Search

Using ultrasound, we investigated whether carotid parameters differed among subtypes of ischemic stroke and evaluated the usefulness of these parameters in discriminating among subtypes. Patients with ischemic stroke admitted to Nippon Medical School Hospital were consecutively recruited and grouped into 3 subtypes based on the Trial of Org 10172 in Acute Stroke Treatment (TOAST) classification: cardioembolism (group CE), large-artery atherosclerosis (group LAA), and small-vessel occlusion (group SVO). All subjects underwent carotid ultrasonography to determine maximum intima-media thickness (IMT), maximum systolic velocity (Vmax), minimum diastolic velocity (Vmin), mean velocity, and pulsatility index (PI). Carotid parameters that differed among subtypes were statistically identified. A total of 138 patients were enrolled. Intergroup comparisons revealed that the Vmin of the affected side was significantly lower in group LAA than in group SVO (mean±SD, 0.12±0.05 m/s vs 0.15±0.05 m/s; P=.02) and the Vmin of the mean of both sides was lower in group LAA than in group SVO (0.12±0.04 vs 0.16±0.05; P=.03). Multivariate analysis showed that the PI of the affected side was a useful adjunct to discriminate between groups SVO and LAA (odds ratio=2.94; P=.03, group SVO as control). Receiver operating characteristic curve analysis found that the Vmin of the affected side was the most useful parameter for discriminating between group SVO and group LAA. The PI and the Vmin of the affected side were found to differ among stroke subtypes, and thus these may be useful parameters for discriminating among ischemic stroke subtypes. [ABSTRACT FROM AUTHOR]

Examines the central nervous system action of JTP-2942, a novel thyrotropin-releasing hormone analogue. Role of JTP-2942 in the reduction of local cerebral blood flow and glucose utilization; Importance of JTP-2942 for improving cerebral circulation and metabolism; Measurement of local cerebral glucose utilization.

Introduction: Febrile neutropenia (FN) is an oncologic emergency requiring immediate empiric antibiotic therapy. Although carboplatin plus etoposide combination chemotherapy is associated with a relatively high frequency of FN, the risk factors are unclear. Hence, this retrospective study aimed to identify predictive markers of carboplatin/etoposide-induced FN., Methods: We conducted a retrospective cohort analysis of patients with previously untreated small-cell lung cancer (SCLC) who received combination chemotherapy with carboplatin (area under the concentration curve: 5 mg/mL·min, day 1) and etoposide (80 or 100 mg/m2, days 1-3) between July 2007 and June 2022. FN was assessed during the 21 days after initiation of carboplatin and etoposide therapy according to the Japanese Society of Medical Oncology's definition. Fisher's exact test for categorical variables and Mann-Whitney U test for continuous variables were used to compare the two groups. Statistical significance was set at p values <0.05. Explanatory variables with p values <0.05 in the univariate analysis were included in the multivariate logistic regression analysis., Results: Among the 176 eligible patients, the incidence of FN during the first cycle of chemotherapy was 25.0% (44/176). Multivariate analysis revealed that co-administration of proton pump inhibitors (PPIs) or potassium-competitive acid blockers (PCABs) and body mass index (BMI) were significantly associated with FN (p = 0.0035 and 0.0011, respectively). Patients with both co-administration of PPIs or PCABs and a BMI ≤22.509 kg/m2 presented with significantly higher frequencies of FN compared with the other patients (13/24 [54.2%] vs. 31/152 [20.4%] patients; odds ratio: 4.56, 95% confidence interval: 1.70-12.48; p = 0.00147)., Conclusion: Patients who received carboplatin plus etoposide for SCLC with co-administration of PPIs or PCABs and a BMI ≤22.509 kg/m2 more frequently present with FN than those without the two factors., (© 2023 S. Karger AG, Basel.)

Background/aim: Lenvatinib (LEN) has been approved as an oral tyrosine kinase inhibitor for advanced hepatocellular carcinoma (HCC). However, in some patients, LEN does not provide adequate therapeutic benefits. In this study, we examined the factors that affect the therapeutic response to LEN., Patients and Methods: This retrospective cohort study involved patients with HCC who received LEN therapy at Osaka Metropolitan University Hospital. We used the delivered dose intensity to body surface area ratio for 60 days (2M-DBR) as an index of the therapeutic response., Results: This study included 83 patients divided into two groups, the high 2M-DBR group (47 patients, 56.7%) and low 2M-DBR group (36 patients, 43.4%). Univariate analysis showed that Child-Pugh class, C-reactive protein, and prognostic nutrition index (PNI) were significant factors for high 2M-DBR. Furthermore, multivariate logistic regression analysis revealed that a PNI>39.15 was significantly associated with high 2M-DBR., Conclusion: A PNI cut-off value of less than 39.15 may indicate a poor response to LEN therapy. PNI, an easy, simple, and inexpensive tool, may be useful in identifying patients in need of early intervention., (Copyright © 2022 International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Introduction: Health care educators are challenged with helping clinicians develop competencies beyond their foundational training. In health care systems where continuing professional development is not integral to practice, clinicians may have few opportunities. We describe the design, implementation, and evaluation of a professional development program in patient safety for Japanese clinical educators to acquire simulation instructional skills and become Patient Safety Champions at their organizations., Methods: Mixed methods were used in a longitudinal pre/post study design. The Kirkpatrick evaluation model was used to evaluate outcomes of a workshop, overall program, on-site training experiences, and impact as Patient Safety Champions. Self-assessment data on skills and knowledge of patient safety, simulation instructional methods, interprofessional collaboration, and leadership were collected and analyzed., Results: Eighty-nine percent of participants facilitated on-site patient safety training within 6 months of workshop completion. Skills and knowledge improvement were observed immediately postworkshop in four categories: patient safety, simulation instructional methods, interprofessional collaboration and communication, and leadership as a patient safety champion. Skills and knowledge increased at 6 months after facilitation of on-site safety training. Program mean satisfaction scores ranged from 84% to 92%. Mean Patient Safety Champion in-facility evaluations were 4.2 to 4.7 on a 5-point scale., Discussion: High levels of knowledge, skill retention, and behavior change are attributed to goal setting, outcome-oriented pedagogy, and reflective sessions. The Patient Safety Champion model and experiential learning approach gave Japanese clinical educators in medicine, nursing, and pharmacy an opportunity to learn from each other in simulations reflecting the practice environment., Competing Interests: Disclosures: The authors declare no conflicts of interest. This project did not receive any grant from funding agencies in the public, commercial, or not-for-profit sectors., (Copyright © 2022 The Alliance for Continuing Education in the Health Professions, the Association for Hospital Medical Education, and the Society for Academic Continuing Medical Education.)

Although axitinib shows a good objective response rate and acceptable tolerability for advanced renal cell carcinoma, substantial differences in drug concentrations among individuals have hampered the reliable administration of the drug in a neoadjuvant setting. This study aimed to evaluate the relationship between axitinib pharmacokinetics and clinical efficacy in patients with advanced renal cell carcinoma treated in a neoadjuvant setting. We retrospectively reviewed 16 patients who underwent neoadjuvant axitinib treatment from prospective phase 2 study cohorts treated with axitinib and assessed whether the drug concentration was associated with clinical efficacy for primary tumors of advanced metastatic/oligometastatic clear cell renal cell carcinoma. Axitinib was administered orally at a starting dose of 5 mg twice daily for 2 months in principle before the operation, and the axitinib pharmacokinetics were examined. Best response, reduction rate, adverse events (AEs), and surgical complication were assessed. Four patients (25.0%) showed a partial response, and 12 (75.0%) had stable disease, with a mean reduction rate of 22.8%. No progressive disease was noted, and 9 of the 16 patinets (56.3%) showed downstaging. The trough level of axitinib significantly correlated with the objective response rate (P = .0052) and best tumor reduction (P = .0128). All AEs could be safely managed until termination of the dosing period. With respect to perioperative complications, grade 2 anemia was observed. Neoadjuvant axitinib treatment showed acceptable antitumor activity and safety profile for advanced renal cell carcinoma. The pharmacokinetics of neoadjuvant axitinib influenced the efficacy in patients with advanced renal cell carcinoma., (© 2019, The American College of Clinical Pharmacology.)

We investigated the relationship between axitinib pharmacogenetics and clinical efficacy/adverse events in advanced renal cell carcinoma (RCC) and established a model to predict clinical efficacy and adverse events using pharmacokinetic and gene polymorphisms related to drug metabolism and efflux in a phase II trial. We prospectively evaluated the area under the plasma concentration-time curve (AUC) of axitinib, objective response rate, and adverse events in 44 consecutive advanced RCC patients treated with axitinib. To establish a model for predicting clinical efficacy and adverse events, polymorphisms in genes including ABC transporters ( ABCB1 and ABCG2 ), UGT1A , and OR2B11 were analyzed by whole-exome sequencing, Sanger sequencing, and DNA microarray. To validate this prediction model, calculated AUC by 6 gene polymorphisms was compared with actual AUC in 16 additional consecutive patients prospectively. Actual AUC significantly correlated with the objective response rate ( P = 0.0002) and adverse events (hand-foot syndrome, P = 0.0055; and hypothyroidism, P = 0.0381). Calculated AUC significantly correlated with actual AUC ( P < 0.0001), and correctly predicted objective response rate ( P = 0.0044) as well as adverse events ( P = 0.0191 and 0.0082, respectively). In the validation study, calculated AUC prior to axitinib treatment precisely predicted actual AUC after axitinib treatment ( P = 0.0066). Our pharmacogenetics-based AUC prediction model may determine the optimal initial dose of axitinib, and thus facilitate better treatment of patients with advanced RCC., Competing Interests: CONFLICTS OF INTEREST None declared.

Objectives: To avoid the chelate formation between levofloxacin (LVFX) and aluminium hydroxide in gastrointestinal tract, an ethoxycarbonyl 1-ethyl hemiacetal ester of levofloxacin (LVFX-EHE) was synthesised as a prodrug., Methods: The effects of aluminium hydroxide on the bioavailability of LVFX following oral administration of LVFX-EHE were investigated in rats. Furthermore, the effects of aluminium hydroxide on small intestinal absorption of LVFX and LVFX-EHE when subjected to a hydrolysis experiment using in situ everted gut sac were investigated, and the minimal inhibitory concentrations (MICs) of LVFX and LVFX-EHE for various intestinal bacteria were measured., Key Findings: When LVFX-EHE was co-administered with and without aluminium hydroxide, the AUC 0-4 h values of LVFX hydrolysed from LVFX-EHE were similar to that of LVFX alone. In everted gut sac experiments, LVFX-EHE was efficiently absorbed even in the presence of aluminium ions after 1 h of incubation, whereas the absorption of LVFX decreased significantly in the presence of aluminium ions. MIC values of LVFX-EHE were far higher than LVFX., Conclusions: This study suggests the benefit of ethoxycarbonyl 1-ethyl hemiacetal esterification of the carboxyl group of new quinolone as a prodrug which is able to avoid chelate formation., (© 2016 Royal Pharmaceutical Society.)

Since 2005, the Japanese pharmacy education program has changed to a six year-system from a four year-system with the aim to help students obtain a higher level of clinical knowledge and skill regarding humanity and morality. Under the new pharmacy education system, the correct assessment of vital signs is observed in pharmacy practice so that pharmacists can sell "over the counter drugs (OTC)" safely. From this point of view, we started a pharmacy practice that recognizes a series of vital signs, i.e., blood pulse, blood pressure, respiratory sound, and electrocardiogram, using a physical figure subjecting to 4th-year students (n=142) . After the practice, a questionnaire was conducted in order to assess the satisfaction of the practice. The results suggested that students could successfully learn physical assessment by using physical figures (ratios more than grade 4: 60%). Students could also evaluate the necessity of physical assessment (ratios more than grade 4: 70%), suggesting the practice of using physical figures was accepted by most students.

Objective: Antidepressants have been recommended for the treatment of post-stroke depression (PSD). The purpose of this study was to evaluate the effect of fluvoxamine maleate, a selective serotonin re-uptake inhibitor (SSRI), on depressive state, sleep disturbance, and serum melatonin levels in patients with depressive state after cerebral infarction., Methods: Nineteen patients who were hospitalized for cerebral infarction and scored 40 points or higher on the Self Depression Scale (SDS) were enrolled in this study. Nine of the 19 patients received fluvoxamine as a treatment group and the other 10 patients were used as untreated controls. Before and after commencing the drug therapy, the patients were assessed by the SDS, Pittsburgh Sleep Quality Index (PSQI), Japan Stroke Scale for Depression (JSSD), and Japan Stroke Scale for Emotional Disturbance (JSSE), and their serum melatonin levels were measured. The control group underwent the same evaluations as the treatment group., Results: The SDS score improved in the treatment group at 1 week after the start of drug treatment, and in the control group at 1 and 2 weeks into the observation period. In the treatment group, the JSSD and PSQI scores improved and serum melatonin levels increased., Conclusion: The administration of fluvoxamine to patients with depressive state after cerebral infarction alleviated both the depressive state and sleep disturbances. Increased melatonin levels by the administration of fluvoxamine may contribute to improvement in sleep disturbance, one of the major symptoms of depression.

The six-year pharmacist education course has begun, and now first-year students receive clinical training. Interdisciplinary problem-solving capabilities covering chemistry, biology, molecular biology, pharmacology, pathology, and pharmacokinetics are necessary for new pharmacists. However, the conventional pharmaceutical science education was so separate from other fields that education for interdisciplinary cooperative capability was insufficient. This was especially true of elemental science courses, because they are not directly connected with clinical knowledge, and there is a problem of low student interest in those courses. As a result, students acquired only recall-level knowledge in clinical courses and their problem-solving capabilities in clinical treatment and drug development deteriorated. Therefore we offered a trial lecture aimed to help students recognize the important relationship between elemental science courses and clinical courses and increase their motivation to enroll in these courses. Specifically, the trial lecture covered cancer therapy, in reference to mechanisms of carcinogenesis, epidemiology, physiology of cancer, anticancer drugs with explanations of the mechanism of action of carcinogens, anticancer drugs, and molecular-targeted drugs from the viewpoints of organic chemistry and biochemistry by a specialized teacher. This paper reports on this experimental lecture with evaluations from students.

Objective: We investigated whether a correlation exists between insulin resistance and the severity of cerebral white matter lesions among non-diabetic patients with ischemic stroke., Methods: The subjects were 105 consecutive patients without diabetes who were hospitalized due to non-cardioembolic stroke. The insulin resistance was evaluated by a homeostasis model assessment of insulin resistance (HOMA-IR). The degrees of periventricular hyperintensity (PVH) and deep and subcortical white matter hyperintensity (DSWMH) were evaluated by the brain MRI. The HOMA-IR values >or=2.5 were indicative of the insulin resistance., Results: The presence of PVH and DSWMH were 86.7 and 83.8%, respectively. The ratio of insulin resistance increased with higher grades of PVH and DSWMH. The HOMA-IR level in grade 3 PVH was significantly higher than those in grades 0 and 1. The HOMA-IR level in grade 3 DSWMH was significantly higher than those in grades 0-2. Multiple linear regression analysis showed that HOMA-IR was significantly associated with PVH or DSWMH., Conclusion: It was found that insulin resistance correlated with white matter lesions among non-diabetic patients with non-cardiogenic ischemic stroke.

The objective of this study was to determine whether the long-term administration of an HMG-CoA reductase inhibitor, atorvastatin, confers protective effects against stroke events in stroke-prone spontaneously hypertensive rats (SHRSPs). Atorvastatin (2 mg/kg, 20 mg/kg) or vehicle was orally administered to 8-week-old SHRSPs for 11 weeks. The survival ratio and stroke incidence were calculated, and plasma lipids and plasma levels of asymmetric dimethylarginine (ADMA), a circulating endogenous competitive inhibitor of NO synthase, were measured after sacrifice. The effect of atorvastatin on local cerebral blood flow (l-CBF) was also determined in 13-week-old SHRSPs after treatment with 20 mg/kg atorvastatin daily for 5 weeks. The survival ratios at 19 weeks of age were 15, 30, and 50% in the vehicle, low-dose (2 mg/kg), and high-dose groups (20 mg/kg), respectively. The survival ratio was significantly higher in the high-dose group than in the vehicle group. The incidence of stroke was significantly lower in the high-dose group than in the vehicle group. The levels of ADMA were 0.81+/-0.18 (mean+/-S.D.), 0.62+/-0.09, and 0.61+/-0.06 micromol/l in the vehicle, low-dose, and high-dose groups, respectively. Atorvastatin administration significantly reduced the ADMA levels without affecting the levels of plasma lipids. The level of l-CBF tended to be higher in the treated group, but not to a significant extent. Thus, atorvastatin was determined to confer a protective effect against hypertension-based stroke. The data suggest that the efficacy of the statin for stroke protection may be partially involved in the improvement of endothelial function via NO production and reduction of ADMA. Statins may confer useful protection against not only atherosclerosis-based stroke, but also hypertension-based stroke.

Ischemic tolerance, the phenomenon where a sublethal ischemic preconditioning protects the brain against a subsequent lethal ischemia, has been widely studied. Studies have been done on cerebral blood flow levels prior to the lethal ischemia, but the hemodynamic pattern after global ischemia with ischemic preconditioning has not been reported. Sequential changes in regional cerebral blood flow (rCBF) in gerbil hippocampus after 5 min global ischemia with or without 2 min ischemic preconditioning were studied to determine if ischemic preconditioning affects rCBF. Four different treatments were given: (1) sham-operated, (2) 2 min ischemia, (3) non-preconditioned, and (4) preconditioned. Groups (1) and (2) (both groups n = 5) were given a 24-h recovery period and the rCBF was measured for baseline values. 24 h after sham-operation (3) and 2 min ischemia (4), gerbils were subjected to 5 min ischemia followed by 1 h, 6 h, 1-day or 7-day reperfusion periods (all groups n = 5). Although no regional difference was observed in the recovery pattern of rCBF, the values of rCBF were significantly higher in the preconditioned group throughout whole brain regions including hippocampus. These results indicate that ischemic preconditioning facilitated the recovery of rCBF after 5 min global ischemia. It needs further study to determine whether the protecting effects of preconditioning relate to the early recovery of rCBF or not. However, our results could be interpreted that the early recovery of rCBF may lead to benefits for cell survival in the CA1 neuron, probably facilitating other protecting mechanisms.

Previous studies have demonstrated that traumatic brain injury (TBI) increases the vulnerability of the brain to an acute episode of hypoxia-ischemia. The objective of the present study was to determine whether TBI alters the vulnerability of the brain to a delayed episode of ischemia and, if so, to identify contributing mechanisms. Sprague-Dawley rats were subjected to lateral fluid-percussion (FP) brain injury (n = 14) of moderate severity (2.3-2.5 atm), or sham-injury (n = 12). After recovery for 24 h, all animals underwent an 8-min episode of forebrain ischemia, followed by survival for 6 days. Ischemic damage in the hippocampus and cerebral cortex of the FP-injured hemisphere was compared to that in the contralateral hemisphere and to that in sham-injured animals. Remarkably, the number of surviving CA(1) neurons in the middle and lateral segments of the hippocampus in the FP-injured hemisphere was significantly greater than that in the contralateral hemisphere and sham-injured animals (p < 0.05). Likewise, in the cerebral cortex the number of damaged neurons tended to be lower in the FP-injured hemisphere than in the contralateral hemisphere. These results suggest that TBI decreased the vulnerability of the brain to a delayed episode of ischemia. To determine whether TBI triggers protective metabolic alterations, glycogen levels were measured in cerebral cortex and hippocampus in additional animals 24 h after FP-injury (n = 13) or sham-injury (n = 7). Cortical glycogen levels in the ipsilateral hemisphere increased to 12.9 +/- 6.4 mmol/kg (mean +/- SD), compared to 6.4 +/- 1.8 mmol/kg in the opposite hemisphere and 5.7 +/- 1.3 mmol/kg in sham-injured animals (p < 0.001). Similarly, in the hippocampus glycogen levels in the FP-injured hemisphere increased to 13.4 +/- 4.9 mmol/kg, compared to 8.1 +/- 2.4 mmol/kg in the contralateral hemisphere (p < 0.004) and 6.2 +/- 1.5 mmol/kg in sham-injured animals (p < 0.001). These results demonstrate that TBI triggers a marked accumulation of glycogen that may protect the brain during ischemia by serving as an endogenous source of metabolic energy.

1. Rat bilateral common carotid artery occlusion (BCAO) was used as a chronic cerebral hypoperfusion model. We observed autoradiographically the long-term changes in regional cerebral blood flow (rCBF) and regional cerebral glucose utilization (rCGU) after 2 days and 1, 4 and 8 weeks of BCAO and in controls. Regions evaluated included the cerebral cortex, white matter and basal ganglia. Pathological changes were also observed with Klüver-Barrera and haematoxylin-eosin staining. 2. After 2 days, rCBF was significantly reduced to 33-58% in the cortex, white matter and amygdala and similar reductions were observed after 1 week. 3. After 4 weeks, rCBF recovered; however, rCBF remained significantly reduced in the occipital cortex, white matter, globus pallidus and substantia nigra. 4. After 2 days, rCGU was mostly maintained but, after 1 week, rCGU was reduced significantly to 40-70% in the cortex, white matter, basal ganglia and thalamus. Four weeks later, these reductions were no longer seen. 5. Rarefaction of the white matter was observed from 1 week. 6. These results showed that the BCAO in rats is an appropriate model for chronic cerebral hypoperfusion and that uncoupling of rCBF and rCGU was observed from 2 days until 4 weeks in the white matter.

Cortical spreading depression (CSD) has previously been shown to induce tolerance to a subsequent episode of transient cerebral ischemia. The objective of the present study was to determine whether CSD also induces tolerance to permanent focal ischemia and, if so, whether tolerance may be mediated by alterations in cerebral blood flow (CBF). Sprague-Dawley rats were preconditioned by applying potassium chloride to one hemisphere for 2 hours, evoking 19 +/- 5 episodes of CSD (mean +/- SD, n = 19). Three days later, the middle cerebral artery (MCA) was permanently occluded using an intraluminal suture. In a subset of animals, laser Doppler blood flow (LDF) was monitored over the parietal cortex before and during the first 2 hours of MCA occlusion. Preconditioning with CSD reduced the hemispheric volume of infarction from 248 +/- 115 mm3 (n = 18) in sham-conditioned animals to 161 +/- 81 mm3 (n = 19, P< 0.02). Similarly, CSD reduced the neocortical volume of infarction from 126 +/- 82 mm3 to 60 +/- 61 mm3 (P < 0.01). Moreover, preconditioning with CSD significantly improved LDF during MCA occlusion from 21% +/- 7% (n = 9) of preischemic baseline in sham-conditioned animals to 29% +/- 9% (n = 7, P< 0.02). Preconditioning with CSD therefore preserved relative levels of CBF during focal ischemia and reduced the extent of infarction resulting from permanent MCA occlusion. To determine whether CSD may have altered preischemic baseline CBF, [14 C]iodoantipyrine was used in additional animals to measure CBF 3 days after CSD conditioning or sham conditioning. CSD, but not sham conditioning, significantly reduced baseline CBF in the ipsilateral neocortex to values 67% to 75% of those in the contralateral cortex. Therefore, CSD causes a long-lasting decrease in baseline CBF that is most likely related to a reduction in metabolic rate. A reduction in the rate of metabolism may contribute to the induction of tolerance to ischemia after preconditioning with CSD.

Purpose: We developed a new photostimulation system for electroretinograms (ERGs). This apparatus is capable of varying the transients during stimulus on and off (up-slope and down-slope times, respectively) as well as the stimulus and background intensities. We applied this system to evaluate the photopic ERG in human eyes., Cases and Methods: Seven normal volunteers and a 43-year-old patient with pre-proliferative diabetic retinopathy served as test subjects. We recorded ERGs with relatively long intervals of up-slope and down-slope times for photostimulation., Results: As the up-slope and down-slope times were increased, the amplitude was reduced and the peak latency was prolonged for both the on and off responses. When the normal subjects' retinas were stimulated with a 341-msec up-slope time and a 34.1-msec down-slope time, the ERG waves showed complete flattening in both on and off responses. The ERGs of a patient with pre-proliferative diabetic retinopathy showed much longer peak latency than that of the normal volunteers as the up-slope time was prolonged. When the patient's retina was stimulated for a prolonged up-slope time, the average of the amplitude of ERG waves was similar to that of the normal volunteers., Conclusion: These results suggest that this new type of photostimulation might potentially uncover some novel aspect of clinical ERGs and might be useful for testing retinal functions of patients with diabetic retinopathy.

Purpose: Contraction of the scar tissue during corneal wound healing changes the shape of the cornea and corneal refraction. In a previous study, it was found that corneal epithelial cells secrete the factor that stimulates collagen gel contraction by keratocytes in vitro. The purpose of the present study was to purify and identify the contraction-stimulating factor derived from corneal epithelial cells., Methods: The cultured medium of rabbit corneal epithelial cells was collected and used as an epithelial cell-conditioned medium (ECCM). Subcultured rabbit keratocytes were embedded in a collagen gel, and collagen gel contraction was investigated. The contraction-stimulating factor in the ECCM was purified through acetone precipitation, affinity chromatography (heparin Sepharose), gel filtration, and reversed-phase chromatography. The amino acid sequence of a contraction-stimulating factor was analyzed., Results: Collagen gel contraction by keratocytes was enhanced by the addition of ECCM in a dose-dependent manner. The amino acids sequence of the contraction-stimulating factor was homologous to a 32-kDa glycoprotein, a secreted protein that is acidic and rich in cysteine (SPARC). Western blot analysis confirmed that SPARC was contained in the ECCM. Collagen gel contraction by keratocytes was enhanced by the addition of purified SPARC in a dose-dependent manner. SPARC was found in the basal layer of the migrating epithelium and activated keratocytes adjacent to the wound 3 days and 1 week after perforating injury in rabbit corneas., Conclusions: Epithelial cells secrete SPARC, which modulates the contraction of scar tissue in the corneal stroma.

Clinical perimetry has made a remarkable advance in the last 40 years and many new automated perimeters and sophisticated techniques are now available. We developed a Central Field Screener, O-II, an automated perimeter, ATS-85, and a central critical fusion frequency (CFF)-meter. First, reappraisal of the clinical significance of manual kinetic perimetry was made in comparison with automated light threshold perimetry in the diagnosis of retinitis pigmentosa and chiasmal tumors. We concluded that manual kinetic perimetry was the basis of clinical perimetry and superior to automated static light threshold perimetry in some cases. Secondly, automated static flicker perimetry was performed with Octopus 1-2-3 and a software package developed by us. We found that automated static flicker perimetry was far superior to automated static light threshold perimetry in the detection of early changes of glaucomatous field defects, especially in cases of immature senile cataract. We also confirmed that measurement of CFF was essential in the diagnosis of optic neuritis and diseases of chiasmal syndrome. Dissociation of central vision and central CFF was observed in many cases of optic neuritis and inverse dissociation of central vision and central CFF was seen in some cases of Leber's disease.

To understand the mechanism of the contraction of cicatricial tissues after corneal stromal wounding, we investigated the effect of corneal epithelial cells on the collagen gel contraction by keratocytes. Subcultured rabbit keratocytes were embedded in type I collagen gel and cultured. Rabbit corneal epithelial cells were cultured separately and the culture medium was collected and used as an epithelial cell-conditioned medium (ECCM). The collagen gel contraction by keratocytes was determined by measuring the diameters of the collagen disks once a day for 5 days. In the presence of fetal calf serum, the diameter of the collagen gel decreased in linear proportion to the number of keratocytes, but in inverse proportion to the concentration of collagen. The addition of the ECCM enhanced the collagen gel contraction by keratocytes in a dose-dependent manner. This stimulatory activity was found in the ECCM obtained from 3- to 7-day cultures of epithelial cells. These findings suggest that corneal epithelial cells secrete factor(s) that stimulate the collagen gel contraction by keratocytes.

To study the mechanism of collagen degradation by keratocytes, we developed the new in vitro model in which keratocytes were cultured three-dimensionally in a collagen matrix. Subcultured rabbit keratocytes were embedded in a type I collagen matrix and cultured in serum-free medium. Collagenolytic activity of the cells was determined by measuring the amount of hydroxyproline released into the medium from degraded collagen. Activities of collagenase in the medium were also measured, using collagen labeled with fluorescein isothiocyanate as a substrate. The presence of plasminogen was required for collagen degradation by keratocytes. In the presence of plasminogen, the amount of collagen degradation depended on both the cultivation period and the number of cells. The addition of interleukin-1 (IL-1) stimulated the collagen degradation in a dose-dependent manner. This stimulatory effect of IL-1 was completely inhibited by the addition of IL-1 receptor antagonist (IL-1ra). Collagenase activity in the medium was stimulated by the addition of IL-1, and IL-1ra antagonized this stimulatory effect. These findings indicate that our present model may be useful for investigating the mechanism of collagen degradation by keratocytes.

Objective: Although the pathology of chronic hypoperfusion in rat following bilateral carotid artery occlusion has been documented, long term changes in cerebral blood flow and metabolism have not been reported. In this study regional cerebral blood flow (rCBF) and regional glucose utilization (rCGU) were analyzed by autoradiographical methods, pathological observation of the brains was also conducted., Methods: Male Wistar rats aged 12 weeks were anesthetized and the bilateral carotid arteries were occluded. Physiological parameters, ABG, MABP, and rectal temperature were measured before and through occlusion. After 2 days, 1, 4, and 8 weeks and controls of rCBF (Sakurada) and rCGU. (Sokoloff) were measured (n = 6). Evaluated regions included frontal cortex (Fcor), parietal cortex (Pcor), temporal cortex (Tcor), occipital cortex (Ocor), genu corpus callosum (gCC), corpus callosum (CC), splenium corpus callosum (sCC), caudate putamen (CPu), globus pallidus (GP), internal capsule (IC), thalamus (Thal), hippocampus CA 1 (CA 1), hypothalamus (HypoTh), amygdal (Amygd), and substantia nigra (SNR). In separate animals (n = 3) tissue sections were stained using Klüver-Barrera (KB) and Hematoxylin-Eosin (HE), and pathological changes were observed., Results: After 2 days the rCBF values were significantly reduced to 33-58% of control values in the Fcor. Pcor, Tcor, Ocor, gCC, CC, sCC, CPu, GP, JC, and Amygd. The reductions were observed from the 2nd day to the 1st week. From the 1st week to the 4th week values began to recover to control levels. However, after 4 weeks they were still significantly reduced in the Ocor, gCC, CC, sCC, GP, IC, and SNR (51-63%). After 8 weeks, the rCBF values in the areas except white matter, CPu, GP, and Ocor, etc., recovered to approximately 90% of control levels. However, in the Ocor, gCC, CC, sCC, CPu, GP, and IC, they were still 70-89% of control levels. After 2 days the rCGU values were reduced to 56-95% (except Amygd) in the measured regions, although not as reduced as the rCBF levels. From the 2nd day to the 1st week, the rCGU values were reduced further and after 1 week the rCGU values were significantly reduced to 39-69% in the Fcor, Pcor, Tcor, Ocor, gCC, CC, sCC, CPu, GP, IC (Lt), Thal (Rt), Amygd, and SNR From the 1st week to the 4th week, the values began to recover to control levels. After 4 weeks, rCGU improved to approximately 90-100% of controls, and remained at that level through 8 weeks occlusion. Rarefaction of the myelinated fibers was observed in the white matter from the 1st week to the 4th week in the KB stained sections, while little change in cortex was observed throughout 1st to 8th week., Conclusion: In this chronic hypoperfusion model rCBF remained depressed after 8 weeks in the Ocor, white matter, and basal ganglia, and rarefaction of the white matter was observed. These results indicate that this model is suitable for the study of chronic cerebral hypoperfusion.

To investigate the role of Interleukin-1 (IL-1) in the pathobiology of the cornea, we measured IL-1 beta concentration in tear fluid samples from patients with corneal disease. Twenty patients with unilateral corneal disease were included in the study. Tear fluid samples were collected during the active stages of the disease and following resolution. The fellow (unaffected) eyes served as controls. The concentration of IL-1 beta in the tear fluid samples was measured using a sandwich ELISA method. IL-1 beta was detected in tear fluid from five eyes (three eyes with chemical burns, one with a Pseudomonas aeruginosa corneal ulcer, and one with a peripheral corneal ulcer) at concentrations between 29 and 218 pg/mL. IL-1 beta was not detected in tear fluid from the remaining 15 affected eyes, nor from the control eyes. The detection of IL-1 beta in tear fluid correlated with limbal conjunctival involvement, but did not correlate with the type of disease, size of epithelial defect, or degree of stromal involvement. IL-1 beta in the tear fluid may be one of the factors modifying the complex inflammatory process of the anterior ocular surface.

Purpose: To understand the biological activity of entactin, a component of the basement membrane of the corneal epithelium, we investigated the ability of rabbit corneal epithelial cells to attach to an entactin matrix and the effect of entactin on the cells' attachment to other corneal basement proteins., Methods: Multiwell plastic plates were coated with bovine serum albumin (BSA), alone or with BSA and entactin, laminin, fibronectin or collagen type IV. Cultured rabbit corneal epithelial cells were seeded on the plates. After incubation (usually 90 min), the cells were fixed and stained with 1% crystal violet. The number of attached cells was counted under a light microscope., Results: The numbers of attached cells increased in proportion to both the incubation period and the concentration of entactin coated. Furthermore, the number of cells attached to the entactin-coated plate was greater than the number attached to the BSA-coated plate for each incubation period (30 to 120 min). Likewise, when laminin-coated plates were treated with entactin, the number of the attached cells increased in proportion to the concentration of entactin. However, entactin did not affect the cellular attachment of fibronectin or type IV collagen. Cellular attachment to entactin was partially inhibited by the cells' preincubation with the synthetic peptide (GRGDSP)., Conclusions: The present results showed that cultured corneal epithelial cells adhere to entactin and that entactin stimulated the attachment of these cells to the laminin matrix. These findings suggest that entactin plays a specific role in maintaining the normal integrity of the corneal epithelium.

1. Malignant or precocious stroke-prone spontaneously hypertensive rats (M-SHRSP) showed hypertensive ocular fundus changes with severe hypertension, but various anti-hypertensive drugs given over a proper period improved funduscopic findings. 2. We treated a M-SHRSP with SQ29,852 (an angiotensin converting enzyme inhibitor, ACEI) or manidipine (a calcium antagonist) and observed hypertensive vascular changes in the fundus. 3. The M-SHRSP treated with anti-hypertensive drugs lived longer and the hypertensive fundic changes improved in both groups. But there were some differences of histochemical staining reaction on the endothelial cell-surface and mucopolysaccharides accumulations between the treated group of ACEI and the group treated with the calcium antagonist. 4. In the treated group with calcium antagonist, the anionic ion functions of the endothelial cell-surface were impaired and organic retinal or choroidal damages appeared to have deteriorated.

To understand the mechanism of corneal stromal wound contraction, we investigated the effect of corneal epithelial cells on the collagen gel contraction by keratocytes. Subcultured rabbit keratocytes embedded in type I collagen gel were cultured. Rabbit corneal epithelial cells were also cultured and the cultured medium was collected and used as an epithelial cell conditioned medium (ECCM). The collagen gel contraction by keratocytes was estimated by measuring the diameter of the collagen disc once a day for 5 days. The diameter of the collagen gel decreased in proportion to the number of keratocytes in the presence of fetal calf serum (FCS), but it decreased in inverse proportion to the concentration of collagen. The collagen gel contraction by keratocytes was enhanced by the addition of ECCM in a dose dependent manner. This stimulatory activity was found in ECCM obtained from 3 to 7 day cultivation of corneal epithelial cells. These findings demonstrated that keratocytes contracted collagen gel in the presence of FCS. It also appeared that corneal epithelial cells secrete a factor or factors that stimulate the collagen gel contraction by keratocytes.

Interleukin-6 (IL-6) has been shown to promote the attachment of rabbit corneal epithelial cells to fibronectin-coated substratum and ex vivo migration of the cells on the corneal stroma. To examine whether IL-6 promotes cell attachment through up-regulation of expression of integrin alpha 5 beta 1, i.e., the major cell surface fibronectin receptor, we quantified the levels of both alpha 5 and beta 1 subunit transcripts by reverse transcription-polymerase chain reaction in cultured rabbit corneal epithelial cells pretreated with various concentrations of IL-6. The levels of both alpha 5 and beta 1 mRNAs were dose-dependently elevated by IL-6, attaining 1.5- and 1.8-fold increases, respectively, at 10 ng/ml. The stimulatory effect of IL-6 was transient; the levels of both subunit mRNAs reached a maximum 1 h after the addition of IL-6 and returned to the basal levels after 6 h. The IL-6-induced up-regulation of integrin alpha 5 and beta 1 mRNAs was also confirmed by Northern blot analysis. These results indicate that the increased attachment of corneal epithelial cells to fibronectin and enhanced ex vivo migration on corneal stroma by IL-6 is, at least in part, due to the temporal up-regulation of integrin alpha 5 beta 1 expression in corneal epithelial cells.

We wished to determine whether hyaluronan would affect the attachment of epithelial cells to extracellular matrix proteins. Multiwell tissue culture plates were coated with human plasma fibronectin, laminin, or collagen type IV (0.01-10.0 micrograms/ml). Single-cell suspensions of rabbit corneal epithelial cells were placed in the wells, and after 45 minutes incubation the cells adhering to the matrix proteins were stained and counted. Cells attached to all three types of proteins. Preincubation of the matrix proteins with hyaluronan (0.1-1.0 mg/ml) significantly increased the number of cells attached to the fibronectin matrix, but it did not increase the numbers of cells attached to laminin or collagen type IV. Hyaluronidase inhibited this stimulatory effect. Glycosaminoglcyans other than hyaluronan (chondroitin sulfate, keratan sulfate, or heparan sulfate) failed to increase the numbers of attached cells. Treatment of the fibronectin matrix with monoclonal antibodies against the cell-binding domain of fibronectin (FN12-8 or FN30-8, 0.03-0.3 mg/ml, for 1 hour), before or after hyaluronan treatment, significantly decreased the numbers of attached cells. Monoclonal antibody against the fibrin- and heparin-binding domain at the N-terminal (FN9-1), however, significantly decreased the number of attached cells only when this antibody treatment preceded the hyaluronan treatment. Preincubation of the cells with hyaluronan had no effect; preincubation with GRGDSP (1 mg/ml), a synthetic peptide that blocks the cell surface receptor for fibronectin, significantly decreased cell attachment whether the fibronectin matrix was treated with hyaluronan or not. Further studies demonstrated that monoclonal antibody against the fibrin- and heparin-binding domain at the N-terminal of plasma fibronectin prevented radiolabeled hyaluronan from binding to fibronectin; likewise, the isolated N-terminal fragment, coupled with Sepharose 4B, bound to hyaluronan in columns. We conclude that hyaluronan binds to a fibrin- and heparin-binding domain at the N-terminal of plasma fibronectin and facilitates the attachment of epithelial cells.

We investigated the combined effects of hyaluronan and fibronectin on corneal epithelial wound closure in vivo. When fibronectin eye drops (1 mg/ml) were administered for the first 4 hours after chemical cautery and hyaluronan eye drops (1 mg/ml) were administered for following 32 hours, the mean healing rate was significantly higher than in control eyes treated with phosphate buffered saline instead of fibronectin or with initial fibronectin alone. These results demonstrated that hyaluronan and fibronectin have a synergistic effect, with fibronectin pretreatment augmenting hyaluronan-stimulated corneal epithelial wound closure.

The influence of target blurring and simulated opacity of the ocular media on the sensitivity of the central visual field in quantitative static perimetry was studied using a SARGON program of the automated perimeter Octopus 201. Ten eyes of 10 normal subjects aged from 21 to 35 were examined with target sizes 1, 3, and 5; 4 spherical plus lenses of +1.0D, +2.0D, +3.0D, and +5.0D; 5 kinds of occlusion diffusers which corresponded to the visual acuities of 1.0, 0.8, 0.6, 0.4, and 0.1; and 4 neutral density filters of 0.1, 0.3, 0.6, and 1.0 log units. The studies suggested that there was little difference between the sensitivity for target size 1 and that for target size 3 in spite of slight blurring of the target and simulated opacity of the ocular media. We concluded that target size 1 was as useful as target size 3 in quantitative static perimetry.

The slowed healing rates observed by some investigators may be caused by vehicles or preservatives in the antimicrobials preparations tested. To determine whether antimicrobials directly inhibit corneal epithelial wound healing, we cultured blocks of the rabbit cornea in media containing various concentrations of antibiotics or antimicrobials (at 1, 10, or 100 micrograms/ml); after 24 hours, we measured the distance of epithelium that had migrated down the side of each block. The higher concentrations of fluoroquinolones (ofloxacin; 74 +/- 5.8% of control at 100 micrograms/ml, p < 0.05, ciprofloxacin; 4.4 +/- 1.5% of control at 100 micrograms/ml, p < 0.01, or norfloxacin; 71 +/- 7.0% at 10 mu g/ml, p < 0.01, and 1.5 +/- 0.4% of control at 100 mu g/ml, p < 0.01) and the highest concentrations of peptides (polymyxin B; 64 +/- 3.0% of control at 100 micrograms/ml, p < 0.01, or colistin; 67 +/- 5.7% of control at 100 micrograms/ml, p < 0.01) or fosfomycin (79 +/- 6.2% of control at 100 micrograms/ml, p < 0.05) had an inhibitory effect on corneal epithelial migration. Among aminoglycosides tested, sisomicin (85 +/- 10.0% of control, not significant), dibekacin (76 +/- 11.6% of control, p < 0.05) and streptomycin (77 +/- 9.4% of control, not significant) were inhibitory at 100 micrograms/ml, but tobramycin had no effect. Penicillins (aspoxicillin, sulbenicillin or ampicillin), cephalosporins (cefmenoxime or cefminox), oxytetracycline, erythromycin and chloramphenicol did not affect epithelial migration at all. These results demonstrate that some antimicrobials are inhibitory at high concentrations, but penicillins, cephalosporins, oxytetracycline, erythromycin or chloramphenicol has no inhibitory effect on corneal epithelial migration.

The role of plasminogen activator (PA) in the migration of corneal reepithelialization was studied. Rabbit corneal blocks were cultured, and both the extent of epithelial migration over the exposed corneal stroma and the activity of PA released into the culture media were measured. A significant, direct correlation between epithelial migration and PA activity in the medium was observed, even when the migration was stimulated by fibronectin or EGF, or was inhibited by cytochalasin B or cycloheximide. Zymography confirmed that the PA released into the culture medium was of the urokinase type (u-PA). Immunohistochemical studies showed that u-PA and plasmin(ogen) were present at the leading edge of the migrating epithelium. Studies of corneal cell cultures indicated that epithelial cells rather than endothelial cells or fibroblasts were the source of the u-PA. The addition of antihuman u-PA IgG or protease inhibitors retarded the migration of the corneal epithelium in a dose-dependent manner, indicating that u-PA activity is essential for the migration of the corneal epithelium. These findings suggest that the migration of corneal epithelial cells requires not only cell attachment to the extracellular matrix through the fibronectin but also degradation of the fibronectin by the release of cellular u-PA.

The peak latency in 30-Hz flicker electroretinography (ERG) was determined in patients with diabetic retinopathy, using a newly developed contact lens electrode with built-in light-emitting diode. A good correlation was obtained between the peak latency of flicker ERG and that of oscillatory potentials of flash ERG. The peak latency of the flicker ERG was only slightly prolonged in background retinopathy, but markedly prolonged in proliferative retinopathy. The stage of pre-proliferative retinopathy with normal peak latency in flicker ERG may be a compensatory state of the retina, which is the most appropriate stage for laser treatment of diabetic retinopathy.

We investigated the effect of interleukin 6 (IL-6) on the migration of rabbit corneal epithelium in vitro and on the attachment of dissociated corneal epithelial cells to a fibronectin matrix. When corneal blocks were cultured with IL-6 for 24 hours, the length of the path of epithelial migration over exposed corneal stroma increased significantly (p less than 0.005 at the concentration of 10 ng/ml) in proportion to the concentrations of IL-6 (0.1-10.0 ng/ml). The addition of antiserum against fibronectin or of GRGDSP abolished the stimulatory effect of IL-6 on epithelial migration. When corneal epithelial cells were cultured with various concentrations of IL-6, suspended, and plated on wells coated with fibronectin (10 micrograms/ml), the number of cells attached to the wells increased in a dose-dependent manner. The presence of antibody against fibronectin or of GRGDSP during the attachment assay decreased the number of cells attached to the fibronectin matrix, regardless of the fact that the cells had been cultured with IL-6 or not. IL-6 stimulated the attachment of corneal epithelial cells to collagen type IV and to laminin matrices. However, the presence of GRGDSP did not affect the cell attachment to collagen type IV and to laminin. These findings strongly indicate that IL-6 stimulates epithelial migration in the cornea by a fibronectin-dependent mechanism, presumably the increased expression of fibronectin receptors.