RESUMO
OBJECTIVES: To determine if a policy of universal fetal echocardiography (echo) in pregnancies conceived by in-vitro fertilization (IVF) is cost-effective as a screening strategy for congenital heart defects (CHDs) and to examine the cost-effectiveness of various other CHD screening strategies in IVF pregnancies. METHODS: A decision-analysis model was designed from a societal perspective with respect to the obstetric patient, to compare the cost-effectiveness of three screening strategies: (1) anatomic ultrasound (US): selective fetal echo following abnormal cardiac findings on detailed anatomic survey; (2) intracytoplasmic sperm injection (ICSI) only: fetal echo for all pregnancies following IVF with ICSI; (3) all IVF: fetal echo for all IVF pregnancies. The model initiated at conception and had a time horizon of 1 year post-delivery. The sensitivities and specificities for each strategy, the probabilities of major and minor CHDs and all other clinical estimates were derived from the literature. Costs, including imaging, consults, surgeries and caregiver productivity losses, were derived from the literature and Medicare databases, and are expressed in USA dollars ($). Effectiveness was quantified as quality-adjusted life years (QALYs), based on how the strategies would affect the quality of life of the obstetric patient. Secondary effectiveness was quantified as number of cases of CHD and, specifically, cases of major CHD, detected. RESULTS: The average base-case cost of each strategy was as follows: anatomic US, $8119; ICSI only, $8408; and all IVF, $8560. The effectiveness of each strategy was as follows: anatomic US, 1.74487 QALYs; ICSI only, 1.74497 QALYs; and all IVF, 1.74499 QALYs. The ICSI-only strategy had an incremental cost-effectiveness ratio (ICER) of $2 840 494 per additional QALY gained when compared to the anatomic-US strategy, and the all-IVF strategy had an ICER of $5 692 457 per additional QALY when compared with the ICSI-only strategy. Both ICERs exceeded considerably the standard willingness-to-pay threshold of $50 000-$100 000 per QALY. In a secondary analysis, the ICSI-only strategy had an ICER of $527 562 per additional case of major CHD detected when compared to the anatomic-US strategy. All IVF had an ICER of $790 510 per case of major CHD detected when compared with ICSI only. It was determined that it would cost society five times more to detect one additional major CHD through intensive screening of all IVF pregnancies than it would cost to pay for the neonate's first year of care. CONCLUSION: The most cost-effective method of screening for CHDs in pregnancies following IVF, either with or without ICSI, is to perform a fetal echo only when abnormal cardiac findings are noted on the detailed anatomy scan. Performing routine fetal echo for all IVF pregnancies is not cost-effective. © 2020 International Society of Ultrasound in Obstetrics and Gynecology.
Assuntos
Fertilização in vitro , Cardiopatias Congênitas/diagnóstico por imagem , Injeções de Esperma Intracitoplásmicas , Análise Custo-Benefício , Árvores de Decisões , Ecocardiografia/economia , Feminino , Cardiopatias Congênitas/economia , Humanos , Gravidez , Qualidade de Vida , Ultrassonografia Pré-Natal/economia , Estados UnidosRESUMO
OBJECTIVE: Mycoplasma pneumoniae (M. pneumoniae) pneumonia is the second-most common cause of community-acquired pneumonia (CAP). This study aimed at investigating into the prevalence of macrolide-resistant M. pneumoniae (MRMP) with respiratory virus co-infection and the antibiotic prescriptions in children with CAP in four provinces in Korea, and to assess the variations in the findings across regions and throughout the year. PATIENTS AND METHODS: This prospective study was conducted in 29 hospitals in Korea between July 2018 and June 2020. Among the enrolled 1,063 children with CAP, all 451 patients with M. pneumoniae underwent PCR assays of M. pneumoniae and respiratory viruses, and the presence of point mutations of residues 2063 and 2064 was evaluated. RESULTS: Gwangju-Honam (88.6%) showed the highest prevalence of MRMP pneumonia, while Daejeon-Chungcheong (71.3%) showed the lowest, although the differences in prevalence were not significant (p=0.074). Co-infection of M. pneumoniae pneumonia and respiratory virus was observed in 206 patients (45.4%), and rhinovirus co-infection (101 children; 22.2%) was the most frequent. The prevalence of MRMP pneumonia with respiratory virus co-infection and the antibiotic prescriptions differed significantly among the four provinces (p < 0.05). The monthly rate of MRMP pneumonia cases among all cases of M. pneumoniae pneumonia and tetracycline or quinolone prescriptions did not differ significantly among the four regions (trend p > 0.05) during the study period. CONCLUSIONS: The prevalence of M. pneumoniae pneumonia with virus co-infection and antibiotic prescriptions could differ according to region, although the MRMP pneumonia rate showed no difference within Korea.
Assuntos
Coinfecção , Infecções Comunitárias Adquiridas , Pneumonia por Mycoplasma , Viroses , Vírus , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Criança , Coinfecção/complicações , Coinfecção/tratamento farmacológico , Coinfecção/epidemiologia , Farmacorresistência Bacteriana , Humanos , Macrolídeos/uso terapêutico , Mycoplasma pneumoniae/genética , Pneumonia por Mycoplasma/tratamento farmacológico , Pneumonia por Mycoplasma/epidemiologia , Prescrições , Estudos Prospectivos , Viroses/tratamento farmacológicoRESUMO
BACKGROUND: Recent findings suggesting the involvement of adventitial cells in coronary repair have raised questions regarding the phenotypic "plasticity" of medial smooth muscle cells (SMCs). Accordingly, the aims of the present study were to examine the characteristics of coronary medial and adventitial cells and to compare the responses of coronary and noncoronary SMCs to stimulation. METHODS AND RESULTS: Enzymatically isolated coronary SMCs (human and porcine) were distinct from noncoronary SMCs, showing poor adhesion and spreading, as well as lower proliferation, collagen synthesis, and LDL degradation. Several extracellular matrix components (Matrigel, collagen I and IV, laminin, vitronectin, fibronectin) or growth factors (epidermal growth factor, platelet-derived growth factor-BB, insulin growth factor-1, interleukin-1alpha) failed to augment the adhesion or proliferation of coronary SMCs to the levels observed in noncoronary SMCs. Unlike coronary SMCs, coronary fibroblasts demonstrated high adhesion, proliferation, collagen synthesis, and avid LDL metabolism. Limited responses of coronary SMCs were associated with sustained expression of differentiation markers (alpha-smooth muscle actin, h-caldesmon, and smooth muscle myosin heavy chain), whereas noncoronary SMCs showed marked phenotypic heterogeneity. CONCLUSIONS: Coronary SMCs appeared to maintain highly differentiated phenotype in response to stimulation, whereas coronary adventitial fibroblasts demonstrated several characteristics that are essential during vascular repair. Coronary SMCs, however, were distinct from noncoronary medial cells, which displayed greater phenotypic heterogeneity and versatility in culture. We postulate that the mechanism of vascular repair may differ among vascular beds, pointing to the importance of coronary artery-specific investigations in vascular biology.
Assuntos
Vasos Coronários/citologia , Fibroblastos/citologia , Músculo Liso Vascular/citologia , Biomarcadores , Adesão Celular , Diferenciação Celular , Divisão Celular , Tamanho Celular , Colágeno/biossíntese , Vasos Coronários/metabolismo , Fibroblastos/metabolismo , Fibroblastos/fisiologia , Humanos , Técnicas In Vitro , Lipoproteínas LDL/metabolismo , Músculo Liso Vascular/metabolismoRESUMO
A monoclonal antibody, 4G10.3, was developed that preferentially binds limbal basal cells in adult rat, rabbit, and human corneas. These cells were hypothesized to be the stem cells for the corneal epithelium. The antibody 4G10.3 was localized by immunofluorescence microscopy in rats 1 d and 1, 1.5, 2, 3, 4, and 6 wk of age. Until 1.5 wk, 4G10.3 bound intensely to all basal cells in the cornea and the limbus. At 2 wks, the basal cells at the central cornea abruptly changed their shape from flattened or ovoid to large and cuboidal and bound 4G10.3 with greatly reduced intensity. Increased stratification of epithelium also was seen. Cells binding 4G10.3 gradually became sequestered to the limbal area after 2 wk, concomitant with increased stratification. At 4 and 6 wk, 4G10.3 binding was identical to that in adult corneas with only limbal basal cells showing positive binding. Basal cells in the limbal epithelium did not decrease their intense binding of 4G10.3 or change their ovoid cellular shape from 1 d through adult life. These results suggest that, during development, stem or stem-like cells are localized throughout the basal layer of the corneal and limbal epithelium. As the cornea matures, these cells are sequestered in the limbus at the same time that stratification of the epithelium and shape changes occur in the basal cells.
Assuntos
Córnea/citologia , Células-Tronco/citologia , Animais , Anticorpos Monoclonais , Córnea/crescimento & desenvolvimento , Células Epiteliais , Epitélio/crescimento & desenvolvimento , Proteínas do Olho , Imunofluorescência , Limbo da Córnea/citologia , Limbo da Córnea/crescimento & desenvolvimento , Microscopia de Fluorescência , Ratos , Ratos EndogâmicosRESUMO
PURPOSE: To examine the expression of the glycolytic enzyme alpha-enolase after limbus-to-limbus epithelial debridement in the rabbit. METHODS: Corneas were debrided, leaving limbal epithelium intact, and were allowed to heal from 2 days to 8 weeks. Immunofluorescence microscopy was used to observe the expression of alpha-enolase. To quantitate changes in alpha-enolase levels 2 days to 4 weeks after wounding, epithelium was harvested, homogenized, and assayed using anti-alpha-enolase in immunoslot blots. RESULTS: Expression of alpha-enolase appeared to increase in the limbus and the central cornea during epithelial migration (2-day time point) with intense labeling of all basal cells. These levels were maintained until wound closure (1 week). By 2 weeks, expression in the limbal basal cells decreased to levels present in unwounded corneas. Expression in the corneal epithelium decreased after 2 weeks, progressing from central cornea to the periphery. At 4 weeks, antibody binding decreased concomitantly with a change in the shape of the basal cells from flattened or ovoid to columnar. At 8 weeks, expression of alpha-enolase was similar to that in control corneas. Immunoslot blot data indicated that alpha-enolase made up 0.28% of the total soluble protein in unwounded corneal epithelium and 0.73%, 1.22%, 0.96%, and 0.49% at 2 days, 1 week, 2 weeks, and 4 weeks after debridement, respectively. CONCLUSIONS: These data indicate that expression of alpha-enolase is elevated during corneal epithelial migration initiating from the stem (limbal basal) cell population and that expression is linked to active migration. Furthermore, it appears that limbal basal cells are metabolically active during the period of epithelial sheet movement, whereas peripheral corneal basal cells remain activated as long as 4 weeks after wounding.
Assuntos
Córnea/fisiologia , Limbo da Córnea/fisiologia , Fosfopiruvato Hidratase/biossíntese , Regeneração/fisiologia , Animais , Anticorpos Monoclonais , Diferenciação Celular , Divisão Celular , Movimento Celular , Córnea/citologia , Córnea/enzimologia , Desbridamento , Epitélio/enzimologia , Epitélio/fisiologia , Imunofluorescência , Limbo da Córnea/citologia , Limbo da Córnea/cirurgia , Coelhos , Células-Tronco/metabolismo , Cicatrização/fisiologiaRESUMO
PURPOSE: It has been demonstrated that cells migrating to cover an epithelial débridement wound exit the cell cycle and that the cell-cycle inhibitor p15(INK4b) is upregulated in these cells. TGF-beta signaling has been implicated in both of these processes, and this study was conducted to determine whether the expression and localization of TGF-beta receptor (TbetaR)-I and -II are altered during corneal epithelial wound repair. METHODS: Three-millimeter superficial keratectomy wounds and 3-mm débridement wounds were made in central rat cornea and allowed to heal in vivo for 1 to 48 hours. Immunofluorescence microscopy and Western blot analysis were used to determine the localization and expression of TbetaR-I and -II. Unwounded rat corneas served as control samples. To determine the effect of epidermal growth factor (EGF) and TGF-beta1 on p15(INK4b) and TbetaR-I and -II expression, human corneal epithelial cells were grown in culture to 50% to 60% confluence, and EGF (5 ng/ml) and/or TGF-beta1 (2 ng/ml) were added for 6 hours. Cells were harvested and p15(INK4b) and TBR-I and -II levels were assayed by using Western blot analysis. RESULTS: In unwounded corneas, TbetaR-I and TbetaR-II were present at low levels across the cornea, with higher levels in limbal epithelium. Both TbetaR-I and -II were upregulated after wounding. However, levels of TbetaR-II appeared to increase in the epithelial cells that had migrated to cover the wound area, whereas TbetaR-I was upregulated in the entire corneal epithelium. Western blot analysis indicated that both TbetaR-I and -II were upregulated threefold after wounding. In cultured cells, EGF and TGF-beta1 stimulated TbetaR-II; however, neither one stimulated TbetaR-I expression. TGF-beta1 stimulated p15(INK4b) protein levels threefold. CONCLUSIONS: After wounding, TbetaR-I and TbetaR-II were both expressed at high levels in cells migrating to cover a corneal wound, suggesting that TGF-beta signaling is involved in blocking migrating cells from progressing through the cell cycle. This blockage, at least in part, involves the inhibitor p15(INK4b). In addition, although both TbetaR-I and TbetaR-II are upregulated during wound repair, they appear to be differentially regulated.
Assuntos
Receptores de Ativinas Tipo I , Epitélio Corneano/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Receptores de Fatores de Crescimento Transformadores beta/metabolismo , Cicatrização , Animais , Western Blotting , Técnicas de Cultura de Células/métodos , Epitélio Corneano/lesões , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Masculino , Ratos , Ratos Sprague-Dawley , Receptor do Fator de Crescimento Transformador beta Tipo I , Receptor do Fator de Crescimento Transformador beta Tipo II , Regulação para CimaRESUMO
PURPOSE: This study's intention was to examine the progression of ocular surface epithelium through the G1/S transition of the cell cycle after corneal epithelial debridement. METHODS: Three-millimeter debridements were made in central rat cornea and allowed to heal 4 to 48 hours in vivo. Unwounded contralateral eyes served as controls. Two hours before the animals were killed, 5-bromo-2-deoxyuridine (BrdU) was injected to detect S-phase cells. Incorporated BrdU was visualized by indirect immunofluorescence microscopy, and expression of G1 cell-cycle markers cyclins D and E was examined by indirect immunofluorescence and immunoblotting. RESULTS: The number of BrdU-labeled cells in conjunctival, limbal, and peripheral epithelium peaked at 28 hours after wounding (3.9-, 4.5-, and 3.2-fold increases, respectively). In unwounded eyes, cyclin D showed diffuse cytoplasmic localization with occasional basal cells exhibiting a nuclear localization, while anti-cyclin E showed intense localization in limbal and conjunctival basal cells but only minimal labeling in corneal epithelium. Within 8 to 12 hours after wounding, the nuclei of most corneal basal cells outside the wound area were bound intensely by anti-cyclins D and E. Immunoblotting revealed that cyclin D and E protein levels increased 4.5- and 12.1-fold after wounding, respectively. Epithelium migrating into the wound area did not incorporate BrdU and did not exhibit nuclear localization of cyclins D and E. CONCLUSIONS: Corneal epithelial debridement stimulates basal cells outside the wound area to synchronously enter the cell cycle. However, cells migrating to cover the wound area do not progress through the cell cycle. These data suggest a compartmentalization of the proliferative and migratory phases of wound repair.
Assuntos
Desbridamento , Epitélio Corneano/citologia , Fase G1/fisiologia , Fase S/fisiologia , Cicatrização/fisiologia , Animais , Western Blotting , Divisão Celular , Movimento Celular , Túnica Conjuntiva/citologia , Túnica Conjuntiva/metabolismo , Ciclina D , Ciclina E/metabolismo , Ciclinas/metabolismo , Replicação do DNA , Epitélio/metabolismo , Epitélio Corneano/metabolismo , Epitélio Corneano/cirurgia , Técnica Indireta de Fluorescência para Anticorpo , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
PURPOSE: To determine the expression patterns of the retinoblastoma protein and the E2F transcription factor families in limbal and corneal epithelia and in corneal keratocytes in situ during corneal development and differentiation. METHODS: Retinoblastoma protein (pRb) and its family members p107 and p130; E2F-1, -2, and -4, members of the E2F family of transcription factors; and Ki67, a marker of actively cycling cells, were localized by indirect immunofluorescence microscopy, in corneas of neonatal, juvenile, and adult rats. Presence of mRNA for pRb, p107, p130, and E2F types 1 to 5 in adult corneal epithelium was determined by reverse transcription-polymerase chain reaction. RESULTS: mRNA for all members of pRb and E2F families was present in adult corneal epithelium. The greatest number of Ki67-positive corneal and limbal epithelial cells were present at days 13 to 19, and Ki67-positive stromal keratocytes at day 2. pRb and E2F-2 were localized to all cells in neonatal, juvenile, and adult corneas. With age, p130 localization became more intense and nuclear in stromal keratocytes and suprabasal cells of corneal and limbal epithelia; p107, initially nuclear in limbal and corneal epithelia, became increasingly cytoplasmic in corneal epithelium. E2F-1 was initially nuclear in keratocytes and diminished after day 10. E2F-1 was localized in the basal cell layer of limbal and corneal epithelia after day 10. E2F4 was always nuclear in limbal epithelium and cytoplasmic in corneal epithelium. CONCLUSIONS: Expression patterns of pRb and E2F family proteins vary with corneal cell differentiation, but are most apparent with p130 and p107. Nuclear localization of p130 appears to correlate with terminal differentiation in epithelium and entrance into a quiescent state by keratocytes. In contrast, p107 is nuclear in the undifferentiated limbal basal cells and is cytoplasmic in the remainder of the corneal epithelial cells.
Assuntos
Proteínas de Transporte , Proteínas de Ciclo Celular , Córnea/crescimento & desenvolvimento , Córnea/metabolismo , Proteínas de Ligação a DNA , RNA Mensageiro/biossíntese , Proteína do Retinoblastoma/genética , Fatores de Transcrição/genética , Envelhecimento/fisiologia , Animais , Animais Recém-Nascidos/metabolismo , Ciclo Celular , Diferenciação Celular , Córnea/citologia , Primers do DNA/química , Fatores de Transcrição E2F , Fator de Transcrição E2F1 , Técnica Indireta de Fluorescência para Anticorpo , Expressão Gênica , Antígeno Ki-67/metabolismo , RNA/isolamento & purificação , Ratos , Ratos Sprague-Dawley , Proteína do Retinoblastoma/biossíntese , Proteína 1 de Ligação ao Retinoblastoma , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator de Transcrição DP1 , Fatores de Transcrição/biossínteseRESUMO
In total, 18 of 26 double high-dose chemotherapies (HDCT) in pediatric solid tumors were rescued with peripheral blood stem cells collected during a single leukapheresis round (single-harvest group, SHG). In the remaining eight HDCT, additional leukapheresis were necessary after the first HDCT (HDCT1) to rescue the second HDCT (HDCT2) (double-harvest group, DHG). Stem cell collection after HDCT1 was inefficient and delayed in patients who had received prior chemotherapy before HDCT1. The interval between HDCT1 and HDCT2 was shorter in SHG than in DHG (median 62.5 days vs 178.5 days, P-value=0.002). Hematologic recovery in HDCT2 was delayed compared to HDCT1. However, there was no difference in hematologic recovery between SHG and DHG. A high rate of treatment-related mortality (TRM) was recorded during HDCT2, but there was no evidence that the shorter interval caused a higher rate of TRM (P-value=0.454). The probability of disease-free survival at 2 years after HDCT2 in the SHG and DHG were 66.7 and 25.0%, respectively (P-value=0.031). Therefore, to administer the second HDCT earlier in double HDCT, and thus to improve the survival of patients with high-risk solid tumors, the single-harvest approach is recommended rather than the double-harvest approach.
Assuntos
Antineoplásicos/administração & dosagem , Transplante de Células-Tronco Hematopoéticas , Leucaférese , Neoplasias do Sistema Nervoso/tratamento farmacológico , Neuroblastoma/tratamento farmacológico , Adolescente , Adulto , Idoso , Plaquetas/citologia , Neoplasias Encefálicas/tratamento farmacológico , Neoplasias Encefálicas/mortalidade , Neoplasias Cerebelares/tratamento farmacológico , Neoplasias Cerebelares/mortalidade , Terapia Combinada , Feminino , Glioma/tratamento farmacológico , Glioma/mortalidade , Humanos , Masculino , Meduloblastoma/tratamento farmacológico , Meduloblastoma/mortalidade , Pessoa de Meia-Idade , Neoplasias do Sistema Nervoso/mortalidade , Neuroblastoma/mortalidade , Fatores de Risco , Taxa de Sobrevida , Transplante Autólogo , Resultado do TratamentoRESUMO
Neural stimulation of the cornea induces conjunctival goblet cell mucous secretion. Immunofluorescence microscopy was used to determine if nerves are present near conjunctival goblet cells and what types of nerves are present. In euthanized rats, the local anesthetic lidocaine (1%) was placed topically on the ocular surface for 10 min to prevent goblet cell mucous secretion. The ocular surface tissues were removed and either fixed in formaldehyde and then frozen, or frozen first and then post-fixed in formaldehyde. Tissue was sectioned and nerves localized by indirect immunofluorescence microscopy, using antibodies to synaptophysin (indicates nerve, independent of type), vasoactive intestinal peptide (VIP, indicates parasympathetic nerves), tyrosine hydroxylase (TH, indicates sympathetic nerves), dopamine beta-hydroxylase (DBH, indicates sympathetic nerves), phenylethanolamine-N-methyltransferase (PNMT, indicates sympathetic nerves), and calcitonin gene-related peptide (CGRP, indicates sensory nerves). Goblet cells were identified by phase-contrast microscopy. Synpatophysin-containing nerves were present in the basolateral region of conjunctival goblet cells clusters. Nerve fibers immunoreactive to VIP were found in the conjunctiva along the epithelial-stromal junction and around the basolateral aspect of goblet cell clusters. Nerve fibers immunoreactive to TH and DBH were detected surrounding goblet cells and in the conjunctival stroma. Nerve fibers immunoreactive to CGRP were detected in the epithelium and at the epithelial stromal junction, but were not localized near goblet cell clusters. CGRP-containing nerve fibers were also detected in the conjunctival stroma under the epithelium. We conclude that efferent parasympathetic and sympathetic, but not afferent sensory, nerves appear to be located adjacent to conjunctival goblet cell clusters. Activation of efferent parasympathetic and sympathetic nerves could directly stimulate conjunctival goblet cell mucous secretion. Antidromic activation of afferent sensory nerves releasing neurotransmitters could stimulate goblet cell secretion by a paracrine mechanism.
Assuntos
Túnica Conjuntiva/inervação , Sistema Nervoso Parassimpático/ultraestrutura , Sistema Nervoso Simpático/ultraestrutura , Animais , Túnica Conjuntiva/citologia , Túnica Conjuntiva/ultraestrutura , Dopamina beta-Hidroxilase/análise , Células Epiteliais , Epitélio/inervação , Epitélio/ultraestrutura , Técnica Indireta de Fluorescência para Anticorpo , Masculino , Proteínas do Tecido Nervoso/análise , Sistema Nervoso Parassimpático/química , Ratos , Ratos Sprague-Dawley , Sistema Nervoso Simpático/química , Tirosina 3-Mono-Oxigenase/análiseRESUMO
The present study was performed to evaluate the effect of tricalcium phosphate implant in human osseous defects. 4 intraosseous defects in 2 patients were treated with mucoperiosteal flap and tricalcium phosphate (TCP) implant and another 5 defects in the same patients were debrided only via mucoperiosteal flap. The healing response was evaluated clinically 10-12 weeks after treatment. More reduction in probing depth and more gains in probing attachment levels were observed in implanted sites than in control sites.
Assuntos
Fosfatos de Cálcio , Implantação Dentária/instrumentação , Doenças Periodontais/cirurgia , Reabsorção Óssea/cirurgia , Implantes Dentários , Inserção Epitelial , HumanosRESUMO
We have been reported that the repeated cerebral ischemia induced more severe disruption of spatial cognition than single ischemia without any other motor disturbance in 8-arm radial maze task in rats. And we have been clarified that it is corresponding with 60% of selective cell injury of the CA1 pyramidal cells in the hippocampus. Recently, characteristics of apoptosis such as internucleosomal DNA fragmentation have been found in excitotoxic neuronal death. In the present study, we investigated how necrosis and apoptosis following repeated ischemia involve to the cell death. Repeated cerebral ischemia (10 min x 2, 1 hr interval) induced significant disruption of spatial cognition not only 24 hrs but also 7 days after reperfusion. The decrease of H.E-positive neurons was found in the hippocampus CA1 area and frontal cortex within 3 days after reperfusion, while an DNA fragmentation and TUNNEL-positive neurons in the same areas were found afterward. Furthermore repeated cerebral ischemia-induced disruption of spatial cognition and apoptosis in the hippocampal CA1 area were inhibited by YM-90 K(15 mg/kg,i.p.), which is a selective AMPA/KA receptor antagonist, but not by MK-801. These results suggested that the apoptotic cell death may be occurred via non-NMDA receptor mechanism in relatively late phase of the reperfusion period and it may relate to the incidence of the disruption of spatial cognition in the rat.
Assuntos
Apoptose/fisiologia , Ataque Isquêmico Transitório/patologia , Percepção Espacial/fisiologia , Animais , Hipocampo/patologia , Ataque Isquêmico Transitório/fisiopatologia , Masculino , Necrose , Ratos , Ratos Wistar , RecidivaRESUMO
We have developed an in vitro and in vivo method to determine if VIP-stimulates conjunctival goblet cell secretion in the rat as nerves which contain vasoactive intestinal peptide (VIP) and are most likely parasympathetic are localized around these cells. For the in vitro method, pieces of rat conjunctiva were incubated for 1 hr with no additions or increasing concentrations of VIP (10(-10)-10(-6)M). Goblet cell secretion was measured by determining the amount of Helix pomatia agglutinin (HPA)-detectable glycoconjugates secreted into the medium. HPA-detectable glycoconjugates were assayed using an enzyme-linked lectin assay. For the in vivo method, drops of buffer containing no additions or varying concentrations of VIP (10(-10)-10(-6) M) were placed on the ocular surface of anesthetized rats for 60 min. The rats were killed, the ocular surface chemically fixed, and a button of conjunctiva removed. Mucin-containing goblet cells were stained by Alcian blue-periodic acid Schiff's reagent and the number of cells per 0.16 mm2 was quantified. A decrease in the number of mucin-containing goblet cells indicated an increase in mucous secretion. By immunofluorescent histochemistry, we found that the lectin HPA was localized predominantly in the secretory granules of rat conjunctival goblet cells with little binding present in the remainder of the conjunctiva. Nerves containing VIP surrounded goblet cells labelled with HPA. In pieces of conjuctiva, in vitro VIP (10(-8)-10(-6) M) stimulated HPA-detectable glycoconjugate secretion in a concentration dependent manner. When applied topically to the ocular surface, in vivo VIP AT 10(-8) M stimulated mucous secretion from conjunctival goblet cells. We conclude that VIP is present in nerves around conjunctival goblet cells and stimulates glycoconjugate secretion from these cells.
Assuntos
Túnica Conjuntiva/metabolismo , Glicoconjugados/metabolismo , Peptídeo Intestinal Vasoativo/farmacologia , Animais , Túnica Conjuntiva/efeitos dos fármacos , Relação Dose-Resposta a Droga , Imunofluorescência , Hemaglutininas , Técnicas In Vitro , Lectinas , Masculino , Ratos , Ratos Sprague-Dawley , Estimulação QuímicaRESUMO
This study was performed to elucidate the mechanism of improved oxygenation after surfactant replacement therapy in respiratory distress syndrome (RDS) of the newborn infants. In 26 newborns with RDS, end tidal-CO2 tension (PetCO2), arterial blood gas analysis and pulmonary function tests were measured at baseline, 30 min, 2 hr and 6 hr after surfactant administration. The changes in dead space/tidal volume ratio (VD/VT ratio=(PaCO2-PetCO2)/PaCO2), oxygenation index and arterial-alveolar partial pressure difference for oxygen ((A-a)DO2) were elucidated and correlated with pulmonary mechanics. Oxygenation index and (A-a)DO2 improved, and VD/VT ratio decreased progressively after surfactant administration, becoming significantly different from the baseline at 30 min and thereafter with administration of surfactant. Pulmonary mechanics did not change significantly during the observation period. VD/VT ratio showed close correlation with OI and (A-a)DO2, but not with pulmonary mechanics. These results suggest that decreased physiologic dead space resulting from the recruitment of atelectatic alveoli rather than improvement in pulmonary mechanics is primarily responsible for the improved oxygenation after surfactant therapy in the RDS of newborn.
Assuntos
Pulmão/fisiopatologia , Surfactantes Pulmonares/uso terapêutico , Espaço Morto Respiratório , Síndrome do Desconforto Respiratório do Recém-Nascido/tratamento farmacológico , Volume de Ventilação Pulmonar , Resistência das Vias Respiratórias , Humanos , Recém-Nascido , Complacência Pulmonar , Troca Gasosa Pulmonar , Síndrome do Desconforto Respiratório do Recém-Nascido/fisiopatologiaRESUMO
Many transcriptional regulators function in homo- or heterodimeric combinations. The same protein can carry out distinct regulatory functions depending on the partner with which it associates. Here, we describe a mutant of the Escherichia coli cAMP receptor protein (CRP) that has an altered dimerization specificity; that is, mutant/mutant homodimers form preferentially over wild-type/mutant heterodimers. CRP dimerization involves the formation of a parallel coiled-coil structure, and our CRP mutant bears an amino acid substitution affecting the first "d" position residue within the alpha-helix that mediates CRP dimerization. The genetic strategy we used to isolate this CRP altered dimerization specificity (ADS) mutant is generalizable and could be utilized to isolate ADS mutants of other dimeric transcriptional regulators.
Assuntos
Escherichia coli/genética , Receptores de AMP Cíclico/química , Receptores de AMP Cíclico/genética , Sequência de Aminoácidos , Bacteriófagos , Sequência de Bases , Eletroforese , Escherichia coli/metabolismo , Modelos Moleculares , Dados de Sequência Molecular , Mutação , Conformação Proteica , Receptores de AMP Cíclico/isolamento & purificação , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/isolamento & purificação , Alinhamento de Sequência , Transcrição Gênica , beta-Galactosidase/genéticaRESUMO
This study was performed to investigate the etiologic agents, age distribution, clinical manifestations and seasonal occurrence of acute viral lower respiratory tract infections in children. We confirmed viral etiologies using nasopharyngeal aspirates in 237 patients of the ages of 15 years or younger who were hospitalized for acute lower respiratory tract infection (ALRI) from March 1996 to February 1998 at Samsung Seoul Hospital, Seoul, Korea. The overall isolation rate was 22.1%. The viral pathogens identified were adenovirus (12.7%), influenza virus type A (21.1%), -type B (13.9%), parainfluenza virus type 1 (13.5%), -type 2 (1.3%), -type 3 (16.0%) and respiratory syncytial virus (21.5%). The occurrence of ALRIs was highest in the first year of life, although parainfluenza virus type 1 infection occurred predominantly in the second year of life and influenza virus caused illnesses in all age groups. The specific viruses are frequently associated with specific clinical syndromes of ALRI. The respiratory agents and associated syndromes frequently have characteristic seasonal patterns. This study will help us to estimate the etiologic agents of ALRI, and establish a program for the prevention and treatment. An annual nationwide survey is necessary to understand the viral epidemiology associated with respiratory illnesses in Korea.