Applications of magnetic and electromagnetic forces in micro-analytical systems.

Novel applications of magnetic fields in analytical chemistry have become a remarkable trend in the last two decades. Various magnetic forces have been employed for the migration, orientation, manipulation, and trapping of microparticles, and new analytical platforms for separating and detecting molecules have been proposed. Magnetic materials such as functional magnetic nanoparticles, magnetic nanocomposites, and specially designed magnetic solids and liquids have also been developed for analytical purposes. Numerous attractive applications of magnetic and electromagnetic forces on magnetic and non-magnetic materials have been studied, but fundamental studies to understand the working principles of magnetic forces have been challenging. These studies will form a new field of magneto-analytical science, which should be developed as an interdisciplinary field. In this review, essential pioneering works and recent attractive developments are presented.

Life-Long Wheel Running Attenuates Age-Related Fiber Loss in the Plantaris Muscle of Mice: a Pilot Study.

The purpose of this study was to investigate whether long-term wheel running would attenuate age-related loss of muscle fiber. Male ICR mice were divided into young (Y, n=12, aged 3 months), old-sedentary (OS, n=5, aged 24 months), and old-exercise (OE, n=6, aged 24 months) groups. The OE group started spontaneous wheel running at 3 months and continued until 24 months of age. Soleus and plantaris muscles were fixed in 4% paraformaldehyde buffer. The fixed muscle was digested in a 50% NaOH solution to isolate single fiber and then fiber number was quantified. The masses of the soleus and plantaris muscles were significantly lower at 24 months than at 3 months of age, and this age-related difference was attenuated by wheel running (P<0.05). Soleus muscle fiber number did not differ among the groups. In the plantaris muscle, the fiber number in the OS group (1 288±92 fibers) was significantly lower than in the Y group (1 874±93 fibers), and this decrease was attenuated in the OE group (1 591±80 fibers) (P<0.05). These results suggest that age-related fiber loss occurs only in the fast-twitch fiber-rich muscle of mice, and that life-long wheel running exercise can prevent this fiber loss.

Brain-derived neurotrophic factor treatment increases the skeletal muscle glucose transporter 4 protein expression in mice.

The purpose of the present study was to investigate whether peripheral brain-derived neurotrophic factor (BDNF) treatment induced metabolic adaptations in mouse skeletal muscle. BDNF (20 mg/kg/day) was injected subcutaneously for successive 14 days. BDNF treatment significantly reduced the total food intake and inhibited the weight gain in comparison to the control group. The glucose transporter 4 (GLUT4) protein expression in the gastrocnemius muscle was significantly increased by BDNF treatment in comparison to the control and pair-fed groups. Neither the oxidative nor the glycolytic enzyme activities in the gastrocnemius muscle changed after the BDNF treatment. These results suggest that the peripheral BDNF treatment promotes the skeletal muscle GLUT4 protein expression as well as hypophagia.

Study on risk management of heavy metals for reuse of biosolids.

The behaviour of heavy metals was investigated at 22 wastewater treatment plants (WWTPs). In addition, the survey of heavy metal balance was conducted in detail at one WWTP. For the measurement, 22 types of heavy metals were selected from the chemical materials of pollutant release and transfer register (PRTR). There were some heavy metals, which were detected not in wastewater but in dewatered sludge. By means of the detailed survey at one WWTP, 60 to 80% of some heavy metals, such as B, Mn, Co, Ni and Mo, were discharged with treated water. According to the results of PRTR, Zn, B and Mn accounted for a large part of the discharge into the water course. To estimate the behaviour of heavy metals in the environment, leaching tests were applied to the products made of biosolids. During a series of leaching tests for building materials, it was observed that the concentration of heavy metals was very small, but the ratio of increase keeps a constant value. Therefore, it was considered that the acid extractable contents of heavy metal would be important.

Turbulence of glomerular hemodynamics involved in progressive glomerulosclerosis.

There is increasing evidence that changes of glomerular hemodynamics or glomerular growth responses may promote the development of glomerulosclerosis. Major problems retarding research progress include lack of suitable experimental animal models, with the exception of the ablation model, and the need for in vivo real-time analysis of glomerular hemodynamics. This study examined the sequence of pathological changes from the viewpoints of microcirculation and histopathology, from the acute stage to the chronic course and the final stage of glomerulosclerosis, using the confocal laser scanning microscope system. There is a marked difference in prognosis between sham-operated (two-kidney) and nephrectomized (one-kidney) rats after injection with anti-Thy-1 antibody. The former reversibly returns to normal and the latter irreversibly go to progressive sclerosis, respectively. The turning point determining the progression of glomerulosclerosis in both groups seemed to be the period from 7 to 14 days after disease induction, when disturbance of local intraglomerular blood flow continued in the one-kidney groups. In conclusion, this study provides the first hemodynamic-based evidence showing that disturbance of intraglomerular microcirculation is a critical marker for progressive glomerulosclerosis.

The contribution of cardiorespiratory fitness and visceral fat to risk factors in Japanese patients with impaired glucose tolerance and type 2 diabetes mellitus.

It is still unclear as to how cardiorespiratory fitness and visceral fat accumulation contribute to coronary heart disease (CHD) risk factors in patients with diabetes mellitus. The purpose of the present study was to investigate whether cardiorespiratory fitness contributes to such risk factors independently of visceral fat accumulation. Two hundred Japanese patients (137 men and 63 women, aged 22 to 81 years) with impaired glucose tolerance (IGT) and type 2 diabetes mellitus (type 2 DM) without any intervention and pharmacological therapy participated in a cross-sectional study. The levels of fasting insulin, triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and resting blood pressure were assessed. Maximal oxygen uptake (V.o(2max)), an index of cardiorespiratory fitness, was predicted by a graded exercise test using a cycle ergometer. Visceral fat area (VFA) was measured by computed tomography scan. The criteria for abnormalities of the risk factors were determined according to the standard values for Japanese. All subjects were divided equally into the following 3 groups according to their fitness level: low-fit (V.o(2max) < 32 mL/kg/min in men, V.o(2max) < 26 mL/kg/min in women), mid-fit (32 < or = V.o(2max) < 36 in men, 26 < or = V.o(2max) < 30 in women), and high-fit (V.o(2max) > or = 36 in men, V.o(2max) > or = 30 in women). The association between fitness level and the prevalence of abnormal values for these parameters was analyzed by a multiple logistic regression model adjusted for age and VFA. The odds ratio (OR) and 95% confidence interval (CI) for the prevalence of hyperinsulinemia were significantly lower in the mid-fit (OR = 0.35, 95% CI, 0.16 to 0.78) and in the high-fit groups (OR = 0.40, 95% CI, 0.16 to 0.98) compared with the low-fit group. In addition, ORs for the prevalence of low HDL-C in the mid-fit and high-fit groups were significantly lower (OR = 0.35, 95% CI, 0.14 to 0.86; and OR = 0.19; 95% CI, 0.08 to 0.60, respectively) than in the low-fit group. These results suggested that cardiorespiratory fitness might be one of the predictors of metabolic abnormalities, especially in patients with hyperinsulinemia and low HDL-C, independent of visceral fat accumulation in Japanese patients with IGT and type 2 DM.

Control of Cryptosporidium with wastewater treatment to prevent its proliferation in the water cycle.

The outbreak of Cryptosporidiosis in 1996 in Japan is thought to have been enlarged by the proliferation of Cryptosporidium in the water cycle from wastewater to drinking water through the river system. From this experience, the wastewater system must have functions to remove Cryptosporidium oocysts effectively. Efficiencies of wastewater treatment processes to remove oocysts were investigated using pilot plants receiving municipal wastewater. An activated sludge process and a following sand filter showed removal efficiencies of 2 log and 0.5 log, respectively. Poly-aluminium chloride dosage improved the efficiencies by 3 log for the activated sludge process and by 2 log for the sand filter. Chemical precipitation of raw wastewater with poly-aluminium chloride could achieve 1 to 3 log removal according on the coagulant concentration.

Dietary obesity-resistance and muscle oxidative enzyme activities of the fast-twitch fibre dominant rat.

OBJECTIVES: To clarify whether the muscle fibre composition and/or muscle oxidative enzyme activity are related to dietary body weight gain and abdominal fat accumulation. METHODS: Genetically fast-twitch fibre dominant rats (FFDR) and control rats (CR) were divided into low-fat (20% of energy from fat) or high-fat (60% of energy from fat) diet groups: CR with a low-fat diet (CL); CR with a high-fat diet (CH); FFDR with a low-fat diet (FL); and FFDR with a high-fat diet (FH). After 6 weeks of following such diets, the body weight gain, abdominal fat content, food intake, muscle fibre composition and oxidative enzyme activities were estimated. RESULTS: The total body weight gain in CH was from 18 to 62% higher than in the other groups (P<0.05) and percentage abdominal fat in CH was also from 26 to 61% higher than in the other groups (P<0.05), while the energy intake did not differ among the groups. The percentage of type IIX fibres of M. gastrocnemius in FL (33.4%) and FH (36.3%) were higher than in CL (16.8%) and CH (19.8%; P<0.05), and the type IIA fibres of M. soleus in FL (14.1%) and FH (11.8%) were higher than in CL (2.0%) and CH (3.5%; P<0.05). The citrate synthase (CS) activity of of M. plantaris in FL and FH were higher than CL (46 and 54%, respectively, P<0.05). beta-Hydroxyacyl CoA dehydrogenase (HAD) activity in FL and FH were higher than in CL (21 and 31%, respectively, P<0.05) and that in FH was higher than CH (23%, P<0.05). On the other hand, the enzyme activities of M. gastrocnemius and soleus were identical among the groups. CONCLUSION: The FFDR was more obesity-resistant than the CR after a high-fat diet. These results suggest that the muscle oxidative capacity rather than muscle fibre composition is a possible determinant of obesity.

Magnetophoretic velocimetry of manganese(II) in a single emulsion droplet at the femtomole level.

We developed a new experimental technique named magnetophoretic velocimetry to determine a small amount of paramagnetic species in a single microdroplet. The magnetophoretic velocity of an aqueous droplet containing paramagnetic metal ion dispersed in an organic medium could response to a very small amount of the metal ion under an inhomogeneous magnetic field. The paramagnetic droplet (2 approximately 8 microm diam) used as a test sample in this study was the aqueous droplet of manganese(II) chloride dispersed in ethylbenzoate whose density was nearly equal to water. A pair of small Nd-Fe-B magnets placed with a gap of 400 microm generated an inhomogeneous magnetic field between the edges, at which the product of the magnetic flux density and the gradient, B(dB/dx), was as high as 410 T2 m(-1). When a silica capillary containing the emulsion was inserted into the gap between the magnets, the magnetophoretic migration of the droplets was observed with a video microscope. The magnetophoretic velocity divided by the squared radius of the droplet was proportional to the MnCl2 concentration in the droplet, as predicted by a theoretical calculation. The estimated detection limit in this simple method was lower than 10(-16) mol for manganese(II).

Role of transesophageal echocardiography in the prediction of thromboembolism in patients with chronic nonvalvular atrial fibrillation.

The purpose of this study was to determine whether parameters derived from transesophageal echocardiography (TEE) could predict thromboembolism in patients with chronic nonvalvular atrial fibrillation (AF). Eighty-nine patients, mean age 66+/-9 years, who underwent TEE in 1996 to 1999 were studied. The clinical endpoint was a thromboembolic event, including transient ischemic attack (TIA). Sixty-seven patients (75%) were anticoagulated with warfarin after TEE. After a follow-up period of 29+/-10 months, 1 patient died suddenly, 4 had a thromboembolism, and 3 had a TIA; the annual embolic event rate was 3.3%. Left atrial appendage (LAA) thrombus (86% vs 17%, p<0.001), LAA dysfunction (LAA velocity <20 cm/s; 71% vs 25%, p=0.009), and severe LA spontaneous echo contrast (29% vs 2%, p=0.002) were more prevalent in patients with an embolic event than in those without. In patients with LAA thrombus, the annual event rate was 11% as compared with 1.2% in those without (p=0.004). On the Cox proportional hazards model analysis, LAA thrombus (chi-square 7.0, p=0.008), severe LA spontaneous echo contrast (chi-square 7.0, p=0.008), and LAA dysfunction (chi-square 5.9, p=0.015) were significantly related to thromboembolism. Multivariate analysis revealed that LAA thrombus (chi-square 5.5, p=0.019) and LAA dysfunction (chi-square 4.0, p=0.045) were the independent predictors. In conclusion, TEE parameters, particularly the presence of LAA thrombus, can be used to assess thromboembolic potential in patients with chronic nonvalvular AF.

Identification and quantification of tamoxifen-DNA adducts in the liver of rats and mice.

A new HPLC gradient system was developed for (32)P-postlabeling analysis to identify and quantify hepatic tamoxifen-DNA adducts of rats and mice treated with tamoxifen. Four stereoisomers of alpha-(N(2)-deoxyguanosinyl)tamoxifen (dG(3')(P)-N(2)-TAM), alpha-(N(2)-deoxyguanosinyl)-N-desmethyltamoxifen (dG(3')(P)-N(2)-N-desmethyl-TAM), and alpha-(N(2)-deoxyguanosinyl)tamoxifen N-oxide (dG(3')(P)-N(2)-TAM N-oxide) were prepared by reacting either alpha-acetoxytamoxifen, alpha-acetoxy-N-desmethyltamoxifen or alpha-acetoxytamoxifen N-oxide with 2'-deoxyguanosine 3'-monophosphate, and used as standard markers for (32)P-postlabeling/HPLC analysis. Our HPLC gradient system can separate the above 12 nucleotide isomers as nine peaks; six peaks representing two each trans epimers (fr-1 and fr-2) of dG(3')(P)-N(2)-TAM, dG(3')(P)-N(2)-N-desmethyl-TAM and dG(3')(P)-N(2)-TAM N-oxide, and three peaks representing a mixture of two cis epimers (fr-3 and fr-4) of nucleotides. Tamoxifen was given to female F344 rats and DBA/2 mice by gavage at doses of 45 mg/kg/day and 120 mg/kg/day, respectively, for 7 days. Totally 15 and 17 tamoxifen-DNA adducts were detected in rats and mice, respectively; among them 13 adducts were observed in both rats and mice. trans-dG-N(2)-TAM (fr-2) and trans-dG(3')(P)-N(2)-N-desmethyl-TAM (fr-2) were two major adducts in both animals. Except for these two adducts, trans-dG-N(2)-TAM N-oxide (fr-2) was the third abundant adduct that accounted for 6.4% of the total adducts in mice, while this accounted for only 0.3% in rats. A trans-isomer (fr-1) and cis-isomers (fr-3 and -4) of dG(3')(P)-N(2)-TAM, dG(3')(P)-N(2)-N-desmethyl-TAM and dG(3')(P)-N(2)-TAM N-oxide were also detected as minor adducts in both animals except for cis-form of dG-N(2)-TAM N-oxide in rats. Although the administered dose for rats was 2.7-fold less than that for mice, the total adduct level of rats (216 adducts/10(8) nucleotides) were 3.8-fold higher than mice (56.2 adducts/10(8) nucleotides). Thus, these three types of tamoxifen adducts accounted for 95.0 and 92.5% of the total DNA adducts of the rats and mice, respectively. The formation of tamoxifen adducts primarily resulted from alpha-hydroxylation of tamoxifen.

Quantification of left atrial appendage spontaneous echo contrast in patients with chronic nonalvular atrial fibrillation.

OBJECTIVES: The left atrial (LA) appendage is the most common site of thrombus formation in patients with atrial fibrillation, and integrated backscatter allows the quantiative assessment of LA spontaneous echo contrast (SEC). Integrated backscatter was used to examine the significance of measuring appendage SEC, specifically in relation to echocardiographic variables implying thromboembolism, in patients with chronic nonvalvular atrial fibrillation. METHODS: Fifty-two patients with chronic nonvalvular atrial fibrillation and no prior anticoagulant therapy (35 men, 17 women, mean age 66 +/- 7 years) underwent transesophageal echocardiography with integrated backscatter analysis. The LA and LA appendage integrated backscatter intensity were measured with the regions of interest placed in the LA cavity and the appendage, respectively. The integrated backscatter intensity values for these two chambers (corrected "LA" and "LA appendage" integrated backscatter intensity, respectively) were corrected using values from the left ventricular cavity. RESULTS: The LA appendage integrated backscatter intensity values were available in 44 patients (85%). Overall, the corrected LA appendage integrated backscatter intensity was significantly increased compared with the corrected LA integrated backscatter intensity (2.8 +/- 2.2 vs 2.0 +/- 1.8 dB, p < 0.001). The corrected LA appendage integrated backscatter intensity was inversely correlated with the LA appendage velocity (r = -0.37, p < 0.05), but not with the LA dimension, appendage size, or left ventricular function. The corrected appendage integrated backscatter intensity (4.5 +/- 2.3 vs 2.4 +/- 1.9 dB, p < 0.01) and LA integrated backscatter intensity (3.2 +/- 2.1 vs 1.7 +/- 1.7 dB, p < 0.05) were higher in patients who had LA appendage thrombus (n = 8) than those who did not. With the corrected appendage integrated backscatter intensity set at > or = 2.5 dB, the sensitivity for the presence of appendage thrombus was 88% and the specificity was 64%. CONCLUSIONS: Patients with chronic nonvalvular atrial fibrillation had a denser SEC in the LA appendage compared with SEC in the main LA cavity. The severity of the appendage SEC was influenced by the LA appendage function rather than its size. Quantification of SEC in the appendage, rather than main LA cavity, was more valuable for assessing embolic potential.

Doppler evaluation of systolic and diastolic heart failure in patients with cardiomyopathy.

BACKGROUND AND OBJECTIVES: Analysis of transmitral flow with Doppler echocardiography can currently evaluate left ventricular diastolic function noninvasively and easily. In addition, measurement of pulmonary venous flow and left atrial appendage flow clarifies not only left ventricular diastolic function but also left atrial systolic function in patients with left ventricular systolic failure and diastolic failure. METHODS AND RESULTS: Doppler echocardiography was used to investigate the differences in left ventricular diastolic dysfunction. In patients with systolic heart failure, left atrial dysfunction due to afterload mismatch to the left ventricle was related to the reduction of transmitral atrial filling wave in a restrictive pattern. In contrast, in patients with diastolic heart failure, transmitral flow seldom indicated a restrictive pattern because left atrial systolic function was preserved even with high pulmonary wedge pressure. CONCLUSIONS: Left ventricular diastolic dysfunction is somewhat distinct between patients with left ventricular systolic failure and patients with left ventricular diastolic failure. Atrial reversal flow in pulmonary vein has the potential to predict left ventricular filling pressure in patients with diastolic failure.

A case of cardiomyopathy showing progression from the hypertrophic to the dilated form: association of Mt8348A-->G mutation in the mitochondrial tRNA(Lys) gene with severe ultrastructural alterations of mitochondria in cardiomyocytes.

This report describes a case of cardiomyopathy with a novel point mutation of mitochondrial DNA coding lysine tRNA in association with severe ultrastructural alterations of the mitochondria in the cardiomyocytes. Abnormalities of energy production and/or abnormal protein synthesis because of the mutation of mitochondrial DNA may have played an important role in the pathogenesis of this case, which showed severe cardiomyocyte degeneration and deterioration from hypertrophic cardiomyopathy to severe dilated cardiomyopathy.

Occurrence of Cryptosporidium in Japan and counter-measures in wastewater treatment plants.

The outbreak of cryptosporidiosis in Ogose in 1996 forced the wastewater treatment authorities to rethink the level of contamination by Cryptosporidium of wastewater and waters in the watersheds and counter-measures in wastewater treatment plants. A survey of Cryptosporidium concentrations in wastewater and treated wastewater conducted nationwide showed relatively low levels. Also, evaluation of wastewater treatment showed a 2 log oocyst removal with an activated sludge process and an additional 1 log removal with coagulant dosing.

HUNT: launch of a full-length cDNA database from the Helix Research Institute.

The Helix Research Institute (HRI) in Japan is releasing 4356 HUman Novel Transcripts and related information in the newly established HUNT database. The institute is a joint research project principally funded by the Japanese Ministry of International Trade and Industry, and the clones were sequenced in the governmental New Energy and Industrial Technology Development Organization (NEDO) Human cDNA Sequencing Project. The HUNT database contains an extensive amount of annotation from advanced analysis and represents an essential bioinformatics contribution towards understanding of the gene function. The HRI human cDNA clones were obtained from full-length enriched cDNA libraries constructed with the oligo-capping method and have resulted in novel full-length cDNA sequences. A large fraction has little similarity to any proteins of known function and to obtain clues about possible function we have developed original analysis procedures. Any putative function deduced here can be validated or refuted by complementary analysis results. The user can also extract information from specific categories like PROSITE patterns, PFAM domains, PSORT localization, transmembrane helices and clones with GENIUS structure assignments. The HUNT database can be accessed at http://www.hri.co.jp/HUNT.

Prediction of mean pulmonary wedge pressure using doppler pulmonary venous flow variables in hypertrophic cardiomyopathy.

We examined whether pulmonary venous flow variables, assessed by transthoracic Doppler echocardiography, could predict mean pulmonary wedge pressure in hypertrophic cardiomyopathy. Forty-four patients with no left ventricular systolic dysfunction (left ventricular fractional shortening > or =25%) were studied. Forty patients with systolic dysfunction (dilated cardiomyopathy group) served as control. Mitral and pulmonary venous flow velocity curves were recorded with the pulsed-Doppler method and were related to mean pulmonary wedge pressure obtained by right heart catheterization. In hypertrophic cardiomyopathy group, the systolic (r=-0.15, P=0.335) and diastolic (r=0.35, P=0.022) forward flow velocity were poorly related to mean pulmonary wedge pressure, whereas the velocity of atrial reversal (r=0.68, P<0.001) correlated well with mean pulmonary wedge pressure. In dilated cardiomyopathy group, the systolic (r=-0.51, P=0.001) and diastolic (r=0.60, P<0.001) forward flow velocity were strongly related to mean pulmonary wedge pressure. With the cut-off value set at the velocity of atrial reversal >30 cm/s in hypertrophic cardiomyopathy group, the sensitivity for predicting mean pulmonary wedge pressure >15 mmHg was 79% and the specificity was 73%. In conclusion, the atrial component of the pulmonary venous flow can be used to predict mean pulmonary wedge pressure in hypertrophic cardiomyopathy.

A novel measure characterized by a polar energy surface approximation for recognition and classification of transmembrane protein structures.

A new theoretical method has been developed for recognition and classification of membrane proteins. The method is based on computation of a polar energy surface that can reveal characteristic interaction patterns for individual helices even if crystal or NMR structure coordinates are not available. A protein with N transmembrane helices is described as a set of N vectors that are derived from a Fourier analysis of this polar energy surface computed for each helix. We then derive a polarity difference score (PDS) for any two proteins computed as the root mean square deviation between the respective vector coordinate sets. The score was found to correlate with the degree of structural similarity between the following three protein families for which tertiary structures have been determined: bacteriorhodopsin, rhodopsin, and the cytochrome c oxidase III subunit.