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In this study, sex determination using polymerase chain reaction (PCR) on tooth material was evaluated from the viewpoint of forensic medicine. The sensitivity of PCR for detection of the Y chromosome-specific alphoid repeat sequence and the X chromosome-specific alphoid repeat sequence was 0.5 pg of genomic DNA. Sex could be determined by PCR of DNA extracted from the pulp of 16 freshly extracted permanent teeth and dentine including the surface of the pulp cavity of 6 freshly extracted milk teeth. Sex could be determined using the pulp in all 20 teeth (10 male and 10 female) preserved at room temperature for 22 years. For the pulp of teeth stored in sea water, the sex could be determined in all 8 teeth immersed for 1 week and in 5 of 6 teeth immersed for 4 weeks. In the remaining 1 tooth, in which sex determination based on the pulp failed, the sex could be determined correctly when DNA extracted from the tooth hard tissue was examined. For teeth stored in soil, the sex could be determined accurately in all 8 teeth buried for 1 week, 7 of 8 teeth buried for 4 weeks, and in all 6 teeth buried for 8 weeks. When teeth were heated for 30 min, sex determination from the pulp was possible in all teeth heated to 100, 150, and 200 degrees C, and even in some teeth heated to 250 degrees C. When this method was applied to actual forensic cases, the sex of a mummified body estimated to have been discovered half a year to 1 year after death could be determined readily by examination of the dental pulp. In the skeletons of 2 bodies placed under water for approximately 1 year and approximately 11 years and 7 months, pulp tissues had been dissolved and lost, but sex determination was possible using DNA extracted from hard dental tissues. These results indicate that this method is useful in forensic practices for sex determination based on teeth samples.

The effect of exogenous thyroxine (T4) administration on the activity of rhodanese, cystathionase, and 3-mercaptopyruvate sulfurtransferase (MPST) in the mitochondrial and cytosolic fractions of mouse liver was investigated. Three groups of mice were treated for 6 consecutive days with subcutaneous injections of T4 (50 micrograms, 100 micrograms, and 250 micrograms per 100 g of body wt, respectively). The other 3 groups were given 100 micrograms of T4 per 100 g of body wt for 1, 2, or 3 days. The dose of 100 micrograms T4 per 100 g of body wt given for 6 days exerted the strongest effect on the activity of all of the investigated enzymes. In comparison to the control, rhodanese activity diminished in the mitochondrial fraction by 40% (P < 0.05), cystathionase activity diminished in the cytosolic fraction by 15% (P < 0.05), and MPST activity in the mitochondrial fraction was reduced by 34% (P < 0.05), whereas cytosolic MPST activity was unaltered. Simultaneously, in the liver homogenate, elevated levels of ATP and sulfate were observed after 6 days of T4 administration. Thus, the present results seem to suggest that in the mouse liver, after 6 days of administration of 100 micrograms T4 per 100 g of body wt, the desulfuration metabolism of L-cysteine is diminished, which is probably accompanied by an increase in oxidative L-cysteine metabolism. The dose of 100 micrograms per 100 g of body wt administered for a shorter period, and the use of a lower dosage (50 micrograms T4 per 100 g of body wt) for 6 days had a stimulatory effect upon MPST activity level, and an increased level of sulfane sulfur was observed.

The influences of emotional changes induced by being exposed to a new environment on the pharmacokinetics of plasma drug concentration were studied in male Wistar rats. Transfer from a familiar home cage to a new home cage was considered to induce psychological (non-physical) emotional changes. First, nicorandil and zonisamide, drugs that act on the peripheral system and central nervous systems, were used, respectively. Immediately after oral administration of nicorandil (10 mg/kg) or zonisamide (50 mg/kg), the animals were transferred to new home cages. Plasma nicorandil and zonisamide concentrations were determined by high-performance liquid chromatography at 1 and 4 h after administration. Plasma nicorandil concentration in the group transferred to new home cages was significantly decreased relative to levels in the non-transferred control group. However, zonisamide concentrations were unchanged. These findings suggest that the pharmacokinetics of nicorandil, but not those of zonisamide, tend to be influenced by non-physically induced emotional changes.

In order to evaluate the relationship between Helicobacter pylori (H. pylori) infection and dietary preference, a cross-sectional study was performed among 626 residents in a rural area of Japan. Seropositive rates were 88.7% in males and 71.4% in females, and these increased with age for both sexes [male P < 0.05 and female P < 0.01]. The relationship between H. pylori-seropositivities and salted-food intake, after adjustment for age, demonstrated a significant result in the "almost every day" group in males with an odds ratio (OR) of 8.39 and with 95% confidence intervals (CI) of 1.02-69.30. As regards an association between seropositivities of H. pylori and levels of serum pepsinogens for the screening of chronic atrophic gastritis (low pepsinogen values used were a pepsinogen I level below 70 ng/ml and a pepsinogen I/pepsinogen II ratio below 3.0), the ORs of H. pylori-seropositivities for low pepsinogen cases were 6.32 [95% CI: 1.42-28.03] in males and 12.72 [95% CI: 4.57-35.46] in females. With regard to the relationship between low pepsinogen cases and light-colored vegetables intake, a significant low OR for the low pepsinogen cases was obtained in the "almost every meal" group in females after adjustment for age and seropositivities of H. pylori with an OR of 0.37 and with 95% CI of 0.15-0.92.

In order to clarify the effects of medical education on attitudes towards mental illness among medical students, a follow-up study was conducted. All 100 students entering Kochi Medical School in 1988 were subjects. The initial questionnaire survey was conducted in 1988, and followed up in 1993. Response rate was 69% in the initial survey, and 83% in the follow-up study. By the time of the follow-up, all of the students had completed their medical education, including courses in psychiatry and mental health. Results were as follows: At the follow-up study, 1) a significantly higher percentage of students replied that they accepted the mentally ill as co-workers; 2) significantly favorable changes were observed in attitudes towards psychiatric services; 3) optimism about the effectiveness of treatment for mental illness at an early stage and prevention of mental illness had decreased; and 4) no change was observed in attitudes toward human rights of the mentally ill, except in the case of one item stating that the mentally ill should not have children in order to avoid hereditary handicaps, with which a lower percentage agreed. Conclusively, medical education can play an important role in attitudes towards mental illness.

Levodopa is the gold standard for the treatment of Parkinson's disease (PD) because of its outstanding clinical efficacy. However, the majority of patients experience various adverse reactions, including the wearing-off phenomenon, the on-off phenomenon, dyskinesia and psychiatric symptoms. The response to levodopa depends not only on the intrinsic responsiveness of the patients, but also on various other important factors including the type of levodopa preparation, its absorption/metabolism, the blood-brain barrier, age at onset of disease and concomitant use of anti-parkinsonian drugs. This review summarizes factors which influence the effects of levodopa in PD. To minimize levodopa-induced adverse reactions and to relieve long-term parkinsonian symptoms, levodopa therapy should be conducted by taking these factors into consideration.

To investigate whether an association exists between vascular endothelial growth factor (VEGF) expression and tumor prognosis in primary lung carcinoma, we used immunohistochemical techniques to analyze microvessel density and VEGF expression in lung carcinoma tissue from 98 patients. Tissue had been fresh-frozen at the time of operation and preserved for more than 5 years. The results indicated that VEGF expression was positive for 50 of the 98 patients (51.0%), with 27 (27.6%) being weakly positive and 23 (23.5%) being strongly positive. The microvessel density in tissue showing weakly positive and strongly positive VEGF expression was significantly higher than that in VEGF-negative tumor tissue (P < 0.05: negative vs. weakly positive, P < 0.01: negative vs. strongly positive), we showed demonstrating that VEGF expression was significantly associated with intratumoral microvessel density. The 5-year survival rates were 8.7% for strongly VEGF-positive patients, 43.9% for weakly VEGF-positive patients and 79.2% for VEGF-negative patients, respectively (P < 0.01: negative vs. weakly positive or strongly positive). Furthermore, multivariate analysis employing multiple regression analysis indicated that VEGF expression correlates highly with the overall survival rates of patients with primary lung carcinoma. Two variables, N status and VEGF expression, were found to be significant prognostic factors (P < 0.01). The results of this study suggest that VEGF expression is associated with intratumoral microvessel density. VEGF expression may constitute important independent prognostic evidence that can help us in predicting the outcomes of patients with primary lung carcinomas.

The present study showed that many neurons in the adult rat brain possessed a perineuronal sulfated proteoglycan surface coat which reacted to cationic iron colloid and aldehyde fuchsin. This surface coat was stained supravitally with Ehrlich's methylene blue and doubly stained with Ehrlich's methylene blue and aldehyde fuchsin. The surface coat was also stained with Gomori's ammoniacal silver and doubly stained with Gomori's ammoniacal silver and cationic iron colloid. The surface coat was usually expressed together with a nerve cell surface glycoprotein net detectable with lectin Wisteria floribunda agglutinin. These findings indicate that the perineuronal proteoglycan surface coat is identical to Cajal's superficial reticulum and contains some collagenous elements. It was further demonstrated that collagenase digestion erased Gomori's ammoniacal silver impregnation within the perineuronal proteoglycan surface coat.

While heat shock protein (HSP) 72 is known as a stress protein, there have been no reports of HSP 72 expression in patients who have undergone surgery for congenital heart disease. Fourteen patients (7 males and 7 females) who had undergone surgery for congenital heart disease were studied. The ages of the patients ranged from 2 months to 43 years old (mean 6.5 +/- 10.8 years old; median 3.0 years old). The diagnoses were Tetralogy of Fallot in seven, pulmonary atresia with ventricular septal defect (VSD) in three, complex anomalies in three, and VSD in one patient. Histological study and HSP analysis using Western blots and immunostaining with anti-HSP 72 monoclonal antibody were performed for right ventricular muscle samples resected during the surgery. The histological findings showed hypertrophic changes of ventricular cardiomyocytes in all samples studied. Western blots detected HSP 72 expression of various degrees in all specimens. Immunostaining using monoclonal antibody against HSP 72 showed that the protein was present in the nuclei and cytoplasm of cardiomyocytes. In conclusion, although it is difficult to determine the cause of the "stress" that triggers HSP 72 expression in cardiomyocytes, low O2 saturation and pressure overload might act as a "stress", and the only common factor that induced HSP 72 in every sample was hypertrophy.

We conducted Myanmar-Japan cooperation studies on hepatitis B and hepatitis C virus markers in patients with thalassemias and those with liver diseases. Among the 102 patients with liver diseases, 92% had a history of hepatitis B virus infection (antibody to hepatitis B core antigen positive), 35% were hepatitis B surface antigen positive, 39% were positive for anti-HCV. Among 28 patients with hepatocellular carcinoma, 46% had hepatitis B surface antigen, 21.4% had antibody to hepatitis C virus, and 7% were positive for both hepatitis B surface antigen and anti hepatitis C virus. The history of HCV infection among blood recipients at the Haematology Department of the Yangon General Hospital and at the Yangon Children's Hospital was found to be 55.5% and 46.7%, respectively, which is comparable to the history of hepatitis B infection (66.7% and 46.7%, respectively). This preliminary survey also encountered 2 cases positive for anti-HCV among 34 voluntary blood donors. This survey is the first one to report that hepatitis C is at the epidemic stage in Myanmar. As there is no effective treatment for hepatitis C in this country, a screening program for blood used in transfusion should be started immediately.

The effect of Blastocystis hominis (B. hominis) in both immunocompetent and immunocompromised subjects has been the subject of debate in recent years, mostly in response to its unknown pathogenicity and frequency of occurrence. We performed a non-randomised, open labelled, single institute study in our hospital in order to investigate the clinical significance and frequency of B. hominis in patients suffering from hematological malignancy (HM) who displayed symptoms of gastrointestinal diseases during the period of chemotherapy-induced neutropenia. The presence and potential role of other intestinal inclusive of parasites were also studied. At least 3 stool samples from each of 206 HM patients with gastrointestinal complaints (the HM group) were studied. These were compared with stool samples from a control group of 200 patients without HM who were also suffering from gastrointestinal complaints. Samples were studied with saline-lugol, formalin-ether, and trichome staining methods. Groups were comparable in terms of gender, age and type of gastrointestinal complaints. In the HM group, the most common parasite was B. hominis. In this group, 23 patients (13%) had B. hominis, while in the control group only 2 patients (1%) had B. hominis. This difference was statistically significant (P < 0.05). Symptoms were non-specific for B. hominis or other parasites in the HM group. The predominant symptoms in both groups were abdominal pain (87-89.5%), diarrhea (70-89.5%), and flatulence (74-68.4%). Although all patients with HM were symptom-free at the end of treatment with oral metranidazol (1,500 mg per day for 10 days) 2 patients with HM had positive stool samples containing an insignificant number of parasites (< 5 cells per field). In conclusion, it appears that B. hominis is not rare and should be considered in patients with HM who have gastrointestinal complaints while being treated with chemotherapy. Furthermore, metranidazol appears to be effective in treating B. hominis infection.

Conventional therapy for colorectal carcinoma using 5-fluorouracil (5-FU) has shown limited antitumor action. The purpose of our study was to investigate synergistic antitumor effects of the streptococcal preparation of OK-432 and 5-FU, and to elucidate the mechanisms of interaction between the 2 agents in mice. Biochemical modulation of OK-432 and 5-FU were determined in vivo against colon-26 carcinoma. The concentration of 5-FU and its metabolites, and the activity of thymidylate synthase and thymidine kinase, respectively, were measured using cytosolic extracts of the tumors. Combination treatment with OK-432 produced a significant increase in intratumor 5-FU and 5-FU in RNA (F-RNA) concentrations, increased the thymidylate synthetase inhibition rate, and decreased thymidine kinase activity, as compared with the results observed in the control mice. These additive antitumor effects are obtained by use of the 2 agents; the mechanism of action is considered to be the suppression of both the de novo and the salvage pathway for DNA synthesis, along with the suppression of RNA synthesis.

The majority of the t(14;18) chromosome translocations that occur in non-Hodgkin centroblastic-centrocytic follicular lymphoma can be detected by various methods. During the translocation process the bcl-2 gene located on chromosome 18 (18q21) is translocated to the JH region of the immunoglobulin gene of chromosome 14 (14q32). The most frequent type of bcl-2 translocations is the mbr type, whereas the immunoglobulin gene breaks mainly at the JH1-6 exons. About one of the 10(5) cells bearing the translocation can already be detected by using nested polymerase chain reaction (PCR). Eight patients suffering from follicular lymphoma were included in this study, which considered the usefulness of the PCR method. The results are in good agreement with those obtained by conventional diagnostic methods. Translocation can be detected, however, in patients with non-malignant diseases such as Sjogren's syndrome (about 5% of the patients) and in a patient with Whipple disease. In addition, translocation was detected in lymphocytes of peripheral blood of a healthy donor. Since lymphomas are detected in patients with Sjogren's syndrome with a relative high frequency, an early diagnosis of the translocation could improve the treatment of the disease. Nevertheless, a diagnosis of lymphoma is valid only in cases of bone marrow translocation-positivity.

Fracture of the penis is an unusual blunt traumatic injury of the erect penis. Twenty-five such cases, treated at the 2 main urological departments in the town of Varna between 1984 and 1999, were analyzed retrospectively. The average annual incidence in this series was 0.33 per 100,000 inhabitants. Most of the injuries occurred during vigorous sexual intercourse. Clinical diagnosis was mainly based on the patient's history and the physical findings. The diagnosis was verified by ultrasonography. In a few cases retrograde urethrography and cavernosography were used as additional diagnostic tools. Cavernosography was complicated by postprocedural priapism in 1 case. Unilateral tear of the corpora cavernosa was found in 24 cases. In 3 cases, partial (in 2) or complete (in 1) disruption of the corpus spongiosum and the urethra was found as well. The trauma was bilateral in 1 case in which 2/3 of the entire circumference of the penis was ruptured, including both corpora cavernosa and the urethra. Seventeen patients received immediate surgical repair. Most of them experienced an uneventful postoperative period with no serious consequences affecting their sexual function. Surgery was delayed in 6 and rejected in 2 cases. In all cases in which conservative (nonsurgical) management was the first treatment option, late complications (penile aneurysm, induration, penile curvature, erectile dysfunction) were observed and the final results were assessed as satisfactory or poor. We recommend immediate surgical treatment of all cases of penile fracture. Emergency surgical repair offers a chance for complete recovery and is the best method for providing a good functional prognosis.

Tacrolimus (FK-506) and cyclosporin A (CsA) are calcineurin antagonists used widely as T-cell immunosuppressants; however, their relative efficacy on the production of interleukin-18 (IL-18) remains undefined. We have examined the effects of FK-506 and CsA on the cytokine generation of human peripheral blood mononuclear cells (PBMCs) in mixed lymphocyte reaction (MLR) with lipopolysaccharide (LPS). We studied the levels of interleukin-18 (IL-18), IL-12, IL-10, IL-6, IL-2 and interferon-gamma (IFN-gamma) in the supernatant in allo-MLR by ELISA assay. Supernatant levels of IFN-gamma, IL-2, IL-6, IL-10 and IL-12 were detected 12 h after MLR and markedly increased thereafter. In contrast, production of IL-18 was detected at 12 h, reached a near maximum level at 24 h and decreased at 72 h. These results suggested that IFN-gamma production depended on IL-18, IL-12 and IL-2 in the early phase of MLR and depended mainly on IL-12 and IL-2 in the late phase. Both calcineurin antagonists inhibit the generation of IL-18, which plays a large role in allogeneic cell interactions, in macrophages and they also promote an equivalent down-regulation of T helper 1 (Th1) and Th2 responses in a concentration-dependent manner. About 90% of IFN-gamma production induced by MLR was inhibited by an anti-IL-18 antibody, showing that IL-18 can trigger IFN-gamma production in MLR. These results suggest that dual signaling consisting of antigen-driven nuclear factor of activated T cells (NFAT) activation and LPS-mediated NF-kappaB activation is crucial for IL-18 production in macrophages, and that IL-18 can trigger IFN-gamma production in T-cells by MLR.

It has been reported that Epstein-Barr virus (EBV) resides in resting B cells in vivo. However, an ideal in vitro system for studying EBV latent infection in vivo has not yet been established. In this study, a mantle cell lymphoma line, SP53, was successfully infected with a recombinant EBV containing a neomycin-resistant gene. The EBV-carrying SP53 cells were obtained by selection using G418. They expressed EBER-1, EBNAs, and LMP1; this expression pattern of the EBV genes was similar to that in a lymphoblastoid cell line (LCL). However, proliferation assay showed that the EBV-carrying SP53 cells have a doubling time of 73 h, compared with 57 h of SP53 cells. Transplantation of 10(8) SP53 cells to nude mice formed tumors in 4 of 10 mice inoculated, but the EBV-carrying SP53 cells did not. Unexpectedly, EBV infection reduced the proliferation and tumorigenicity of SP53 cells. However, the EBV-carrying SP53 cells showed higher resistance to apoptosis induced by serum starvation than did the SP53 cells. The inhibition of proliferation and the resistance to apoptosis induced in SP53 cells by EBV infection indicate that this cell line might to some extent provide a model of in vivo EBV reservoir cells.

Endoscopic biopsy is an important tool for histological diagnosis of lesions residing in gastrointestinal tracts. However, it is less useful in submucosal lesions due to the existence of normal overlying mucosa. We developed a new and safe technique for the diagnosis of submucosal tumor using Stiegmann-Goff endoscopic ligator. After removing surface mucosa to expose submucosal tissue by this method, conventional secured histological diagnosis could be performed. To determine definitive histological diagnosis, this technique is useful as well as Endoscopic Ultrasound (EUS) with fine needle aspiration biopsy and other modalities.

To find a new anti-malarial medicine derived from natural resources, we examined the leaves of 13 common Japanese plants in vitro. Among them, a leaf-extract of Hydrangea macrophylla, a common Japanese flower, inhibited the parasitic growth of Plasmodium falciparum. The IC50 of Hydrangea macrophylla leaf extract to Plasmodium falciparum was 0.18 microg/ml. The IC50 to NIH 3T3-3 cells, from a normal mouse cell line, was 7.2 microg/ml. Thus, selective toxicity was 40. For the in vivo test, we inoculated Plasmodium berghei, a rodent malaria parasite, to ddY mice and administered the leaf-extract of Hydrangea macrophylla (3.6 mg/0.2 ml) orally 3 times a day for 3 days. Malaria parasites did not appear in the blood of in the treated mice, but they did appear in the control group on day 3 or 4 after inoculation with the parasites. When leaf extract was administered to 5 mice 2 times a day for 3 days, malaria parasites did not appear in 4 of the mice but did appear in 1 mouse. In addition, the leaf-extract was administered orally 3 times a day for 3 days to Plasmodium berghei infected mice with a parasitemia of 2.7%. In the latter group, malaria parasites disappeared on day 3 after initiating the treatment, but they appeared again after day 5 or 6. Although we could not cure the mice entirely, we confirmed that the Hydrangea macrophylla leaf extract did contain an anti-malarial substance that can be administered orally.

In order to examine fatigue of the knee flexor and extensor muscles and to investigate the characteristics of muscular fatigue in different sports, a Cybex machine was used to measure muscle fatigue and recovery during isokinetic knee flexion and extension. Eighteen baseball players, 12 soccer players and 13 marathon runners were studied. Each subject was tested in the sitting position and made to perform 50 consecutive right knee bends and stretches at maximum strength. This was done 3 times with an interval of 10 min between each series. The peak torque to body weight ratio and the fatigue rate were determined in each case. In all subjects, the peak torque to body weight ratio was higher for extensors than flexors. Over the 3 trials, the fatigue rate of extensors showed little change, while that of flexors had a tendency to increase. In each subject, knee extensors showed a high fatigue rate but a quick recovery, while knee flexors showed a low fatigue rate but a slow recovery. As the marathon runners had the smallest fatigue rates for both flexors and extensors, we concluded that marathon runners had more stamina than baseball players and soccer players.