Assessing the infection risk of enteropathogens from consumption of raw vegetables washed with contaminated water in Kathmandu Valley, Nepal.

AIMS: To assess diarrhoeal risks from enteropathogenic Escherichia coli, Giardia and Cryptosporidium from consuming raw spinach, cabbage, carrots and tomatoes in Kathmandu Valley, Nepal. METHODS AND RESULTS: The annual infection risk was quantified using the probabilistic Quantitative Microbial Risk Assessment approach, which considered 12 vegetable washing combinations. A new model was used to estimate dose of pathogens per exposure comprising parameters such as pathogen concentration in vegetable wash water before selling and eating, vegetable consumption rate, remaining pathogen ratio after washing, remaining water on vegetables after washing and water treatment removal efficiency. When all washing combinations were considered, high infection risks above the acceptable level of -4 log10 infection per person per year were obtained, whereas the risk was reduced when other sources excluding river water were used. Assuming use of water treated with ceramic filters by all consumers, a 0-2 log10 reduction in the estimated risks was obtained, which was insufficient to achieve the required risk level. CONCLUSION: High risk of diarrhoea prevails among raw vegetable consumers in the valley. SIGNIFICANCE AND IMPACT OF THE STUDY: It is needed to protect vegetable washing water sources and establish advanced water treatment methods to achieve the required level of public health risk.

Comparative morphology of the Papillae Linguales and their connective tissue cores in the tongue of the greater japanese shrew-mole, Urotrichus talpoides.

The external morphology of the papillae linguales (papillae filiformes, papillae fungiformes and papillae vallatae) and their connective tissue cores (CTCs) of the greater Japanese shrew-mole (Urotrichus talpoides) were analysed by optical and scanning electron microscopy. Papillae filiformes were distributed over the dorsal surface of the apex linguae, and on the rostral and caudal regions of the corpus linguae but were less numerous in the mid-region. They were absent from the radix linguae. A pair of oval papillae vallatae was situated at the border between the corpus linguae and the radix linguae. Papillae foliatae were absent. The epithelial surface of each papilla filiformis consisted of a circular concavity, a ring-like wall and either a single thumb-like process or 2-3 slender pointed processes, depending on their location. The morphology of the CTCs of the papillae filiformes also varied regionally. The papillae linguales of the Japanese shrew-mole were morphologically similar to those of other Talpidae and Soricidae, including the common shrew, particularly with respect to the papillae filiformes in the mid- and caudal regions of the corpus linguae.

Influence of leukotriene pathway polymorphisms on clinical responses to montelukast in Japanese patients with asthma.

WHAT IS KNOWN AND OBJECTIVE: Montelukast, a cysteinyl leukotriene receptor 1 antagonist, is safe and efficacious in patients with asthma. The mechanisms underlying the significant interpatient variability in response to montelukast are not clear but are believed to be, in part, because of genetic variability. METHODS: To examine the associations between polymorphisms in candidate genes in the leukotriene pathway and outcomes in patients with asthma on montelukast for 4-8 weeks, we evaluated the changes in peak expiratory flow (PEF), forced expiratory volume in 1 s (FEV(1·0) ) and patients' subjective symptom before and after montelukast treatment. DNA was collected from 252 Japanese participants. RESULTS AND DISCUSSION: Two single-nucleotide polymorphisms (SNPs) in the ALOX5 (rs2115819) and LTA4H (rs2660845) genes were successfully typed. There was no difference between members of the general population (n = 200) and patients (n = 52) in each genotype frequency. Significant associations were found between SNP genotypes in the LTA4H gene and changes in PEF and FEV(1·0) . The PEF and FEV(1·0) responses to montelukast in the A/A genotypes (n = 4) for the LTA4H SNP were significantly higher than those in the G allele carriers (A/G+G/G) (n = 17). WHAT IS NEW AND CONCLUSION: Despite the small sample size, our results suggest that genetic variation in leukotriene pathway candidate genes contributes to variability in clinical responses to montelukast in Japanese patients with asthma.

Growth of human squamous cell carcinoma xenografts in mice is inhibited by local angiostatin gene therapy.

The possibility of inhibiting tumor growth by blocking the formation of new tumor vessels has recently received attention. Antiangiogenic tumor therapies have recently attracted intense interest because of their direct endothelial targeting and the absence of drug resistance. Local antiangiogenic gene therapy for cancer offers a potential way to achieve sustained therapeutic release of antiangiogenic substances. As a step toward this goal, we used liposomes complexed to angiostatin cDNA and targeted to human squamous cell carcinoma cell lines in vivo. Tumor cells expressing angiostatin after local gene transfer showed markedly reduced vascularity and contained many apoptotic tumor cells. These results demonstrate the potential utility of liposome-derived angiostatin for adjuvant therapy of oral cancer in humans.

Stereo-structural study of the lingual papillae and their connective tissue cores in relation to ageing changes in the human tongue.

This study describes the stereo structure of human lingual papillae along lifespan with particular emphasis to ageing. The following results were obtained. Numerous slender protrusions extend from the basal column of each filiform papilla of young human tongues and decrease in number with age. Many cases having an entirely flat area on the surface of the tongue without papillae were found in old age (Loss of protrusions and flattened epithelium). The number, thickness and length of each secondary protrusion of connective tissue core (CTC) of the filiform papilla decreases roughly proportionally with age (Ramification at the end of protrusions, and fused patterns of CTC of the filiform papilla). The CTC of fungiform papillae which have a corolliform structure, become thin with age. Numerous small rod-shaped secondary CTC are distributed under the epithelium of the vallate papillae in the young subjects and decrease in number and become irregularly distributed in older age. The fact that there is a great deal of individual variation of human lingual papillae regardless of age must be emphasized.

Increased plasminogen activator inhibitor-1 and apolipoprotein (a) in coronary atherectomy specimens in acute coronary syndromes.

BACKGROUND: Although increased tissue factor expression is known in vulnerable plaques, there is no reported study to compare plaque fibrinolysis in stable and unstable plaques. This study investigates the extent of plasminogen activator inhibitor-1 (PAI-1) and apolipoprotein (a) [apo(a)] in the plaques of different types of coronary artery disease as well as the correlation between these molecules and infiltration of macrophages to plaques. METHODS: Using immunohistochemical staining, we examined PAI-1 expression and apo(a) deposition in coronary atherosclerotic specimens obtained by directional coronary atherectomy from 19 patients with acute myocardial infarction (AMI), 12 with unstable angina pectoris (UAP), and 13 with stable angina pectoris (SAP). The percentages of the total areas of specimens stained with PAI-1 or apo(a) were estimated by an NIH image program. The proportion of macrophages as a percentage of all cells in plaques was calculated as the macrophage density. RESULTS: We found significantly higher percentages of total areas of specimens stained with PAI-1 in AMI (25.5 +/- 8.6%, P < 0.001) and UAP (22.2 +/- 10.4%, P < 0.005) than in SAP (9.5 +/- 5.0%), as well as with apo(a) (AMI; 11.7 +/- 7.1%, P < 0.005, UAP; 11.1 +/- 5.5%, P < 0.01 versus SAP; 3.9 +/- 1.5%). Linear regression analysis of all the samples showed a correlation between PAI-1 or apo(a) and macrophage density (PAI-1: r = 0.75, P < 0.001 and apo(a): r = 0.56, P < 0.001). CONCLUSIONS: Our results suggest a possible contribution of increased PAI-1 and apo(a) in plaques to the pathogenesis of acute coronary syndromes including impaired fibrinolysis.

Disseminated histoplasmosis in a sea otter (Enhydra lutris).

Disseminated histoplasmosis was diagnosed in a 4.75-year-old, captive female sea otter (Enhydra lutris). At necropsy, the liver was found to be markedly swollen, with many nodules (4-12 mm in diameter). Histologically, macrophages containing numerous intracellular yeast-like organisms were noted in the liver, spleen, lung and kidney. These organisms were labelled immunohistochemically with anti-histoplasma yeast antibody. Ultrastructurally, the yeast-like organisms, 2-4 microm in diameter, were found within membranous structures in the cytoplasm of macrophages. This is the first confirmed report of disseminated histoplasmosis in sea otters.

Angiostatin gene therapy inhibits the growth of murine squamous cell carcinoma in vivo.

Tumor growth is an angiogenesis-dependent process and therapeutic strategies aimed at inhibiting angiogenesis are theoretically attractive. Angiostatin has been shown to potently inhibit endothelial proliferation in vitro and tumor growth in vivo. We now show that a shift in the balance of tumor angiogenesis by gene transfer of a cDNA coding for mouse angiostatin into mouse squamous cell carcinoma NRS-1 and SCC-VII cells suppresses tumor growth in vivo. The inhibition of an angiostatin-transfected tumor was accompanied by a marked reduction in vascularity and the presence of many apoptotic tumor cells. However, transfected-angiostatin cDNA does not affect the expression of the vascular endothelial growth factor (VEGF) and VEGF-R2 in the vascular endothelium. The inhibition mechanisms of neovascularization may be mediated independent of VEGF:VEGF-R2 complex. Our data may provide a useful approach for human oral cancer therapy by gene therapy with angiostatin.

Input-output budget of nitrogen and the effect of experimentally changed deposition in the forest ecosystems in central Japan.

To evaluate the current nitrogen (N) status in Japanese forests, field measurements of rainfall, throughfall, litter layer percolation, and soil solution percolation were conducted in a red pine stand (Kannondai) and a deciduous stand (Yasato) located in central Japan. N input via throughfall was 31 and 14 kg ha(-1) year(-1) and output below rooting zone was 9.6 and 5.5 kg ha1 year(-1) in Kannondai and in Yasato, respectively. Two thirds of input N were retained in plant-soil systems. Manipulation of N input was carried out. Ionic constituents were removed from throughfall with ion exchange resin at removal sites and ammonium nitrate containing twice the N of the throughfall was applied at N addition sites periodically. SO4(2-) output below 20-cm soil layer changed depending on the input, while NO3- output was regulated mainly by the internal cycle and effect of manipulation was undetected. These Japanese stands were generally considered to have a larger capacity to assimilate N than NITREX sites in Europe. However, N output fluxes had large spatial variability and some sites in Kannondai showed high N leaching below rooting zone almost balanced with the input via throughfall.

Myasthenia gravis associated with reduced masticatory function.

This paper describes a case of myasthenia gravis in a 38-year-old woman who first consulted a dentist and then an oral surgeon because of chewing difficulty. Although myasthenia gravis commonly presents with diplopia, ptosis, or both as initial symptoms, chewing difficulty is rare. The patient was given steroid therapy and underwent thymectomy. Changes in bite force were monitored during treatment. The bite force was low when a high titer of anti-acetylcholine receptor antibody (11.0 nmol/l, normal <0.2) was found in the blood, but increased after the titer had decreased (1.5 nmol/l) in response to therapy.

Ectopic pheochromocytoma arising in the spermatic cord 5 years after removal of bilateral carotid body tumors and adrenal pheochromocytomas.

A patient with ectopic pheochromocytoma that developed in the spermatic cord about 5 years after successive resections of bilateral carotid body tumors and bilateral adrenal pheochromocytomas is reported. This is thought to be the first case of pheochromocytoma of the spermatic cord in a setting of multiplicity.

Technical aspects of venous stenting in high-grade stenoses using a long guidewire between dual venous access sites.

We describe the technical benefits of the dual-access technique for venous stenting in high-grade stenosis. Stents were implanted successfully with this technique in all patients, although the preceding transfemoral interventions had failed. The dual-access technique is useful to facilitate the intervention when the stenoses are too severe to place the stent through just a single access point.

Adhesion mediated by LFA-1 is required for efficient IL-12-induced NK and NKT cell cytotoxicity.

Interleukin 12 (IL-12)-activated NK1.1+TCRalpha beta+ (NKT2) and NK1.1+TCRalpha beta- (NK) cells exhibit cytotoxic activity against a wide variety of tumor cells in the absence of prior sensitization. Here we demonstrate that the integrin adhesion receptor LFA-1 (CD11a/CD18) regulates the cytotoxic activity of IL-12-activated NKT and NK cells against YAC-1 and EL-4 tumor cells. Differentiation in vivo and the expression of the cytolytic effector molecules perforin and Fas-L were comparable in both IL-12-activated NKT and NK cells from LFA-1-/ - and LFA-1+/+ mice. However, LFA-1-/-IL-12-activated NKT and NK cells showed impaired conjugate formation with target cells. These results provide the first genetic evidence for a role for an adhesion receptor in killing by IL-12-activated NK cells.

Expression of cytokine and adhesion molecule mRNA in atherectomy specimens from patients with coronary artery disease.

Coronary arteriosclerosis is an underlying condition in acute myocardial infarction (AMI), unstable angina pectoris (UAP) and stable angina pectoris (SAP), and is also related to restenosis (RS) following coronary intervention. To investigate the pathogenesis of this condition, a quantitative reverse transcriptase polymerase chain reaction was used to determine relative levels of mRNA for interleukin (IL)-1beta, IL-6, IL-8, transforming growth factor beta (TGF-beta), intercellular adhesion molecule (ICAM)-1, E-selectin and vascular cell adhesion molecule (VCAM)-1 using directional coronary atherectomy (DCA) specimens. Eleven patients with AMI, 7 with UAP, 10 with SAP and 6 with RS following a previous coronary intervention underwent DCA. The mRNA intensity for each molecule was expressed by comparing it with that of beta-actin mRNA. The AMI and UAP patients showed high frequencies of mRNA for IL-1beta, IL-8, TGF-beta, and ICAM-1 together with strong intensities of expression, whereas SAP patients showed decreased mRNA expression for these molecules. Increased IL-6 mRNA expression was observed only in AMI samples. Specimens from RS patients revealed an accumulated expression of proinflammatory cytokines, except for IL-6, as well as of TGF-beta. The study suggests that variation in mRNA expression may reflect the pathophysiology of specific types of coronary artery disease, and remodeling following vascular injury.

Granulocyte-macrophage colony-stimulating factor prevents the progression of atherosclerosis via changes in the cellular and extracellular composition of atherosclerotic lesions in watanabe heritable hyperlipidemic rabbits.

BACKGROUND: A cytokine network is involved in atherogenesis. This study was conducted to investigate the effects of granulocyte-macrophage colony-stimulating factor (GM-CSF) on the development and composition of atherosclerotic lesions in Watanabe heritable hyperlipidemic (WHHL) rabbits. METHODS AND RESULTS: GM-CSF (10 microg. kg-1. d-1) was administered to 4-month-old WHHL rabbits (n=9) 5 days a week for 7.5 months, whereas an equal dose of human serum albumin was administered to controls (n=9). The cholesterol levels were not changed significantly by the treatment. Age-matched 4-month-old rabbits (n=7) had atheromatous plaques over 30.7+/-5.7% of the inner surface area of the aortic arch. After treatment, the percentages of surface atheromatous plaques to total aortic arch area were 45.0+/-12.6% in the GM-CSF group and 74.3+/-11.0% in controls (P<0.0001). Histological examination demonstrated that GM-CSF reduced the ratio of intima to media (P<0.01) and cross-sectional areas of atherosclerotic lesions (P<0.0001). Quantitative analysis indicated a marked decrease in the areas of smooth muscle cells (P=0.0001), collagen (P=0.0001), and extracellular lipid deposits (P<0.05) of atheromatous plaques in GM-CSF-treated rabbits compared with controls. The terminal deoxynucleotidyltransferase-mediated dUTP-digoxigenin nick end-labeling (TUNEL) method and immunohistochemistry were performed to examine the relationship between decreased atherosclerotic lesions and apoptosis. The percentage of TUNEL-positive cells increased in the GM-CSF group (GM-CSF, 24.1+/-4.4% versus control, 11.6+/-3.2%; P<0.0001). GM-CSF enhanced the apoptosis of smooth muscle cells in the shoulder region and the fibrous cap (P<0.0001), suggesting one of the mechanisms for the antiatherogenic effect. CONCLUSIONS: GM-CSF altered the composition of atherosclerotic lesions and reduced the atherosclerosis in WHHL rabbits.

[Anesthesia for tracheobronchial stent insertion].

We experienced nine cases of general anesthesia for tracheobronchial stent insertion. As far as possible, we anesthetized the patients under spontaneous respiration with inhalation anesthesia, depending on the types of the tracheobronchial stent (Dumon stent or expandable metallic stent), or respiratory status of the patient. Having experienced a case requiring PCPS (percutaneous cardiopulmonary support) to recover from severe ventilatory insufficiency due to tracheal perforation, we, in advance, inserted 18- or 20-gage cannulas into the femoral artery and vein for PCPS standby, and could manage general anesthesia safely for tracheobronchial stent insertion.

Oligodontia: a radiographic comparison of subjects with Down syndrome and normal subjects.

Panoramic radiographs and clinical records were used to investigate developmentally absent permanent teeth in 98 subjects with Down syndrome (trisomy-21). This retrospective study was based on the records and panoramic radiographs of subjects from approximately five years of age (the age at which mineralization of the permanent tooth germ could be identified) through to their most recent records. The time period covered by records ranged from 6 to 28 years. The majority of subjects with Down syndrome (63%) exhibited oligodontia, and many subjects were missing two or more teeth (53%). The most frequently absent teeth were the lower lateral incisors (23.3%), the upper second premolars (18.2%), the upper lateral incisors (16.5%), and the lower second premolars (15.3%). In general, the distribution of the developmentally absent teeth was similar for teeth in homologous positions (i.e., left and right canines, etc.) on either side of the midline or between the maxilla and the mandible. The only significant exceptions to this pattern were seen with the central incisors and the second molar. This study's findings suggest a high risk of oligodontia in subjects with Down syndrome.