Case report: Electron microscopic evaluation of bone from a patient treated with cinacalcet hydrochloride, maxacalcitol, and alfacalcidol for hyperparathyroid bone disease with secondary hyperparathyroidism.

Evaluation of bone is of great importance in chronic kidney disease patients, as these patients are at an increased risk for fractures. We treated a hemodialysis patient suffering from hyperparathyroid bone disease with cinacalcet hydrochloride and concurrent administration of maxacalcitol and alfacalcidol for a year. Hyperparathyroid bone disease is characterized by cortical thinning, increased cortical porosity, reduced trabecular bone volume, and increased hypomineralized matrix volume, and there is little information to date about the effects of treatment with cinacalcet hydrochloride on the bone fragility in patients with hyperparathyroid bone disease. In the present study, histological and backscattered electron microscopic evaluation of this combination treatment revealed an excellent improvement of both bone volume and bone morphology. This treatment improved cortical thinning, cortical porosity, and trabecular thinning. Furthermore, the treatment also reduced hypomineralized matrix volume, indicative of improved mineralization by osteocytes. We speculate that the intermittent maxacalcitol administration may have effectively stimulated the vitamin D receptors expressed on osteocytes and osteoblasts, resulting in increased mineralization. Our approach for evaluating the bone in patients with chronic kidney disease by backscattered electron microscopy is novel.

Associations between the levels of sclerostin, phosphate, and fibroblast growth factor-23 and treatment with vitamin D in hemodialysis patients with low intact PTH level.

UNLABELLED: Serum sclerostin levels could be closely associated with serum phosphate and fibroblast growth factor-23 levels in hemodialysis patients with low intact parathyroid hormone (PTH) levels. Further study is required to indicate whether these close associations are present in patients with spontaneously low PTH levels without any vitamin D treatment. INTRODUCTION: Intact parathyroid hormone (iPTH) is involved in the interaction between sclerostin and phosphate/fibroblast growth factor-23 (FGF23) in animal models. However, their relationship in patients on hemodialysis (HD) is unclear. METHODS: Data of 102 HD patients were collected regarding clinical and laboratory parameters and mineral bone disorder medications. The patients were divided into subgroups according to the iPTH level (A, <70 pg/mL; B, 70-150 pg/mL; C, 150-300 pg/mL; and D, ≥ 300 pg/mL). RESULTS: The sclerostin level was significantly and positively correlated with phosphate and log of FGF23 levels in subgroups A, B, and combined A and B. Multiple linear regression analysis in the combined A and B subgroup revealed that male sex (t = 3.24, P = 0.01; 95% confidence interval [CI] 11.78 to 50.43) and phosphate level (t = 2.13, P = 0.04; 95% CI, 1.08 to 36.91) were independent factors for serum sclerostin level. The log of serum FGF23 level (t = 1.90, P = 0.06, 95% CI -1.85 to 63.50) appeared to be an important factor for serum sclerostin level. The frequency of patients using vitamin D treatment was not significantly different among subgroups A (93.1%), B (88.0%), C (85.2%), and D (90.5%). CONCLUSION: Serum sclerostin levels were associated with serum phosphate and FGF23 levels in patients with low iPTH levels. Further study is required to indicate whether these close associations are present in patients with spontaneously low iPTH levels without vitamin D treatment.

Reliability of predicted renal function in Japanese patients on cisplatin therapy.

Cisplatin, cis-Dichlorodiammine platinum (II) (CDDP) remains a major antineoplastic drug for the treatment of solid tumors. Its chief dose-limiting side effect is nephrotoxicity. To make a safe and effective dosing regimen of a drug excreted mainly by the renal route, evaluation of patients' renal function is essential. Creatinine clearance (CLcr) or glomerular filtration rate (GFR) is considered to be a standard renal-function test. Several equations have been used in clinical settings, to predict CLcr and GFR using serum creatinine concentration. We carried out a retrospective analysis of the correlation between 24-hour CLcr measured by a urine collection method; and the predicted CLcr and GFR estimated by various equations such as Jelliffe, Yasuda, Orita, Mawer, Mawer, MDRD and modified MDRD, and Cockcroft-Gault. This study used data from Japanese head-and-neck cancer patients, before and after chemotherapy with CDDP. Slopes of regression lines of scatter plots between measured CLcr and predicted renal function in post-CDDP patients were less compared to pre-CDDP patients. On the other hand, Y-intercepts were noted in the scatter plots on renal function from all equations. These results suggest that evaluation of renal function using predictive formulae may have been over-/under-estimated after CDDP administration.

Oxidised LDL/LDL-cholesterol ratio and coronary artery calcification in haemodialysis patients.

BACKGROUND AND AIMS: Serum malondialdehyde-modified low-density lipoprotein (MDA-LDL) and MDA-LDL/LDL-cholesterol (LDL-c) ratio are risk factors for arteriosclerosis and cardiovascular disease (CVD). However, no information is available on these parameters or their associations with coronary artery calcification (CAC) in haemodialysis (HD) patients. METHODS AND RESULTS: Fifty-seven HD patients and 26 control subjects were included in this cross-sectional study. Serum MDA-LDL concentrations and MDA-LDL/LDL-c ratios were examined. HD patients had significantly higher MDA-LDL/LDL-c ratios than the controls (105.1 ± 27.5 vs. 81.4 ± 18.9 mU/mg, P < 0.001); however, there was no significant difference in serum MDA-LDL levels between the 2 groups. CAC scores were examined only in HD patients and their possible associations with the clinical/laboratory data were analysed. Analysis of HD patients showed that MDA-LDL/LDL-c ratio has an association with presence of CVD, CAC score, HD duration, MDA-LDL, or haemoglobin A1C. In addition, the CAC score was positively correlated with serum MDA-LDL level (P = 0.048) and MDA-LDL/LDL-c ratio (P = 0.006). Furthermore, multivariate logistic regression analysis showed that MDA-LDL/LDL-c ratio (ß = 0.04, P = 0.003) and HD duration (ß = 0.16, P = 0.007) were independently associated with CAC score. CONCLUSION: The MDA-LDL/LDL-c ratio of HD patients was significantly higher than that of non-HD subjects and was independently associated with the CAC score. Therefore, this ratio could be an important risk factor for CAC in HD patients.

Moderate and high endemicity of schistosomiasis is a predictor of the endemicity of soil-transmitted helminthiasis: a systematic review.

The authors conducted a systematic literature review with the following aims: to investigate how frequently soil-transmitted helminthiasis (STH) infections are endemic where schistosomiasis is present; and to assess the correlation between the risk level of schistosomiasis and that of STH. Among 155 sites on which data were collected and analyzed, schistosomiasis was present in 130, all of which were also co-endemic for STH, whereas 25 sites were endemic only for STH. Ninety percent (117 out of 130) of the areas eligible for preventive chemotherapy (PC) against schistosomiasis are also eligible for PC against STH. This fact provides managers of control programmes with the operationally important indication that use of available information on endemicity of schistosomiasis is a valid tool to predict the presence of STH in the same geographical area and to estimate the need of PC for STH. The implementation of this tool is expected to save financial and human resources and help accelerate the scale-up of PC throughout the world.

Assessment of E. coli and Salmonella spp. infection risks associated with different fecal sludge disposal practices in Thailand.

The proper management of fecal sludge (FS), to block the transmission pathways of pathogens, is rarely enforced in many parts of the world. Health risks associated with different disposal practices of FS in peri-urban settings of a large metropolis in Thailand were assessed; Tha Klong sub-district with indiscriminate FS dumping, and Klong Luang sub-district which has an FS treatment system. The study showed that indiscriminate FS dumping from along the canal banks and discharge of market waste were likely the major sources of E. coli and Salmonella spp. in contamination of the canal water. The increased microbial pathogen concentrations near the FS treatment facility also indicated contamination risks from poorly designed treatment facilities. Quantitative microbial risk assessment (QMRA) indicated very high water-related infection risk levels compared to the actual locally recorded disease occurrences. These results indicated that the QMRA model needs to be modified to take account of immunological differences between populations in developed countries, where the model was developed, and developing countries. In addition, further sensitivity factors are needed to reflect different societal behavior patterns, and therefore contact with potentially contaminated water, in different sub-populations of many less developed communities.

A quinol peroxidase inhibitor prevents secretion of a leukotoxin from Aggregatibacter actinomycetemcomitans.

BACKGROUND AND OBJECTIVE: Quinol peroxidase (QPO) catalyzes peroxidase activity using quinol in the respiratory chain as a substrate. Quinol peroxidase is essential for the secretion of leukotoxin (LtxA), which destroys leukocytes and erythrocytes in humans and is one of the major virulence factors of Aggregatibacter actinomycetemcomitans, which is associated with localized aggressive periodontitis. In the present study, we aimed to find a highly potent QPO inhibitor to attenuate the virulence of A. actinomycetemcomitans. MATERIAL AND METHODS: For screening of QPO inhibitors, QPO activity was measured kinetically by SpectraMax Plus with 96-well UV plates. Three hundred compounds in the Kitasato Institute for Life Sciences Chemical Library were screened. Secretion of LtxA in the culture supernatant was examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Cytotoxicity against human promyelocytic leukemia cell line (HL-60) cells from the culture supernatant was measured by Trypan Blue exclusion test. RESULTS: The present study characterized ascofuranone as a highly potent inhibitor of QPO (K(i) = 9.557 +/- 0.865 nm). Ascofuranone inhibited secretion of LtxA by A. actinomycetemcomitans in a dose-dependent manner, making A. actinomycetemcomitans less pathogenic to HL-60 cells. CONCLUSION: Quinol peroxidase inhibitors are promising candidates as alternative drugs for the treatment and prevention of the onset of localized aggressive periodontitis. Using ascofuranone as a seed compound, further study of QPO inhibitors could provide novel chemotherapeutic strategies for controlling localized aggressive periodontitis.

Participation of the secreted dipeptidyl and tripeptidyl aminopeptidases in asaccharolytic growth of Porphyromonas gingivalis.

BACKGROUND AND OBJECTIVE: Porphyromonas gingivalis secretes gingipains, endopeptidases essential for the asaccharolytic growth of this bacterium. P. gingivalis also secretes dipeptidyl aminopeptidases (DPPIV and DPP-7) and a tripeptidyl aminopeptidase (PTP-A), although their role in asaccharolytic growth is unclear. The present study was carried out to elucidate the role of these dipeptidyl/tripeptidyl aminopeptidases on the asaccharolytic growth of P. gingivalis. MATERIAL AND METHODS: Knockout mutants for the DPPIV (dpp), dpp7 and/or PTP-A genes were constructed. Brain-heart infusion medium supplemented with sterile hemin and menadione (BHIHM) was used as a complex medium, and the minimal medium used was GA, in which the sole energy source was a mixture of immunoglobulin G and bovine serum albumin. Growth of P. gingivalis was monitored by measuring the optical density of the culture. RESULTS: All knockout mutants for DPPIV, dpp7 and PTP-A grew as well as strain W83 in BHIHM. In GA, growth of single-knockout and double-knockout mutants was similar to that of W83, whereas growth of a triple-knockout mutant (83-47A) was reduced. We purified recombinant DPPIV and recombinant PTP-A from recombinant Escherichia coli overproducers, and purified DPP-7 from the triple-knockout mutant 83-4A. GA supplemented with the three purified dipeptidyl/tripeptidyl aminopeptidases supported the growth of 83-47A. CONCLUSION: DPPIV, DPP-7 and PTP-A contribute to the normal growth of P. gingivalis by cleaving substrate peptides into short-chain polypeptides that are efficient energy sources for P. gingivalis.

Epinastine inhibits eosinophil chemotaxis and adhesion molecules in atopic dermatitis.

PURPOSE: To investigate the effects of epinastine on eosinophil chemotaxis and changes in eosinophil adhesion molecules induced by epinastine and three other antiallergic agents, using eosinophils of atopic dermatitis (AD) patients. RESULTS: Epinastine reduced eosinophil chemotaxis toward eotaxin when the eosinophils had been prestimulated with interleukin (IL)-5, but given alone it did not alter eosinophil chemotaxis toward IL-5. CD11b expression was inhibited when peripheral blood was prestimulated with IL-5, but eosinophil adhesion molecule expression was not altered. CONCLUSIONS: Epinastine suppresses allergic inflammation not only through its strong antihistamine and antimediator effects, but also by inhibiting eosinophilic chemotaxis and the expression of adhesion molecules involved in chemotaxis, especially CD11b.

Development of low-turnover bone diseases after parathyroidectomy and autotransplantation.

Parathyroidectomy and immediate autotransplantation (PTX-AT) has been shown to decrease bone pain and increase bone mineral density. However, adynamic bone disease (ABD) has been predicted to develop if the serum intact parathyroid hormone (i-PTH) level remains lower than normal for a long period of time. Therefore, we investigated the bone histology of patients whose serum i-PTH levels did not increase over 70 pg/mL for 1 year after PTX-AT. Four chronic hemodialysis patients were investigated. The serum intact osteocalcin (i-OC) level was measured and histomorphometry for cancellous bone was performed 1 year after the operation. Tetracycline hydrochloride was administered in the 12 weeks after PTX-AT. The serum i-PTH levels were 20.5 +/- 15.0 pg/mL and i-OC levels were 19.5 +/- 0.9 ng/mL. Histomorphometric analyses showed the osteoclast surface to be 0.1% in two cases and 0% in the other two cases, the eroded surface was 7.7 +/- 6.1%, and the fibrosis volume and osteoblast surface were 0% in all four cases. Osteoid volume, osteoid surface and osteoid thickness were lower in cases 1-3, but higher in case 4. All tetracycline labelings were in contact with the mineralization front in cases 1 and 3, but some were not in cases 2 and 4. Serum i-PTH and i-OC levels indicated that ABD developed in these four cases. Histomorphometric analyses revealed that ABD developed in case 1, while either ABD or low-turnover osteomalacia developed in cases 2 and 4, and low-turnover osteomalacia was observed in case 3 after PTX-AT. In conclusion, i-PTH should not be maintained at lower levels to avoid low-turnover bone diseases.

Detection of HTLV-I proviral DNA in sarcoidosis.

'Sarcoidosis-lymphoma syndrome' is known as an association of sarcoidosis with malignant lymphoma. We report a 56-year-old woman with systemic sarcoidosis who was seropositive for antibody against human T cell lymphoma/leukemia virus type I (HTLV-I). This patient showed integration of HTLV-I proviral DNA within cutaneous sarcoid nodules, but not in peripheral blood mononuclear cells. Neither atypical lymphocytes nor a T cell receptor beta1 gene rearrangement were observed in peripheral blood mononuclear cells or in cutaneous nodules, indicating that the patient did not have a smouldering type of adult T cell lymphoma/leukemia. Detection of integration of HTLV-I proviral DNA in cutaneous sarcoid nodules could suggest that the sarcoid nodules might have been generated as a protective response to chronic stimuli of HTLV-I.

Early changes of bone histology and circulating markers of bone turnover after parathyroidectomy in hemodialysis patients with severe hyperparathyroidism.

AIMS: There have so far been no reports on the changes in bone histology in the early period after parathyroidectomy and autografting (PTX-AG). We investigated the effects of PTX-AG on bone histology during the initial 12 weeks after undergoing these surgical procedures. MATERIALS AND METHODS: We performed bone histomorphometry 3 times (before as well as 4 and 12 weeks after PTX-AG) in 6 patients and 2 times (before and 4 weeks after PTX-AG) in 3 hemodialysis patients. In addition, the circulating parameters of bone metabolism were also assessed before and after PTX-AG in all 9 patients. The changes in the histomorphometric (static) parameters between pre-surgery and 4 weeks after surgery and those between 4 weeks and 12 weeks after surgery were assessed by the t-test while changes in the circulating parameters of bone metabolism were analyzed by Friedman's test. RESULTS: Bone formation parameters including carboxy terminal propeptide of human type I procollagen (PICP), alkaline phosphatase (ALP) and intact osteocalcin (i-OC) were all extremely high before surgery. These parameters initially increased after PTX-AG and thereafter gradually declined. In contrast, the circulating bone resorption parameters including tartrate-resistant acid phosphatase (TRAP) and deoxypyridinoline (Dpyr) were also extremely high at baseline but markedly declined after operation. Osteoid-related parameters including osteoid volume (OV/BV), osteoid surface (OS/BS), and osteoid thickness (O.Th) all initially increased at 4 weeks after PTX-AG. In contrast, osteoblast surface (Ob.S/BS), osteoclast surface (Oc.S/BS), eroded surface (ES/BS), and fibrosis volume (Fb.V/TV) all decreased at 4 weeks after surgery, while Ob.S/BS decreased further at 12 weeks in cases 1-6. Although bone mineralization was ongoing at 4 weeks after surgery, both the mineral apposition rate (MAR) and bone formation rate (BFR) remained below the mean for normal individuals. CONCLUSIONS: The circulating bone formation parameters and osteoid-related parameters showed an initial increase after PTX-AG. The concomitant decline in the circulating bone resorption parameters reflected the reduction in bone resorption. BFR decreased, but bone mineralization did not stop after PTX-AG.

Evaluating the accuracy of uterine cancer screening with the regional cancer registration system.

OBJECTIVE: To evaluate the effectiveness of uterine cancer screening by analyzing the accuracy of cervical and endometrial cytodiagnoses as screening methods. STUDY DESIGN: During the year of April 1, 1991-March 31, 1992, 186,161 and 5,697 women underwent cervical and endometrial cytodiagnoses, respectively, and their cytodiagnostic results were computer registered at the Miyagi Cancer Society. By comparison of these examinees with 753 cancer patients who were registered at the regional cancer registry between 1991 and 1993, 133 individuals who were assumed to be identical between the two systems were selected, and of these cases, 83 patients, including test-positive cases, were found within one year. The sensitivity and specificity of each screening method were investigated. RESULTS: Regarding examinees diagnosed as having cancer by the same month in the following year after diagnosis on screening as false negative, the sensitivity, specificity and false negative rates of cervical cytodiagnosis were 94.7%, 98.9% and 5.3%, respectively, and those of endometrial cytodiagnosis were 83.3%, 96.7% and 16.7%, respectively. CONCLUSION: In comparison with the accuracy of cancer examinations for other organs performed by the health care administration, the accuracy of cervical and endometrial cytodiagnoses was sufficient to designate them screening methods.

Involvement of granule-mediated apoptosis in the cyclic changes of the normal human endometrium.

Our objective is to investigate the involvement of granule-mediated apoptosis in the cyclic changes of the endometrium. We demonstrated the localization of CD56, perforin, granzyme B and caspase-3 in the endometrium by immunohistochemistry. We also confirmed the localization of perforin by immuno-electron microscopy, and demonstrated apoptosis in endometrial glandular cells by TdT-mediated dUTP-biotin nick end labeling (TUNEL) and electron microscopy. Uterine CD56-positive natural killer (NK) cells expressed perforin and granzyme B in its cytoplasm. Uterine NK cells increased significantly in the endometrial stroma during the secretory phase, and peaked during the late secretory phase. These cells started decreasing in number during the menstrual period. In endometrial glandular cells, caspase-3 and TUNEL-positive cells increased significantly from the late secretory phase, with apoptosis reaching a peak during the menstrual period. Using electron microscopy, we observed uterine NK cells with chromatin rich, segmented nuclei and intracytoplasmic granules in the stroma obtained from late secretory phase endometria. These cells extended projections to the lining of endometrial glandular cells and attached to form a cell-to-cell contact. In addition, nuclear chromatin was observed to have already cohered and small cytoplasmic organelles were beginning to disappear, suggesting that these endometrial glandular cells were undergoing apoptosis. Utilizing immuno-electron microscopy, intracytoplasmic granules in uterine NK cells were stained with anti-perforin antibody. The findings of this study suggest that granule-mediated apoptosis in endometrial glandular cells induced by NK cells expressing perforin and granzyme B may be associated with the onset of menstruation.

Synthesis and absolute configuration of MQ-A3 [1-(14'-methylhexadecanoyl) pyrrolidine], a novel aliphatic pyrrolidine amide from the tropical convolvulaceous species.

A novel pyrrolidine amide (MQ-A3) isolated from the tropical convolvulaceous species was synthesized in 5 steps by starting from commercially available 12-bromododecanol and (S)-2-methylbutylbromide. The absolute configuration of the natural product was confirmed by a comparison of the specific rotation values.

Prognostic value of thymidine phosphorylase immunostaining in patients with uterine cervical cancer treated concurrently with doxifluridine, radiotherapy and immunotherapy.

Thymidine phosphorylase (dThdPase) is reportedly identical to platelet-derived endothelial cell growth factor (PD-ECGF). We conducted immunohistochemical staining of dThdPase to assess correlation between its expression in cancer tissue and efficacy of a combination therapy with 5'-DFUR, radiotherapy and sizofilan (SPG) in uterine cervical cancer patients. No difference in response rates was observed between dThdPase positive and negative tumor and stromal cells. Survival curves significantly differed between stromal dThdPase positive and negative groups (p=0.032). Results showed that dThdPase immunostaining is possibly prognostic and predictive in determining success of the combination therapy.

Effects of concomitant use of doxifluridine, radiotherapy and immunotherapy in patients with advanced cervical cancer.

Clinical effects of doxifluridine (group A, 600 mg/body/day; group B, 800 mg/body/day) combined with radiotherapy and immunotherapy were evaluated in patients with advanced cancer of the uterine cervix. Response rates were 84.2% (16/19 patients) in group A and 100% (18/18 patients) in group B, respectively (p=0.230). There was no significant difference in adverse reaction incidence between the methods but significantly higher grade adverse reaction were observed in group B than in group A (p=0.048). Time to progression (TTP) was longer in group B than in group A (p=0.081). The optimal 5'-DFUR dose was 800 mg/body (group B), by which higher grade adverse reactions were fully controlled and TTP was prolonged.

Observed versus expected rates of unbalanced fetal karyotype at second trimester amniocentesis when one parent carries a balanced translocation.

Our purpose is to evaluate a model proposed by Stene et al. for estimating the risk of unbalanced progeny at the time of second trimester amniocentesis for 114 pregnancies of 71 couples, who are carriers of a reciprocal translocation. The overall estimated risk was 18%, 1.26 times higher than the observed risk (14%). There were no unbalanced fetuses in the 19 pregnancies of the no-risk group. In the low-risk group, the observed risk was highly correlated with the estimated risk. However, the estimated risks in the medium- and high-risk groups were much higher than the observed risks (2- and 1.5-fold). In the group ascertained through spontaneous abortions, the average estimated risk was 1.9 times higher than the observed risk. Although the model is useful in estimating individual empirical risks, it may be necessary to consider the type of ascertainments and add some modifications to calculate the risks in the second trimester.

Analysis of estrogen receptor alpha and beta in endometrial carcinomas: correlation with ER beta and clinicopathologic findings in 45 cases.

Estrogens play important roles in the pathogenesis of the great majority of endometrial endometrioid adenocarcinoma. Recently, a novel estrogen receptor (ER), ER beta, has been characterized, but little is known about the status of ER beta in endometrial carcinoma. We therefore examined expression of both ER alpha and ER beta in 45 cases of endometrioid endometrial adenocarcinoma using mRNA in situ hybridization, reverse transcription and polymerase chain reaction (RT-PCR), and immunohistochemistry. We also correlated the findings with various clinicopathologic parameters in these cases to examine their possible biologic significance. Accumulation of mRNA hybridization signals for both ER alpha and ER beta was detected predominantly in the cytoplasm of carcinoma cells, and to a lesser extent in some stromal cells. ER beta mRNA was detected in 16/45 cases (35.6%), and ER alpha mRNA hybridization signals were detected in 36/45 cases (80.0%). Among the 16 ER beta positive cases, 15 cases also had ER alpha mRNA hybridization signals. In the cases that expressed both ER alpha and ER beta, ER alpha mRNA hybridization signals were more widely distributed than ER beta mRNA. In 21 cases, carcinoma cells had ER alpha mRNA hybridization signals but not ER beta mRNA. There was a statistically significant positive correlation between the results of mRNA in situ hybridization and semiquantitative RT-PCR or immunohistochemistry for both ER alpha and ER beta. There were no significant correlations between ER beta mRNA expression and PR labeling index, Ki67 LI, age, or histologic grade. The results from our study indicate that ER beta is coexpressed with ER alpha, and that the estrogenic effects occur predominantly through ER alpha in endometrial carcinomas.