastic syndrome.
Asunto(s)
Síndromes Mielodisplásicos/sangre , Trombopoyetina/sangre , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Biomarcadores/sangre , Células Sanguíneas/metabolismo , Médula Ósea/patología , Femenino , Proteínas Ligadas a GPI/deficiencia , Proteínas Ligadas a GPI/metabolismo , Humanos , Inmunosupresores/uso terapéutico , Masculino , Persona de Mediana Edad , Síndromes Mielodisplásicos/diagnóstico , Síndromes Mielodisplásicos/tratamiento farmacológico , Pronóstico , Curva ROC , Trombocitopenia/sangre , Resultado del Tratamiento , Adulto JovenRESUMEN
Acquisition of new genetic traits by horizontal gene transfer is a bacterial strategy for adaptation to the environment. We previously showed that Escherichia coli can transmit non-conjugative plasmids laterally in a co-culture containing strains with and without the plasmid. In this study, using the Keio collection, a comprehensive library of E. coli knock-out mutants for non-essential genes, we screened for genes responsible for the execution and promotion of cell-to-cell plasmid transfer in recipient cells. By stepwise screening of 'transfer-down' mutants, two essential genes and six promoting genes were obtained. One of the essential genes was priA, which is involved in DNA replication. This priA mutant was also unable to be transformed by artificial transformation methods, probably due to the deficiency of the plasmid maintenance function. The other essential gene was rodZ (yfgA), a gene involved in the regulation of rod-shaped structure of E. coli cells. This rodZ mutant was transformable by all three methods of artificial transformation tested, suggesting that this gene is essential for cell-to-cell plasmid transfer but not for artificial transformation. These are the first data that suggest that rodZ plays an essential role in DNA acquisition.
Asunto(s)
Proteínas del Citoesqueleto/fisiología , Proteínas de Escherichia coli/fisiología , Escherichia coli/genética , Transferencia de Gen Horizontal/genética , Estudio de Asociación del Genoma Completo , Plásmidos/genética , Transformación Bacteriana/genética , Conjugación Genética/genética , Proteínas del Citoesqueleto/genética , ADN/genética , Proteínas de Escherichia coli/genéticaRESUMEN
Acquiring new genetic traits by lateral gene transfer is a bacterial strategy for environment adaptation. We previously showed that Escherichia coli could laterally transmit non-conjugative plasmids in co-cultures containing strains with and without the plasmid. In this study, using the Keio collection, a comprehensive library of E. coli knock-out mutants for non-essential genes, we screened for genes responsible for repressing cell-to-cell plasmid transfer in recipient cells. By stepwise screening, we identified 55 'transfer-up' mutants that exhibited approximately 2- to 30-fold increased activities. We confirmed plasmid acquisition by these 'up' mutants and revealed that there were no significant changes in antibiotic resistance in the original Keio strains. The presumed functions of these gene products covered a wide range of activities, including metabolism and synthesis, transport, transcription or translation and others. Two competence-gene homologues (ybaV and yhiR) were identified from among these genes. The presumed localizations of these 55 gene products were estimated to be 34 cytoplasmic proteins, 20 in or around the cell surface and 1 unknown location. Our results suggest that these 55 genes may be involved in repressing plasmid uptake during cell-to-cell plasmid transfer.
Asunto(s)
Conjugación Genética/genética , Escherichia coli/genética , Transferencia de Gen Horizontal/genética , Plásmidos/genética , Técnicas de Inactivación de Genes , Mutación , Análisis de Secuencia de ADN , Transformación GenéticaRESUMEN
To characterize bone marrow failure with del(13q), we reviewed clinical records of 22 bone marrow failure patients possessing del(13q) alone or del(13q) plus other abnormalities. All del(13q) patients were diagnosed with myelodysplastic syndrome-unclassified due to the absence of apparent dysplasia. Elevated glycosylphosphatidylinositol-anchored protein-deficient blood cell percentages were detected in all 16 with del(13q) alone and 3 of 6 (50%) patients with del(13q) plus other abnormalities. All 14 patients with del(13q) alone and 2 of 5 (40%) patients with del(13q) plus other abnormalities responded to immunosuppressive therapy with 10-year overall survival rates of 83% and 67%, respectively. Only 2 patients who had abnormalities in addition to the del(13q) abnormality developed acute myeloid leukemia. Given that myelodysplastic syndrome-unclassified with del(13q) is a benign bone marrow failure subset characterized by good response to immunosuppressive therapy and a high prevalence of increased glycosylphosphatidylinositol-anchored protein-deficient cells, del(13q) should not be considered an intermediate-risk chromosomal abnormality.
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Anemia Aplásica/genética , Anemia Refractaria/clasificación , Anemia Refractaria/genética , Deleción Cromosómica , Cromosomas Humanos Par 13/genética , Síndromes Mielodisplásicos/genética , Adulto , Anciano , Anciano de 80 o más Años , Anemia Aplásica/mortalidad , Anemia Aplásica/terapia , Anemia Refractaria/mortalidad , Anemia Refractaria/terapia , Anticuerpos Monoclonales/inmunología , Femenino , Citometría de Flujo , Estudios de Seguimiento , Humanos , Hibridación Fluorescente in Situ , Masculino , Persona de Mediana Edad , Síndromes Mielodisplásicos/mortalidad , Síndromes Mielodisplásicos/terapia , Pronóstico , Tasa de Supervivencia , Organización Mundial de la Salud , Adulto JovenRESUMEN
We investigated whether the feeding of high H2-generating dietary fibre and resistant starch (RS) could suppress hepatic ischaemia-reperfusion (IR) injury, which results from oxidative stress, in rats fed a pectin (Pec) or high-amylose maize starch (HAS) diet. Male Sprague-Dawley rats were fed a control (C) diet, with or without Pec (0-5 % Pec) or HAS (0-30 % HAS) supplementation for 7 d. Portal H2 concentration showed a significant dose-dependent increase with the amount of Pec or HAS supplementation. Plasma alanine and aspartate aminotransferase activities remarkably increased in the C rats (5 % cellulose) due to IR treatment, while it decreased significantly or showed tendencies to decrease in 5 % Pec and 20 % HAS diet-fed rats. The hepatic oxidised glutathione (GSSG):total glutathione ratio increased significantly in IR rats maintained on the C diet compared with sham-operated rats. On the other hand, reduced glutathione (GSH):total glutathione and GSH:GSSG ratios decreased significantly. The GSSG:total glutathione ratio that increased due to IR treatment decreased significantly on HAS and Pec intake, while GSH:total glutathione and GSH:GSSG ratios increased significantly. Hepatic sinusoids of IR rats fed the C diet were occluded, but those of IR rats fed the Pec diet were similar to those in the sham-operated rats. In conclusion, we found that Pec or HAS, which enhance H2 generation in the large intestine, alleviated hepatic IR injury. The present study demonstrates another physiological significance of dietary fibre and RS.
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Hidrógeno/sangre , Isquemia/fisiopatología , Hígado/patología , Pectinas/uso terapéutico , Prebióticos , Daño por Reperfusión/dietoterapia , Almidón/uso terapéutico , Amilosa/administración & dosificación , Amilosa/análisis , Amilosa/uso terapéutico , Animales , Ciego/microbiología , Fermentación , Glutatión , Enfermedad Veno-Oclusiva Hepática/etiología , Enfermedad Veno-Oclusiva Hepática/fisiopatología , Hidrógeno/metabolismo , Hígado/irrigación sanguínea , Hígado/metabolismo , Hígado/fisiopatología , Masculino , Oxidación-Reducción , Estrés Oxidativo , Pectinas/administración & dosificación , Prebióticos/análisis , Ratas , Ratas Sprague-Dawley , Daño por Reperfusión/metabolismo , Daño por Reperfusión/patología , Daño por Reperfusión/fisiopatología , Semillas/química , Almidón/administración & dosificación , Almidón/química , Almidón/metabolismo , Zea mays/químicaRESUMEN
Tongue function is regarded as a primary factor in the etiology of malocclusion, but details of the relationship remain unknown. The purpose of the present study was to investigate maximum tongue pressure, in preschool children to examine its relationship with dental occlusion. A total of 477 healthy children (248 boys, 229 girls, aged 4-6 years) were recruited. Dental occlusion was assessed visually to record sagittal, vertical, and transverse malocclusion, and space discrepancies. Maximum tongue pressure was measured using a balloon-based tongue pressure measurement device. Additionally, 72 children (37 boys, 35 girls, aged 4-5 years) were recruited for a 1-year follow-up study. Approximately half of the children (53.5%) showed some type of malocclusion in the present study. Maximum tongue pressure was highest in the 6-year-old children. The results of a two-way ANCOVA show that the effect of age was significant (p < 0.001); however, the effects of sex and dental occlusion, or the interactions among these variables, did not reach significance. Additionally, maximum tongue pressure increased significantly in the 1-year follow-up study (p < 0.001), especially in the normal occlusion group. Maximum tongue pressure increases markedly with growth in the preschool years and can be associated with some types of malocclusion in preschool children.
RESUMEN
PURPOSE: To gain new insights into the etiology of blepharitis, we investigated the causative bacteria in patients with blepharitis and the effects of 1% azithromycin ophthalmic solution. STUDY DESIGN: A multicenter, prospective observational study. METHODS: After the subjects were diagnosed as having blepharitis they were administered 1% azithromycin ophthalmic solution for up to 14 days. Bacterial cultures and smear microscopic examinations of the eyelid margin were conducted at the initial visit, after administering eye drops, and 1 month after the end of eye drop administration. The minimum inhibitory concentrations (MICs) of azithromycin were measured. RESULTS: At the initial visit, the bacterial morphology determined by smear microscopic examinations coincided with that of strains isolated by culture taken from 22 of 45 patients. All detected bacteria were gram-positive, and Corynebacterium spp., Cutibacterium acnes, Staphylococcus epidermidis, Streptococcus spp., and Enterococcus faecalis were isolated most commonly. Compared with the initial visit the number of isolated strains per eye decreased significantly at 7 days after the start of eye drop administration and 1 month after the end of eye drop administration. The azithromycin MICs were temporarily increased after the start of eye drops but then decreased. CONCLUSION: Our study suggests that in blepharitis pathogenicity is characterized by increased strain numbers and amounts of indigenous bacteria. Administering a 1% azithromycin ophthalmic solution suppresses the number of bacterial strains within 1 month after the end of eye drop administration without increasing drug resistance.
Asunto(s)
Azitromicina , Blefaritis , Humanos , Azitromicina/uso terapéutico , Soluciones Oftálmicas , Antibacterianos/uso terapéutico , Resultado del Tratamiento , Blefaritis/diagnóstico , Blefaritis/tratamiento farmacológico , Bacterias , Protocolos Clínicos , Estudios Observacionales como Asunto , Estudios Multicéntricos como AsuntoRESUMEN
PURPOSE: To optimize prophylactic antibiotic administration, antibiotic susceptibility before cataract surgery was investigated using ocular bacteria isolated preoperatively. DESIGN: Retrospective cross-sectional study. METHODS: In 204 eyes of 102 patients who underwent routine bilateral cataract surgery, conjunctival sac scrapings were collected 1-2 weeks before surgery. A total of 192 major pathogens among the 470 isolated bacteria were subjected to susceptibility testing. The major pathogens included Staphylococcus aureus, Staphylococcus epidermidis, coagulase-negative staphylococci (CNS) other than S. epidermidis, Enterococcus faecalis, and Streptococcus spp. The following antibiotics were tested: cefmenoxime (CMX), ceftazidime (CAZ), tobramycin (TOB), vancomycin (VAN), erythromycin (EM), moxifloxacin (MFLX), gatifloxacin (GFLX), levofloxacin (LVFX), chloramphenicol (CP), and imipenem (IP). RESULTS: The proportions of isolates with minimum inhibitory concentration (MIC) of S. epidermidis (N = 82), exceeding 4 µg/ml were high for CAZ (95.1%), EM (32.9%), LVFX (39.0%), and CP (82.9%). Susceptible (S) proportion was high for CMX (98.8%), VAN (100%), CP (93.9%), and IP (97.6%) but relatively low for MFLX (59.8%), GFLX (54.9%), and LVFX (54.9%). The MIC90 values were high for CMX (16 µg/ml), CAZ (64 µg/ml), TOB (32 µg/ml), EM (128 µg/ml), LVFX (16 µg/ml), and CP (8 µg/ml). The MIC of quinolonesof pathogenic bacteria other than S.epidermidis (N = 108), exceeded 4 µg/ml for 11 isolates, including two Methicillin-resistant Staphylococcus aureus. CONCLUSIONS: The increase in resistance of resident bacteria present in the conjunctival sac to antibiotics indicates that systemic and topical antibiotics are no longer effective, especially against external organisms affecting the eye.
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Bacterias/aislamiento & purificación , Extracción de Catarata , Conjuntiva/microbiología , Anciano , Anciano de 80 o más Años , Antibacterianos/uso terapéutico , Profilaxis Antibiótica , Bacterias/efectos de los fármacos , Estudios Transversales , Farmacorresistencia Bacteriana , Endoftalmitis/prevención & control , Femenino , Humanos , Masculino , Pruebas de Sensibilidad Microbiana , Estudios RetrospectivosRESUMEN
PURPOSE: To verify the possibility that preoperative topical antibiotics are not essential as long as iodine disinfection is performed during surgery. STUDY DESIGN: Crossover equivalence trial. PATIENTS AND METHODS: In 204 eyes of 102 patients who underwent routine bilateral cataract surgery, 1 eye was treated with intraoperative iodine, and the other, with preoperative topical antibiotics. For the intraoperative iodine eyes, 5 mL of 0.25% povidone-iodine was applied at 2 stages: (1) just after the placement of the speculum and (2) before intraocular lens (IOL) insertion. For the contralateral eyes, preoperative topical antibiotics were administered 3 days before surgery without intraoperative iodine. Conjunctival samples for culture were obtained at 3 time points: (a) presurgery, (b) beginning of surgery, and (c) postsurgery. Real-time polymerase chain reaction (PCR) samples were obtained at the beginning of surgery and before IOL insertion. Intracameral moxifloxacin was applied in all the cases. RESULTS: The respective positive bacterial culture rates for intraoperative iodine eyes and preoperative topical antibiotics eyes were 95.1% and 98.0% at (a), 7.8% and 5.9% at (b), and 60.8% and 62.7% at (c). A significant difference in the positive bacterial culture rate was not found at any time point. For the intraoperative iodine eyes, the bacterial DNA copy number at (b) was significantly lower than that for the preoperative topical antibiotics eyes. CONCLUSIONS: The cleanliness of the operative field without using topical antibiotics was revealed to be equivalent to that of the conventional method (using preoperative antibiotics without intraoperative iodine) as long as intraoperative iodine was used.
Asunto(s)
Antiinfecciosos Locales/administración & dosificación , Desinfección/métodos , Implantación de Lentes Intraoculares , Facoemulsificación , Povidona Yodada/administración & dosificación , Administración Oftálmica , Anciano , Anciano de 80 o más Años , Antibacterianos/administración & dosificación , Bacterias/aislamiento & purificación , Conjuntiva/microbiología , Estudios Cruzados , ADN Bacteriano/genética , Endoftalmitis/microbiología , Endoftalmitis/prevención & control , Infecciones Bacterianas del Ojo/microbiología , Infecciones Bacterianas del Ojo/prevención & control , Femenino , Humanos , Periodo Intraoperatorio , Levofloxacino/administración & dosificación , Masculino , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
The molecular mechanisms involved in the terminal differentiation of erythroblasts have been elucidated by comparing enucleation and cell division. Although various similarities and differences between erythroblast enucleation and cytokinesis have been reported, the mechanisms that control enucleation remain unclear. We previously reported that dynein and microtubule-organizing centers mediated the polarization of nuclei in human erythroblasts. Moreover, the accumulation of F-actin was noted during the enucleation of erythroblasts. Therefore, during enucleation, upstream effectors in the signal transduction pathway regulating dynein or actin, such as cell division control protein 42 homolog (Cdc42), may be crucial. We herein investigated the effects of the Cdc42 inhibitor, CASIN, on cytokinesis and enucleation in colony-forming units-erythroid (CFU-Es) and mature erythroblasts (day 10). CASIN blocked the proliferation of CFU-Es and their enucleation in a dose-dependent manner. Dynein adopted an island-like distribution in the cytoplasm of non-treated CFU-Es, but was concentrated near the nucleus as a dot and co-localized with γ-tubulin in CASIN-treated cells. CASIN blocked the accumulation of F-actin in CFU-Es and day 10 cells. These results demonstrated that Cdc42 plays an important role in cytokinesis, nuclear polarization and nuclear extrusion through a relationship with dynein and actin filament organization during the terminal differentiation of erythroblasts.
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Actomiosina/metabolismo , Diferenciación Celular , Eritroblastos/citología , Eritroblastos/metabolismo , Proteína de Unión al GTP cdc42/genética , Biomarcadores , Diferenciación Celular/genética , Núcleo Celular/metabolismo , Células Precursoras Eritroides/citología , Células Precursoras Eritroides/metabolismo , Eritropoyesis/genética , Técnica del Anticuerpo Fluorescente , Expresión Génica , Humanos , Inmunohistoquímica , Proteína de Unión al GTP cdc42/metabolismoRESUMEN
More than 2million human erythroblasts extrude their nuclei every second in bone marrow under hypoxic conditions (<7% O2). Enucleation requires specific signal transduction pathways and the local assembly of contractile actomyosin rings. However, the energy source driving these events has not yet been identified. We examined whether different O2 environments (hypoxic [5% O2] and normoxic [21% O2] conditions) affected human CD34+ cell erythroblast differentiation. We also investigated the regulatory mechanisms underlying energy production in erythroblasts during terminal differentiation under 5% or 21% O2 conditions. The results obtained revealed that the enucleation ratio and intracellular levels of adenosine triphosphate (ATP), lactate dehydrogenase (LDH) M3H, and hypoxia-inducible factor 1α in erythroblasts during terminal differentiation were higher under the 5% O2 condition than under the 21% O2 condition. We also found that the enzymatic inhibition of glyceraldehyde 3-phosphate dehydrogenase and LDH, key enzymes in anaerobic glycolysis, blocked the proliferation of colony-forming units-erythroid and enucleation of erythroblasts, and also reduced ATP levels in erythroblasts under both hypoxic and normoxic conditions. Under both conditions, phosphorylation of the Ser232, Ser293, and Ser300 residues in pyruvate dehydrogenase (inactive state of the enzyme) in erythroblasts was involved in regulating the pathway governing energy metabolism during erythroid terminal differentiation. This reaction may be mediated by pyruvate dehydrogenase kinase (PDK) 4, the major PDK isozyme expressed in erythroblasts undergoing enucleation. Collectively, these results suggest that ATP produced by anaerobic glycolysis is the main source of energy for human erythroblast enucleation in the hypoxic bone marrow environment.
Asunto(s)
Adenosina Trifosfato/biosíntesis , Eritroblastos/metabolismo , Glucólisis/fisiología , Anaerobiosis/fisiología , Antígenos CD34/metabolismo , Eritroblastos/citología , Humanos , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Lactato Deshidrogenasa 5/metabolismo , Fosforilación/fisiología , Piruvato Deshidrogenasa Quinasa Acetil-Transferidora/metabolismoRESUMEN
The hydrogen molecule (H2), which has low redox potential, is produced by colonic fermentation. We examined whether increased hydrogen (H2) concentration in the portal vein in rats fed high amylose maize starch (HAS) helped alleviate oxidative stress, and whether the transplantation of rat colonic microbiota with high H2 production can shift low H2-generating rats (LG) to high H2-generating rats (HG). Rats were fed a 20% HAS diet for 10 days and 13 days in experiments 1 and 2, respectively. After 10 days (experiment 1), rats underwent a hepatic ischemia-reperfusion (IR) operation. Rats were then categorized into quintiles of portal H2 concentration. Plasma alanine aminotransferase activity and hepatic oxidized glutathione concentration were significantly lower as portal H2 concentration increased. In experiment 2, microbiota derived from HG (the transplantation group) or saline (the control group) were orally inoculated into LG on days 3 and 4. On day 13, portal H2 concentration in the transplantation group was significantly higher compared with the control group, and positively correlated with genera Bifidobacterium, Allobaculum, and Parabacteroides, and negatively correlated with genera Bacteroides, Ruminococcus, and Escherichia. In conclusion, the transplantation of microbiota derived from HG leads to stable, high H2 production in LG, with the resultant high production of H2 contributing to the alleviation of oxidative stress.
Asunto(s)
Amilosa/administración & dosificación , Colon/microbiología , Trasplante de Microbiota Fecal , Microbioma Gastrointestinal , Hidrógeno/metabolismo , Almidón/administración & dosificación , Animales , Bacteroidetes , Bifidobacterium , Colon/metabolismo , Dieta Alta en Grasa , Firmicutes , ARN Ribosómico 16S/aislamiento & purificación , Ratas , Ruminococcus , Análisis de Secuencia de ADN , Zea mays/químicaRESUMEN
BACKGROUND: Little information is available on periodontopathic bacterial infection in childhood. We assessed the prevalence by age of 10 putative periodontopathic microorganisms in periodontally healthy children using a polymerase chain reaction (PCR) assay. METHODS: Plaque samples were collected from the buccal-mesial sulcus of the first molar or second primary molar in the right upper quadrant of 144 children (2 to 13 years old, 12 subjects from each year of age) who showed negligible periodontal inflammation. Using species-specific primers of Porphyromonas gingivalis, Bacteroides forsythus, Prevotella intermedia, Prevotella nigrescens, Campylobacter rectus, Eikenella corrodens, Actinobacillus actinomycetemcomitans, Capnocytophaga ochracea, Capnocytophaga sputigena, and Treponema denticola, PCR amplification was performed with bacterial genomic DNA from plaque samples. RESULTS: The results indicated that C. rectus, E. corrodens, A. actinomycetemcomitans, C. ochracea, and C. sputigena were found in about 50% of the plaque samples from all age groups, while B. forsythus and P. intermedia were detected less frequently, and P. gingivalis and T. denticola were not found. In contrast, the percentage of P. nigrescens-positive subjects increased with age in primary dentition, and reached about 50% at 7 years of age and older. Subject-based analyses suggested that the number of bacterial species in the plaque samples increased gradually with age until 5 years old, and then reached a plateau after the mixed dentition period. CONCLUSIONS: The colonization of many putative periodontopathic microorganisms can occur quite early in childhood without clinical signs of periodontal disease. However, colonization by P. gingivalis and T. denticola was not detected in periodontally healthy children.