Effect of Donor Age on Graft Function and Pathologic Findings in Living Donor Transplantation.

BACKGROUND: The frequency of renal transplants from elderly living donors has increased because of a shortage of donors. However, the results of renal transplantation using aged kidney grafts have yet to be determined conclusively. METHODS: We evaluated 45 patients who underwent living donor kidney transplantation at our institution. The patients were categorized according to donor age at the time of the transplant: ≥ 60 years (elderly donor group, n = 21) and <60 years (young donor group, n = 24). We reviewed the renal function of the recipients and pathologic findings of the graft including interstitial fibrosis score, tubular atrophy score, tubular atrophy and interstitial fibrosis grades, and arteriosclerosis up to 2 years posttransplantation. RESULTS: Significant differences were observed in the preoperative creatinine clearance of the donor, prevalence of hypertension in the donor, and age of the recipient. Serum creatinine levels in the elderly donor group were significantly higher from 2 months to 1 year posttransplantation, and the estimated glomerular filtration rate was significantly lower from 7 days to 1 year posttransplantation. However, the decline in estimated glomerular filtration rate from 14 days to up to 2 years posttransplantation was similar in the 2 groups. There was no significant difference in the renal biopsy findings between the 2 groups except for arteriosclerosis 1 year posttransplantation. CONCLUSION: Kidney grafts from elderly living donors were not associated with a deterioration in renal function, and their pathologic findings were comparable with those of young donors for up to 2 years posttransplantation.

Karyotypic changes associated with loss of prolactin dependency of rat Nb2 node lymphoma cell cultures.

The parent line of cultured "Nb2 node" lymphoma cells is dependent on the hormone prolactin (PRL) for growth and is widely used for the in vitro bioassay of lactogenic hormones. As reported previously, PRL-independent sublines have been developed in vitro from the parental line by lactogen deprivation. The present study describes the G-banded karyotypes of the Noble (Nb) strain of rat (in which the original lymphoma developed), the PRL-dependent cell line (157th generation), and two of its PRL-independent sublines (1220th and 2372nd generations). The karyotype of the Nb rat was determined to be the same as that of Rattus norvegicus. The stemline karyotype of the PRL-dependent cells contains a number of well-defined chromosomal abnormalities. The PRL-independent sublines examined have the same chromosomal abnormalities as the PRL-dependent cells plus a few additional changes indicative of clonal evolution from the PRL-dependent stemline. The development of PRL independence (as seen in the 1220th generation) was associated with only two karyotypic changes, i.e., loss of the Y chromosome and a translocation involving chromosomes 14 and 17. The recently reported mapping of the rat PRL gene and other PRL-related genes to chromosome 17 suggests that rearrangement of chromosome 17 could be involved in the development of the PRL independence. The PRL-dependent and PRL-independent Nb2 cell lines provide a useful system for studying chromosomal and molecular genetic events associated with the malignant progression of polypeptide hormone-dependent cancers.

A new thiazolidinedione, NC-2100, which is a weak PPAR-gamma activator, exhibits potent antidiabetic effects and induces uncoupling protein 1 in white adipose tissue of KKAy obese mice.

Thiazolidinediones (TZDs) reduce insulin resistance in type 2 diabetes by increasing peripheral uptake of glucose, and they bind to and activate the transcriptional factor peroxisome proliferator-activated receptor-gamma (PPAR-gamma). Studies have suggested that TZD-induced activation of PPAR-gamma correlates with antidiabetic action, but the mechanism by which the activated PPAR-gamma is involved in reducing insulin resistance is not known. To examine whether activation of PPAR-gamma directly correlates with antidiabetic activities, we compared the effects of 4 TZDs (troglitazone, pioglitazone, BRL-49653, and a new derivative, NC-2100) on the activation of PPAR-gamma in a reporter assay, transcription of the target genes, adipogenesis, plasma glucose and triglyceride levels, and body weight using obese KKAy mice. There were 10- to 30-fold higher concentrations of NC-2100 required for maximal activation of PPAR-gamma in a reporter assay system, and only high concentrations of NC-2100 weakly induced transcription of the PPAR-gamma but not PPAR-alpha target genes in a whole mouse and adipogenesis of cultured 3T3L1 cells, which indicates that NC-2100 is a weak PPAR-gamma activator. However, low concentrations of NC-2100 efficiently lowered plasma glucose levels in KKAy obese mice. These results strongly suggest that TZD-induced activation of PPAR-gamma does not directly correlate with antidiabetic (glucose-lowering) action. Furthermore, NC-2100 caused the smallest body weight increase of the 4 TZDs, which may be partly explained by the finding that NC-2100 efficiently induces uncoupling protein (UCP)-2 mRNA and significantly induces UCP1 mRNA in white adipose tissue (WAT). NC-2100 induced UCP1 efficiently in mesenteric WAT and less efficiently in subcutaneous WAT, although pioglitazone and troglitazone also slightly induced UCP1 only in mesenteric WAT. These characteristics of NC-2100 should be beneficial for humans with limited amounts of brown adipose tissue.

MAT-1, a monoclonal antibody that specifically recognizes human tyrosinase.

There is no available monoclonal antibody which reacts specifically recognizes human tyrosinase. Employing a synthetic peptide, MEKEDYHSLYQSHL, corresponding to the carboxyl terminus of human tyrosinase as an immunogen, we produced a mouse monoclonal antibody MAT-1 of the IgG1 isotype. The epitope for MAT-1 was determined to be EDYH, the sequence of which is not present in human tyrosinase-related protein-1 (TRP-1) or tyrosinase-related protein-2 (TRP-2). By transient expression assays and immunofluorescence technique, we show that MAT-1 reacts specifically with cells expressing human tyrosinase cDNA but not with cells expressing TRP-1 or TRP-2 cDNA. The results of immunohistochemical staining also confirmed that MAT-1 reacts specifically with epidermal melanocytes in human skin sections. MAT-1 should be invaluable for studying the interaction between tyrosinase and TRPs and for detecting the changes in the levels of tyrosinase expression. In addition, MAT-1 should be useful as a sensitive immunohistochemical tool for investigation of various pigmentary disorders and possibly for the diagnosis of melanoma.

Localization of rat FGF-5 protein in skin macrophage-like cells and FGF-5S protein in hair follicle: possible involvement of two Fgf-5 gene products in hair growth cycle regulation.

It has been reported that the gene for murine fibroblast growth factor-5 (Fgf-5) is expressed in the rat hair follicle and that this expression may be associated with catagen induction (Hebert et al, 1994). In this study, we analyzed the Fgf-5 gene product in skin because the gene generates two mRNA that translate into the FGF-5 protein and a short form of the FGF-5 protein (FGF-5S) as a result of an alternative splicing (Hattori et al, 1996; Ozawa et al, 1996). Indeed, we detected both types of FGF-5 mRNA in rat skin samples. Two monoclonal anti-FGF-5 antibodies, one (E723) being specific for FGF-5 long-form protein and the other (B2B6) being reactive with both FGF-5 and FGF-5S proteins, were used to locate these proteins by immunohistochemistry. Staining of the rat skin revealed that only the B2B6 antibody reacted with hair follicles and that both antibodies reacted with macrophage-like round cells, suggesting that the product of the Fgf-5 gene in the hair follicle is FGF-5S. The immunoreactivity of the FGF-5S protein increased during early anagen VI and decreased rapidly during catagen. The density of FGF-5-positive macrophage-like cells in the dermis increased during anagen and decreased during catagen and telogen, whereas the density of these cells in the panniculus adiposus did not change during anagen and increased during catagen and telogen. There was no apparent association between the density of FGF-5-positive macrophage-like cells and that of FGF-5-negative, dendritic macrophage-like cells. Thus, the results suggest the possible involvement of FGF-5S in the hair follicle in anagen VI and catagen development and that the density of FGF-5-positive macrophage-like cells may also be associated with the hair growth cycle.

Astrocytoma with a unique telomere association.

Many primary pediatric brain tumors are characterized by nonrandom cytogenetic abnormalities involving specific structural rearrangements and loss or gain of specific chromosomes. We describe a low-grade cerebellar astrocytoma with telomeric association of chromosomes 18 and 21 in a 7-year old girl. The nonrandom telomeric association of these chromosomes represents a unique cytogenetic finding in this type of pediatric brain tumor.

Cytogenetic and molecular genetic analysis of a pediatric pleomorphic sarcoma reveals similarities to adult malignant fibrous histiocytoma.

Cytogenetic and molecular genetic studies were performed on a pleomorphic sarcoma removed from the left atrium of a 15-year-old girl. Histologic analysis was consistent with a storiform-pleomorphic malignant fibrous histiocytoma (MFH). Although MFH is the most common soft-tissue sarcoma of late adulthood. It is extremely rare in childhood and its existence in the pediatric population remains controversial. Cytogenetic analysis revealed several alterations previously associated with adult MFH, including abnormalities of chromosomal bands 11p11 and 19p13. Moreover, the tumor demonstrated homogeneously staining regions (HSR) and double minute chromosomes (dmin) suggestive of gene amplification. We therefore screened the case for amplification of genes localized to chromosomal bands 12q13-14, including the putative protooncogenes MDM2, CDK4, SAS, CHOP, and CLI, which are frequently amplified and overexpressed in adult MFH. Southern and Northern blot analysis confirmed the coamplification of MDM2, CDK4, SAS, and CHOP. To our knowledge, such coamplification studies of the 12q13-14 amplicon have not been previously detected in pediatric MFH. Our results provide cytogenetic and molecular genetic evidence that pediatric and adult MFH are histogenetically related entities.

groE-homologous operon of Wolbachia, an intracellular symbiont of arthropods: a new approach for their phylogeny.

Wolbachia, a member of rickettsia found in the cells of many arthropod species, are cytoplasmically inherited bacteria which interfere with host's sexuality and reproduction. Wolbachia strains have been phylogenetically divided into A and B groups based on the nucleotide sequences of their ftsZ genes. In an attempt to further define the phylogenetical relationship among these endosymbionts, we cloned and sequenced the entire length of the groE operon of a Wolbachia harbored by a cricket. The operon encoded two heat shock proteins, which represented the third and fourth proteins of any Wolbachia ever characterized. Also, 800 bp stretches of the groE operons of several other Wolbachia were sequenced, and a phylogenetic tree was constructed based on the results. The groE tree defined the relationship among A group Wolbachia strains that had not been successfully resolved by the ftsZ tree, and suggested unexpected horizontal transmission of these bacteria.

Optical resolution and absolute configuration of N-benzyloxycarbonyl-alpha-alkoxyglycines

Optical resolution of racemic N-benzyloxycarbonyl-protected alpha-alkoxyglycines, (+/-)-Cbz-Gly(OR)-OH (R = Et and Pr(i)), was achieved by means of fractional crystallization of diastereomeric salts with (+)-(1S,2S)-2-amino-1-phenyl-1,3-propanediol or diastereomeric esters of (+)- or (-)-menthol. The D- and L-configurations were assigned to the (+)- and (-)-Cbz-Gly(OR)-OH, respectively, based on L-enantioselective enzymatic hydrolysis of (+/-)-Cbz-Gly(OR)-OR' (R = Me, Et, and Pr(i); R' = CH(2)CF(3) and Me) using porcine pancreatic lipase and papain. Chiroptical properties and HPLC retention times of D- and L-Gly(OR)-residue (R = Me and Pr(i))-containing peptides were compared in relation to their configurational assignments. Copyright 1999 Wiley-Liss, Inc.

[Development and validation of a self-management skill scale].

OBJECTIVES: The aims of this study were: 1) to develop a scale for self-reported assessment of self-management skills (SMS), and to confirm its reliability and validity; 2) to verify its applicability to health services including health education by analyzing the relationship between the SMS score and behavioral changes among smoking cessation campaign participants. METHODS: A 20-item preliminary SMS scale was developed based on the previous findings of Rosenbaum's self-control schedule, social skills and life skills. A 10-item final SMS scale was developed by testing internal consistency and test-retest reliability among 55 subjects who were second to fourth-year college students (mean age: 20.9) in 1997. A questionnaire, including the SMS scale, self-efficacy for quitting smoking, and smoking cessation behavior was administered to 501 smokers (mean age: 46.3) who participated in the smoking cessation campaign that began in December 1997 and ended in June 1998. RESULTS: Among the participants of the smoking cessation campaign, the SMS scale showed a high level of internal consistency (Cronbach's alpha = 0.75) and positive correlations with age and self-efficacy for quitting smoking. The SMS scale also showed positive correlations with the outcome of the program, which means that those who had low SMS tended to drop out. CONCLUSION: The findings indicate that the SMS scale has acceptable levels of reliability and validity. They also suggest that it might be a useful instrument in health education and health services because the score correlates with people's health behavior.

[The relationship between stages and biochemical markers of smoking. Workplace-based cross-sectional and longitudinal studies].

In order to clarify smokers' characteristics by "Stages of Change" based on Prochaska's transtheoretical model, we conducted cross-sectional and logitudinal studies with biochemical markers of smoking and smoking habits. In a workplace-based sample of 277 male smokers, we examined cross-sectionally the relationships between stages and biochemical markers of smoking which include expired carbon monoxide concentrations and urinary nicotine metabolite concentrations, and smoking habits which include the number of cigarettes smoked per day, yields of cigarettes, inhalation patterns, time to first morning cigarette, and quit attempts in the past. Additionally we examined longitudinally the relationship between stages and expired carbon monoxide concentrations, the number of cigarettes, and yields of cigarettes. In the cross-sectional study there were significant differences among stages on expired carbon monoxide concentrations (P = 0.006), urinary nicotine metabolite concentrations (P = 0.049), the number of cigarettes smoked per day (P = 0.001), and yields of cigarettes (P = 0.042) using analyses of variance. There were also significant differences among stages on time to first morning cigarette (P = 0.018) and quit attempts in the past (P < 0.001) using chi-square tests. In the longitudinal study for each level of elevation in stage during a one-year period, expired carbon monoxide concentrations decreased on an average of 2.3 ppm (P = 0.125) and the number of cigarettes smoked per day decreased on an average of 2.8 cigarettes per day (P = 0.07). However, the yields of cigarettes did not change during the one-year period.

[Development of a smoking cessation program during health checkups. Preliminary study to evaluate the usefulness of this program].

Development of a simple and effective smoking cessation program is needed to provide cessation counseling during health checkups. A new cessation program, which consists of brief individual counseling and 4 follow-up telephone calls, was developed based on the stage model for life-style change. This program was performed during health checkups in the town of Nose to evaluate its usefulness. Smoking status questionnaires were completed to assess the smoking habits of subjects and to evaluate their smoking stage before the counseling session. Then, stage-matched cessation counseling was provided using a self-help guide. During the counseling, carbon monoxide measurement of expired air and Health Risk Appraisal feedback were performed to enhance self-perception of smoking. Follow-up calls were provided for only those clients who set a quit date during the individual counseling. It was easy to implement this program, and it required between 15 and 20 minutes to conduct. The cessation rate was 19% at 8 months after the health checkups. This result was more effective than data for other programs reported previously. Therefore, this program was effective and could be used at health checkups. This trial had no control group, so further studies are needed to clarify the efficacy and effectiveness of this program. In addition, training courses for health professionals must be developed to disseminate this program into general use.

[The effect of smoking cessation counseling at health checkup].

Smoking cessation counseling is an important element of tobacco control in the workplace, but it is not easy to persuade workers to stop smoking. We performed a controlled intervention trial to evaluate the effectiveness of a new cessation program developed by Nakamura et al., which consisted of one brief individual counseling session and 4 follow-up telephone calls. Two hundred and twenty-eight smokers who visited our center for an annual health checkup were randomly divided into two group: 117 were assigned to the intervention group, and 111 were controls. Smoking status questionnaires were administered to assess the smoking habit of each subject and to evaluate their stages of change toward smoking cessation before the counseling session. Stage-matched cessation counseling was then provided to the intervention group by nurses who had completed training courses for this program. During the counseling session, carbon monoxide in expired air and nicotine metabolites in urine were measured to enhance self-perception of smoking. Only those clients who set a quit date during their counseling sessions received follow-up telephone calls. It was easy to implement this program (15 to 20 minutes long) during a health checkup. No significant differences were observed in the baseline characteristics of the two groups. The cross-sectional smoking cessation rates at 6 months and 1 year of follow-up were 6.2 times higher in the intervention group than in the control group. The continuous smoking cessation rate at 1 year of follow-up was 7.6 times higher in the intervention group than in the control group. In the intervention group, the lower level of nicotine metabolites in urine and higher smoking stage were related to cessation success, but other baseline characteristics were similar in those who quit smoking and those who did not. The effectiveness and easy applicability of this cessation program was proved in the present study. Further examinations in various settings are expected to clarify the effectiveness of this program.

[The genetic architecture of emotionality in rats using Mendelian cross analysis].

Two inbred strains of Tsukuba Emotionality Rat (THE and TLE) and four hybrids derived from them were observed for ambulatory response in a novel environment as a measure of emotionality. The Runway Test (modified "timidity test") was run for successive three days. Mean transformed (square root) scores of ambulation for generation were subjected to Mendelian cross analysis. Total ambulation scores showed significant additive genetic effect, maternal effect and environmental effect. Directional dominance toward low level of response was present only for males. Daily ambulation scores showed changes in the genetic architecture over three days and the difference between males and females. For females, directional dominance occurred toward low level on Day 1, toward high level on Day 3, but no directional dominance was observed on Day 2. Males' directional dominance toward low level, on the other hand, declined with days. These findings on the genetic architecture of emotionality were discussed with reference to evolutionary adaptation.

Usefulness of monitoring cell-mediated immunity for predicting post-kidney transplantation viral infection.

BACKGROUND: Monitoring cell-mediated immunity (CMI) can be used to estimate the risk of viral infections in kidney transplant recipients. The Immuknow (IMK) assay measures CD4(+) T-cell adenosine triphosphate activity, assesses patient CMI status, and assists clinicians in determining the risk of viral infection. METHODS: We retrospectively analyzed 224 IMK values in 39 kidney transplant recipients at our institution from April 2012 to January 2013. We analyzed the relationship between IMK value and viral infection during the early and late post-transplantation periods. Multiple regression analyses were performed, to determine which factors impacted the results of the IMK assay. RESULTS: Eight patients developed viral infections, including BK virus, cytomegalovirus, herpes simplex, and shingles. Five infections occurred in the early post-transplantation period (<50 d) and 3 in the late period (>120 d). The IMK levels in patients who developed an infection in the early period were within normal limits; however, those in the late period were significantly lower than 200 ng/mL (421.0 ± 062.6 for early vs 153.7 ± 72.7 for late; P = .02). Our multiple regression analyses indicated that peripheral white blood cell and neutrophil counts affected IMK values (P = .03 and P = .02, respectively). CONCLUSIONS: The IMK assay is a useful test for identifying patients at risk for post-transplantation viral infections in the late transplant period.

Induction of cells with cancer stem cell properties from nontumorigenic human mammary epithelial cells by defined reprogramming factors.

Cancer stem cells (CSCs), a small and elusive population of undifferentiated cancer cells within tumors that drive tumor growth and recurrence, are believed to resemble normal stem cells. Although surrogate markers have been identified and compelling CSC theoretical models abound, actual proof for the existence of CSCs can only be had retrospectively. Hence, great store has come to be placed in isolating CSCs from cancers for in-depth analysis. On the other hand, although induced pluripotent stem cells (iPSCs) hold great promise for regenerative medicine, concern exists over the inadvertent co-transplantation of partially or undifferentiated stem cells with tumorigenic capacity. Here we demonstrate that the introduction of defined reprogramming factors (OCT4, SOX2, Klf4 and c-Myc) into MCF-10A nontumorigenic mammary epithelial cells, followed by partial differentiation, transforms the bulk of cells into tumorigenic CD44(+)/CD24(low) cells with CSC properties, termed here as induced CSC-like-10A or iCSCL-10A cells. These reprogrammed cells display a malignant phenotype in culture and form tumors of multiple lineages when injected into immunocompromised mice. Compared with other transformed cell lines, cultured iCSCL-10A cells exhibit increased resistance to the chemotherapeutic compounds, Taxol and Actinomycin D, but higher susceptibility to the CSC-selective agent Salinomycin and the Pin1 inhibitor Juglone. Restored expression of the cyclin-dependent kinase inhibitor p16INK4a abrogated the CSC properties of iCSCL-10A cells, by inducing cellular senescence. This study provides some insight into the potential oncogenicity that may arise via cellular reprogramming, and could represent a valuable in vitro model for studying the phenotypic traits of CSCs per se.