Antiproteinuric Effects of Green Tea Extract on Tacrolimus-Induced Nephrotoxicity in Mice.

INTRODUCTION: It has been reported that proteinuria is an early predictive marker in detection of tacrolimus (TAC) nephrotoxicity. The aim of this study was to investigate the antiproteinuric effects of green tea extract (GTE) on TAC-induced acute nephrotoxicity in mice. METHODS: The mice (n = 20) were divided into 4 groups (n = 5 per group); control group mice were intraperitoneally (IP) injected with 0.9% saline, TAC group mice were IP injected with TAC 1 mg/kg, and inducible nitric oxide synthase (iNOS) inhibitor group mice were given in addition NG-nitro-L-arginine-methyl ester 12 mmol/L by subcutaneous injection. TAC-GTE group mice were given TAC by IP injection and GTE 100 mg/kg by subcutaneous injection. RESULTS: The 24-hour urine protein amounts were significantly increased in TAC group mice (36.1 ± 9.9 mg/d) compared with control group mice (13.3 ± 5.4 mg/d) and significantly decreased in TAC-GTE group mice (19.1 ± 6.9 mg/d, P < .01) compared with TAC group mice. The nitric oxide (NO) production by TAC was significantly suppressed by GTE and iNOS inhibitor injection. Renal tissue malondialdehyde (MDA) level was significantly increased in the TAC group compared with the control group and was significantly decreased in the TAC-GTE group compared with that of the TAC group. The antioxidant enzyme activities of superoxide dismutase and catalase were significantly suppressed in the TAC group compared with the control group and were restored in the GTE injection group. CONCLUSIONS: GTE treatment has beneficial antiproteinuric effects on TAC-induced acute renal injury in mice.

Pore volume is most highly correlated with the visual assessment of skin pores.

BACKGROUND/PURPOSE: Many studies have been focused on evaluating assessment techniques for facial pores amid growing attention on skin care. Ubiquitous techniques used to assess the size of facial pores include visual assessment, cross-section images of the skin surface, and profilometric analysis of silicone replica of the facial skin. In addition, there are indirect assessment methods, including observation of pores based on confocal laser scanning microscopy and the analysis of sebum secretion and skin elasticity. The aim of this study was to identify parameters useful in estimating pore of surface in normal skin. METHODS: The severity of pores on the cheek area by frontal optical images was divided on a 0-6 scale with '0' being faint and small pore and '6' being obvious and large pore. After the photos of the frontal cheek of 32 women aged between 35 and 49 were taken, the size of their pores was measured on a 0-6 scale; and the correlation between visual grading of pore and various evaluations (pore volume by 3-D image, pore area and number by Optical Image Analyzer) contributing to pore severity investigated using direct, objective, and noninvasive evaluations. RESULTS: The visual score revealed that the size of pores was graded on a 1-6 scale. Visual grading of pore was highly correlated with pore volume measured from 3-D images and pore area measured from 2-D optical images in the order (P < 0.01). Visual grading of pore was also slightly correlated with the number of pores in size of over 0.04 mm(2) (P < 0.05). CONCLUSION: High correlation between pore score and pore volume can be explained by 3-D structural characteristics of pores. It is concluded that pore volume and area serve as useful parameters in estimating pore of skin surface.

A serosurvey of Orientia tsutsugamushi from patients with scrub typhus.

Many countries where scrub typhus is endemic use their own cutoff values for antibody titres to differentiate between cured cases and current infections. To establish an antibody titre cutoff value, one needs to investigate the seroprevalence in endemic areas, and the duration of the increase in titre after complete cure. We conducted a follow-up study of anti-Orientia tsutsugamushi antibody titres using indirect immunofluorescence assays (IFA) and passive haemagglutination assays (PHA) in patients with scrub typhus. After the onset of symptoms, IgM antibody titres increased gradually over 2-3 weeks, peaked at about 4 weeks, and started to decrease rapidly between 4 and 5 weeks. At 1-year follow-up, the median IgM value was 1:10. Out of 77 patients who were tested at that time, 36 (47%) had IgM titres > or =1:20, and none had titres exceeding 1:80. Over the first 2 weeks, IgG antibody titres increased sharply, peaked at about 4 weeks and decreased rather gradually thereafter, with a median titre of 1:128 maintained up to the 18th month. At 1-year follow-up, five out of 77 patients (6.5%) had titres > or =1:1,024 and 57% had titres > or =1:128. Based on these results, a cutoff value of > or =1:160 for IgM antibody should differentiate between previous and current infections in endemic areas such as Korea and Japan, where scrub typhus occurs mainly in the autumn.

Analysis on donor and isolation-related factors of successful isolation of human islet of Langerhans from human cadaveric donors.

We analyzed the preexisting donor factors and isolation variables that affected isolation of human islets of Langerhans. Sixty-nine pancreata from cadaveric donors were analyzed for donor factors of age, gender, body mass index, cause of death as well as graft factors of cold ischemia time, pancreas status, distensibility during intraductal collagenase distension and time of collagenase expansion and digestion. Islet isolations that recovered >100,000 IEQ (n = 53) were compared to those generating less than 100,000 IEQ (n = 16) to analyze the factors affecting islet yield during donor harvest and isolation procedures. The mean islet recovery was 216.0 x 10(3) (IEQ) or 2840 (IEQ) per gram of pancreas. Mean purity was 54%. The success rate of islet isolation was 76%. Mean age was 31 years, and mean cold ischemia time was 6.9 hours. In univariate analysis, the status of the pancreas was the only significant factor for successful isolation, and gender, time of collagenase expansion and digestion were marginal factors. In stepwise multivariate logistic regression analysis of donor and isolation-related factors, donor gender, pancreas status and digestion time were significant factors. During the same period we performed three cases of clinical islet allotransplantation and one autotransplantation. This study confirmed that the same donor factors and variables in the isolation process can affect the ability to obtain successful human islet isolation. Enough experience and pertinent review of donor and isolation factors can make islet isolation consistent, supporting clinical islet transplantation without unnecessary cost.

Comparative study on biologic and immunologic characteristics of the pancreas islet cell between 24 degrees C and 37 degrees C culture in the rat.

The aim of this study was to investigate the effect of culture at 24 degrees C on cell viability, cellular function, immunogenicity, and cytokine profiles of rat pancreatic islets. Pancreatic islets were isolated from Lewis rats and cultured at either 24 degrees C or 37 degrees C for 14 days. Islet recovery was counted as islet equivalents; islet viability was examined with fluorescent vital staining. Islet function was measured with a glucose stimulation test. Annexin V, and MHC class I and II expression were measured using flow cytometric assay for apoptosis and immunogenicity, respectively. Lymphocyte cell proliferation was examined with WST-1 proliferation assay. Cytokine profiles were analyzed with quantitative real time RT-PCR. All these parameters were measured on 1, 3, 5, 7 and 14 culture days after islet isolation. Islet recovery was higher in islets cultured at 24 degrees C than 37 degrees C without a change in viability. Insulin secretion after glucose stimulation was more effective in 24 degrees C culture conditions. Decreased apoptotic cell death was demonstrated in 24 degrees C cultured islets. Both MHC class I and II expression on islets and lymphocyte proliferation upon coculture with islets were less prominent in 24 degrees C cultured islets. TNF-alpha expression was lower in islets cultured at 24 degrees C than in islets cultured at 37 degrees C. Both IL-1beta and IL-10 cytokine expressions were similar under both culture conditions. This study demonstrated that cell recovery and function are increased in islets cultured at 24 degrees C than those at 37 degrees C with decreased antigenicity and proinflammatory cytokine expression.

Experimental islet isolation in porcine pancreas with new enzyme Liberase PI.

The aim of this study was to investigate the results of 20 consecutive porcine islet isolations using a new enzyme Liberase PI. Twenty pancreata were procured for islet isolation, which was performed using modified Ricordi's method with Liberase PI. Quantitation of islet viability staining, insulin stimulation assay, intracellular insulin content/DNA, and in vivo transplantability into diabetic nude mice were examined for quality control. The results were compared between a high-yield group (>2500 IEQ/g pancreas) and a low-yield group (<2500 IEQ/g pancreas). Sufficient amount of purified islets (3000 IEQ/g pancreas) were obtained using the new brand enzyme Liberase PI. These islets showed good quality in structure and functions, which were demonstrated by in vitro and in vivo standard assays. Isolation index (IEQ/number) of the low-yield group was lower than that of high-yield group (0.75 vs 0.86), which means more fragmentation of islets in the low-yield group. There were no differences in function between the two groups. In conclusion, we obtained sufficient numbers of viable, functional islets from porcine pancreas using a new brand enzyme Liberase PI and low-temperature isolation technique. However, overdigestion of islets during the isolation remains to be overcome. Advance in porcine islet isolation technique will in the future make the porcine islet xenotransplantation a reality for the cure of diabetes mellitus.

Biochemical characterization of a novel hypoxanthine/xanthine dNTP pyrophosphatase from Methanococcus jannaschii.

A novel dNTP pyrophosphatase, Mj0226 from Methanococcus jannaschii, which catalyzes the hydrolysis of nucleoside triphosphates to the monophosphate and PPi, has been characterized. Mj0226 protein catalyzes hydrolysis of two major substrates, dITP and XTP, suggesting that the 6-keto group of hypoxanthine and xanthine is critical for interaction with the protein. Under optimal reaction conditions the k(ca)(t) /K(m) value for these substrates was approximately 10 000 times that with dATP. Neither endonuclease nor 3'-exonuclease activities were detected in this protein. Interestingly, dITP was efficiently inserted opposite a dC residue in a DNA template and four dNTPs were also incorporated opposite a hypoxanthine residue in template DNA by DNA polymerase I. Two protein homologs of Mj0226 from Escherichia coli and Archaeoglobus fulgidus were also cloned and purified. These have catalytic activities similar to Mj0226 protein under optimal conditions. The implications of these results have significance in understanding how homologous proteins, including Mj0226, act biologically in many organisms. It seems likely that Mj0226 and its homologs have a major role in preventing mutations caused by incorporation of dITP and XTP formed spontaneously in the nucleotide pool into DNA. This report is the first identification and functional characterization of an enzyme hydrolyzing non-canonical nucleotides, dITP and XTP.