Characteristic oscillatory motion of a camphor boat sensitive to physicochemical environment.

A self-propelled camphor boat on water was investigated from the viewpoint of characteristic features of motion and mode-bifurcation depending on the diffusion length of camphor molecules. When a camphor disk was connected to the bottom of a larger plastic plate and then was placed on water, either oscillatory motion (repetition between rest and motion) or continuous motion was observed. In this paper, we report the novel features of this motion and mode-bifurcation as a function of the diffusion length of camphor molecules, e.g., multiple accelerations during oscillation, period-2 or irregular oscillatory motion, and reciprocating oscillation. These characteristic motion and mode-bifurcation are discussed in relation to the diffusion length of camphor molecules under the camphor boat and the development of camphor molecules from the camphor boat on water.

Expression of Forkhead box P3 in tumour cells causes immunoregulatory function of signet ring cell carcinoma of the stomach.

BACKGROUND: It was recently reported that the transcription factor Forkhead box P3 (FoxP3) is expressed not only in regulatory T cells (Tregs) but also in cancer cells. The aim of this study was to clarify the clinical significance of FoxP3 expression in gastric carcinoma. METHODS: We performed immunohistochemical staining of FoxP3 to examine the association of FoxP3 expression with clinicopathological features of 194 patients with gastric cancer who underwent surgical resection from 2000 to 2010. We also investigated the immunosuppressive function of FoxP3 using gastric cancer cell lines. RESULTS: Immunohistochemical staining indicated FoxP3-positive cells within tumour tissue including both Tregs and tumour cells. Forkhead box P3-positive tumour cells were observed in 79.3% of signet ring cell carcinoma patients, and the expression of FoxP3 showed a significant correlation with lymph node metastasis. We showed that transforming growth factor-ß augmented FoxP3 mRNA expression in cell lines derived from signet ring cell carcinoma. Indoleamine-2,3-dioxygenase and galectin-1, key effectors of Treg-mediated immunosuppression, were downregulated by FoxP3 knockdown. CONCLUSION: Our findings suggested that FoxP3 expression by tumour cells might have important roles in immune escape of gastric carcinoma, and be associated with the malignant potential of scirrhous gastric carcinoma.

Association of insulin-like growth factor-1 receptor gene polymorphisms with left ventricular mass and geometry in essential hypertension.

Stimulation of insulin-like growth factor (IGF)-1 receptor by IGF-1 and insulin strongly induces cardiomyocyte hypertrophy. In this study, we assessed the hypothesis that genetic variations of the IGF-1 receptor may be linked to the diversity of left ventricular (LV) structure in hypertensive patients. Genotypes in 12 single nucleotide polymorphisms (SNPs) of the IGF-1 receptor gene identified by direct sequencing were determined in 795 Japanese patients with essential hypertension. In echocardiographic examinations, LV mass index (LVMI) and relative wall thickness (RWT) were measured. Among 12 SNPs, promoter -328C>T and intron-13 275124A>C polymorphisms were significantly associated with LV hypertrophy (LVMI> or =125 g m(-2)) and concentric change (RWT> or =0.44), respectively. In allele frequencies, the C allele of -328C>T was related to LV hypertrophy, and the A allele of 275124A>C was related to LV concentric change. In fact, LVMI and prevalence of LV hypertrophy increased in CC genotype of -328C>T. RWT and prevalence of LV concentric change increased in AA genotype of 275124A>C. A multiple logistic regression analysis revealed that the presence of CC genotype of -328C>T or AA genotype of 275124A>C was an independent determinant for LV hypertrophy or concentric change, respectively. Furthermore, the combination of CC of -328C>T and AA of 275124A>C genotypes was significantly associated with abnormal LV geometry, especially concentric hypertrophy. Our findings show that two SNPs of the IGF-1 receptor gene are related to LV hypertrophy in patients with essential hypertension, suggesting that the genetic variation of the IGF-1 receptor may be involved in the diversity of LV structure in hypertensives.

Pathological changes in pigs experimentally infected with porcine teschovirus.

Nonsuppurative encephalomyelitis with neurological signs expressed as flaccid paralysis of the hindlimbs was experimentally induced in three-week-old piglets by a single intravenous injection of the Toyama 2002 strain of porcine teschovirus (PTV) isolated from field pigs in Japan. Lesions characterized by perivascular cuffing of mononuclear cells, focal gliosis, neuronal necrosis and neuronophagia were observed, mainly in the ventral horn of the spinal cord. Nonsuppurative ganglionitis of the spinal ganglion and neuritis of the spinal root were also detected. PTV antigens were detected immunohistochemically and the distribution of these antigens corresponded closely with the distribution of brain lesions. PTV antigens were observed in the ganglion cells before the appearance of the inflammatory changes 3 days post-inoculation (dpi) and were present in the dorsal root and spinal cord on 9 dpi. No lesions of the central nervous system were induced in pigs by oral or intranasal inoculation of this strain of PTV.

Brain lesions induced by experimental intranasal infection of Japanese encephalitis virus in piglets.

Non-suppurative encephalitis was experimentally induced in three-week-old piglets by a single intranasal inoculation of Japanese encephalitis virus (JEV) isolated from field pigs. The lesions consisted of glial cell aggregates and perivascular cuffing throughout the olfactory tract and pyriform cortex. JEV antigens were detected in the cytoplasm and neuronal processes of small nerve cells in the granule cell layer of the olfactory bulb, in the neuronal processes of the olfactory tract and in the cytoplasm of neurons in the pyriform cortex. The distribution of the antigens corresponded closely with the distribution of brain lesions. These findings suggest that JEV may enter the brain by the olfactory pathway in addition to via haematogenous spread in piglets.

Evaluation of unexpected positive results from a commercial ELISA for antibodies to PRRSV.

Unexpected positive results from the widely used IDEXX ELISA for the detection of antibodies to porcine reproductive and respiratory syndrome virus (PRRSV) may confound investigations of the disease. Supplementing the ELISA with blocking agents and the use of IgG purified from serum samples had no effect on the unexpected positive results, suggesting that they were due to an antibody-antigen reaction. Simple competitive and blocking ELISAs were developed by modifying the IDEXX ELISA, and they and an indirect fluorescent antibody test (IFAT) were used to examine PRRSV antibodies in 33 antibody-negative, 88 antibody-positive and 73 unexpectedly positive sera. All the unexpectedly positive sera were negative by IFAT, and 89.0 per cent were negative by both the competitive and blocking ELISAs. The competitive ELISA (97.7 per cent) and the blocking ELISA (96.5 per cent) detected more positive sera than the IFAT (90.9 per cent). These results show that both ELISAs are capable of distinguishing positive and unexpectedly positive sera, and suggest that most of the unexpected positive signals are false-positives.

Changes in the myosin heavy chain 2a and 2b isoforms of the anterior belly of the digastric muscle before and after weaning in mice.

During the process of growth and development, the digastric muscle is subjected to marked functional changes, including the change from suckling to mastication. In particular, because the anterior belly of the digastric muscle, which is one of the suprahyoid muscles, plays an important role in mastication. Therefore, this muscle seems to undergo a marked functional change before and after weaning. However, the details remain unknown. Here, to clarify the changes in the muscle fibre characteristics of the anterior belly of the digastric muscle before and after weaning, we examined myosin heavy chain isoforms at the protein (immunohistochemistry) and mRNA (transcription) levels. As a control, the changes in the muscle fibre characteristics of the sternohyoid muscle, which is anatomically aligned in the same direction as the anterior belly of the digastric muscle, were analyzed. The results showed that, in the anterior belly of the digastric muscle that is involved in mandibular movements in mice, the ratio of a fast-contraction isoform with strong contractile force increased after weaning. We believe that this occurred in response to a functional change from suckling to mastication. On the other hand, there was little change in the composition of sternohyoid muscle.

Immunohistochemical detection of porcine teschovirus antigen in the formalin-fixed paraffin-embedded specimens from pigs experimentally infected with porcine teschovirus.

Porcine teschovirus (PTV) antigens were detected by a streptavidin-biotin complex method in formalin-fixed paraffin-embedded tissues of 3-week-old pigs that had been inoculated intravenously with PTV Talfan strain. PTV antigens were detected in cytoplasm of nerve cells, glial cells and endothelial cells in the cerebellar nuclei, the grey matter of the midbrain, pons and medulla oblongata and the ventral horn of the spinal cord and of ganglion cells in the spinal ganglion corresponding to those lesions characterized as non-suppurative encephalomyelitis and ganglionitis. The results of this study suggest that nerve cells of the brain stem and spinal cord and ganglion cells of the spinal ganglion permit PTV replication and represent the main target cell population of PTV. This is the first study to demonstrate PTV antigen by immunohistochemistry in formalin-fixed paraffin-embedded tissue specimens from pigs infected with PTV.

Association of single nucleotide polymorphisms in endothelin family genes with the progression of atherosclerosis in patients with essential hypertension.

Endothelin-1 (ET-1) is a potent vasoconstrictive peptide and its activity is mediated by the receptors ET type A (EDNRA) and ET type B (EDNRB). Although ET-1 is thought to play an important role in the development of atherosclerosis, it remains unclear whether polymorphisms of ET-1 family genes, including the ET-1 gene (EDN1), EDNRA, EDNRB and the genes for endothelin converting enzymes 1 and 2 (ECE1 and ECE2), are associated with the progression of atherosclerosis. We investigated the relationship between 11 single nucleotide polymorphisms (SNPs) of ET-1 family genes (including three in EDN1, one in EDNRA, two in EDNRB, four in ECE1 and one in ECE2) and atherosclerotic changes assessed using pulse wave velocity (PWV) and carotid ultrasonography in 630 patients with essential hypertension (EHT). In male subjects, we found significant differences in brachial-ankle PWV (baPWV) in additive and recessive models in EDNRB-rs5351 after Bonferroni correction. Also in male subjects, there were significant differences in mean intima-media thickness (IMT) in additive and recessive models in EDNRA-rs5333 after Bonferroni correction. We found no significant correlation between any SNPs in the ET family genes and baPWV, IMT and Plaque score (PS) in female subjects. Furthermore, after multiple logistic regression analysis, only EDNRB-rs5351 indicated as an independent risk of atherosclerosis in male hypertensive subjects. Of the endothelin-related genes, EDNRB-rs5351 was the most susceptible SNP associated with atherosclerosis in male hypertensives, and the genetic background may be involved in the progression of atherosclerosis in EHT patients.

Optic nerve hyperintensity on T2-weighted images among patients with pituitary macroadenoma: correlation with visual impairment.

PURPOSE: Visual acuity (VA) disturbance other than field defect is important in evaluating patients with pituitary macroadenoma. The purpose of this study was to evaluate MR imaging appearances of optic nerves in patients with pituitary macroadenoma and to ascertain whether visual impairment was correlated with abnormality in optic nerve signal intensity. PATIENTS AND METHODS: Twenty-seven patients with pituitary macroadenoma were examined. Optic nerves were evaluated on T2-weighted images and correlations of signal intensity abnormality with VA disturbance, visual field disturbance, degree of optic chiasm compression, pathologic findings of surgical specimen, and disease duration were statistically analyzed. Correlations between recovery of VA after treatment and the above-mentioned factors were also determined. RESULTS: Coronal T2-weighted images demonstrated unilateral optic nerve hyperintensity lesions in 9 patients. Bilateral signal intensity abnormality of the optic nerve was seen in 5 patients. Signal intensity abnormality of the optic nerve was seen at the site of compression and in the ventral side of the tumor. These patients did not demonstrate signal intensity abnormality posterior to the tumor. Presence of such signal intensity abnormalities was correlated with the degree of optic chiasmal compression and with VA disturbance. Recovery of VA after treatment was correlated with disease duration. CONCLUSION: Hyperintensity of the optic nerves ventral to the pituitary macroadenoma was associated with VA impairment. Recovery of VA after treatment was correlated with disease duration. MR imaging of the optic nerves can provide valuable information for management of pituitary macroadenoma.

Genetic variation and geographic distribution of porcine reproductive and respiratory syndrome virus in Japan.

Porcine reproductive and respiratory syndrome virus (PRRSV) has two genotypes, the North American-type (NA-type) and the European-type (EU-type), and each genotype is also genetically diverged. We sequenced the ORF5 gene of 30 PRRSVs isolated from 23 prefectures of Japan during 1992 and 1993 and during 2000 and 2001. All of the isolates were of the NA-type. Phylogenetic analysis of the overall NA-type viruses isolated from around the world identified five major genetic clusters. The 1992-1993 Japanese samples belonged to only two genetic clusters, while the 2000-2001 samples included more diverged ORF5 genomes. One genetic cluster, which included 63% (20/32) of Japanese isolates, one Taiwanese isolate and one Chinese isolate, was mainly found in the eastern part of Japan. Another genetic cluster, which was found in various areas around the world, was distributed in the western part of Japan.

Nonsuppurative encephalitis in piglets after experimental inoculation of Japanese encephalitis flavivirus isolated from pigs.

Nonsuppurative encephalitis was experimentally induced in 3-week-old piglets by a single intravenous inoculation of either of two strains (IB 2001 or AS-6) of Japanese encephalitis flavivirus (JEV) isolated from field pigs. The lesions, which consisted of neuronal necrosis, neuronophagia, glial nodules, and perivas-cular cuffing, were distributed in the cerebrum, midbrain, pons, medulla oblongata, and cerebellum, particularly in the gray matter of the frontal and temporal lobes and thalamus. The gray matter of the spinal cord of piglets that were given the AS-6 strain also was affected. JEV antigen was immunohistochemically detected in the cytoplasm of the nerve cells in the cortex of the frontal and temporal lobes and in the gray matter of the thalamus and midbrain. Two JEV strains isolated from field pigs exhibited neurovirulence, inducing nonsup-purative encephalitis in piglets.

Diagnostic value of carotid intima-media thickness and plaque score for predicting target organ damage in patients with essential hypertension.

Carotid intima-media thickness (IMT) assessed by ultrasonography is regarded as an early predictor of general arteriosclerosis in patients with essential hypertension. However, the methods of measuring IMT have not been globally standardized, and it remains unclear whether conventional measurement of IMT represents the prevalence of hypertensive target organ damage. In this study, we verified the association between several commonly used carotid ultrasonographical parameters and the severity of hypertensive target organ damage (retinal arteriosclerosis, microalbuminuria, left ventricular hypertrophy (LVH)). Carotid ultrasonography, echocardiography, urinalysis, and funduscopy were performed in 184 patients (64 +/- 12 years, 96 males and 88 females) with various stages of essential hypertension. Carotid arteriosclerosis was assessed using four methodologically different methods: conventional-IMT, maximum-IMT (Max-IMT), Mean-IMT, and Plaque Score (the sum of all plaque thicknesses). Age and all carotid ultrasonographical parameters were significantly associated with albuminuria, retinal arteriosclerosis, and left ventricular mass index. High-sensitivity CRP was significantly correlated with retinopathy and LVH. Carotid parameters in patients with histories of cardiovascular events were significantly greater in those without events. Among all carotid parameters, Max-IMT showed the highest correlation coefficient of the severity of target organ damage, and showed significant association with CRP. Stepwise regression analysis revealed that Max-IMT was the independent factor for predicting target organ damage. Max-IMT is suggested to be the most reliable and simplest parameter for predicting hypertensive target organ damage including microangiopathy in patients with essential hypertension.

Positive association of CYP11B2 gene polymorphism with genetic predisposition to essential hypertension.

Predispositions to essential hypertension and cardiovascular diseases are possibly associated with gene polymorphisms of the renin-angiotensin system. Gene polymorphisms of angiotensinogen and angiotensin-converting enzyme genes have been suggested to be risk factors for hypertension and myocardial infarction. Concerning the polymorphism of aldosterone synthase (CYP11B2) gene, earlier studies have shown inconsistent results in terms of its relation to hypertension. In the present case-control study, we investigated the association of -344T/C polymorphism in the promoter region of human CYP11B2 gene with genetic predisposition to hypertension. The genotype of -344T/C polymorphism was determined in essential hypertension subjects (n=250) and normotensive subjects (n=221). The distributions of three genotypes (TT, TC, and CC) were significantly different between the hypertensive and the normotensive groups (chi(2)=9.61, P=0.008). Namely, the frequency of C allele was higher in the hypertensive patients than in the normotensive subjects (34.2 vs 26.5%, P=0.010). Our data suggest that the -344C allele of CYP11B2 gene polymorphism is associated with the genetic predisposition to develop essential hypertension.

Comparison of the effects of amlodipine and losartan on 24-hour ambulatory blood pressure in hypertensive patients.

Effects of amlodipine (AML), a long-acting calcium antagonist, and losartan (LOS), an angiotensin II receptor antagonist, on 24-hr blood pressure profile were compared in 15 patients with essential hypertension. After 4 weeks of placebo period, the patients were treated with AML or LOS in a random crossover design for 12-16 weeks each. Either drug was given once daily at 0800 and the doses were titrated so that the office blood pressure was reduced lower than 140/90mmHg. At the end of each period, 24-hr blood pressure was monitored. Average office blood pressure was lowered from 158 +/- 2/ 98 +/- 2 mmHg to 134 +/- 1/87 +/- 1 mmHg by AML and 134 +/- 2/88 +/- 1 mmHg by LOS. Average 24-hr blood pressure was also reduced from 144 +/- 3/ 92 +/- 2 mmHg to 131 +/- 2/84 +/- 2 mmHg by AML and 135 +/- 3/85 +/- 2 mmHg by LOS. The averaged 24-hr systolic blood pressure was significantly lower in AML than in LOS (p < 0.05). Then, the 24-hr blood pressure was analyzed for four segments; morning (0530-0900 h), daytime (0930-1800 h), evening (1830-2300 h) and night (2330-0500 h). Although the daytime blood pressure was comparable between AML and LOS, systolic blood pressure in the evening and morning hours were lower in AML than in LOS (133 +/- 2 vs. 138 +/- 3mmHg,p<0.01; 129 +/- 3 vs. 134 +/- 4,p<0.05). Troughtopeakratio of antihypertensive effect on systolic blood pressure was significantly greater in AML than in LOS (62 +/- 5% vs. 55 +/- 4%, p < 0.05). Either drug did not cause reflective increase in pulse rate over 24 hours. These results suggest that both AML and LOS are equally effective in lowering daytime blood pressure without eliciting reflex tachycardia, however, the antihypertensive effect of AML lasts longer than that of LOS. Such information seems important to achieve 24-hr blood pressure control using these drugs.

[A case of fundus albipunctatus with a retinol dehydrogenase 5 gene mutation in a child].

BACKGROUND: We examined a family with fundus albipunctatus in which mutation of the retinol dehydrogenase 5(RDH 5) gene was suspected to be the cause of this disease. CASE: An 8-year-old girl had diffuse multiple white dots in her fundus except for the macula. She had good central vision. The amplitude of her electroretinogram wave was low, but it recovered after three hours of dark adaptation. Dark adaptometry showed an elevated threshold for rod adaptation. No visual field loss was observed. A homozygous missense mutation was found in exon 5 of the RDH 5 gene that substituted histidine for arginine at codon 280(Arg 280 His). Her mother had a normal fundus but was heterozygous for the same mutation. CONCLUSION: A missense mutation of RDH 5(Arg 280 His) was found in a Japanese family with fundus albipunctatus.

[Comparisons of factors affecting voiding disorders between patients with benign prostate hyperplasia and volunteers].

PURPOSE: The prostate size and motivation to visit clinics were investigated in patients with prostate hyperplasia. OBJECTS AND METHODS: One hundred ninety-five patients who had urinary symptoms and visited our outpatient clinic between September 1994 and October 1999 and 268 age-matched volunteers in Mitaka City who underwent a medical examination of the prostate in June 1997 were compared. International Prostate Symptom Score (IPSS), Quality Of Life Score (QOL score), residual urine volume, prostate volume and urinary flow rate were measured. RESULTS: The prostate volume of the volunteers was 20-25 cm3 irrespective of the age. The prostate size of the outpatients was larger than that of the volunteers for every age group. IPSS and QOL score were significantly higher in the outpatients than in the volunteers. Diurnal urinary frequency and sense of residual urine contributed to the discrimination index of the two groups more significantly than the other scores. There was a significant correlation between prostate volume and residual urine volume. The score of weak urinary steam was inversely and significantly correlated with peak urinary flow rate. CONCLUSIONS: There was no age-related enlargement of the prostate gland. The prostate gland was significantly larger in the patients than in the volunteers even in those in their fifties. Urinary frequency and sense of residual urine are important factors for men to seek and receive medical care.

Analysis of second-order kernel response components of multifocal electroretinograms elicited from normal subjects.

PURPOSE: It has been reported that the second-order kernel response components of multifocal electroretinograms (mERGs) reflect the electrical activity of the inner retinal layers. In this study, we have investigated whether the amplitudes of the second-order kernel response components correlate with the spatial distribution of human retinal ganglion cells. METHODS: Multifocal electroretinograms were recorded using the Veris III system from 5 healthy subjects with different stimulus and recording parameters. The mERGs were analyzed using the Veris Science software programs. The stimuli consisted of densely arranged arrays of 103, 61, 37 or 19 hexagonal elements. Four minutes were required to record one set of mERG responses using 8 sessions, and 8 minutes using 16 sessions. The second-order kernel response components were extracted and analyzed using the Veris Science program. RESULTS: The signal-to-noise ratio of the first-order kernel response components was improved considerably by the summation of the nine reproducible responses from the same subject but the second-order kernel response components were not. The summation of the nine reproducible responses was insufficient to identify an array of the second-order kernel response components. Both the first- and second-order kernel response components were larger when fewer hexagonal elements were used. There was no significant difference in the individual responses between the 4-minute and the 8-minute recordings. A response density analysis revealed a weak correlation between the amplitude distribution of the second-order kernel response components and the spatial distribution of human retinal ganglion cells. CONCLUSIONS: The distribution of the amplitudes of the second-order kernel response components of the mERGs elicited from normal subjects did not correlate with the distribution of human ganglion cells. This suggests that the theory that second-order kernel response components arise from the activity of retinal ganglion cells should be reconsidered.

[The significance of early detection for prostate cancer in mass screening].

PURPOSE: In Mitaka city, mass screening for prostate cancer was conducted for 3 years from 1995 to 1997. Clinical stages were compared between patients found by screening and those diagnosed at our clinic during the same time. The significance of serum-free prostate specific antigen (PSA) in mass screening for prostate cancer was examined. MATERIAL AND METHODS: A prospective clinical trial was conducted on men aged 50 years or older. The primary examination consisted of taking the international prostate symptom score, quality of life score, PSA (Tandem-R) and digital rectal examination (DRE). If PSA was greater than 4.0 ng./ml and/or if DRE suggested cancer, transrectal ultrasound-guided sextant prostate biopsies were indicated. RESULTS: Of the men screened, 23.2% (320/1375) had serum PSA greater than 4.0 ng./ml. and/or suspicious findings on DRE. Biopsy was performed in 199 of 320 (62.1%). Cancer was detected in 21 (1.5%, 21/1375). Prostate cancer was found in one case among 154 males (0.65%, 1/154) who were screened twice or more. The cancer stage found by screening was significantly earlier than that diagnosed at the outpatient clinic (Wilcoxon's rank-sum test: p = 0.0047). Receiver operating characteristics analysis showed that the optimal free PSA-to-PSA ratio was 12%. Positive predictive value increased from 18% to 50% when free PSA-to-PSA ratio was combined with PSA. CONCLUSION: 1. Cancer detection rate was 1.5% in the mass screening in Mitaka City. 2. Cancer stage found by screening was significantly earlier than that diagnosed at the outpatient clinic. 3. Free PSA determination might eliminate unnecessary biopsies in men with PSA above 4.0 ng./ml with minimal loss of cancer detection.