Identification of independent APP locus duplication in Japanese patients with early-onset Alzheimer disease.

BACKGROUND: The occurrence of duplications of the amyloid precursor protein gene (APP) has been described in European families with early-onset familial Alzheimer disease (EO-FAD) and cerebral amyloid angiopathy. However, the contribution of APP duplication to the development of AD in other ethnic populations remains undetermined. METHODS: The occurrence of APP duplication in probands from 25 families with FAD and 11 sporadic EO-AD cases in the Japanese population was examined by quantitative PCR and microarray-based comparative genomic hybridisation analyses. APP expression level was determined by real-time quantitative reverse-transcription (RT) PCR analysis using mRNA extracted from the peripheral blood of the patients. RESULTS: We identified APP locus duplications in two unrelated EO-FAD families. The duplicated genomic regions in two patients of these families differed from each other. No APP duplication was found in the late-onset FAD families or sporadic EO-AD patients. The patients with APP duplication developed insidious memory disturbance in their fifties without intracerebral haemorrhage and epilepsy. Quantitative RT-PCR analysis showed the increased APP mRNA expression levels in these patients compared with those in age- and sex-matched controls. CONCLUSIONS: Our results suggest that APP duplication should be considered in patients with EO-FAD in various ethnic groups, and that increased APP mRNA expression level owing to APP duplication contributes to AD development.

Arthroscopic management of tibial plateau fractures--comparison with open reduction method.

BACKGROUND: Arthroscopic treatment of tibial plateau fractures were compared with open traditional methods retrospectively. METHODS: Twenty-eight patients were included in this comparative study based on fracture patterns and divided into two groups. Using Schatzker's classification system, patients with type II (split depression), type III (isolated depression) fractures, or other fracture types were included. Nineteen of these patients were treated by arthroscopically assisted management (group S), and the remaining 9 underwent the conventional open method (group O). RESULTS: There was no significant difference between both groups in terms of duration of operation, postoperative flexion, and clinical results. In group S, however, the postoperative rehabilitation was easier and faster (the time to obtain 120 degrees of flexion was 4.6+/-3.9 weeks in group S, 9.1+/-8.6 weeks in group O). Furthermore, 16 of 19 patients (84%) in group S obtained an anatomical reduction (defined as <2 mm of residual displacement after surgery), whereas in group O, only 5 of 9 patients (55%) had an anatomical reduction. Also, accurate diagnosis and treatment of any associated joint pathology was possible with arthroscopic management. CONCLUSION: The arthroscopic procedure is recommended in selected tibial plateau fractures.

Increase in doxorubicin cytotoxicity by inhibition of P-glycoprotein activity with lomerizine.

Acquired resistance to chemotherapy is a major problem during cancer treatment. One mechanism for drug resistance is overexpression of the MDR (multidrug resistance)1 gene encoding the transmembrane efflux pump, P-glycoprotein (P-gp). Calcium channel blockers such as verapamil, nifedipine and nicardipine have been shown to reverse cellular drug resistance by inhibiting P-gp drug efflux. This study evaluated whether a new calcium channel blocker, lomerizine, influenced doxorubicin (Dox) cytotoxicity and P-gp activity in a P-gp-expressing cell line compared to a non-expressing subline. Verapamil, and even more markedly, lomerizine, increased cellular uptake of calcein transported by P-gp in a P-gp-expressing erythroleukemia cell line, K562-Dox. Ten microM of lomerizine reduced the IC50 of doxorubicin in the K562-Dox from 60000 ng/ml to 800 ng/ml, whereas the IC50 of doxorubicin in the K562 subline was only marginally affected by these drugs. Lomerizine showed greater reduction in P-gp efflux than verapamil at an equimolar concentration. These results suggest that lomerizine has the clinical potential to reverse tumor MDR involving the efflux protein P-gp.

Unicompartmental knee arthroplasty for lateral gonarthrosis: midterm results.

Thirty-eight lateral compartment arthroplasties were performed in 22 years. Eighteen joints in 17 patients with an average age of 64.5 years were evaluated with minimum 5-year follow-up. Using the Hospital for Special Surgery knee score, 16 of 18 joints had satisfactory results in terms of function and pain relief. The average preoperative alignment on standing was 14.9 degrees of valgus angulation, which was corrected to 6.9 degrees of valgus. On radiographic analysis, no radiolucent lines were seen under the tibial component, but the femoral component was loose in 1 joint. Although 5 joints showed slight deterioration of osteoarthritic change in the medial compartment, lateral compartment arthroplasty is a reliable and successful option in the treatment of patients with a low level of physical activity.

[Pharmacokinetics of cisplatin and methotrexate in a patient suffering from advanced ureteral tumor accompanied by chronic renal failure, undergoing combined hemodialysis and systemic M-VAC chemotherapy].

The pharmacokinetics of cisplatin and methotrexate were determined in a patient suffering from advanced ureteral tumor accompanied by chronic renal failure undergoing 4 consecutive cycles of M-VAC chemotherapy and hemodialysis. No significant difference was observed in t1/2, AUC or CLtot of total platinum between the patient with the chronic renal failure and patients with normal renal function. The AUC and CLtot of free platinum in the patient with the chronic renal failure were higher and lower, respectively, than in the patients with normal renal function. The free cisplatin rebounded remarkably after the end of dialysis, which may be partly attributed to an increase in the AUC and decrease in CLtot. However, the dialysis index was about 75 and 85% in the 3rd and 4th cycles, respectively. The t1/2 and CLtot of methotrexate in the patient with the chronic renal failure tended to be longer and smaller than those in patients with normal renal function, respectively. Seventy-two hours after administration, the methotrexate level was 0.02 microM, which was not at the high-risk level of high-dose therapy. After four cycles of M-VAC therapy, the rest of the right ureteral tumor was extirpated and the clinical response was CR. In conclusion, it is considered that cisplatin and methotrexate can be given to a patient with chronic renal failure. However, the cisplatin and methotrexate serum levels must be monitored, even after very low doses.

Effects of patient demands on satisfaction with Japanese hospital care.

OBJECTIVE: The objective of this study was to detect whether there was any difference among the characteristics of patient satisfaction between two patient emphasis groups: patients demanding technical elements of hospital care and patients demanding interpersonal elements. DESIGN AND SETTING: The sample for this study was drawn from in-patients discharged from 77 voluntarily participating hospitals throughout Japan. The relationship between overall satisfaction with hospital care and patient satisfaction, and the evaluation of a hospital's reputation, was explored by stepwise multiple regression analysis of 33 variables relevant to aspects of hospital care for each patient group. RESULTS: In the interpersonal emphasis (IE) group, 'nurse's kindness and warmth' was associated significantly with overall satisfaction, while 'skill of nursing care' and 'nurse's explanation' were significant predictors of overall satisfaction in the technical emphasis (TE) group. On the other hand, 'doctor's clinical competence', 'recovery from distress and anxiety', and items pertaining to the hospital's reputation were significantly related to overall satisfaction in both emphasis groups. CONCLUSION: For overall patient satisfaction, it is essential to satisfy specific items related to the aspect of hospital care emphasized by the patient. Specific significant predictors of overall satisfaction (e.g. 'doctor's clinical competence') were indispensable measures of professional performance in hospital care, irrespective of the patients' emphasis. A positive perception of hospital reputation items might increase overall patient satisfaction with Japanese hospitals.

Neutralization of feline infectious peritonitis virus: preparation of monoclonal antibody that shows cell tropism in neutralizing activity after viral absorption into the cells.

Feline infectious peritonitis virus (FIPV) infection of feline macro-phages is enhanced by mouse anti-FIPV monoclonal antibody (MAb). This anti-body-dependent enhancement (ADE) of FIPV infection is dependent on mouse MAb subclass, and MAb of IgG2a subclass has a strong ADE activity. Furthermore, MAb showing strong neutralizing activity in Felis catus whole fetus (fcwf-4) cells and Crandell feline kidney (CrFK) cells shows strong enhancing activity in feline macrophages, indicating that the neutralizing epitope and the enhancing epitope are closely related. In this study, we prepared MAb FK50-4 that showed a strong neutralizing activity in feline macrophages, despite the fact that the MAb belonged to the IgG2a subclass. However, MAb FK50-4 did not exhibit neutralizing activity in CrFK cells or fcwf-4 cells, thus showing a very unusual property. MAb FK50-4 recognized FIPV small integral membrane glycoprotein (M protein). Even when feline macrophages were pretreated with MAb FK50-4 prior to FIPV inoculation, this antibody prevented FIPV infection. This reaction disappeared after treatment of FK50-4 with protein A. The neutralizing activity of FK50-4 was also effective on feline macrophages after the cells were inoculated with FIPV. These findings indicated that the FIPV replication mechanism differs between feline macrophages and CrFK/fcwf-4 cells and that a neutralizing epitope that can prevent FIPV infection of feline macrophages after viral absorption is present on M protein.

[Evaluation of effectiveness of community mental health system in Kawasaki City--a comparison with national statistics].

UNLABELLED: The pioneering community mental health system of Kawasaki was evaluated by comparison with national statistics using the data from a citywide survey of patients with schizophrenia. METHOD: Subjects were citizens of Kawasaki who were receiving medical treatment for schizophrenia (ICD-9) in the city. All inpatients (565 cases) and a random sample of outpatients (685 cases; extraction rate was 1/3) who met the criteria were asked to complete the questionnaires by themselves. The responsible medical staffs of the subjects were also asked to complete the questionnaires. In order to calculate the total prevalence rate in the city, the number of patients who received medical treatment outside the city was estimated by extrapolation. RESULTS: Higher outpatient rate and lower inpatient rate of Kawasaki were indicated despite the almost equal prevalence (i.e. 32.7 per 10,000 population) compared to the national rate. The outpatient rate was 70.6% in Kawasaki, which was 16% higher than the national average. And the rate of outpatients with schizophrenia living alone in Kawasaki was three times higher. While national outpatient rates declined significantly when patients became 40 years old, such a decline was not evident in Kawasaki. On the other hand, as patients with schizophrenia got older, the rate of outpatients living alone or living with spouse increased in Kawasaki due to the change of caregivers' generation. CONCLUSION: The higher outpatient rate especially of people over 40 years old and the higher rate of patients who lived alone may be explained by the effective community care system in Kawasaki. However, more an intensive support system which provides' daily services such as home-help and group home services should be developed, as the living conditions of outpatients who lived alone were very difficult.

Protective effects of betamipron on renal toxicity during repeated cisplatin administration in rats and protective mechanism.

The protective effects of betamipron (BP, N-benzoyl-beta-alanine) against nephrotoxicity induced by repeated cisplatin injections were examined. The ratio of the kidney weight to body weight and the lipid peroxide level after treatment with cisplatin plus BP tended to be larger and lower than those after treatment with cisplatin plus alkaline solution, respectively. The blood urea nitrogen, serum creatinine and glutathione levels in the animals treated with cisplatin plus BP differed significantly from those in the animals treated with cisplatin plus alkaline solution. Furthermore, the mechanism of the preventive effects of BP was analyzed for cisplatin-induced nephrotoxicity. The concentration of cisplatin in the renal cortex significantly decreased with concomitant BP. BP inhibited the uptake of cisplatin into the renal cortex in a competitive manner in the same way as an anionic transport inhibitor, probenecid. The treatment with BP appears to be useful for the renal toxicity induced by repeated cisplatin administration.

Betamipron reduces cisplatin nephrotoxicity in rodents without modifying its antileukemic activity in mice.

Protective effects of betamipron (BP, N-benzoyl-beta-alanine), one of a series of N-acyl amino acids, on cisplatin-induced nephrotoxicity were examined. Since the damage observed in the kidney is localized to the proximal tubule cells, we investigated the influence of BP on urinary enzymes and excreta. Male Wistar rats and ddY mice were injected i.p. with 6 mg/kg and 16 mg/kg, respectively, of cisplatin combined with an i.p. 250 mg/kg BP dose. The toxicity of cisplatin as indicated by body weight gain, blood urea nitrogen, and serum creatinine levels was significantly (p < 0.05) suppressed by administration of BP after cisplatin treatment. The increase in urinary N-acetyl-beta-D-glucosaminidase activity, increase and subsequent decrease in gamma-glutamyl transferase activities, and increase in beta 2-microglobulin level observed after treatment with cisplatin were suppressed by administration of BP after cisplatin treatment. The combination of cisplatin and BP had no apparent effect on the efficacy of cisplatin against P388 leukemic cells in mice.

Effects of betamipron on cisplatin nephrotoxicity and its pharmacokinetics in rats.

The protective effects of betamipron (BP, N-benzoyl-beta-alanine) on the nephrotoxicity of cisplatin were examined as indicated by body weight gain, ratio of kidney weight to body weight, blood urea nitrogen and serum creatinine levels in tumor-bearing rats. The results showed clearly that administration of BP 1 h after cisplatin treatment affords protection against the nephrotoxicity of cisplatin. Furthermore, the addition of BP to cisplatin had no apparent effect on the efficacy of cisplatin against Walker 256 carcinosarcoma cells in rats. In addition, no observed significant difference in plasma cisplatin concentration between cisplatin with alkaline solution and cisplatin with BP may be partly attributed to the decrease in the cisplatin exsorption to the intestine and its excretion to bile, and to an increase in cisplatin excretion to the urine.

Protective effects of N-benzoyl amino acids on cisplatin nephrotoxicity in rats.

The protective effects of N-benzoyl amino acids (NAAs) and piperacillin (PIP), anionic transport inhibitors, against the nephrotoxicity of cisplatin were examined in rats. Male Wistar rats were injected i.p. with 6 mg/kg of cisplatin combined with i.p. NAAs or PIP. Rats were sacrificed on day 5 after cisplatin injection to weigh the kidney and liver, and to determine blood urea nitrogen (BUN) and serum creatinine (serum Cr) levels. Treatments with NAAs were an effective means of protection against cisplatin-induced nephrotoxicity. The combination of cisplatin with NAAs containing a short and straight chain significantly suppressed (p < 0.05) the changes in body, kidney and liver weights, BUN and serum Cr. Further, betamipron (BP) at a 2000 mg/kg dose showed no apparent effect on the body, kidney and liver weights, BUN and serum Cr levels in rats. The combination of cisplatin with PIP caused a loss in body weight. The protective effects of PIP against cisplatin toxicity are inferior to those of BP when compared at 250 mg/kg doses.

Protective effect of N-benzoyl-beta-alanine against cisplatin nephrotoxicity in rats.

Prophylactic effects of N-benzoyl-beta-alanine (betamipron, BP), one of a series of N-acyl amino acids, were examined against cisplatin-induced nephrotoxicity. Male Wistar rats were injected i.p. with 6 mg/kg of cisplatin combined with an i.p. BP dose given at various times and various doses. Rats were sacrificed 5 days after cisplatin injection to weigh the kidney and liver, and to determine blood urea nitrogen (BUN) and serum creatinine (serum Cr) levels. Preliminary results suggest that treatment with BP is an effective means of protection against cisplatin-induced nephrotoxicity. Combination with BP reduced the weight loss following treatment with cisplatin. The ratios of the kidney and liver weights to the body weight in the animals treated with cisplatin followed later with BP are significantly different (p < 0.05) from those in the animals that received only cisplatin. The BUN and serum Cr levels in the animals treated with cisplatin followed from -1 to 4 hr, and from -4 to 4 hr later with 250 mg/kg BP dose and followed 1 hr later with from 250 to 1000 mg/kg, and from 250 to 2000 mg/kg BP doses differed significantly (p < 0.05) from those in the animals that received only cisplatin. Histological analysis of the kidneys confirmed the protective effect of BP.

DMI-2 and DMI-3, DNA methyltransferase inhibitors produced by Streptomyces sp. strain no. 560.

Streptomyces sp. strain No. 560 produces several types of DNA methyltransferase inhibitors in the culture filtrate. Two of them, DMI-2 and DMI-3, were distinguished from the previously reported DMI-1 by their inhibitory spectrum and inhibition characteristics against DNA methyltransferase. The molecular weights of DMI-2 and DMI-3 were 854 and 435, respectively. The structure of DMI-2 was determined to be 4"'R,6aR,10S,10aS-8-acetyl-6a, 10a-dihydroxy-2-methoxy-12-methyl-10-[4'-[3"-hydroxy-3",5"-dimethyl-4" (Z-2"',4"'-dimethyl-2"'-heptenoyloxy) tetrahydropyran-1"-yloxy]-5'-methylcyclohexan-1'-yloxy ]-1,4,6, 7,9-pentaoxo-1,4,6,6a,7,8,9,10,10a,11-decahydronaphthacene. The chemical structure of DMI-2 was established as a tautomer of dutomycin which is an antitumor antibiotic produced by Streptomyces sp. 1725. DMI-2 and DMI-3 showed strong inhibition against N6-methyladenine-DNA methyltransferase (M. Eco RI). DMI-2 inhibited M. Eco RI in a competitive manner with respect to plasmid pUC19 used as DNA substrate and in an uncompetitive manner with respect to S-adenosylmethionine (SAM) used as methyl donor. DMI-3 inhibited M. Eco RI in a competitive manner with respect to plasmid pUC19 and SAM. The inhibitory activities of both inhibitors depended upon the pH and temperature in the assay media.

Inhibition of DNA methyltransferase by microbial inhibitors and fatty acids.

Streptomyces sp. strain No. 560 produces four kinds of DNA methyltransferase inhibitors in the culture filtrate. One of them, DMI-4 was distinguished from DMI-1, -2 and -3 previously reported with respect to certain properties, DMI-4 is considered to be a triglyceride consisting of the fatty acids anteisopentadecanoic acid (C15:0), isopalmitic acid (C16:0) and isostearic acid (C18:0) from the results of gas chromatography analysis. Since DMI-4 contains three molecules of fatty acid, and the previously reported DMI-1, 8-methylpentadecanoic acid, is analogous to a fatty acid, the inhibitory activity has been examined of various fatty acids and their methyl esters against Eco RI DNA methyltransferase (M. Eco RI). Oleic acid (C18:1) was found to be a potent inhibiton of M. Eco RI. The inhibitory activity of oleic acid was shown to be pH- and temperature-dependent and inhibited M. Eco RI in a noncompetitive manner with respect to DNA or S-adenosylmethionine (SAM). The number of carbon atoms and double bonds in the fatty acid molecule affected the inhibitory activity, but their methyl esters were not inhibitors. Our results suggest that the length of the carbon chain, the number of double bonds and the presence of a carboxyl group and branched methyl group in the fatty acid molecule may play an important role in the inhibition of DNA methyltransferase.

DMI-1, a new DNA methyltransferase inhibitor produced by Streptomyces sp. strain No. 560.

A new inhibitor of DNA methyltransferase named DMI-1 has been discovered in the culture filtrate of Streptomyces sp. strain No. 560. DMI-1 was purified by extraction with ethyl acetate followed by Diaion HP-20SS and silica gel column chromatography. The structure of DMI-1 was determined to be 8-methylpentadecanoic acid (C16H32O2). DMI-1 is a novel inhibitor of methyltransferase isolated from microorganisms and is structurally different from sinefungin and A9145C which are structural analogs of S-adenosylmethionine (methyl donor). DMI-1 was a strong inhibitor of N6-methyladenine-DNA methyltransferase (M. Eco RI, EC 2.1.1.72) in a noncompetitive manner and its inhibition depended on the pH and temperature in the assay media.

The role of IgG subclass of mouse monoclonal antibodies in antibody-dependent enhancement of feline infectious peritonitis virus infection of feline macrophages.

Antibody-dependent enhancement (ADE) of feline infectious peritonitis virus (FIPV) infection was studied in feline alveolar macrophages and human monocyte cell line U937 using mouse neutralizing monoclonal antibodies (MAbs) directed to the spike protein of FIPV. Even among the MAbs that have been shown to recognize the same antigenic site, IgG 2a MAbs enhanced FIPV infection strongly, whereas IgG 1 MAbs did not. These IgG 2a MAbs enhanced the infection even when macrophages pretreated with the MAb were washed and then inoculated with the virus. Immunofluorescence flow cytometric analysis of the macrophages treated with each of the MAbs showed that the IgG 2a MAbs but not the IgG 1 MAbs bound to feline alveolar macrophages. Treatment of the IgG 2a MAb with protein A decreased the binding to the macrophages and, in parallel, diminished the ADE activity. Although no infection was observed by inoculation of FIPV to human monocyte cell line U937 cells, FIPV complexed with either the IgG 2a MAb or the IgG 1 MAb caused infection in U937 cells which are shown to express Fc gamma receptor (Fc gamma R) I and II that can bind mouse IgG 2a and IgG 1, respectively. These results suggest that the enhancing activity of MAb is closely correlated with IgG subclass and that the correlation is involved in binding of MAb to Fc gamma R on feline macrophage.

[A case of Kennedy-Alter-Sung syndrome associated with external ophthalmoplegia--therapeutic efficacy of fluoxymesterone].

We reported a case with Kennedy-Alter-Sung syndrome (KAS) associated with bilateral external ophthalmoplegia. The patient had movement disturbance of bilateral infra-oblique muscles. The doll's eye phenomenon was not noted. It was suggested that the external ophthalmoplegia was due to the involvement of the oculomotor nucleus that innervated infra-oblique muscle. The serum levels of testosterone and gonadotropin were high, suggesting that the feminization of KAS patients was caused by androgen insensitivity. The feminization of KAS patients is similar to the incomplete form of testicular feminization syndrome except that they do not have feminization of genitals. Therefore, we proposed that abnormalities of androgen receptors might play a role in the pathogenesis of KAS. Fluoxymesterone therapy significantly improved the muscle weakness of the extremities of the patient, even though the therapeutic efficacy was shown temporarily. The therapeutic efficacy of fluoxymesterone for muscle weakness supports our hypothesis.

Macroconidial development and germination in Trichophyton mentagrophytes.

Trichophyton Mentagrophytes was investigated for macroconidial development with particular emphasis on the conidial ageing by light and scanning electron microscopy. Macroconidial germination was also studied under various conditions. Sabouraud glucose agar supplemented with 3% NaCl was used to enhance production of macroconidia. After a long-term cultivation macroconidial compartments changed to spherical thick-walled structure. Some 12-month-old macroconidia were still capable of germination. A wide range of temperature (15-37 degrees C), and inoculum of less than 1 X 10(5) conidia per ml of rich media were appropriate for macroconidial germination. The germination process of macroconidia was highly tolerant to NaCl. A small fraction of the conidia were able to germinate even in distilled water without activation. Effect of freeze-thaw or ultraviolet irradiation on macroconidial germination was determined.

Germ tube-forming cells of Candida albicans are more susceptible to clotrimazole-induced killing than yeast cells.

Yeast and germ tube-forming cells of Candida albicans were compared with respect to their susceptibility to killing induced by the imidazole antifungal clotrimazole. Cultures consisting largely of germ tube-forming cells or exclusively yeast cells were prepared by incubating cells of a germ tube-proficient strain in a proline-containing phosphate buffer at 37 degrees C or 25 degrees C, respectively. When treated with clotrimazole at 37 degrees C, the cultures of germ tube cells lost colony-forming ability much more rapidly than those of yeast cells. However, this difference was diminished in the cells preincubated at 37 degrees C but prevented from forming germ tubes by 5 mM cysteine, a suppressor of germ tube formation. In another C. albicans isolate showing a very poor capacity to form germ tubes at 37 degrees C, such a difference in killing rate was much smaller than that for the germ tube-proficient strain. Furthermore, when an isogenic pair of strains, one proficient and the other deficient in germ tube formation, were compared with each other, germ tube-forming cultures of the former were found to be more sensitive than yeast cell cultures of the latter. It is inferred from these results that the germ tube-forming cell of C. albicans is more sensitive to clotrimazole-induced killing than the yeast cell.