Nuclear localization of activated STAT6 and STAT3 in epidermis of prurigo nodularis.

BACKGROUND: Prurigo nodularis (PN) is a chronic dermatitis characterized by discrete, raised, and firm papulonodules with intense pruritus. The pathogenesis still remains to be elucidated. OBJECTIVES: To clarify the role of Th1 and Th2 cytokines in the pathogenesis of PN. METHODS: We examined the cytokine signatures, such as phosphorylation of STAT1, STAT3 and STAT6, HLA-DR and hyaluronan accumulation, to reveal the Th1 and Th2 cytokine influence on the lesional epidermis of PN. RESULTS: We first optimized antigen retrieval methods to detect these signatures with antibodies for phospho-STAT1 (pSTAT1), phospho-STAT3 (pSTAT3), phospho-STAT6 (pSTAT6), HLA-DR and hyaluronic acid binding protein (HABP) on the formalin-fixed paraffin-embedded sections of psoriasis, lichen planus and atopic dermatitis biopsy samples. Activation of STAT1 and STAT6 in epidermis by Th1 and Th2 cytokines was further confirmed in a cultured skin equivalent model treated with interferon-γ or interleukin (IL)-4/IL-13. With the relevant immunostaining methods, we examined the cytokine signatures in 22 cases of PN. The results revealed that (i) the entire epidermis of 19 cases was stained with anti-pSTAT6 antibody, (ii) 21 cases demonstrated nuclear staining with anti-pSTAT3 antibody, (iii) the entire epidermis of 21 cases was stained with HABP, (iv) the epidermis of eight cases showed scattered staining with anti-pSTAT1 antibody, and (v) six cases were positive for HLA-DR membrane expression. CONCLUSIONS: These data indicated that Th2 cytokines related to STAT6 activation together with some unknown stimuli that activate STAT3 play a principal role in the pathogenesis of PN.

Association between a cytochrome P450 CYPIA1 genotype and incidence of lung cancer.

We analysed 267 samples from patients with lung cancer and 54 samples from patients with pancreatic cancer and classified them into three types (A, B and C) based on their P450CYPIA1 genotypes. This was performed by restriction fragment length polymorphism (RFLP) analysis of DNA (that had been amplified) using the polymerase chain reaction (PCR). Our results reconfirmed previous studies with RFLP analyses by revealing significantly higher frequencies of C type in lung cancer patients, particularly light smokers with squamous cell carcinoma (p < 0.001). Also when we compared the appearance rate of P450CYPIA1 genotype in lung cancer patients between metastasis group and no metastasis group, lymph node or distant metastasis was more frequently observed among the patients of squamous cell carcinoma, especially the light smoking group, with C genotypes (p < 0.05). These results suggested that this genotype is not only associated with the etiology of lung cancer but also with the status of metastasis. Furthermore, genotyping could be useful in predicting the incidence of lung cancer in the smokers and the general population as well as the prognosis of individuals who develop this disease.

Altering the course of cataracts in diabetic rats.

A potent new aldose reductase (AR) inhibitor was effective in preventing cataractous changes in diabetic rats. Untreated diabetic rats developed early lens changes by 3 weeks and dense nuclear opacities by 6 to 9 weeks. In contrast, diabetic rats treated with the AR inhibitor showed no lens changes during the 5-month period of the experiment.

Interaction of poly(rC)-binding protein 2 with the 5'-terminal stem loop of the hepatitis C-virus genome.

The 5' noncoding region (NCR) of hepatitis C virus (HCV) contains an internal ribosome entry site for translation initiation. Cellular proteins (e.g. La, polypyrimidine tract-binding protein, and p25) that interact with HCV 5' NCR have been implicated in facilitating efficient internal initiation. The 5' NCR may also contain RNA structures and specific RNA sequences that interact with cellular proteins to promote RNA replication. UV crosslinking experiments revealed a 43-kDa cellular protein (p43) also interacts with the HCV 5' NCR. Further UV crosslinking experiments with deletion mutants of HCV 5' NCR demonstrated that p43 bound specifically to the 5'-terminal stem-loop of the HCV 5' NCR. Achromobactor proteinase I digests, competition experiments, and immunoprecipitation confirmed that p43 was identical to human poly(rC)-binding protein 2 (PCBP2). We prepared a PCBP2-immunodepleted rabbit reticulocyte lysate with an anti-PCBP2 antibody. Translation activity promoted by the HCV internal ribosome-entry site was the same in PCBP2-depleted lysates as in mock-depleted lysates. In conclusion, PCBP2 specifically interacted with the 5' terminus of HCV genome but had no effect on HCV translation. We speculate that PCBP2's interaction with HCV 5' NCR may be involved in the replication-initiation complex of HCV.

Aldose reductase in diabetic complications of the eye.

Aldose reductase (AR) appears to initiate the cataractous process in galactosemic and diabetic animals. Sugars in excess are converted to polyols by lens AR. In sugar cataracts, polyols accumulate to levels substantial enough to cause a hypertonicity leading to lens fiber swelling. All other changes appear secondary to polyol accumulation and lens swelling. The development of sugar cataracts can be duplicated in organ culture. In culture, the various changes that occur were minimized or did not occur when inhibitors of AR were included in the medium. Moreover, AR inhibitors were shown to effectively delay the onset of sugar cataract development in animals. A defect in the corneal epithelium of diabetics became apparent in vitrectomy. One manifestation of this problem was the delay in the reepithelialization of denuded corneas. In examining this problem experimentally, the epithelium was removed from the corneas of diabetic and normal rats. The regeneration of epithelium in corneas of diabetic rats required a longer period than in the normal. The possibility that AR, active in the epithelium, was involved in this phenomenon was investigated. The corneal epithelium was removed from both eyes of a diabetic rat. One eye was treated topically with the AR inhibitor CP-45,634 while the other served as control. The eye treated with CP-45,635 regenerated epithelium much more quickly than the untreated eye. Other AR inhibitors had similar beneficial effects.

Multicyclic reverse transcription-polymerase chain reaction assay system for quantification of GB virus-C/hepatitis G virus RNA in serum.

A new quantitative reverse transcription-polymerase chain reaction (RT-PCR) method is described for analyzing the amount of GB virus-C (GBV-C)/hepatitis G virus (HGV) RNA in serum. This multicyclic RT-PCR (MRT-PCR) method used oligonucleotide primers deduced from the 3' noncoding region (3'NCR) that is highly conserved among GBV-C/HGV isolates. Quantitation of GBV-C/HGV RNA using MRT-PCR ranged between 10(2) and 10(10) copies/ml when PCR cycle number was regulated at exponential amplification of the products. Competitive RT-PCR (CRT-PCR) was carried out with mutant RNA and sample that had been measured by MRT-PCR. Quantitation of GBV-C/HGV RNA using both methods agreed. MRT-PCR detected viral RNA in a single step PCR, and demonstrated a high degree of sensitivity that was equal to that of the RT-PCR procedure, which used nested primers deduced from the non-structural (NS) 3 region. The MRT-PCR method for quantitation of GBV-C/HGV RNA in serum may prove useful for diagnosis.

Cultured epithelial autografting on meshed skin graft scars: evaluation of skin elasticity.

Many patients with meshed skin graft scars complain of the scars' unsightly appearance and hardness. Since 1989 we have shaved away meshed skin graft scars and then resurfaced the area with autologous cultured epithelium in nine patients. This method improved the disfigurement of meshed skin graft scars, with minimal sacrifice of normal donor skin. Furthermore, autologous cultured epithelium grafted areas had high skin elasticity compared with meshed skin graft scars, as measured with a noninvasive suction device.

[Calculation of left ventricular volume and ejection fraction from ECG-gated myocardial SPECT: automatic detection of endocardial borders by threshold method].

A new method which calculate end-diastolic volume (EDV), end-systolic volume (ESV) and ejection fraction (LVEF) of the left ventricle from myocardial short axis images of ECG-gated SPECT using 99mTc myocardial perfusion tracer has been designed. Eight frames per cardiac cycle ECG-gated 180 degrees SPECT was performed. Threshold method was used to detect myocardial borders automatically. The optimal threshold was 45% by myocardial SPECT phantom. To determine if EDV, ESV and LVEF can also be calculated by this method, 12 patients were correlated ventriculography (LVG) for 10 days each. The correlation coefficient with LVG was 0.918 (EDV), 0.935 (ESV) and 0.900 (LVEF). This method is excellent at objectivity and reproductivity because of the automatic detection of myocardial borders. It also provides useful information on heart function in addition to myocardial perfusion.

[Anesthetic experience of emergency cesarean section for a patient with myotonic dystrophy].

We report an anesthetic experience of a 28-year-old female patient complicated with myotonic dystrophy who underwent emergency Cesarean section due to threatened abortion. Anesthesia was induced with intravenous thiopental followed by topical spray of 4% lidocaine 5 ml to intubate trachea and maintained with neuroleptanesthesia with droperidol, fentanyl and nitrous oxide in oxygen. No muscle relaxant was used. The course of anesthesia and emergence from anesthesia were uneventful. However, the female infant, with Apgar score of 1 point at five minutes after delivery, died due to multiple organ failure three weeks after the delivery. Anesthetic management of a patient with myotonic dystrophy was also discussed with a literature review.

[Preanesthetic meals in elective surgical patients].

This study investigated the effect of preanesthetic meals on the volume and pH of gastric contents in forty elective surgical patients ranging in ages from 20 to 60 years. Twenty patients who were given either isotonic beverage 250 ml or apple juice 250 ml on the morning of the operative day were subjected as control group and twenty patients of the breakfast group took two slices of bread with the above drink. About seven hours following drinking and feeding, the mean values of gastric volume were 20.9 +/- 18.3 ml in the control group and 19.2 +/- 16.3 ml in the breakfast group. The mean values of gastric pH were 4.3 +/- 2.3 in the drink group and 4.6 +/- 2.3 in the breakfast group. There were no significant differences in the gastric volume and pH between the two groups. However, very small amount of the bread was detected in the gastric fluid of three patients in the breakfast group. As preanesthetic drinking and feeding are advantageous for reducing the anxieties of preoperative patients and also for their nutrition during operation, it is encouraging that eating two slices of bread did not induce a significant effect of gastric volume or pH. The minute fragment of bread seems to have no clinically significant effect.