RESUMO
In this article we develop an algorithm for the efficient simulation of electrolytes in the presence of physical boundaries. In previous work the discrete ion stochastic continuum overdamped solvent (DISCOS) algorithm was derived for triply periodic domains, and was validated through ion-ion pair correlation functions and Debye-Hückel-Onsager theory for conductivity, including the Wien effect for strong electric fields. In extending this approach to include an accurate treatment of physical boundaries we must address several important issues. First, the modifications to the spreading and interpolation operators necessary to incorporate interactions of the ions with the boundary are described. Next we discuss the modifications to the electrostatic solver to handle the influence of charges near either a fixed potential or dielectric boundary. An additional short-ranged potential is also introduced to represent interaction of the ions with a solid wall. Finally, the dry diffusion term is modified to account for the reduced mobility of ions near a boundary, which introduces an additional stochastic drift correction. Several validation tests are presented confirming the correct equilibrium distribution of ions in a channel. Additionally, the methodology is demonstrated using electro-osmosis and induced-charge electro-osmosis, with comparison made to theory and other numerical methods. Notably, the DISCOS approach achieves greater accuracy than a continuum electrostatic simulation method. We also examine the effect of under-resolving hydrodynamic effects using a "dry diffusion" approach, and find that considerable computational speedup can be achieved with a negligible impact on accuracy.
RESUMO
The hippocampus plays a critical role in contextual fear conditioning. Population activity in the hippocampal CA1 encoding the surrounding environment is thought to be responsible for retrieval of contextual fear memory. However, the characteristics of CA1 neuronal ensemble activity during retrieval of contextual fear memory remain unclear. Here, we examined CA1 ensemble activity during contextual fear memory expression in male C57Bl/6J mice, using Arc cellular compartment analysis of temporal activity by fluorescence in situ hybridization. The "Shock" group was conditioned with a footshock in two separate chambers, whereas the "No shock" group was not exposed to shocks in the chamber. Animals were then re-exposed to either the same chamber twice or two different conditioning chambers. In the No shock group, exposure to the same chamber twice activated a more significantly overlapping neuronal population than exposure to two different chambers. In the Shock group, exposure to the same conditioning chamber twice activated a similarly overlapping neuronal population as exposure to two different chambers, with overlap smaller than in nonshocked mice exposed to the same chamber twice. Thus, population activity in the hippocampal CA1 encoding the surrounding environment is detected during spatial exploration, but absent during contextual fear memory expression. Even the variable ensemble activity of CA1 may contribute to retrieval of contextual fear memory.
Assuntos
Região CA1 Hipocampal/fisiologia , Condicionamento Clássico/fisiologia , Medo/fisiologia , Memória/fisiologia , Animais , Hibridização in Situ Fluorescente , Masculino , Camundongos , Camundongos Endogâmicos C57BLRESUMO
OBJECTIVES: In our previous study, scores determined via a multiple linear regression method (EN-MLR) involving an electronic nose provided objective halitosis-related measurements; however, this model afforded only relative expression exclusively. The objective of this investigation was to assess clinically oral malodor intensity expressed as an absolute value using an electronic nose. SUBJECTS AND METHODS: Sixty-six subjects were evaluated based on results of an actual organoleptic test (OLT), measurements of volatile sulfur compound (VSC) concentrations, a score representing malodor intensity (EN-MI) as the absolute value and EN-MLR measured with an electronic nose system. Oral health parameters were also examined. RESULTS: The OLT score served as a benchmark. The area under the receiver-operating characteristic (ROC) plots of EN-MI score (0.975) was significantly larger than that of log VSC (0.896) (P = 0.036); however, the area did not differ significantly from that of EN-MLR score (0.932). Percentage of teeth with pocket depth greater than or equal to 4 mm, tongue coating score and plaque control record displayed meaningful association with EN-MI score in multiple logistic regression analyses. CONCLUSION: Oral malodor intensity expressed as an absolute value employing an electronic nose may be a suitable method for clinical evaluation of oral malodor.
Assuntos
Testes Respiratórios/instrumentação , Halitose/diagnóstico , Adulto , Eletrônica Médica , Gases/análise , Humanos , Curva ROC , Análise de Regressão , Compostos de Enxofre/análiseRESUMO
A recently developed electronic nose has not yet been clinically applied to evaluations of oral malodor. This investigation sought to determine whether an electronic nose could clinically assess oral malodor. Twenty-nine healthy adults and 49 patients were assessed by results of an actual organoleptic test, a score representing malodor strength with an electronic nose in "top-note" mode (top-note score), and measurements of volatile sulfur compound (VSC) concentrations. The correlation coefficient between top-note and actual organoleptic scores (r = 0.71) was comparable with the log VSC and actual organoleptic scores (r = 0.63). However, the area under the receiver-operating characteristic plots for top-note score was significantly larger than that for log VSC. In logistic regression analyses with top-note score as a dependent variable, probing depth, tongue coating, and plaque control record each had independent associations. Our findings suggest that the top-note score from an electronic nose examination may be useful for the clinical evaluation of oral malodor.
Assuntos
Técnicas Biossensoriais/métodos , Halitose/diagnóstico , Saúde Bucal , Compostos de Enxofre/análise , Adulto , Eletrônica , Estudos de Avaliação como Assunto , Feminino , Halitose/etiologia , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Índice Periodontal , Compostos de Enxofre/efeitos adversosRESUMO
BACKGROUND: It is well known that selectin is involved in the development of endotoxin induced uveitis (EIU), and has a major role in leucocyte infiltration. Recently, a novel selectin inhibitor (SKK-60060) that can block P and L selectins in vitro has been developed. This study was designed to investigate the anti-inflammatory effects of SKK-60060 on the inflammatory reaction during EIU in rats by studying leucocyte-endothelium interactions. METHODS: EIU was induced in Lewis rats by footpad injection of lipopolysaccharide (LPS). SKK-60060 was administered 15 minutes before LPS injection, and its suppressive effects on inflammatory leucocyte behaviour were evaluated in vivo with acridine orange digital fluorography; the diameters of retinal arteries and veins were also measured. After these studies, aqueous humour was collected to evaluate leucocyte infiltration and protein leakage. RESULTS: After LPS injection, rolling leucocytes were observed in major retinal veins, followed by leucocyte infiltration into the vitreous cavity. Following treatment with SKK-60060, leucocyte rolling was significantly inhibited in the retinal veins (p <0.01), and subsequent leucocyte infiltration into the vitreous cavity was also significantly suppressed (p <0.01). Retinal vasodilation was also substantially suppressed in SKK-60060 treated rats (p <0.01). Similarly, leucocyte infiltration and protein leakage into the aqueous humour were reduced significantly by SKK-60060 (p <0.01). CONCLUSIONS: SKK-60060 treatment significantly inhibited the inflammatory reaction induced by LPS. Its inhibitory effects on P and L-selectin resulted in suppression of leucocyte infiltration and the subsequent inflammatory reaction caused by accumulated leucocytes. The current findings suggest that SKK-60060 may be useful in the management of uveitis.
Assuntos
Anti-Inflamatórios/farmacologia , Dissacarídeos/farmacologia , Leucócitos/efeitos dos fármacos , Selectinas/fisiologia , Uveíte/tratamento farmacológico , Animais , Humor Aquoso/metabolismo , Movimento Celular/efeitos dos fármacos , Modelos Animais de Doenças , Endotélio Vascular/fisiopatologia , Fluoroscopia/métodos , Processamento de Imagem Assistida por Computador/métodos , Contagem de Leucócitos , Leucócitos/fisiologia , Lipopolissacarídeos , Ratos , Ratos Endogâmicos Lew , Uveíte/metabolismo , Uveíte/patologia , Uveíte/fisiopatologiaRESUMO
alpha(1)-Acid glycoprotein (AGP) is the major transport protein for cationic drugs, endogenous ligands, and some anionic drugs in plasma. Hepatic synthesis and secretion of AGP are altered during acute inflammation as well as by a number of drugs. This alteration could influence the binding of drugs and its biological function. Macrolide antibiotics are widely used in the treatment of a variety of infectious diseases. The effects of macrolide antibiotics have been studied with respect to rat AGP expression in vivo. After the individual administration of six macrolides to rats, with the exception of oleandomycin, five increased AGP levels in serum. Of these five, clarithromycin (CAM) was the most potent inducer of AGP, which reached a maximum level between 3 to 7 days after administration. CAM increased the steady-state level of AGP mRNA in liver as well as protein level in serum in a dose-dependent manner. In addition, CAM increased AGP mRNA levels in primary cultured hepatocytes. In the luciferase promoter assay, CAM potentiated dexamethasone-increased promoter activity of the AGP gene, which contained the glucocorticoid response element, in cultured rat hepatocytes, although CAM itself had no effect on its activity. The effect of CAM and dexamethasone was diminished by glucocorticoid response element deletion or mutation or by adding the antiglucocorticoid, RU486. Further, in the mouse mammary tumor virus (MMTV) promoter containing functional glucocorticoid response element, CAM potentiated dexamethasone-increased promoter activity. In the adrenalectomized rats, CAM did not increase AGP levels in serum. These findings suggest that CAM may cause transcriptional induction of AGP, at least in part, via a glucocorticoid-mediated mechanism.
Assuntos
Antibacterianos/farmacologia , Claritromicina/farmacologia , Fígado/efeitos dos fármacos , Orosomucoide/metabolismo , Regiões 5' não Traduzidas/efeitos dos fármacos , Regiões 5' não Traduzidas/genética , Adrenalectomia , Animais , Células Cultivadas , Dexametasona/farmacologia , Interações Medicamentosas , Expressão Gênica/efeitos dos fármacos , Glucocorticoides/fisiologia , Hepatócitos/efeitos dos fármacos , Hepatócitos/metabolismo , Fígado/metabolismo , Masculino , Orosomucoide/genética , Estabilidade de RNA/efeitos dos fármacos , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Regulação para Cima/efeitos dos fármacosRESUMO
PURPOSE: Recent reports have shown that ischemic preconditioning induces strong protection against retinal damage by subsequent prolonged ischemia and that this protection is mediated by mechanisms involving the adenosine A1 receptor. This study was designed to evaluate quantitatively the effects of ischemic preconditioning on leukocyte-mediated reperfusion injury after transient retinal ischemia and to define the role of the adenosine A1 receptor in these effects. METHODS: Transient retinal ischemia was induced in male rats by temporary ligation of the optic nerve. Ischemic preconditioning (5 minutes of ischemia) was induced 24 hours before 60 minutes of ischemia. The adenosine A1 receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX) was administered intramuscularly immediately after ischemic preconditioning. Leukocyte behavior in the retina after 60 minutes of ischemia was evaluated in vivo with acridine orange digital fluorography. RESULTS: Ischemic preconditioning inhibited leukocyte rolling. The maximum number of rolling leukocytes was reduced to 3.0% at 12 hours after reperfusion (P < 0.01). Subsequent leukocyte accumulation was also decreased with ischemic preconditioning. The maximum number of accumulated leukocytes was reduced to 22.6% at 24 hours after reperfusion (P < 0.01). These inhibitory effects were suppressed by administration of DPCPX (P < 0.0001). The numbers of rolling leukocytes at 12 hours after reperfusion and accumulated leukocytes at 24 hours after reperfusion were 102.7% (NS) and 83.4% (P < 0.01), respectively, compared with the number without ischemic preconditioning. CONCLUSIONS: The present study demonstrates the inhibitory effects of ischemic preconditioning on leukocyte rolling and subsequent leukocyte accumulation during retinal ischemia-reperfusion injury. Furthermore, the adenosine A1 receptor may play an important role in these inhibitory effects.
Assuntos
Adenosina/análogos & derivados , Precondicionamento Isquêmico , Leucócitos/fisiologia , Traumatismo por Reperfusão/metabolismo , Doenças Retinianas/metabolismo , Vasos Retinianos/metabolismo , Laranja de Acridina , Adenosina/farmacologia , Animais , Angiofluoresceinografia , Corantes Fluorescentes , Injeções Intramusculares , Masculino , Modelos Animais , Antagonistas de Receptores Purinérgicos P1 , Ratos , Ratos Long-Evans , Receptores Purinérgicos P1/metabolismo , Xantinas/farmacologiaRESUMO
PURPOSE: Accumulating evidence suggests that platelets play an important role in ischemia-reperfusion injury. To fulfill that role, platelets flowing in the bloodstream would have to interact with retinal endothelial cells and to accumulate in the postischemic retina. This study was designed to investigate quantitatively platelet-endothelial interactions in postischemic retina after transient retinal ischemia. METHODS: Transient retinal ischemia was induced in Long-Evans rats for 60 minutes by temporal ligation of the optic nerve. Isolated platelet samples labeled with carboxyfluorescein diacetate succinimidyl ester were administered intravenously to recipient rats after various reperfusion periods. Platelet-endothelial interactions in postischemic retina were evaluated in vivo with a scanning laser ophthalmoscope. Anti-P-selectin monoclonal antibody (mAb) was administered 5 minutes before the injection of labeled platelets. P-selectin gene expression in the postischemic retina was studied by semiquantitative polymerase chain reaction. RESULTS: Under basal conditions, infused platelets showed minimal interactions with retinal endothelial cells. In contrast, postischemic retinas showed active platelet-endothelial interactions. Many platelets were observed rolling along and adhering to the major retinal veins. The number of rolling and adhering platelets reached a peak (555 +/- 65/mm per min and 25.8 +/- 3.2/mm(2)) 12 hours after reperfusion. However, the interactions between platelets and postischemic retinal endothelial cells were substantially inhibited by neutralizing P-selectin expressed on endothelial cells. In addition, P-selectin gene expression in postischemic retina corresponded with the time course of platelet-endothelial interactions during the reperfusion period. CONCLUSIONS: This study demonstrated that platelets actively interacted with retinal endothelial cells in the postischemic retina through P-selectin expressed on the retinal endothelial cells.
Assuntos
Plaquetas/metabolismo , Endotélio Vascular/metabolismo , Traumatismo por Reperfusão/metabolismo , Doenças Retinianas/metabolismo , Animais , Adesão Celular , Fluoresceínas , Corantes Fluorescentes , Expressão Gênica , Processamento de Imagem Assistida por Computador , Masculino , Oftalmoscopia , Selectina-P/genética , Reação em Cadeia da Polimerase , RNA Mensageiro/biossíntese , Ratos , Ratos Long-Evans , Vasos Retinianos/metabolismoRESUMO
Histamine has been shown to play an important role in the step of leukocyte rolling, the initial step to leukocyte infiltration into an inflamed region. We investigated the roles of histamine in the leukocyte recruitment during endotoxin-induced uveitis (EIU) in vivo using acridine orange digital fluorography. An injection of histamine into the vitreous cavity of a Lewis rat induced leukocyte rolling along the major retinal veins. In other experiments, EIU was induced in Lewis rats by footpad injection of lipopolysaccharide (LPS). Leukocyte rolling was also observed in the retinal veins of EIU rats. To block the histamine H1 receptor, diphenhydramine (DPH) was administered intraperitoneally 15 min before the LPS injection. DPH significantly inhibited leukocyte rolling along the major retinal veins of EIU rats, suppressing leukocyte infiltration into the vitreous cavity. The vasodilation in EIU was also significantly suppressed with DPH. Moreover, leukocyte infiltration into aqueous humor was significantly suppressed in DPH-treated rats. Although the inhibitory effects of DPH was less obvious at later time points, addition of DPH every 12 hr showed prolonged anti-inflammatory effects up to 48 hr after LPS injection. In contrast, protein leakage into the aqueous humor was not suppressed as much as leukocyte infiltration with DPH. These results suggest that histamine would play a pivotal role in leukocyte recruitment during EIU in rats. Blocking the histamine H1 receptor might help to prevent or minimize leukocyte infiltration in uveitis.
Assuntos
Difenidramina/farmacologia , Antagonistas dos Receptores Histamínicos H1/farmacologia , Leucócitos/efeitos dos fármacos , Infiltração de Neutrófilos/efeitos dos fármacos , Uveíte/imunologia , Animais , Humor Aquoso/citologia , Contagem de Células , Feminino , Expressão Gênica , Processamento de Imagem Assistida por Computador , Microscopia Confocal , Selectina-P/efeitos dos fármacos , Selectina-P/genética , Reação em Cadeia da Polimerase , RNA Mensageiro , Ratos , Ratos Endogâmicos Lew , Uveíte/induzido quimicamente , Corpo Vítreo/citologiaRESUMO
The aim of the present study was to investigate whether clinical doses of propofol and thiamylal affect oxygen free radical production and intracellular calcium concentration ([Ca2+]i) in the post-ischemic reperfused heart. Forty-eight rat hearts were perfused with a Langendorff system and loaded with Fura-2 / AM as a [Ca2+]i marker. The hearts were divided into 6 groups as follows (each group: n = 8); Group S (saline), Group TL (thiamylal 100 microM), Group TH (thiamylal 300 microM), Group I (Intralipid), Group PL (propofol 3 microM), and Group PH (propofol 10 microM). All hearts were initially perfused for 5 min as control aerobic perfusion. Afterwards, no-flow ischemia was induced for 15 min, followed by reperfusion for 20 min. The formation of hydroxyl radicals in the coronary effluent was measured with high performance liquid chromatography using salicylic acid. At the beginning of the ischemia and reperfusion periods, increases in systolic and diastolic [Ca2+]i were observed in all groups except Group TH. The high dose of thiamylal significantly suppressed this initial increase in cytosolic [Ca2+]i (Group S 1.30+/-0.15; Group TL 0.99+/-0.17; Group TH 0.70+/-0.09, at 1 min after reperfusion; systolic [Ca2+]i : p < 0.05). Total DHBAs in the coronary effluent of all groups increased significantly 1 min after reperfusion, however, there were no significant differences among the groups. Clinical doses of propofol had no significant effect on myocardial function and [Ca2+]i before and after ischemia, whereas thiamylal suppressed the increase in [Ca2+]i during ischemia and reperfusion. However, free radical formation during reperfusion was unaffected by thiamylal and propofol.
Assuntos
Sequestradores de Radicais Livres/farmacologia , Traumatismo por Reperfusão Miocárdica/tratamento farmacológico , Propofol/farmacologia , Tiamilal/farmacologia , Análise de Variância , Animais , Cálcio/metabolismo , Frequência Cardíaca , Hemodinâmica , Técnicas In Vitro , Contração Miocárdica , Traumatismo por Reperfusão Miocárdica/fisiopatologia , Ratos , Ratos WistarRESUMO
PURPOSE: This study was designed to investigate the suppressive effects of antithrombin (AT)III on inflammatory reactions during endotoxin-induced uveitis (EIU) in rats by studying leukocyte-endothelium interactions. METHODS: EIU was induced in Lewis rats by footpad injection of lipopolysaccharide (LPS). ATIII was administered immediately after or at 6 hours after LPS injection. Its suppressive effects on inflammatory leukocyte behavior were evaluated in vivo with acridine orange digital fluorography. Clinical signs of inflammation were also examined, and aqueous humor (AH) was collected to evaluate leukocyte infiltration and protein leakage. In a separate experiment, P-selectin mRNA expression was studied in the iris-ciliary body (ICB) and the retina. RESULTS: After treatment with ATIII, leukocyte rolling was substantially inhibited along the retinal veins, suppressing subsequent leukocyte infiltration into the vitreous cavity. Similarly, leukocyte infiltration and protein leakage into the AH were significantly reduced with ATIII treatment. The clinical grade of EIU was substantially lower in ATIII-treated rats. In addition, delayed administration of ATIII after EIU induction significantly attenuated these inflammatory reactions. The levels of P-selectin mRNA expression in both ICB and retina, which were upregulated after LPS injection, were substantially lower in the ATIII-treated rats. CONCLUSIONS: ATIII treatment significantly inhibited inflammatory reactions induced with LPS. Its suppressive effects on P-selectin expression could contribute to the attenuation of leukocyte infiltration, possibly by inhibiting leukocyte rolling. The current findings suggest that ATIII may have a role in the management of patients with uveitis.
Assuntos
Antitrombina III/farmacologia , Quimiotaxia de Leucócito/efeitos dos fármacos , Lipopolissacarídeos/toxicidade , Salmonella typhimurium , Inibidores de Serina Proteinase/farmacologia , Uveíte/prevenção & controle , Animais , Corpo Ciliar/metabolismo , Regulação para Baixo , Feminino , Expressão Gênica , Iris/metabolismo , Contagem de Leucócitos , Leucócitos/patologia , Selectina-P/genética , RNA Mensageiro/biossíntese , Ratos , Ratos Endogâmicos Lew , Retina/metabolismo , Uveíte/induzido quimicamente , Uveíte/metabolismo , Uveíte/patologia , Corpo Vítreo/patologiaRESUMO
We developed a method to measure membrane fluidity of living cancer cells in two- and three-dimensional cultures, and found that there was a close relationship between the membrane fluidity of cancer cells and their proliferative and infiltrative ability. Membrane fluidity is thus a promising indicator of the probability of cancer recurrence.
Assuntos
Carcinoma Hepatocelular/patologia , Membrana Celular/fisiologia , Neoplasias Hepáticas/patologia , Fluidez de Membrana , Bromodesoxiuridina , Divisão Celular , Polarização de Fluorescência , Humanos , Microscopia de Contraste de Fase , Modelos Biológicos , Células Tumorais CultivadasRESUMO
A postmortem case of an atypical form of dural graft associated Creutzfeldt-Jakob disease (CJD) is described. A 42 year old man developed progressive spastic paresis 163 months after a cadaveric dura mater graft. He presented with no myoclonus and very late occurrence of periodic synchronous discharges on EEG. The prion protein (PrP) gene was homozygous for methionine at the polymorphic codon 129. Neuropathological examination disclosed plaque-like PrP deposits with atypical distribution of synaptic PrP accumulations in the brain. This patient represents an atypical form of dural graft associated CJD characterised by unusual clinicopathological features.
Assuntos
Encéfalo/patologia , Síndrome de Creutzfeldt-Jakob/etiologia , Síndrome de Creutzfeldt-Jakob/patologia , Dura-Máter/patologia , Dura-Máter/transplante , Transplantes/efeitos adversos , Adulto , Humanos , Masculino , Neoplasias Hipofisárias/cirurgiaRESUMO
We have experienced anesthetic management for mitral valve replacement in a 48-year-old female with idiopathic hypereosinophilic syndrome. Preoperative examination showed mild biventricular dysfunction. Anesthesia was induced and maintained with meticulous administration of fentanyl and midazolam in 66% oxygen. Administration of dopamine, dobutamine and prostaglandin E1 contributed to reducing afterload and maintaining cardiac output. The operative and postoperative courses were uneventful. Hypereosinophilic syndrome is one of the identified causes of restrictive cardiomyopathy. Anesthesia for patients with hypereosinophilic syndrome must be carried out carefully, because heart or respiratory failure is the most dangerous complication. In patients with hypereosinophilic syndrome requiring general anesthesia, perioperative steroid cover is advisable. This may reduce or prevent serious lung complications.
Assuntos
Anestesia Geral , Síndrome Hipereosinofílica/complicações , Insuficiência da Valva Mitral/cirurgia , Assistência Perioperatória , Cardiomiopatia Restritiva/etiologia , Cardiomiopatia Restritiva/prevenção & controle , Feminino , Implante de Prótese de Valva Cardíaca , Humanos , Complicações Intraoperatórias , Metilprednisolona/administração & dosagem , Pessoa de Meia-Idade , Insuficiência da Valva Mitral/complicações , Complicações Pós-Operatórias , Insuficiência Respiratória/etiologia , Insuficiência Respiratória/prevenção & controleRESUMO
PURPOSE: Accumulating evidence has suggested that 17beta-estradiol exerts protective effects against ischemic damage in various organs. In addition, leukocytes that accumulate in postischemic tissues are thought to play a central role in ischemia-reperfusion injury. This study was designed to evaluate quantitatively the inhibitory effects of 17beta-estradiol on leukocyte accumulation during ischemia-reperfusion injury and on subsequent retinal damage after transient retinal ischemia. METHODS: Transient (60 minutes) retinal ischemia was induced in male rats by temporary ligation of the optic nerve. Thirty minutes before induction of ischemia, 17beta-estradiol (0.1 mg/kg) was administered intraperitoneally. At 6, 12, 24, and 48 hours after reperfusion, leukocyte accumulation in the retina was evaluated in vivo by means of acridine orange digital fluorography. Histologic and electroretinographic (ERG) studies were carried out to evaluate retinal damage. RESULTS: Treatment with 17beta-estradiol significantly inhibited postischemic leukocyte accumulation; the maximum number of accumulating leukocytes was reduced by 35.7% at 24 hours after reperfusion (P = 0.01). Histologic examination showed that administration of 17beta-estradiol significantly reduced retinal damage, which was most obvious in the inner retina, 168 hours after reperfusion (P = 0.0001). ERG studies at 12 and 168 hours after reperfusion showed that recovery of the b-wave amplitude was significantly improved with treatment of 17beta-estradiol (P = 0.023). CONCLUSIONS: The present study demonstrated the inhibitory effects of 17beta-estradiol on leukocyte accumulation and subsequent tissue injury during retinal ischemia-reperfusion injury.
Assuntos
Estradiol/farmacologia , Traumatismo por Reperfusão/prevenção & controle , Doenças Retinianas/prevenção & controle , Laranja de Acridina , Animais , Movimento Celular/efeitos dos fármacos , Eletrorretinografia , Estradiol/administração & dosagem , Fluorofotometria , Injeções Intraperitoneais , Contagem de Leucócitos , Leucócitos/fisiologia , Masculino , Ratos , Ratos Long-Evans , Traumatismo por Reperfusão/metabolismo , Doenças Retinianas/metabolismo , Vasos Retinianos/citologia , Vasos Retinianos/fisiologiaRESUMO
PURPOSE: The activity of protein kinase C (PKC), preferentially beta isoform of PKC, has been shown to be elevated in the diabetic retina. Recently, LY333531, a specific inhibitor of PKC-beta, has been reported to improve the decrease of retinal blood flow in early diabetes. Increased leukocyte entrapment has been suggested to be involved in blood flow disturbances in the early diabetic retina. This study was designed quantitatively to evaluate leukocyte entrapment in the retinal microcirculation of diabetic rats and the effect of LY333531 on leukocyte entrapment. METHODS: Diabetes was induced in male Long-Evans rats by intraperitoneal injection of streptozotocin (60 mg/kg). LY333531 (0.1, 1.0, or 10.0 mg/kg/d) was administered orally during a 4-week diabetic period. Leukocyte entrapment in the retinal microcirculation was quantitatively evaluated in vivo with acridine orange digital fluorography. RESULTS: The number of leukocytes trapped in the retinal microcirculation of diabetic rats (mean +/- SEM; 14.3 +/- 1.3 cells/mm2) was significantly increased, compared with nondiabetic control rats (7.5 +/- 0.3 cells/mm2; P < 0.0001). Oral administration of LY333531 significantly decreased the number of leukocytes trapped in the retinal microcirculation of diabetic rats (10.9 +/- 0.6, 11.3 +/- 0.7, and 10.4 +/- 0.4 cells/mm2 with LY333531 0.1, 1.0, and 10.0 mg/kg/d, respectively; P < 0.05). CONCLUSIONS: Treatment with LY333531 attenuated the increase of leukocyte entrapment in the retinal microcirculation during the period of early diabetes. This effect may contribute to the improvement of abnormal retinal blood flow in early diabetes with LY333531. LY333531 might have a therapeutic efficacy in preventing microcirculatory flow disturbances by trapped leukocytes in the early diabetic retina.
Assuntos
Adesão Celular/efeitos dos fármacos , Diabetes Mellitus Experimental/metabolismo , Retinopatia Diabética/metabolismo , Inibidores Enzimáticos/farmacologia , Indóis/farmacologia , Leucócitos/metabolismo , Maleimidas/farmacologia , Proteína Quinase C/antagonistas & inibidores , Vasos Retinianos/metabolismo , Laranja de Acridina , Administração Oral , Animais , Movimento Celular/efeitos dos fármacos , Diabetes Mellitus Experimental/patologia , Retinopatia Diabética/patologia , Inibidores Enzimáticos/administração & dosagem , Corantes Fluorescentes , Fluorofotometria , Indóis/administração & dosagem , Masculino , Maleimidas/administração & dosagem , Microcirculação , Ratos , Ratos Long-Evans , Vasos Retinianos/patologiaRESUMO
Diabetes is associated with increased neural damage after transient cerebral ischemia. Recently, leukocytes, which are thought to play a central role in ischemia-reperfusion injury, have been suggested to be involved in exacerbated damage after transient ischemia in diabetic animals. The present study was designed to clarify whether the anticipated worse outcome after transient cerebral ischemia in diabetic animals was due to augmented leukocyte-mediated neural injury. Using rats with streptozotocin-induced diabetes of 4-wk duration, we investigated leukocyte-endothelial cell interactions during reperfusion after a transient 60-min period of retinal ischemia. Unexpectedly, postischemic diabetic retina showed no active leukocyte-endothelial cell interactions during reperfusion. The maximal numbers of rolling and accumulating leukocytes in diabetic retina were reduced by 73.6 and 41.2%, respectively, compared with those in nondiabetic rats. In addition, neither preischemic insulin treatment of diabetic rats nor preischemic glucose infusion of nondiabetic rats significantly influenced leukocyte-endothelial cell interactions during reperfusion. The present study demonstrated that high blood glucose concentration before induction of ischemia did not exacerbate leukocyte involvement in the postischemic retinal injury. Furthermore, diabetic retina showed suppressed leukocyte-endothelial cells interactions after transient ischemia, perhaps due to an adaptive mechanism that developed during the period of induced diabetes.
Assuntos
Comunicação Celular/imunologia , Retinopatia Diabética/patologia , Endotélio Vascular/citologia , Ataque Isquêmico Transitório/patologia , Leucócitos/citologia , Animais , Glicemia , Movimento Celular/imunologia , Diabetes Mellitus Experimental/imunologia , Diabetes Mellitus Experimental/patologia , Olho/irrigação sanguínea , Ataque Isquêmico Transitório/imunologia , Masculino , Microcirculação/fisiologia , Microscopia de Vídeo , Ratos , Ratos Long-Evans , Traumatismo por Reperfusão/imunologia , Traumatismo por Reperfusão/patologia , Artéria Retiniana/imunologia , Artéria Retiniana/patologia , Estresse Mecânico , Vasoconstrição/fisiologia , Vasodilatação/fisiologiaRESUMO
Platelets and leukocytes are thought to play a leading role in the pathogenesis of many inflammatory conditions. To recruit flowing blood cells to the inflammatory region, it would be necessary for them to interact with vascular endothelial cells. Recently, many reports have indicated the resistance of spontaneous hypertensive rats (SHR) to endotoxic sepsis. Their resistance might be derived from suppressed interaction between these blood cells and endothelial cells. Therefore, SHR and age-matched Wistar-Kyoto rats (WKY) were induced with endotoxic sepsis by intravenous injection of lipopolysaccharide (LPS). At 4, 12, 24, and 48 hours after induction, leukocyte-endothelial interactions in the retina were evaluated in vivo with acridine orange digital fluorography. Fluorescently labeled platelets were also injected to investigate platelet-endothelial interactions in the retina in endotoxic sepsis. Leukocyte rolling in SHR after LPS injection was significantly suppressed; the maximum number of rolling leukocytes was reduced by 80.1% at 12 hours after LPS injection in SHR compared with WKY. Subsequent leukocyte infiltration into the vitreous cavity was significantly inhibited in SHR. Furthermore, platelet-endothelial interactions in the retina were also suppressed in SHR treated with LPS. The maximum numbers of rolling and adherent platelets were reduced by 59.5% and 62.6%, respectively, in SHR compared with WKY. In both strains, leukocyte- and platelet-endothelial interactions were substantially inhibited by the blocking of P-selectin. These suppressed interactions could contribute to the reduction of leukocyte- and platelet-mediated tissue injury in endotoxic sepsis in SHR, resulting in their resistance to endotoxemia.
Assuntos
Células Sanguíneas/citologia , Endotélio Vascular/citologia , Vasos Retinianos/citologia , Sepse/fisiopatologia , Animais , Anticorpos Monoclonais/farmacologia , Células Sanguíneas/efeitos dos fármacos , Plaquetas/citologia , Plaquetas/efeitos dos fármacos , Comunicação Celular , Endotélio Vascular/fisiopatologia , Endotoxemia , Hipertensão/fisiopatologia , Contagem de Leucócitos/efeitos dos fármacos , Leucócitos/citologia , Leucócitos/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , Masculino , Selectina-P/imunologia , Ratos , Ratos Endogâmicos SHR , Ratos Endogâmicos WKY , Vasos Retinianos/efeitos dos fármacos , Vasos Retinianos/fisiopatologia , Especificidade da EspécieRESUMO
PURPOSE: The purpose of this study is to confirm the reliability of scanning laser Doppler flowmetry in the rat retina and optic nervehead, and the validity of measuring changes of retinal blood flow in rats while breathing 100% oxygen. METHODS: We used a commercially available scanning laser Doppler flowmeter. To ascertain reliability, five consecutive and separate perfusion measurements of 12 eyes of 12 anesthetized pigmented rats were performed. To evaluate the validity of the system, repeated measurements were taken in anesthetized rats breathing room air or 100% oxygen. This series of measurements was repeated three times. RESULTS: The reliability coefficients of volume, flow, and velocity in the optic nervehead and the retina ranged from 0.80 to 0.83 and 0. 77 to 0.82, respectively. After the first exposure to oxygen, the measured values of volume, flow, and velocity were reduced by an average of 20.9-24.0%, 21.2-28.2%, and 19.5-24.5%, respectively. After the values returned to the basal condition, the second and third exposures to oxygen yielded measured values that were reduced by the same amounts as at the first exposure. CONCLUSIONS: Scanning laser Doppler flowmetry provided relatively good reliability in measurements of blood flow in the rat retina and optic nervehead. This study has indicated the possibility of applying this system to the rat retina.
Assuntos
Fluxometria por Laser-Doppler , Disco Óptico/irrigação sanguínea , Vasos Retinianos/fisiologia , Animais , Velocidade do Fluxo Sanguíneo/fisiologia , Masculino , Disco Óptico/fisiologia , Ratos , Ratos Long-Evans , Reprodutibilidade dos TestesRESUMO
Hematogenous dissemination is a significant short-coming of colorectal carcinoma treatment. To screen patients with high risk for such blood-borne metastasis, we previously developed a highly sensitive system for the detection of cytokeratin 20 (CK-20) mRNA in blood. For a more practical application, we improved this system by making it quantitative and capable of analyzing peripheral venous blood for the detection of perioperative changes in CK-20 mRNA. CK-20 mRNA was not always detected in the preoperative blood, even in patients in an advanced stage, but it was identified without fail in intra- and post-operative blood. In addition, more copies of CK-20 mRNA were observed in the intra-operative blood than in pre- and post-operative blood. This study suggests that analysis of perioperative changes may provide important information for the precise evaluation of hematogenous dissemination and of the effect of surgical maneuvers on recurrence.