HslO ameliorates arrested ΔrecA polA cell growth and reduces DNA damage and oxidative stress responses.

Chromosome damage combined with defective recombinase activity has been widely considered to render cells inviable, owing to deficient double-strand break repair. However, temperature-sensitive recAts polA cells grow well upon induction of DNA damage and supplementation with catalase at restrictive temperatures. These treatments reduce intracellular reactive oxygen species (ROS) levels, which suggests that recAts polA cells are susceptible to ROS, but not chronic chromosome damage. Therefore, we investigated whether polA cells can tolerate a complete lack of recombinase function. We introduced a ΔrecA allele in polA cells in the presence or absence of the hslO-encoding redox molecular chaperon Hsp33 expression plasmid. Induction of the hslO gene with IPTG resulted in increased cell viability in ΔrecA polA cells with the hslO expression plasmid. ΔrecA polA cells in the absence of the hslO expression plasmid showed rich medium sensitivity with increasing ROS levels. Adding catalase to the culture medium considerably rescued growth arrest and decreased ROS. These results suggest that hslO expression manages oxidative stress to an acceptable level in cells with oxidative damage and rescues cell growth. Overall, ROS may regulate several processes, from damage response to cell division, via ROS-sensitive cell metabolism.

Seed production and storage for endangered Morus boninensis using an ex-situ living collection.

Meaningful conservation techniques for the endangered Morus boninensis require seed production and storage, since in situ purebred seed production is infrequent in Ogasawara Islands (World Natural Heritage Site) in Japan. Ex situ living M. boninensis specimens produced seeds by open- and artificial-pollination in a greenhouse. Seed desiccation tolerance and storage characteristics were examined. Open-pollinated seeds were dried at 75, 33 and 8% relative humidity (RH) for 5 days then cryopreserved at -170 °C. Artificially pollinated seeds were dried at 8% RH then stored cryogenically at -170 °C and in freezers at -80 or -20 °C for 6 months. Germination rates were compared among seeds before and after drying and storage. Germination rates of undried seeds were 83% for open-pollinated seeds and 65% for artificially pollinated seeds. M. boninensis seeds exhibited the characteristic high desiccation tolerance of orthodox seeds and maintained a high germination rate after drying to a moisture content of approximately 4%. Cryopreservation of open-pollinated dry seeds with moisture contents of ca. 12, 7 and 4% maintained germination rates, while cryopreservation of undried seeds (moisture content ca. 44%) resulted in no germinability. Cryogenic and freezer storage of artificially pollinated dry seeds for 6 months resulted in maintenance of germination rates comparable to undried seeds before storage. Seed production was enabled by ex situ M. boninensis collection. The present study also developed a method for low temperature seed storage. Ex situ living tree collection represents a feasible seed resource for reintroduction and restoration programmes for M. boninensis.

A genome-wide association study of coping behaviors suggests FBXO45 is associated with emotional expression.

Individuals use coping behaviors to deal with unpleasant daily events. Such behaviors can moderate or mediate the pathway between psychosocial stress and health-related outcomes. However, few studies have examined the associations between coping behaviors and genetic variants. We conducted a genome-wide association study (GWAS) on coping behaviors in 14088 participants aged 35 to 69 years as part of the Japan Multi-Institutional Collaborative Cohort Study. Five coping behaviors (emotional expression, emotional support seeking, positive reappraisal, problem solving and disengagement) were measured and analyzed. A GWAS analysis was performed using a mixed linear model adjusted for study area, age and sex. Variants with suggestive significance in the discovery phase (N = 6403) were further examined in the replication phase (N = 7685). We then combined variant-level association evidence into gene-level evidence using a gene-based analysis. The results showed a significant genetic contribution to emotional expression and disengagement, with an estimation that the 19.5% and 6.6% variance in the liability-scale was explained by common variants. In the discovery phase, 12 variants met suggestive significance (P < 1 × 10-6 ) for association with the coping behaviors and perceived stress. However, none of these associations were confirmed in the replication stage. In gene-based analysis, FBXO45, a gene with regulatory roles in synapse maturation, was significantly associated with emotional expression after multiple corrections (P < 3.1 × 10-6 ). In conclusion, our results showed the existence of up to 20% genetic contribution to coping behaviors. Moreover, our gene-based analysis using GWAS data suggests that genetic variations in FBXO45 are associated with emotional expression.

Effect of weight loss using formula diet on renal function in obese patients with diabetic nephropathy.

OBJECTIVE: To evaluate the effect and safety of treatment with low-calorie formula diet on renal function and proteinuria in obese patients with diabetic nephropathy. DESIGN: Prospective study on safety and efficacy of a 4-week low-calorie (11-19 kcal/kg/day) normal-protein (0.9-1.2 g/kg/day) diet partly supplemented with formula diet. SUBJECTS: In all, 22 obese patients with diabetic nephropathy (BMI: 30.4+/-5.3 kg/m(2), HbA1c: 7.1+/-1.4%, serum creatinine: 172.4+/-57.5 micromol/l, urinary protein: 3.3+/-2.6 g/day). RESULTS: The mean body weight decreased by 6.2+/-3.0 kg. The mean systolic blood pressure, creatinine, blood urea nitrogen, urinary protein, and 8-hydroxydeoxyguanosine decreased significantly by 7.5+/-12.7 mmHg, 41.6+/-23.9 micromol/l, 1.50+/-1.61 mmol/l, 1.8+/-1.7 g/day, and 3.1+/-3.6 ng/mg creatinine, respectively. No patient had increased serum creatinine and urinary protein. Mean creatinine clearance (40.6+/-17.9 to 46.1+/-14.6 ml/s/1.73 m(2)) and serum albumin showed no significant changes. Delta serum creatinine and Delta urinary protein correlated with Delta body weight (r=0.62 and 0.49, respectively) and Delta visceral fat area (r=0.58 and 0.58, respectively), but did not correlate with Delta systolic blood pressure, Delta fasting blood glucose and Delta subcutaneous fat area. CONCLUSION: These results suggested that weight reduction using formula diet might improve renal function and proteinuria safely for a short term in obese patients with diabetic nephropathy.

Effectiveness of coagulation factor XIII concentrate for reversing loss of tensile strength of rat intestinal anastomoses.

BACKGROUND: Blood coagulation factor XIII (F-XIII) promotes cross-linking of fibrin during blood coagulation. Impaired clot stabilization in patients with genetic deficiencies of F-XIII is associated with marked pathologies of wound healing. METHODS: 60 rats given carbon tetrachloride underwent ileal anastomosis after which they received albumin (Alb animals) or F-XIII concentrate (F-XIII animals) immediately after surgery and daily thereafter until day 2 or 4 and were euthanized on day 3 or 5, respectively, or until day 5 and were euthanized on day 7, 10, or 21. We measured the plasma F-XIII activity and anastomotic tensile strength, followed by immunohistochemical localization of F-XIII subunit A within anastomoses. RESULTS: On day 3, there were no significant differences between Alb and F-XIII animals for plasma F-XIII activity and tensile strength, and both groups of animals showed little immunostaining for F-XIII on anastomoses. Plasma F-XIII activities did not differ between Alb and F-XIII animals on day 5 (115.8 +/- 16.8 vs. 137.3 +/- 14.9%). Although the tensile strength in both groups was increased compared with that of day 3, that in F-XIII animals (129.8 +/- 3.3 gf) was significantly higher than that in Alb animals (100.8 +/- 5.3 gf, p = 0.014). F-XIII animals showed de novo collagen fibers and intense immunoreactivity of F-XIII in the extracellular matrix around the anastomoses. Similar differences occurred on day 7 but not days 10 and 21. CONCLUSIONS: F-XIII concentrate may accelerate the early healing process of intestinal anastomosis because of the protein's accumulation in situ.

Capability of wild Rosa rugosa and its varieties and hybrids to produce sesquiterpene components in leaf glandular trichomes.

The sesquiterpene contents in leaves of wild Rosa rugosa and of sixty-one hybrid rugosas were quantitatively measured by a GC analysis. In this group of samples, the greater the number of glandular trichomes the hybrid rugosas possessed on their leaves, the larger the amount of sesquiterpenes they accumulated. In contrast, those having no leaf glandular hairs contained only a trace amount of sesquiterpene components. The concentrations of bisaborosaol A (1) and carota-1,4-dienaldehyde (2) as representative sesquiterpenes of R. rugosa were positively correlated with the density of the glandular trichomes. Furthermore, an approximately regular correlation was observed between the concentrations of 1 and 2 in most of the sesquiterpene-producing hybrid rugosas, regardless of their productivity. This suggests that a major part of these hybrid rugosas have inherited from R. rugosa the ability to produce two skeletally different sesquiterpenes in parallel with a phenotype to develop leaf glandular trichomes. This investigation also led to discovering 1-dominant (e.g., Amelie Gravereaux and Purple Pavement), 2-dominant (e.g., David Thompson), and other-dominant (e.g., Martin Frobisher) types of sesquiterpene-producing hybrid rugosas.

Peripheral nerve regeneration through a long detergent-denatured muscle autografts in rabbits.

Muscle segments excised from rabbit biceps femoris muscles were treated with detergent sodium dodecyl sulphate to denature cellular constituents, and each was autografted in a 5 cm gap of the sciatic nerve in the same rabbit. Axonal regrowth through the grafts and reinnervation into the host sciatic nerves and muscles were studied morphologically, and electrophysiologically, 4 months after grafting. Regenerating axons accompanied by Schwann cells extended through basal lamina tubes of the grafts into the distal host nerves. Reinnervation of the tibialis anterior muscles by motor nerves was confirmed by recovery of the compound muscle action potentials (CMAP) and the reinnervation of the muscle spindles was demonstrated by electron microscopy. These findings indicated that the basal lamina tubes of denatured muscles were effective scaffolds through which the regenerating nerve fibers grew across as large a gap as 5 cm.

[Biliary tract cancer].

The proportion of mortalities from biliary tract cancer, which includes gallbladder and extrahepatic bile duct cancers, is only 5% of all deaths from malignant neoplasms in Japan. Epidemiologists, however, are concerned about the cancer due to its peculiar geographic distribution. In addition, they have also noted the recent decreasing trends in mortality rates, especially among females. The most probable explanation is either a change in the effects of risk factors or the development of abdominal ultrasound sonography and laparoscopic cholecystectomy, or both. Further research should be directed towards an analysis of the causes of this decreasing trend.

Grafting of choroid plexus ependymal cells promotes the growth of regenerating axons in the dorsal funiculus of rat spinal cord: a preliminary report.

Nerve regeneration in the central nervous system has been studied by grafting various tissues and cells. In the present study, we demonstrated that choroid plexus ependymal cells can promote nerve regeneration when grafted into spinal cord lesions. The choroid plexus was excised from the fourth ventricle of adult rats (Wistar), minced into small fragments, and grafted into the dorsal funiculus at the C2 level in adult rat spinal cord from the same strain. Electron microscopy and fluorescence histochemistry showed that ependymal cells of the grafted choroid plexus intimately interacted with growing axons, serving to support the massive growth of regenerating axons. CGRP-positive fibers closely interacted with grafted ependymal cells. HRP injection at the sciatic nerve showed that numerous HRP-labeled regenerating fibers from the fasciculus gracilis extended into the graft 7 days after grafting. This regenerating axons from the fasciculus gracilis was maintained for at least 10 months, with some axons elongating rostrally into the dorsal funiculus. Evoked potentials of long duration were recorded at a level ca. 5 mm rostral to the lesion in the rats 8 to 10 months after grafting. These findings indicate that choroid plexus ependymal cells have the ability to facilitate axonal growth in vivo, suggesting that they may be a promising candidate as graft for the promotion of nerve regeneration in the spinal cord.

Effect of physical activity during teenage years, based on type of sport and duration of exercise, on bone mineral density of young, premenopausal Japanese women.

In this cross-sectional study, 91 healthy premenopausal women aged 20-39 years were investigated to determine the effect of physical activities during their teenage years on their current bone mineral densities (BMD). We measured whole-body BMD (WBMD), lumbar BMD (LBMD), and radial BMD (RBMD) with dual energy X-ray absorptiometry (DXA). Using a questionnaire, we asked the women about their physical activities during junior and senior high school and at present. We also asked about their current nutritional status and past and current milk intake. After adjusting for age, body mass index (BMI), current total calorie and calcium (Ca) intake, and milk intake when they were teenagers and at present, we determined that subjects who exercised during extracurricular activities at each of the three periods (during junior and senior high school and at present) had significantly higher WBMD and LBMD (P <0.01, respectively) than did those who did not exercise at those times. Subjects who played high-impact sports at each period had significantly higher WBMD and LBMD than did subjects who played low-impact sports (P <0.05, respectively). Subjects who had exercised regularly from their teenage years to the present had significantly higher BMD at all sites than BMD in other subjects after adjusting for the potential confounders described above (P <0.05, respectively). Our data suggest that continuous exercise beginning in junior high school, especially high-impact sports, may be associated with greater current bone mass. It is important to incorporate adequate exercise beginning in the teenage years to lower one's future risk for osteoporosis.

(+)-4-epi-alpha-bisabolol as a major sesquiterepene constituent in the leaves of two Rosa rugosa hybrids, Martin Frobisher and Vanguard.

During an investigation of the sesquiterpene phases and contents in leaves of several Rosa rugosa hybrids (hybrid rugosas), Martin Frobisher and Vanguard were found to accumulate a large amount of (+)-4-epi-alphabisabolol (1) as a single constituent. Although glandular trichomes of Martin Frobisher on the leaves are dense, this R. rugosa hybrid produces none of the carota-1,4dienaldehyde (2) or bisaborosaol A (3) that are both found as representative sesquiterpenes of the carotane and bisabolane classes, respectively, in a glandular trichome exudate of wild-type R. rugosa. Compound 1 was also apparent as a nearly single constituent detectable by GC in the leaf constituents of Vanguard possesses sparse glandular trichomes on the leaf. Martin Frobisher and Vanguard had likely lost their capability to form carotane-type sesquiterpenes and had also lost their activity to oxygenate the C-7 allyl methyl carbon of compound 1 to convert 3. The presence of (+)-4-epi-alphabisabolol-accumulating R. rugosa hybrids is significant when considering the sesquiterpene biogenesis of Rosa rugosa.

Ph-negative non-Hodgkin's lymphoma occurring in chronic phase of Ph-positive chronic myelogenous leukemia is defined as a genetically different neoplasm from extramedullary localized blast crisis: report of two cases and review of the literature.

This report describes two cases of Philadelphia chromosome-negative (Ph(-)) non-Hodgkin's lymphomas (NHLs) recognized in patients with chronic phase Ph-positive (Ph(+)) chronic myelogenous leukemia (CML). Lymph node biopsy of patient 1 was initially diagnosed as diffuse large B cell non-Hodgkin's lymphoma (NHL, T cell rich variant), but at relapse showed immunoblastic features with a marked decrease of admixed lymphocyte components. Patient 2 presented with thickened parietal pleura which revealed a CD30-positive anaplastic large cell lymphoma showing null cell phenotype and genotype with abundant admixed neutrophils and lymphocytes. At the time of lymphoma diagnosis, the patients had CML for 33 and 10 months, respectively. DNA obtained from bone marrow cells at the time of lymphoma diagnosis showed BCR/ABL gene rearrangements by both Southern blot analysis and reverse transcription polymerase chain reaction (RT-PCR), but lacked both immunoglobulin and T cell receptor gene rearrangements. BCR gene rearrangement and BCR/ABL fusion gene were also identified in lymph node and pleural biopsies by Southern blot and RT-PCR analysis, respectively. However, both biopsy specimens also contained reactive lymphocytes and neutrophils, and no fusion signals between BCR and ABL genes were identified in the hyperdiploid lymphoma cells of either case by fluorescence in situ hybridization (FISH). These data suggest the lymphoma cells in both cases were not genetically associated with BCR/ABL. Therefore, these cases were not diagnosed as an extramedullary localized blast crisis in CML, but as Ph(-) NHLs. This represents the first definitive demonstration of peripheral B cell lymphoma occurring by a separate genetic pathway, lacking BCR/ABL, in patients with Ph(+) CML. A review of the literature identified two different subtypes of malignant lymphomas arising in patients with an antecedent or concurrent diagnosis of CML. The most common are T cell lymphomas displaying an immature thymic phenotype, while peripheral B cell lymphomas are more rare. Our study shows, however, that 'Ph(+) NHL' occurring in CML or acute lymphocytic leukemia (ALL) may represent an unrelated neoplasm, even if standard cytogenetic analysis reveals a Ph(+) chromosome, and that FISH is required to confirm whether a localized lymphoid neoplasm is either a true extramedullary localized blast crisis or genetically distinct neoplasm. Leukemia(2000) 14, 169-182.

Vitamin D nutritional status of women living on a solitary island in Japan: A population-based study.

OBJECTIVE: Serum 25-hydroxyvitamin D [25(OH)D] is a vitamin D metabolite and a good indicator of vitamin D nutritional status. Low 25(OH)D levels accelerate age-related bone loss in women. The aim of this study was to assess 25(OH)D levels using population-based samples from women in a community in Japan. SUBJECTS AND METHODS: Of all 187 adult women living on a solitary island (Niigata, Japan), 150 (80.2%) were enrolled in a cross-sectional study in early June 1998. After excluding 6 subjects who were undergoing treatment for osteoporosis, 144 female subjects were analyzed. Serum 25(OH)D(2) and 25(OH)D(3) were determined by high-performance liquid chromatography. The sum of 25(OH)D(2) and 25(OH)D(3) was calculated, yielding 25(OH)D, for which a concentration of less than 30 nmol/L was defined as vitamin D insufficiency. Demographic data such as age, height, weight, and body mass index (BMI) were also recorded. RESULTS: The average age of the subjects was 61.3 years (SD 12.8), ranging from 21 to 87. The average concentrations of 25(OH)D(2) and 25(OH)D(3) were 0.5 nmol/L (SD 3.2) and 64.6 nmol/L (SD 17.6), respectively. The number of subjects with 25(OH)D concentration less than 30 nmol/L was 4 of 149 (2.7%). Serum 25(OH)D concentrations were not significantly correlated with age (r=-0.065, p=0.44l) or BMI (r=0.086, p=0.310). CONCLUSION: The present population-based study confirms adequate levels of 25(OH)D and low prevalence of vitamin D insufficiency in Japanese women. Further research should be directed toward darifying which dietary factors determine vitamin D nutrition.

[Ecological correlation studies on standardized incidence ratios of malignant neoplasms contracted from different sources of drinking water supply].

Ecological correlation analysis was made between the source of drinking water and standardized incidence ratios (SIRs) of malignant neoplasms registered in Niigata, Japan. The purpose of this study was to find out some clues for generating a working hypothesis that the contamination of drinking water with environmental chemicals especially of endocrine disrupting chemicals in big rivers may be related to the occurrence of cancers. Sources of drinking water in the year 1974 were classified into two categories; one from the Shinano and Agano and their down-stream rivers, and the other from small rivers flowing into these big rivers, rivers originating from the mountain areas, underground and reservoirs (The other group). After excluding registered cases under 20 years of age, the registry data (1992-94) were sub-divided into two categories according to the source of drinking water. The SIRs were calculated by using two different standard populations; Niigata total and "The other group". All of the sites were examined in the former standard. In the latter case, however, 12 and 9 sites in males and females, respectively were selected based on the assumption that the expected numbers in each age group totaled 5 or over when there were more than 200 registered patients as a total. Concerning the judgment for the significant association in the former standard, we considered the presence of positive associations when both of the SIRs in the two categories were significantly different from the Niigata total population (p < 0.05 by chi 2 test). Of the 33 sites in males, the following sites were correlated with the source of drinking water; tongue, colon, rectum, gallbladder, etc., prostate, kidney, etc. and all sites. Of 35 sites in females, those were the colon, gallbladder, etc., bronchus and lung, breast, corpus uteri, thyroid and all sites. Of the 12 and 9 respective sites subjected to the second analysis, positive associations were observed in the following: esophagus, colon, rectum, liver, gallbladder, etc., bronchus and lung, prostate, bladder and all sites in males, and colon, rectum, gallbladder, etc., bronchus and lung, breast and all sites in females. Final conclusion, however, should be waived until the consistency of association can be confirmed by using more reliable incidence data, because the difference in the death certificates only (DCO) rate between the two groups might have played as a bias. The use of mortality data may be an alternative way to evaluate the present findings in terms of the consistency of association.

[A neutropenic acute myeloid leukemia patient complicated with chronic otitis media due to Aspergillus niger and yeast-like fungi caused by superinfection].

There have been few reports describing otomycosis in association with compromised hosts. So we report a neutropenic acute myeloid leukemia (AML) patient complicated with otomycosis caused by superinfection. A 51-year-old male was admitted because a third relapse of AML in March 1998. Two years ago, he was diagnosed as having chronic otitis media involving the VII cranial nerve due to Pseudomonas aeruginosa coinciding with AML. Then, he had suffered from a right-sided earache and otic discharge in accord with every myelosuppression, which improved on treatment with otic administration of ofloxacin. After 1 course of induction chemotherapy, he developed a spiking fever with severe earache and otic discharge at a nadir period of WBC. Ear swab cultures yielded Aspergillus niger and yeast-like fungi. So, he was treated with intravenous administration of amphotericin B (AMPH-B): initial dose was 5 mg/day and was gradually increased to 30 mg/day. Thereafter, the otic symptoms subsided and never recurred. Subsequently, he was given another antifungal agent, itraconazole. Although induction chemotherapies resulted in failure, he did not suffer otic symptoms until his death due to cerebral bleeding in January 1999. For neutropenic patients without rapid hematological improvement, we recommend intensive antifungal therapy as the first-line of therapy for otomycosis rather than local therapy.

Left ventricular mass regression after implantation of St. Jude Medical cardiac valves in small aortic roots.

In this study, we analyzed the extent of regression of left ventricular hypertrophy in patients who received small St. Jude Medical (SJM) aortic valves and compared the results with those of another group receiving larger valves. Eighty-eight patients received either 19 or 21 mm valves (Group 1, 25 patients) or either 23 or 25 mm valves (Group 2, 53 patients). Echocardiographic studies were done before the operation and 5 years postoperatively. At follow-up a significant reduction in the left ventricular mass was found for both patient groups (p < 0.0001). Doppler echocardiography derived pressure gradients for both groups were obtained during the follow-up period. As expected, the patients in Group 1 had higher peak pressure gradients than did those in Group 2. However, there was no significant difference between the 2 groups or any significant correlations between peak pressure gradients and body surface area (BSA). Actuarial survival was 84.7% at 15 years for Group 1 and 85.9% at 17 years for Group 2. Actuarial freedom from valve related events was 91.4% at 15 years for Group 1 and 82.7% at 17 years for Group 2. There was no significant difference in survival or valve related event free curves between the 2 groups. After implantations of SJM valves in small aortic roots, significant left ventricular mass regression was obtained, and the results were comparable to those for valves of other sizes. The long-term performance of aortic valve replacement with small valves was satisfactory as judged by improvement in the functional class of patients and survival statistics, the durability of the prosthesis, and valve related morbidity comparable to that of valves of other sizes.