Tetramethylbenzidine method for monitoring the free available chlorine and microbicidal activity of chlorite-based sanitizers under organic-matter-rich environments.

UNLABELLED: Chlorine is a principal disinfectant for food and environmental sanitation. Monitoring of free available chlorine (FAC) is essential for ensuring the efficacy of food disinfection processes that rely on chlorine. N,N-diethyl-p-phenylenediamine (DPD) is commonly used for FAC monitoring. However, here, we show that upon contact with bovine serum albumin (BSA) or broiler carcasses, chlorite (HClO2 )-based sanitizers acquire a pink colour, which can interfere with measurement of oxidized DPD absorbance at 513-550 nm. Alternatively, the pink colour did not interfere with 3,3',5,5'-tetramethylbenzidine (TMB)-based FAC monitoring. The FAC levels of NaClO and weakly acidified chlorous acid water (WACAW) were first adjusted by the TMB method and the killing activity of these sanitizers towards methicillin-resistant Staphylococcus aureus (MRSA) and feline calicivirus (FCV) was compared in the presence or absence of 0·5% BSA. At 200 ppm FAC, NaClO lost its bactericidal activity against MRSA after 10-min incubation with 0·5% BSA. Meanwhile, under the same conditions WACAW reduced the number of bacteria to below the detection limit. Similar results were obtained with FCV, indicating that the chlorite-based WACAW sanitizer is relatively stable under organic-matter-rich conditions. Moreover, TMB is suitable for in situ FAC monitoring of chlorite-based sanitizers in food and environmental disinfection processes. SIGNIFICANCE AND IMPACT OF THE STUDY: For practical applications of chlorine in food processing, monitoring of FAC is critical to validate disinfection efficacy. In this study we found that chlorite-based sanitizers acquired a pink colour upon contact with BSA or broiler carcasses. This pink colour interfered with FAC monitoring by methods that measure oxidized N,N-diethyl-p-phenylenediamine absorbance between 513-550 nm. Alternatively, FAC levels of chlorite-based sanitizers could be monitored using the absorbance of 3,3',5,5'-tetramethylbenzidine at 650 nm, which does not overlap with the acquired pink colour. These data provide valuable information for safety management of disinfection processes that use chlorite-based sanitizers.

Blockade of Th1 chemokine receptors ameliorates pulmonary granulomatosis in mice.

Sarcoidosis is a granulomatous disease of unknown aetiology. We identified immunological targets for the treatment of pulmonary granulomatosis using a murine model generated with Propionibacterium acnes. Sensitisation and challenge using heat-killed P. acnes and dendritic cells (DCs) were performed to produce pulmonary granulomatosis in C57BL/6 mice. Immunological analyses using ELISA as well as cDNA microarray analysis were used to search for cytokines or chemokines associated with the formation of granulomas in the lungs. Co-administration of P. acnes and DCs reproducibly induced the formation of pulmonary granulomas, which resembled sarcoid granulomas. The cDNA microarray assay demonstrated that the gene expression of CXCL9 and CXCL10, ligands for CXCR3, and of CCL4, a ligand for CCR5, was strongly upregulated during granulomatosis. ELISA confirmed that levels of CXCL9 and CXCL10 as well as T-helper (Th)1 cytokines and chemokines including tumour necrosis factor-α and interferon-γ were elevated in bronchoalveolar lavage fluid (BALF). The blockade of Th1 chemokine receptors using TAK-779, a dual blocker for CXCR3 and CCR5, led to reduced numbers of CXCR3+CD4+ and CCR5+CD4+ T-cells in BALF. Furthermore, administration of TAK-779 ameliorated the granulomatosis. The targeted inhibition of Th1 chemokines might be useful for inhibiting Th1-biased granulomatous diseases, including sarcoidosis.

Prevalence of Parkinson's disease and atypical parkinsonian syndromes in a rural Japanese district.

OBJECTIVES: To investigate the prevalence of Parkinson's disease (PD) and atypical parkinsonian syndromes (APS) in a rural Japanese district. METHOD: Collaboration with the medical institutions, the long-term care insurance system facilities, and the public health office. RESULTS: The crude prevalence rates were 175 per 100,000 (95% CI: 143-206) for PD, 18 (8-28) for progressive supranuclear palsy, 17 (7-26) for multiple system atrophy (MSA), and 9 (2-16) for corticobasal degeneration. The age-adjusted prevalence rates were 109 per 100,000 (88-134), 10 (2-17), 13 (4-21), and 6 (0-12), for each condition. There was a preponderance of women with PD and of men with APS. Nine of the 116 PD patients and 7 of the 29 APS patients were newly diagnosed in this study. CONCLUSIONS: There are high prevalence rates for PD and APS and suboptimal recognition of APS. This is the first epidemiological prevalence study of MSA from Japan.

[Myers technique and patch enlargement of ascending aorta for diffuse supravalvular aortic stenosis].

A 6-year-old boy was diagnosed with aggravation of diffuse supravalvular aortic stenosis (SVAS). The pressure gradient between the sinus of Valsalva and ascending aorta was 48 mmHg. The diameter of the sino-tubular junction was 7 mm and the ascending aorta was hypoplastic. We performed Myers operation because 3 sinus reconstructions had resulted in superior hemodynamics and reductions in both mortality rate and need for reoperation. We avoided using autologous pericardium because of the possibility of shrinkage and aneurysm. We could easily perform patch enlargement of the ascending aorta by selective cerebral perfusion. The postoperative course was excellent and there was no SVAS or aortic regurgitation (AR). A catheterization showed the pressure gradient was 5 mmHg with trivial AR upon follow-up at 1 year.

[Stenosis of left coronary artery ostium by a hypoplastic cusp].

Occlusion of a coronary ostium due to fusion of the aortic cusp to the aortic wall is a rare but noteworthy anomaly, because it may cause a sudden death. We report a 9-year-old girl with severe stenosis of the left coronary ostium by the aortic cusp that fused to the aortic wall. The left coronary flow was restored by excision of the adherent left aortic cusp, and aortic valve replacement was performed with the technique of bilateral enlargement of the aortic valve ring (Yamaguchi's method) to prevent prosthesis-patient mismatch in the future. The patient had an uneventful recovery and was discharged 14th postoperative day (POD). Postoperative angiography showed no coronary ostial stenosis.

Adrenomedullin inhibits connective tissue growth factor expression, extracellular signal-regulated kinase activation and renal fibrosis.

Systemic administration of the potent vasodilating peptide adrenomedullin reduces cardiac and renal fibrosis in hypertensive animals. Here, we investigated the effects of kidney-specific adrenomedullin gene delivery in normotensive rats after unilateral ureteral obstruction, an established model of renal tubulointerstitial fibrosis. Overexpression of exogenous adrenomedullin in the renal interstitium following ureteral obstruction significantly prevented fibrosis and proliferation of tubular and interstitial cells. In this model, there is upregulation of connective tissue growth factor (CTGF) mRNA expression and extracellular signal-regulated kinase (ERK) phosphorylation, and adrenomedullin overexpression suppressed both of these activities without altering the blood pressure. In NRK-49F renal fibroblasts, adrenomedullin reduced transforming growth factor-beta-induced CTGF and fibronectin mRNA upregulation through the cyclic AMP/protein kinase A signaling pathway, and suppressed ERK phosphorylation and cell proliferation. In the kidneys with an obstructed ureter, adrenomedullin receptor gene expression was upregulated along with cyclic AMP production in kidney slices. The latter effect was partially blocked by a neutralizing antibody to adrenomedullin, indicating that an endogenous peptide-receptor system was activated. Our results show that overexpression of exogenous adrenomedullin in the ureteral-obstructed kidney prevents tubulointerstitial fibrosis and cell proliferation through the cyclic AMP-mediated decrease of CTGF induction and ERK phosphorylation.

Flow regime classification in air-magnetic fluid two-phase flow.

A new experimental/numerical technique of classification of flow regimes (flow patterns) in air-magnetic fluid two-phase flow is proposed in the present paper. The proposed technique utilizes the electromagnetic induction to obtain time-series signals of the electromotive force, allowing us to make a non-contact measurement. Firstly, an experiment is carried out to obtain the time-series signals in a vertical upward air-magnetic fluid two-phase flow. The signals obtained are first treated using two kinds of wavelet transforms. The data sets treated are then used as input vectors for an artificial neural network (ANN) with supervised training. In the present study, flow regimes are classified into bubbly, slug, churn and annular flows, which are generally the main flow regimes. To validate the flow regimes, a visualization experiment is also performed with a glycerin solution that has roughly the same physical properties, i.e., kinetic viscosity and surface tension, as a magnetic fluid used in the present study. The flow regimes from the visualization are used as targets in an ANN and also used in the estimation of the accuracy of the present method. As a result, ANNs using radial basis functions are shown to be the most appropriate for the present classification of flow regimes, leading to small classification errors.

RNA polymerase beta-subunit gene (rpoB) sequence analysis for the identification of Bacteroides spp.

Partial rpoB sequences (317 bp) of 11 species of Bacteroides, two Porphyromonas spp. and two Prevotella spp. were compared to delineate the genetic relationships among Bacteroides and closely related anaerobic species. The high level of inter-species sequence dissimilarities (7.6-20.8%) allowed the various Bacteroides spp. to be distinguished. The position of the Bacteroides distasonis and Bacteriodes merdae cluster in the rpoB tree was different from the position in the 16S rRNA gene tree. Based on rpoB sequence similarity and clustering in the rpoB tree, it was possible to correctly re-identify 80 clinical isolates of Bacteroides. In addition to two subgroups, cfiA-negative (division I) and cfiA-positive (division II), of Bacteroides fragilis isolates, two distinct subgroups were also found among Bacteroides ovatus and Bacteroides thetaiotaomicron isolates. Bacteroides genus-specific rpoB PCR and B. fragilis species-specific rpoB PCR allowed Bacteroides spp. to be differentiated from Porphyromonas and Prevotella spp., and also allowed B. fragilis to be differentiated from other non-fragilisBacteroides spp. included in the present study.

[Primary biventricular repair for interrupted aortic arch with left ventricular outflow tract obstruction and tricuspid valve regurgitation; report of a case].

An interrupted aortic arch was diagnosed in a 10-day-old girl weighing 3.3 kg, as was perimembranous ventricular septal defect (VSD) and severe tricuspid valve regurgitation (TR). The subaortic diameter was 3.6 mm and the aortic valve (3.7 mm in diameter) was bicuspid. We chose definitive repair, modified Yasui procedure, because of severe TR and no straddling of mitral valve. In primary biventricular repair, we undertook extended aortic arch anastomosis. Left ventricular outflow tract reconstruction consisted of intracardiac rerouting from the VSD to the pulmonary artery by using expanded-polytetrafluoroethylene (ePTFE) and Damus-Kaye-Stansel (DKS) anastomosis. Right ventricular outflow tract reconstruction was performed by the Rastelli procedure with an ePTFE valved conduit. Moreover, we carried out semicircular annuloplasty for severe TR.

[Sandwich technique for closing of the large muscular Swiss-cheese type ventricular septal defects].

A 9-year-old girl, who has had a diagnosis as a Taussig-Bing anomary, underwent an original Jatene procedure (o-J) 2 months after birth. This time, she had a diagnosis of the muscular multiple ventricular septal defects (mVSD) and pulmonary stenosis after o-J. The mVSD was Swiss-cheese type and was large from the proximal of the infundibular septum to the apex and posterior of the septum. It was closed by the sandwich technique using a pair of felt patches, which of one was placed at right ventricular side and the other was at left ventricular side, slightly larger than the whole area of the mVSD. The patch fixation was placed with 1 stitch at the center between the patches and a few stitches around the right ventricular side patch to the ventricular septum. Postoperative cardiac function was uneventful regardless of the very large patches placed at the ventricular septum and the cardiac catheterization after 1 year postoperatively showed the pulmonary/systemic blood flow ratio was 1.0. This technique for the closure of the large Swiss-cheese type mVSD can be considered to be very effective.

Alteration of response to neuropeptide Y in the nucleus tractus solitarius of spontaneously hypertensive rats.

In normotensive rats, microinjections of neuropeptide Y (2.5 to 25 pmol) into the unilateral nucleus tractus solitarius elicited dose-dependent vasodepressor and bradycardiac responses accompanied by an inhibition of sympathetic nerve firing. After microinjections of the alpha 2-adrenergic receptor antagonist yohimbine (100 ng) into the nucleus tractus solitarius, the depressor and bradycardic responses to the injection of neuropeptide Y (25 pmol) into the nucleus tractus solitarius were significantly attenuated. In contrast, pretreatment with the alpha 1-adrenergic receptor antagonist doxazosin (200 ng) injected into the nucleus tractus solitarius did not alter these responses. In spontaneously hypertensive rats, microinjections of neuropeptide Y (25 pmol) into the nucleus tractus solitarius also elicited depressor and bradycardic responses that were significantly less than those of normotensive Wistar-Kyoto rats. However, pretreatment with yohimbine (100 ng) in the nucleus tractus solitarius did not diminish these depressor responses in spontaneously hypertensive rats. Depressor responses to neuropeptide Y, which was administered after yohimbine pretreatment, were also less in Wistar-Kyoto rats than in spontaneously hypertensive rats. The results suggest that the depressor and bradycardic responses elicited by neuropeptide Y were accompanied by the inhibition of sympathetic nerve activity. These responses may be mediated in part by alpha 2-adrenergic receptor in the nucleus tractus solitarius. The impairment of alpha 2-adrenergic receptor-mediated responses to neuropeptide Y in spontaneously hypertensive rats may contribute to the development of hypertension.

Genome analysis of Bacteroides by pulsed-field gel electrophoresis: chromosome sizes and restriction patterns.

The chromosomal DNAs of nine strains of seven Bacteroides species including B. fragilis, the type species of the genus Bacteroides, were digested with rare-cutting restriction enzymes I-Ceu I, Not I, and Asc I and analysed by pulsed-field gel electrophoresis. The genome sizes of B. fragilis, B. distasonis, B. eggerthii, B. ovatus, B. thetaiotaomicron, B. uniformis, and B. vulgatus were determined to be 5.3, 4.8, 4.4, 6.9, 4.8, 4.6, and 5.1 Mbp, respectively. B. distasonis and B. vulgatus, and also B. uniformis and B. eggerthii, showed similar I-Ceu I restriction profiles. I-Ceu I cut B. uniformis and B. eggerthii genomes into four, B. ovatus into five, B. fragilis and B. thetaiotaomicron into six, and B. distasonis and B. vulgatus into seven fragments. On the basis of genome size, restriction profile, and I-Ceu I fragment number, a phylogenetic tree of the Bacteroides species was proposed. This was in overall agreement with the previous phylogenetic tree obtained by 16S rRNA data, with the exceptions of B. distasonis and B. ovatus.

Kidney-specific expression of a novel mouse organic cation transporter-like protein.

Using the signal sequence trap method, we have cloned a novel 12-membrane-spanning transporter-like protein, termed renal-specific transporter (RST), from the mouse kidney. RST is a 553-amino-acid protein highly homologous to recently cloned organic cation transporters, e.g. it is 30% identical to rat organic cation transporter I at the amino acid level. Northern blot analysis has revealed that the RST gene is expressed abundantly and specifically in the kidney. In situ hybridization analysis has shown that RST gene expression is restricted to the renal proximal tubule, where various organic cations such as endogenous catecholamines and choline or clinically used cationic drugs are known to be actively excreted.

Direct evidence of impaired cardiac sympathetic innervation in essential hypertensive patients with left ventricular hypertrophy.

UNLABELLED: Increased sympathetic nervous activity has been proposed as one of the causes of left ventricular hypertrophy (LVH) associated with hypertension. However, the precise relationship is not fully understood. METHODS: To elucidate the relationship between myocardial sympathetic nervous activity and LVH in patients with essential hypertension EHT), we performed 123I-metaiodobenzylguanidine (MIBG) myocardial scintigraphy in 49 patients with EHT and 17 normotensive control subjects. Sympathetic innervation of the left ventricle was evaluated using SPECT, and the whole heart uptake of the tracer was quantitatively assessed as the heart-to-mediastinum uptake ratio on both the early (15-min) and delayed (5-hr) images. Myocardial washout rate (MWR) of the tracer from 15 min to 5 hr after the isotope administration was also calculated. The left ventricular mass index (LVMI) was determined echocardiographically. RESULTS: In 49 hypertensive patients, there was a negative correlation between LVMI and heart-to-mediastinum uptake ratio on both the early and delayed images (r=-0.55, p < 0.0001; r=-0.63, p < 0.0001, respectively). In addition, there was a positive correlation between the LVMI and MWR of 123I-MIBG in these hypertensive patients (r=0.59, p < 0.0001). As for the regional uptake of the tracer, there was no significant difference between control subjects and hypertensive patients without cardiac hypertrophy, but a significant decrease of the uptake in the inferior and lateral regions was observed in hypertensive patients with cardiac hypertrophy. CONCLUSION: Patients with EHT had decreased accumulation and increased MWR of 123I-MIBG in proportion to the degree of LVH. Hypertensive patients with cardiac hypertrophy had impaired sympathetic innervation in the inferior and lateral regions of the left ventricle.

C4d and C4bp deposition along the glomerular capillary walls in a patient with preeclampsia.

Complement (C) 4d and cofactor C4b binding protein (C4bp) are detected in the glomerular capillary walls of a patient with preeclampsia. A 32-year-old nullipara had proteinuria of 1.2 g/d and edema at the 33rd week of pregnancy. Gradually the urinary protein excretion increased, reaching 5.1 g/d at the 37th week. The patient also showed hypertension at this stage. After normal mature delivery, the level of the urinary protein excretion remained at 3 to 4 g/d. Renal biopsy performed by means of light and electron microscopy, 15 days after delivery, showed almost normal glomeruli and modest subendothelial widening. Immunohistochemistry indicated that immunoglobulin (Ig) A, IgG, C1q, C3c, and C4c were not deposited in the glomeruli, whereas weakly positive IgM and fibrin-related antigen (FRA) were observed. Conversely, C4d, C3d, and C4bp were strongly deposited. Protein S (PS) also was observed, with a similar distribution pattern to that of C4bp. Immunoelectron microscopy showed the deposition of C4d along the capillary walls and of C4bp in the subendothelium. These findings suggest that the C4 activation process as well as the regulation process of C system and of the inflammatory coagulation axis by C4bp and PS may play an important role in the pathophysiology of preeclampsia, so-called glomerular capillary endotheliosis (GCE).

Intraglomerular metastasis from pancreatic cancer.

Few case reports have shown the presence of metastatic tumor cells in renal glomeruli. We report one case with intraglomerular metastasis proved at renal biopsy. A 60-year-old man suffered from weight loss and fever of unknown origin. Urinalysis revealed proteinuria with cellular and granular casts. Because vasculitis was suspected, renal biopsy was performed. Presence of tumor cells occupying the glomerular capillary lumina was shown by means of light microscopy and electron microscopy. Laboratory findings revealed elevated leukocyte count (28.9 x 10(3)/mm(3)), serum granulocyte colony-stimulating factor (G-CSF) (77 pg/mL), and serum CA 19-9 (21,885 U/mL). The patient soon developed disseminated intravascular coagulation and died. Autopsy findings revealed pancreatic cancer showing positive staining for G-CSF and CA 19-9. Tumor cells in the glomerular capillary lumina showed positive staining for CA 19-9 and proliferating cell nuclear antigen (PCNA). These results suggest that the pancreatic tumor cells producing G-CSF were entrapped in the glomerular capillary lumina where they proliferated. This may have been the first step in renal metastasis.

Second meiotic nondisjunction of the rearranged chromosome in a familial reciprocal 5/13 translocation.

We describe a 20-week-gestation male fetus with partial dup(5p) and proximal dup(13q), 47,XY,t(5;13)(p15;q21), + der(13)t(5;13)(p15;q21) mat. This finding is attributable to second meiotic nondisjunction of the rearranged chromosome in a maternal balanced reciprocal translocation. To the best of our knowledge, there have been only 3 previous reports of a similar error in the segregation of the rearranged chromosomes. For the first time evidence has been given that this unusual segregation is due to maternal second meiotic nondisjunction, using QFQ banding heteromorphisms. Second meiotic malsegregation should be taken into account in the consideration of reproductive problems in carriers of balanced translocations.

Prosthetic replacement of entire left hemidiaphragm in malignant fibrous histiocytoma of the diaphragm.

A case of primary malignant fibrous histiocytoma of the diaphragm which, to our knowledge, is the first record in the world literature, is presented. It occupied almost the entire left hemidiaphragm and was surgically removed with the left lower pulmonary lobe, stomach, colon, and spleen. The entire left hemidiaphragm then was replaced with Marlex mesh. Prosthetic replacement of the entire hemidiaphragm, which has not been previously reported, was successfully performed with preservation of pulmonary function on the affected side. The patient died of brain metastases of malignant fibrous histiocytoma more than 4 months postoperatively. Diagnosis of diaphragmatic tumors and a technique which involves the use of a prosthesis are discussed.

Left ventricular geometry as an independent predictor for extracardiac target organ damage in essential hypertension.

Left ventricular hypertrophy (LVH) is an independent cardiovascular risk factor. It has not been established, however, whether left ventricular geometry is an independent predictor of extracardiac target organ damage in essential hypertension. Study groups were classified according to relative wall thickness: 27 patients with concentric LVH and 50 patients with eccentric LVH. Age and left ventricular mass indexes of two groups were matched. As indexes of extracardiac target organ damage, retinal funduscopic grade, and serum creatinine level were measured. The severity of hypertensive retinopathy and the renal involvement were more severe in patients with concentric LVH than in patients with eccentric LVH. Extracardiac target organ damage was consistently higher in patients with concentric LVH than in those with eccentric LVH. Systemic hemodynamics paralleled ventricular geometric patterns, with higher peripheral resistance and lower aortic compliance in patients with concentric LVH, whereas end-diastolic volumes and stroke volumes were higher in patients with eccentric LVH than in patients with concentric LVH. In addition, total peripheral resistance was related to retinal fundoscopic grade (r = 0.41, P < .01), and serum creatinine level (r = 0.28, P < .05). Even in the presence of an identical degree of LVH, echocardiographically determined left ventricular geometry may provide a further independent stratification of extracardiac target organ damage in essential hypertension.

Comparative immunochemical studies of sulfite oxidases of vertebrate livers.

1. The sulfite oxidases from various vertebrates, including rat, rabbit, chicken, frog, and eel, were partially purified and their physiochemical, kinetic, and immunochemical properties compared. The physicochemical and kinetic properties of these five enzymes were similar. 2. Antibody against the molybdenum-containing peptide prepared from purified rat liver sulfite oxidase was raised in rabbits. In Ouchterlony double diffusion and quantitative immunoprecipitation tests, the antibody could form precipitates with the enzymes from rats and rabbits, but no reaction was observed with enzymes from other sources. 3. The sulfite-cytochrome c reductase activity of the enzymes from these five animals were inhibited by the antibody, though the enzymes from chicken, frog, and eel were less sensitive to the antibody than those from rat and rabbit. 4. The inhibition of binding between 3H-labeled rat sulfite oxidase and its antibody by unlabeled enzymes from other animals demonstrated that 10 to 20% of the antibody could cross-react with these enzymes.