Calcium-activated CL- current is enhanced by acidosis and contributes to the shortening of action potential duration in rabbit ventricular myocytes

Ca2+-activated Cl- current (I(Cl(Ca))) is activated by Ca2+ transient via Ca2+-induced Ca2+ release from sarcoplasmic reticulum in cardiac myocytes and is supposed to play an important role in the repolarization of action potential. It is not well understood, however, how I(Cl(Ca)) is modulated to affect action potential in normal or pathological conditions. In this study we examined the effects of external acidosis on I(Cl(Ca)) and action potential. A whole-cell patch clamp was performed to record action potential and I(Cl(Ca)), using isolated rabbit ventricular myocytes. In the standard solution at pH 7.4, action potential duration (APD) was markedly prolonged by lowering the extracellular Cl- concentration ([Cl-](o)) or by applying an anion channel blocker, 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS). In the low pH solution at 6.4, APD was markedly shortened and the amplitude of I(Cl(Ca)) was increased at all membrane potentials. At pH 6.4, the apparent steady-state inactivation curves of I(Cl(Ca)) were shifted to more positive potentials compared with those at pH 7.4, but no change in inactivation occurred at a holding potential of -60 mV. The apparent activation curves were not changed between the two sets of conditions. When I(Cl(Ca)) was inhibited at low pH, early afterdepolarizations and triggered activities were induced. The amplitude of I(Cl(Ca)) was suggested to be enhanced by the external acidosis, which may have prevented the induction of early afterdepolarization or triggered activity.     

Treatment of meralgia paresthetica with a deep inferior epigastric perforator adiposal flap: case report

A case of a 40-year-old female with meralgia paresthetica after malignant tumor resection in the right inguinal region is reported. Traditionally, meralgia paresthetica is treated with neurotransection or neurolysis. The therapeutic strategy using neurolysis, and the use of a deep inferior epigastric perforator adiposal flap wrapping as a prophylactic procedure against reentrapment is discussed.     

Expression and function of receptors for advanced glycation end products in bovine corneal endothelial cells

PURPOSE: The corneal endothelium is a target of the aging process. This study was undertaken to reveal the relationship between corneal endothelial cell (CEC) death and the accumulation of advanced glycation end products (AGEs), by investigating the possible mechanism of accumulation of AGE in CECs and its effects on CEC death. METHODS: First, the in vivo expression of the receptor was investigated for AGE (RAGE) and galectin-3, both receptors for AGE, at both the mRNA and protein levels. Second, AGEs were added to the culture media of the cultured CECs, and the uptake of AGEs, the generation of reactive oxygen species, and the induction of apoptosis were investigated. RESULTS: Immunohistochemistry and RT-PCR demonstrated that both RAGE and galectin-3 were expressed in bovine CECs. After administration of AGE-modified bovine serum albumin to the culture medium, uptake of AGE was observed in the cytoplasm of the cultured bovine CECs. In addition, with increasing concentration of AGEs, the generation of reactive oxygen and the number of apoptotic cells also increased. CONCLUSIONS: These results show that the accumulation of AGEs in CECs induced apoptosis, in part, by increasing cellular oxidative stress. The accumulation of AGEs in the CECs of elderly patients may be involved in the loss of CECs during the aging process.     

Pulmonary gene delivery by chitosan-pDNA complex powder prepared by a supercritical carbon dioxide process

Chitosan-plasmid DNA (pDNA) complex powders as a pulmonary gene delivery system were prepared with a supercritical carbon dioxide (CO(2)) process and their in vivo activity was evaluated. The powders with mannitol as a carrier were prepared by dispersing aqueous solutions of a luciferase expression plasmid driven by the cytomegalovirus promoter (pCMV-Luc) with or without chitosan as a cationic vector in a supercritical CO(2)/ethanol admixture. The supercritical CO(2) process with a V-shaped nozzle successfully produced chitosan-pDNA powders. The addition of chitosan suppressed the degradation of pCMV-Luc during the supercritical CO(2) process and increased the yield of powders. The luciferase activity in mouse lung was evaluated after pulmonary administration of the powders or pCMV-Luc solutions. The chitosan-pDNA powders increased the luciferase activity in mouse lung compared with pCMV-Luc powders without chitosan or pCMV-Luc solutions with or without chitosan. The chitosan-pDNA powder with an N/P ratio = 5 increased the luciferase activity to 2700% of that of the pCMV-Luc solution. These results suggest that gene powder with chitosan is a useful pulmonary gene delivery system.     

An autopsy case of bilateral tension pneumothorax after acupuncture

Acupuncture is one of the most popular complementary therapies in the world. Pneumothorax due to perforation of the lungs by needle insertion is one of the most common and serious complications of acupuncture treatment. Although there have been several case studies of pneumothorax induced by acupuncture, as far as we know there have been no reports on the pathological findings of autopsy cases.In this report, we describe the pathological findings of an autopsy case of bilateral tension pneumothorax after acupuncture. The patient suffered dyspnea and chest pain soon the completion of an acupuncture treatment, and died 90 min later. Several ecchymoses were macroscopically observed on the parietal pleura in the left and right thoracic cavity, suggesting that needles were inserted into the thoracic cavity and that the lungs were perforated. The many black spots we observed on the parietal pleura along the vertebral column microscopically consisted of a number of dust-like black pigments and macrophages containing these pigments. These spots seemed to have appeared because of the previous insertion of needles.     

Four-week oral toxicity study of 1-carboxy-5,7-dibromo-6-hydroxy-2,3,4-trichloroxanthone (HXCA), an impurity of Phloxine B, in F344 rats

This study was designed to evaluate and characterize any subacute toxicity of 1-carboxy-5,7-dibromo-6-hydroxy-2,3,4-trichloroxanthone (HXCA), an impurity of Phloxine B (Food Red No. 104 in Japan, D&C Red No. 28 in the USA), when administered to both sexes of F344 rats at dietary levels of 0 (control), 0.005, 0.05 and 0.5%. During the study, the treatment had no effects on clinical signs, survival, urinalysis or ophthalmology. Hematology, blood biochemistry, gross pathology, organ weights, organ to body weight ratios and histopathology exhibited no differences of toxicological significance between control and treated rats. Reactions to treatment may be summarized as follows: there was a tendency for increased food and water consumption and decreased food efficiency in both sexes of the 0.5% group. Thus, these results indicated the no-observed-adverse-effect level (NOAEL) of HXCA to be 0.05% (39.3 mg/kg/day for males, and 41.0 mg/kg/day for females).     

A medium-term rat liver bioassay for rapid in vivo detection of carcinogenic potential of chemicals

A reliable medium-term bioassay system for rapid detection of carcinogenic potential of chemicals in the human environment has been developed. The 8-week-protocol consists of 2 stages; male F344 rats are given a single intraperitoneal injection of diethylnitrosamine (200 mg/kg) for initiation of liver carcinogenesis, followed by a 6-week test chemical treatment starting 2 weeks thereafter. Test chemicals are usually given in the diet or the drinking water and in the 2nd week of test chemical treatment, all rats are subjected to two-thirds partial hepatectomy in order to induce regenerative cell replication. The end-point marker is the glutathione S-transferase placental form (GST-P)-positive hepatic focus, the numbers and sizes of which are analyzed using an image-analyzer and expressed as values per unit liver section (1 cm²). When the yield of GST-P-positive foci is significantly enhanced (P<0.05) over the control value, a chemical is judged to possess carcinogenic or promotion potential for the liver. Among 313 chemicals already tested in this system in our laboratory, 30/31 (97%) mutagenic hepatocarcinogens and 29/33 (88%) non-mutagenic hepatocarcinogens gave positive results. Ten out of 43 (23%) agents known to be carcinogenic in organs other than the liver were also positive. It is particularly important that only one of 48 non-carcinogens gave a very weak positive result, so that the system has a very low false-positivity rate. It is now well documented that the assay system is highly effective for detecting hepatocarcinogens, bridging the gap between traditional long-term carcinogenicity tests and short-term screening assays. At the Fourth International Conference on Harmonization, our medium-term liver bioassay based on an initiation and promotion protocol was recommended in the guidelines as an acceptable alternative to the long-term rodent carcinogenicity test. (Cancer Sci 2003; 94: 3–8)     

Time- and Dose-dependent Induction of Invasive Urinary Bladder Cancers by N-Ethyl-N-(4-hydroxybutyl)nitrosamine in B6C3F1 Mice

A sequential investigation of N-ethyl-N-(4-hydroxybutyl)nitrosamine (EHBN) bladder carcino-genesis was performed in male B6C3F₁mice maintained ad libitum on tap water containing 0.025% EHBN for 4, 12, 20, 28 and 36 weeks. A total of 81 invasive tumors, comprising 55 smiamous cell carcinomas (SCCs) (68%), 25 transitional cell carcinomas (TCCs) (31%) and 1 adenocarcinoma (1%) were found. Of these, 23 (22 SCCs and 1 TCC) demonstrated invasion to the prostate, 3 metastasized to the lung, and 2 spread by peritoneal seeding. The anaplastic grade and extent of invasion of the SCCs significantly exceeded those of the TCCs. The results suggested a histogenetic pathway from simple dysplasia through papillary or nodular dysplasia and/or carcinoma in situ to eventual development of invasive carcinomas     

Aortic pressure augmentation as a marker of cardiovascular risk in obstructive sleep apnea syndrome

Obstructive sleep apnea syndrome (OSAS) is associated with increases in cardiovascular morbidity and mortality. Vascular changes in individuals with OSAS have not been fully elucidated, however. The possible impact of OSAS on the extent of aortic pressure augmentation (AG), an indicator of cardiovascular risk, was investigated. Forty-five consecutive male patients aged 35 to 78 years (56.0+/-9.6 years) who were referred to the sleep clinic of Nagoya University Hospital for screening and treatment of OSAS and 71 age-matched healthy men were enrolled in the study. AG was derived from the pressure waveform measured at the radial artery by applanation tonometry. The number of apnea and hypopnea episodes per hour (apnea-hypopnea index [AHI]) was determined by standard polysomnography. AG was significantly greater in OSAS patients than in controls (9.0+/-4.1 vs. 6.4+/-3.4 mmHg, p<0.001), and it was significantly reduced in 19 OSAS patients treated with continuous positive airway pressure. AG was also significantly correlated with the AHI (r=0.562, p<0.001) and age (r=0.356, p=0.016) but not with the serum concentrations of low and high density lipoprotein-cholesterol, triglyceride, or glycosylated hemoglobin. Stepwise multiple regression analysis revealed that the AHI was the most significant contributing factor to the increased AG in OSAS patients (beta=0.109, r=0.530, p<0.001). OSAS may thus have an adverse effect on vascular function that can be ameliorated by appropriate treatment.     

ENDOSCOPIC COAGULATION THERAPY IS USEFUL FOR IMPROVING ENCEPHALOPATHY IN CIRRHOTIC PATIENTS WITH GASTRIC ANTRAL VASCULAR ECTASIA

Background: Gastric antral vascular ectasia (GAVE) is an uncommon but treatable cause of chronic gastrointestinal bleeding often associated with severe cirrhosis. Efficacy of endoscopic treatment is well known; however, long‐term outcome after endoscopic treatment is not clear. Methods: We studied 16 cases of GAVE patients with liver cirrhosis that were endoscopically treated using heater probe unit or argon plasma coagulator. Endoscopic finding of GAVE and clinical finding of hepatic encephalopathy in particular was evaluated after endoscopic therapy. Results: After endoscopic ablation therapy, mucosal vascular lesion of GAVE and rebleeding occurred in four patients during follow up; however, re‐treatment was effective and long‐term hemostasis was achieved. Moreover, a case report herein revealed the effectiveness of endoscopic therapy on hepatic encephalopathy of GAVE patients with cirrhosis. In the follow‐up study of 13 patients of GAVE with cirrhosis that suffered from encephalopathy, the coma level of nine patients was improved after endoscopic therapy. Conclusion: Endoscopic treatment, such as heater probe coagulation and argon plasma coagulation therapy, are effective and useful for long‐term follow up. Also, the beneficial effect of endoscopic therapy on hepatic encephalopathy for GAVE with cirrhosis encourages us to find and treat the mucosal lesion intensively to improve the quality of life of such patients.