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Background Cerebral malaria (CM) is associated with morbidity and mortality despite the use of potent anti-malarial agents. Brain endothelial cell activation and dysfunction from oxidative and inflammatory host responses and products released by Plasmodium falciparum-infected erythrocytes (IE), are likely the major contributors to the encephalopathy, seizures, and brain swelling that are associated with CM. The development of adjunctive therapy to reduce the pathological consequences of host response pathways could improve outcomes. A potentially protective role of the nuclear factor E2-related factor 2 (NRF2) pathway, which serves as a therapeutic target in brain microvascular diseases and central nervous system (CNS) inflammatory diseases such as multiple sclerosis was tested to protect endothelial cells in an in vitro culture system subjected to tumour necrosis factor (TNF) or infected red blood cell exposure. NRF2 is a transcription factor that mediates anti-oxidant and anti-inflammatory responses. Methods To accurately reflect clinically relevant parasite biology a unique panel of parasite isolates derived from patients with stringently defined CM was developed. The effect of TNF and these parasite lines on primary human brain microvascular endothelial cell (HBMVEC) activation in an in vitro co-culture model was tested. HBMVEC activation was measured by cellular release of IL6 and nuclear translocation of NFκB. The transcriptional and functional effects of dimethyl fumarate (DMF), an FDA approved drug which induces the NRF2 pathway, on host and parasite induced HBMVEC activation was characterized. In addition, the effect of DMF on parasite binding to TNF stimulated HBMVEC in a semi-static binding assay was examined. Results Transcriptional profiling demonstrates that DMF upregulates the NRF2-Mediated Oxidative Stress Response, ErbB4 Signaling Pathway, Peroxisome Proliferator-activated Receptor (PPAR) Signaling and downregulates iNOS Signaling and the Neuroinflammation Signaling Pathway on TNF activated HBMVEC. The parasite lines derived from eight paediatric CM patients demonstrated increased binding to TNF activated HBMVEC and varied in their binding and activation of HBMVEC. Overall DMF reduced both TNF and CM derived parasite activation of HBMVEC. Conclusions These findings provide evidence that targeting the NRF2 pathway in TNF and parasite activated HBMVEC mediates multiple protective pathways and may represent a novel adjunctive therapy to improve infection outcomes in CM.

Purpose We will analyze the PDE5 inhibitor prescription at Kobe-Sannomiya EAST Ekimae Clinic(predecessor). Methods To analyze the type of antihypertensive drug in hypertensive patients receiving pharmacotherapy among male re-examination patients who prescribed PDE5 inhibitors more than 6 months after the initial consultation. Results The target 445 age groups were 5 in their 30s, 35 in their 40s, 144 in their 50s, 177 in their 60s, 79 in their 70s, and 5 in their 80s. The elapsed time from initial consultation to re-examination was 96 for 6 months or more and less than 1 year, 214 for 1 year or more and less than 2 years, and 135 for 2 years or more. The antihypertensive drugs were Ca antagonist 133 (24 other drugs other than ARB), angiotensin Ⅱ receptor antagonist (ARB) 112 (19 other drugs other than Ca antagonist), Ca antagonist and there were 186 patients with ARB (43 with other drugs) and 14 with other drugs. The breakdown of other concomitant drugs and other drugs was β-blockers, α-blockers, αβ-blockers, ACE inhibitors, and diuretics. Very few patients voluntarily complained of ED symptoms associated with taking antihypertensive drugs, and various types of antihypertensive drugs were taken. Discussion Even if a PDE5 inhibitor is more effective, it is necessary to consult with your GP about taking antihypertensive drugs. Disclosure Work supported by industry: no.

Additional file 5. Binding specificity of P. falciparum parasites ITgICAM1 and IT4var19 to HBMVEC. A. Binding of ITgICAM1 (DC17) IE to resting and TNF activated HBMVEC is shown as percent binding to HBMVEC relative to unstimulated HBMVEC. Inhibitory antibodies against ICAM1 (mAb15.2) or IgG isotype control were added to HBMVEC monolayers prior to IE cytoadherence. B. Binding of IT4var19 (DC8) IE to resting HBMVEC is shown as percent binding to HBMVEC. Inhibitory antibodies against EPCR (mAb252) or IgG isotype control were added to HBMVEC monolayers prior to IE cytoadherence (n = 3 independent experiments). Results shown were analysed using analysis of variance followed by post-hoc multiple comparisons using Turkey’s test, ***p

Occurrence of Chagas disease and arbovirus coinfections is unknown, despite the vast co-endemic areas throughout the Americas. This study examined the proportion of individuals positive for Trypanosoma cruzi and coinfections with dengue, chikungunya, and Zika viruses in Machala, Ecuador (January 2014-December 2015). Chagas seropositivity was evaluated with five commercially available assays. Dengue infections were identified by nonstructural protein 1 rapid test and enzyme linked immunosorbent assay (ELISA), immunoglobulin M ELISA, and reverse transcription PCR (RT-PCR); chikungunya and Zika infections were identified by RT-PCR. Of 658 individuals, six were positive for T. cruzi (0.91%), including one T. cruzi/dengue coinfection and one T. cruzi/chikungunya/dengue coinfection. The clinical manifestations of coinfected individuals corresponded to severe dengue and dengue with warning signs, respectively. We observed discrepant results by using the Hemagen Chagas kit and the rapid test Chagas Detect Plus (false positives: 3.9% and 15.4%), highlighting the need to assess diagnostic assays in geographic regions with distinct taxonomic units of T. cruzi.

When dengue virus (DENV)-infected mosquitoes use their proboscis to probe into human skin during blood feeding, both saliva and virus are released. During this process, cells from the epidermis and dermis layers of the skin, along with small blood vessels, may get exposed to or infected with DENV. In these microenvironments of the skin, the presence of DENV initiates a complex interplay among the DENV-infected and non-infected neighboring cells at the initial bite site. Previous studies suggested that DENV-infected human dermal fibroblasts (HDFs) participate in the immune response against DENV by secreting soluble mediators of innate immunity. In the present study, we investigated whether DENV-infected HDFs activate human dermal microvascular endothelial cells (HDMECs) in co-cultures. Our results suggest that co-cultures of DENV-infected HDFs and HDMECs elicit soluble mediators that are sufficient to reduce viral replication, activate HDMECs, and induce leukocyte migration through HDMEC monolayers. These effects were partly dependent on HDF donor and DENV serotype, which may provide novel insights into the natural variation in host susceptibility to DENV disease.

Reference materials are used in every laboratory in order to assure the appropriate quality of analytical results. The production of reference materials is not an easy task especially in the case of gaseous, toxic and malodorous compounds. Thermal decomposition of immobilized compounds is a convenient way for the generation of those kinds of mixtures. Temperature is an important variable in this process. In this case, thermoanalytical techniques (thermogravimetry and differential scanning calorimetry (TG-DSC)) and scanning electron microscopy (SEM) were used for checking the temperature range which had been previously determined using thermal desorber-gas chromatograph-flame ionization detector (TD-GC-FID) system.

A Bombyx EST cDNA database was searched using the Drosophila takeout gene and nine cDNAs were obtained. The homology search suggested that these genes are widespread in insects and organize a large gene family, and that they have hydrophobic ligands. A phylogenetic tree indicated that the genes are first divided into two large groups, juvenile hormone binding protein and other protein genes, and the latter group diversified within a short time at an early stage. The expression study of five Bombyx genes indicated that they are expressed in various tissues and are regulated by development and feeding conditions. The Bombyx genes might have roles related to the regulation of metabolism, growth or development related to nutritional conditions.

There is a need to provide an index of muscle contractility in evaluating myopathies especially in the clinical setting. This study was conducted to investigate if the mechanomyogram post-activation potentiation (MMG-PAP) can be used as an index of muscle contractile force potentiation (force-PAP), if it differs between normal and myopathic muscles, and if it can reflect abnormalities in muscle fiber anatomy.The correlation between MMG-PAP and force-PAP was evaluated in 12 normal subjects after maximum voluntary contraction (MVC) of the biceps brachii muscle. The same method was then applied to study MMG-PAP in 16 patients with myopathies, 16 disease and 25 normal controls. Mean fiber diameters and the proportions of type 1 and 2 fibers in biopsied biceps brachii muscle were determined and compared with MMG-PAP values.There was a significant positive correlation between force-PAP (197 +/- 148%) and MMG-PAP (135 +/- 68%) immediately after MVC (P0.05). The mean MMG-PAP in myopathies (66 +/- 53%) was significantly lower than those of the disease (128 +/- 34%; P0.005) and normal controls (120 +/- 56%; P0.005). Patients with non-dystrophic myopathies, including those with myositis, had significantly lower MMG-PAP values (38 +/- 20%; P0.005) than those with muscular dystrophy (148 +/- 23%). MMG-PAP did not clearly correlate with either type 2 fiber atrophy or type 2 fiber disproportion based on muscle biopsy analysis of myopathic patients.This study shows that MMG-PAP can be used as an index of muscle contractility and that it is significantly lower in non-dystrophic myopathies compared to normal subjects. MMG-PAP does not seem to reflect abnormal muscle fiber anatomy.MMG-PAP may become a valuable non-invasive tool in augmenting routine clinical electrophysiologic studies especially in evaluating muscle contractility in myopathies.

A key enzyme of polyamine catabolism, spermidine/spermine N(1)-acetyltransferase (SSAT), is responsive to antiproliferative agents. The role of SSAT in cellular responses to X-ray irradiation was examined.Exponentially growing HeLa S3 cells were irradiated by X-rays, and mRNA levels for SSAT were measured as a function of post-irradiation time through Northern hybridization. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect alternatively spliced SSAT mRNAs. The intracellular polyamine content was measured by the o-phthalaldehyde method and the enzymatic activity of SSAT by the increased amount of acetylated spermidine after incubation.Not only SSAT mRNA, but also an alternatively spliced mRNA accumulated at the initial stage of growth inhibition after the first or second replication of irradiated cells. The maximum fold increase relative to the level of non-irradiated cells was 3.0-3.5 for both transcripts after 5-Gy irradiation. On the other hand, the mRNA of ornithine decarboxylase, a key enzyme of polyamine synthesis, was little influenced by X-ray treatment. Enzymatic activity of SSAT and the acetylspermidine level were elevated after X-ray irradiation.Activation of SSAT and the induction of alternatively spliced mRNA of the SSAT gene play an important role in regulating growth inhibition and cell death after X-ray irradiation.

This chapter focuses on the pathogenesis of malaria caused by Plasmodium falciparum and Plasmodium vivax. It talks about two factors, parasite multiplication rate and cytoadherence of parasitized Red Blood Cells (RBCs), both of which enable parasites to achieve high densities and induce microvascular inflammation. The chapter considers several cytoadherence interactions between parasitized RBCs and host cells to better understand how parasites cause microvascular obstruction, inflammation, and dysfunction in vivo. Additional evidence for endothelial dysfunction in severe malaria has now been reported from a study of Indonesian adults. In this study, cell-free hemoglobin and arginase levels were elevated in patients with severe malaria, compared to those with moderately severe malaria or those who were healthy. The chapter presents the pathogenesis of vivax malaria, and discusses human genetic resistance to vivax malaria. It also describes working models of malaria pathogenesis which the authors believe provide useful frameworks for future studies. The authors propose that the levels of parasitemia and the strength of pathogen-host cell interactions are critically involved in the pathogenesis of malaria, and that host factors which interfere with parasite multiplication and sequestration are centrally important in ameliorating the severity of malaria. They suggest that RBC polymorphisms other than Duffy-negativity may reduce P. vivax infection of RBCs or impair cytoadherence interactions between P. vivax-infected RBCs and host cells. Like Duffy negativity, RBC polymorphisms that are found to protect against P. vivax may teach the important aspects of the pathogenesis of this fascinating and sometimes deadly parasite.

Malaria parasites grow within vertebrate erythrocytes and increase host cell permeability to access nutrients from plasma. This increase is mediated by the plasmodial surface anion channel (PSAC), an unusual ion channel linked to the conservedclaggene family. Although PSAC recognizes and transports a broad range of uncharged and charged solutes, it must efficiently exclude the small Na+ion to maintain infected cell osmotic stability. Here, we examine possible mechanisms for this remarkable solute selectivity. We identify guanidinium as an organic cation with high permeability into human erythrocytes infected withPlasmodium falciparum, but negligible uptake by uninfected cells. Transport characteristics and pharmacology indicate that this uptake is specifically mediated by PSAC. The rank order of organic and inorganic cation permeabilities suggests cation dehydration as the rate-limiting step in transport through the channel. The high guanidinium permeability of infected cells also allows rapid and stringent synchronization of parasite cultures, as required for molecular and cellular studies of this pathogen. These studies provide important insights into how nutrients and ions are transported via PSAC, an established target for antimalarial drug development.

Condenser microphones (MIC) have been widely used in mechanomyography, together with accelerometers and piezoelectric contact sensors. The aim of the present investigation was to clarify the mechanical variable (acceleration, velocity or displacement) indicated by the signal from a MIC transducer using a mechanical sinusoidal vibration system. In addition, the mechanomyogram (MMG) was recorded simultaneously with a MIC transducer and accelerometer (ACC) during voluntary contractions to confirm the mechanical variable reflected by the actual MMG and to examine the influence of motion artifact on the MMG. To measure the displacement-frequency response, mechanical sinusoidal vibrations of 3 to 300 Hz were applied to the MIC transducer with different sizes of air chambers (5, 10, 15 and 20 mm in diameter and 15, 20 or 25 mm long). The MIC transducer showed a linear relationship between the output amplitude and the vibration displacement, however, its frequency response declined with decreasing diameter and decreasing length of the air chamber. In fact, the cut-off frequency (-3dB) of the MIC transducer with the 5-mm-diameter chamber was 10, 8 and 4 Hz for the length 15, 20 and 25 mm, respectively. The air chamber with at least a diameter of 10 mm and a length of 15 mm is recommended for the MIC transducer. The sensitivity of this MIC transducer arrangement was 92 mV microm(-1) when excited at 100 Hz. During voluntary contraction, the amplitude spectral density function of the MMG from the MIC transducer resembled that of the double integral of the ACC transducer signal. The angle of the MIC transducer was delayed by 180 degrees in relation to the ACC transducer signal. The sensitivity of the MIC transducer was reduced to one-third because of the peculiar volume change of air chamber when the MMG was detected on the surface of the skin. In addition, the MIC transducer was contaminated by a smaller motion artifact than that from the ACC transducer. The maximal peak amplitude of the MIC and ACC transducer signal with the motion artifact was 7.7 and 12.3 times as much as the RMS amplitude of each signal without the motion artifact, respectively. These findings suggest that the MIC transducer acts as a displacement meter in the MMG. The MIC transducer seems to be a possible candidate for recording the MMG during dynamic muscle contractions as well as during sustained contractions.

The piezoelectric contact sensor has been widely utilised in mechanomyography (MMG). The authors aim to clarify the mechanical variables (i.e. acceleration, velocity or displacement) reflected by the MMG signal detected with a piezoelectric contact sensor (PEC), and compare the results with those obtained simultaneously by an accelerometer (ACC). To measure the acceleration-frequency response, a mechanical sinusoidal excitation of 5 to 300 Hz at a constant magnitude of 0.01 G was applied to the two transducers. The acceleration-frequency response of the ACC transducer was confirmed to be almost flat. The PEC without any restriction of the transducer housing (including the combined seismic mass) demonstrated a similar response to the ACC transducer. The PEC transducer output with restricted housing decreased with increasing sinusoidal frequency and an attenuation slope of -40 dB/decade and phase angle of -180 degrees. The voluntary MMG signal during isometric knee extension was recorded simultaneously with the two transducers. The amplitude spectral density distribution of the MMG from the PEC transducer was narrow and the mean frequency was approximately one-half that obtained from the ACC transducer. The amplitude spectral density distribution with the PEC transducer resembled that of the double integral over time of the ACC transducer signal. The phase angle of the PEC transducer signal was different from that of the ACC transducer signal by approximately -180 degrees. These results suggest that the PEC transducer acts as a displacement meter of muscle vibration. In addition, differences in the MMG frequency components relating to the transducer type must be taken into consideration when investigating the mechanical activity of muscle.

Background Inflammatory cytokinemia and systemic activation of the microvascular endothelium are central to the pathogenesis of Plasmodium falciparum malaria. Recently, ‘parasite-derived’ uric acid (UA) was shown to activate human immune cells in vitro, and plasma UA levels were associated with inflammatory cytokine levels and disease severity in Malian children with malaria. Since UA is associated with endothelial inflammation in non-malaria diseases, we hypothesized that elevated UA levels contribute to the endothelial pathology of P. falciparum malaria. Methodology/Principal Findings We measured levels of UA and soluble forms of intercellular adhesion molecule-1 (sICAM-1), vascular cell adhesion molecule-1 (sVCAM-1), E-selectin (sE-Selectin), thrombomodulin (sTM), tissue factor (sTF) and vascular endothelial growth factor (VEGF) in the plasma of Malian children aged 0.5–17 years with uncomplicated malaria (UM, n = 487) and non-cerebral severe malaria (NCSM, n = 68). In 69 of these children, we measured these same factors once when they experienced a malaria episode and twice when they were healthy (i.e., before and after the malaria transmission season). We found that levels of UA, sICAM-1, sVCAM-1, sE-Selectin and sTM increase during a malaria episode and return to basal levels at the end of the transmission season (p

Background Plasmodium falciparum elicits host inflammatory responses that cause the symptoms and severe manifestations of malaria. One proposed mechanism involves formation of immunostimulatory uric acid (UA) precipitates, which are released from sequestered schizonts into microvessels. Another involves hypoxanthine and xanthine, which accumulate in parasitized red blood cells (RBCs) and may be converted by plasma xanthine oxidase to UA at schizont rupture. These two forms of ‘parasite-derived’ UA stimulate immune cells to produce inflammatory cytokines in vitro. Methods and Findings We measured plasma levels of soluble UA and inflammatory cytokines and chemokines (IL-6, IL-10, sTNFRII, MCP-1, IL-8, TNFα, IP-10, IFNγ, GM-CSF, IL-1β) in 470 Malian children presenting with uncomplicated malaria (UM), non-cerebral severe malaria (NCSM) or cerebral malaria (CM). UA levels were elevated in children with NCSM (median 5.74 mg/dl, 1.21-fold increase, 95% CI 1.09–1.35, n = 23, p = 0.0007) and CM (median 5.69 mg/dl, 1.19-fold increase, 95% CI 0.97–1.41, n = 9, p = 0.0890) compared to those with UM (median 4.60 mg/dl, n = 438). In children with UM, parasite density and plasma creatinine levels correlated with UA levels. These UA levels correlated with the levels of seven cytokines [IL-6 (r = 0.259, p

We measured cross sections of target He atoms ([sigma][sub [ital T]]) and those of electron capture into the excited levels of projectile ions ([sigma][sub [ital P]]) for He 3 [sup 1,3][ital S],[ital P],[ital D] levels in He[sup +]-He symmetric collisions in the energy range 20--300 keV. By phenomenological analysis of the excitation functions, we found structures characteristic of the intermediate-velocity regime for both [sigma][sub [ital T]] and [sigma][sub [ital P]]: the magnitudes and the velocities of the peak positions of the structures show notable dependence on the angular-momentum states of the levels. Similar dependence on angular-momentum states is also recognized in the excitation functions for electron capture by protons into H(3,[ital l]) levels in H[sup +]-Li and H[sup +]-He collisions obtained by other investigators. This fact suggests close correlation between the behaviors of the excitation functions and the angular-momentum states of the initial and the final states, because all the processes mentioned above are concerned with electron transfer from the [ital s] level to upper [ital s], [ital p], and [ital d] levels. A simple momentum-matching electron-transfer model explains the observed characteristics qualitatively. Characteristics observed for the structures of [sigma][sub [ital P]] are also observed for the structures of [sigma][submore » [ital T]], although the structure of [sigma][sub [ital P]] appears in a velocity range about 1.4 times larger than that for the structure of [sigma][sub [ital T]]. Thus we expect that the same kind of mechanism that governs electron capture into excited states also causes target excitation accompanied by electron exchange.« less

Wolbachia and Cardinium are bacterial endosymbionts that are widely distributed amongst arthropods. Both cause reproductive alterations, such as cytoplasmic incompatibility, parthenogenesis and feminization. Here we studied differentially expressed genes in Wolbachia- and Cardinium-infected Bm-aff3 silkworm cells using a silkworm microarray. Wolbachia infection did not alter gene expression or induce or suppress immune responses. In contrast, Cardinium infection induced many immune-related genes, including antimicrobial peptides, pattern recognition receptors and a serine protease. Host immune responses differed, possibly because of the different cell wall structures of Wolbachia and Cardinium because the former lacks genes encoding lipopolysaccharide components and two racemases for peptidoglycan formation. A few possibly non-immune-related genes were differentially expressed, but their involvement in host reproductive alteration was unclear.

Relative emission cross sections were measured for all Ne(2p53p) levels (2p1-2p10 in the Paschen notation) excited by He+ ions with incident energy E=0.2-20 keV. The polarization degree was also determined for each emission line. For most of the levels, the energy dependence of both the emission cross sections and the polarization degrees show oscillatory structures in the energy range observed. These were interpreted as Rosenthal oscillations. From the phase correlation between the oscillations of these emission cross sections and those of He I 31P, we conclude that the He I 31P level interferes with 2p2-2p8. From the polarization degree we determine the partial emission cross sections of the magnetic sublevels for J=1 (J is the total angular momentum) levels (2p2, 2p5 and 2p7). For 2p2 and 2p5, the magnetic sublevels with MJ=+or-1 (MJ is the magnetic quantum number) were dominantly excited over the sublevel with MJ=0 and showed oscillations with larger amplitude than that for the Mj=0 level.

Hemodynamic responses and hormonal changes induced by lower body negative pressure (LBNP) were measured in bedridden patients with severe disabilities to identify their cardiovascular function. After a control period of 5 minutes supine, each subject underwent a LBNP test of ‐20 mmHg for 5 minutes. Disabled patients were classified into two groups: Group HI consisted of the members who had a blood pressure response to LBNP exposure similar to normal control subjects, and group H2 patients displayed a greater reduction in blood pressure than the normal response. Both disabled groups demonstrated a significantly higher level of resting heart rate as well as a smaller stroke volume and cardiac output than normal subjects. Exposure to LBNP in disabled patients produced a smaller decrement in stroke volume and cardiac output than in normal subjects. The expected increase in heart rate for the compensatory effect was not observed in the HI group, and a decreased heart rate lower than resting values was measured...

The purpose of this investigation was to identify the cardiovascular response to an orthostatic tolerance test in severely disabled patients during prolonged recumbency in a sitting position and under lower body negative pressure. The disabled subjects were categorized into three groups according to the degree of their exposure to gravitational stress in daily living and their past history of posture and motor ability. The members of one group (ST1) were now supported in a sitting position during present daily living and were able to sit or stand independently in the past. The subjects in the second group (ST2) were never able to sit or stand independently but were also supported in an orthostatic posture during daily living as were ST1 group members. The third group (BR) was bedridden fully from birth and had no experience, ever, of orthostatic forces during daily living. A marked decrease in photoplethysmogram of the big toe in the change from the resting supine to the sitting condition, which represent...

Treatment of chronic hepatitis C virus (HCV) infection with interferon (IFN) and ribavirin improves the rate of eradication of the virus by less than 20% in patients with genotype 1b and a high viral load. In this study we assessed whether IFN-beta induction/IFN-alpha2b plus ribavirin enhances the efficacy of the therapy in patients with chronic hepatitis C. The efficacy of IFN-beta induction/IFN-alpha2b plus ribavirin therapy (group A, n=7) was compared with that of IFN-alpha2b plus ribavirin (group B, n=7) in 14 patients with high levels of HCV-RNA (100 K/U/ml). No significant differences were observed in the clearance of HCV-RNA between the two groups (A and B, respectively) 2 weeks after the start of the treatment (0% and 14.3%), at the end of the treatment (71.4% and 100%) and 6 months after the end of the treatment (28.6% and 14.3%). Recovery was complete in 28.6% and 14.3%, transient in 42.9% and 85.7% and absent in 28.6% and 0% in groups A and B, respectively. Early log changes in the viral load from the baseline after 2 weeks of treatment were 2.41 +/- 0.91 and 2.77 +/- 0.20 in groups A and B, respectively, with no significant difference between the two groups. In the present study, we were not able to demonstrate that IFN-beta induction/IFN-alpha2b plus ribavirin therapy was superior to IFN-alpha2b plus ribavirin therapy in patients with genotype 1b and high viral loads.

New automatic through hole inspection techniques for printed wiring boards (PWBs) were developed. The new method called Leakage Light Detection can detect defects such as cracks and breaks in plating of through holes with aspect ratio as large as ten. A prototype system for automatic through hole inspection is constructed, based on these techniques. This system detects defects in a 250 mm × 250 mm area of a PWB in 8 minutes.

Electromyographic and mechanomyographic estimation of motor unit activation strategy in voluntary force production. In order to determine whether electromyogram (EMG) and mechanomyogram (MMG) are suitable for the noninvasive estimation of the motor unit (MU) activation strategy, the EMG/force and MMG/force relationships were examined simultaneously during isometric ramp contractions in biceps brachii muscle. The highest mean power frequency (MPF) of the EMG, which reflects the full MU recruitment, was determined at 51% MVC. Two obvious inflection points were identified on the MMG-amplitude/force relationship that showed an initial slow increase followed by a rapid increase and a progressive decrease at higher force levels. Our results suggest that the MMG amplitude allows the estimation of the beginning of recruitment of MUs that innervate the first-twitch fibers in addition to identification of the full MU recruitment. The rate coding strategy was qualitatively reflected by the MMG-MPF/force relationship. We conclude that the MU activation strategy is estimated in more detail by the MMG than by the EMG.

We studied microsatellite frequency and distribution in 21.76-Mb random genomic sequences, 0.67-Mb BAC sequences from the Z chromosome, and 6.3-Mb EST sequences of Bombyx mori. We mined microsatellites of ≥15 bases of mononucleotide repeats and ≥5 repeat units of other classes of repeats. We estimated that microsatellites account for 0.31% of the genome of B. mori. Microsatellite tracts of A, AT, and ATT were the most abundant whereas their number drastically decreased as the length of the repeat motif increased. In general, tri- and hexanucleotide repeats were overrepresented in the transcribed sequences except TAA, GTA, and TGA, which were in excess in genomic sequences. The Z chromosome sequences contained shorter repeat types than the rest of the chromosomes in addition to a higher abundance of AT-rich repeats. Our results showed that base composition of the flanking sequence has an influence on the origin and evolution of microsatellites. Transitions/transversions were high in microsatellites of ESTs, whereas the genomic sequence had an equal number of substitutions and indels. The average heterozygosity value for 23 polymorphic microsatellite loci surveyed in 13 diverse silkmoth strains having 2–14 alleles was 0.54. Only 36 (18.2%) of 198 microsatellite loci were polymorphic between the two divergent silkworm populations and 10 (5%) loci revealed null alleles. The microsatellite map generated using these polymorphic markers resulted in 8 linkage groups. B. mori microsatellite loci were the most conserved in its immediate ancestor, B. mandarina, followed by the wild saturniid silkmoth, Antheraea assama.

The sex chromosomes of the silkworm, Bombyxmori, are designated ZW(XY) for females and ZZ(XX) for males. The W chromosome of B. mori does not recombine with the Z chromosome and autosomes and no genes for morphological characters have been mapped to the W chromosome as yet. Furthermore, femaleness is determined by the presence of a single W chromosome, regardless of the number of autosomes or Z chromosomes. To understand these interesting features of the W chromosome, it is necessary to analyze the W chromosome at the molecular biology level. Initially to isolate DNA sequences specific for the W chromosome as randomly amplified polymorphic DNA (RAPD) markers, we compared the genomic DNAs between males and females by PCR with arbitrary 10-mer primers. To the present, we have identified 12 W-specific RAPD markers, and with the exception of one RAPD marker, all of the deduced amino acid sequences of these W-specific RAPD markers show similarity to previously reported amino acid sequences of retrotransposable elements from various organisms. After constructing a genomic DNA lambda phage library of B. mori we obtained two lambda phage clones, one containing the W-Kabuki RAPD sequence and one containing the W-Samurai RAPD sequence and found that these DNA sequences comprised nested structures of many retrotransposable elements. To further analyze the W chromosome, we obtained 14 W-specific bacterial artificial chromosome (BAC) clones from three BAC libraries and subjected these clones to shotgun sequencing. The resulting assembly of sequences did not produce a single contiguous sequence due to the presence of many retrotransposable elements. Therefore, we coupled PCR with shotgun sequencing. Through these analyses, we found that many long terminal repeat (LTR) and non-LTR retrotransposons, retroposons, DNA transposons and their derivatives, have accumulated on the W chromosome as strata. These results strongly indicate that retrotransposable elements are the main structural component of the W chromosome.

The accelerometer is used for mechanomyogram (MMG) recordings of muscle contractions. Although the mechanical characteristics of other MMG transducers have been determined with reference to the accelerometer, mechanical aspects of the accelerometer itself, including the weight of the transducer, have not been verified. This study was designed to reinvestigate the mechanical variable of the MMG signal detected with an accelerometer, with reference to a laser distance sensor (LDS), and then to clarify the influence of the accelerometer weight on the MMG recording during muscle contractions. The study was performed during mechanical sinusoidal vibrations and during voluntary contractions of the quadriceps muscle. Maximum differences in the amplitude spectral density functions between the LDS signal and the double integral of the accelerometer signal were approximately 4 microm. The results verified that the MMG signal from the accelerometer accurately reflected the acceleration of body surface vibration. However, the MMG signal was gradually distorted when weight was added to the accelerometer: the addition of 4.0 g (total 6.0 g, including 2.0 g of accelerometer) substantially attenuated the MMG signal. The results suggest that the appropriate weight for the accelerometer should be less than 5.0 g for measurements of the quadriceps muscle and indicate that the transducer weight must be taken into account for accurate measurement of muscles of different sizes.

This paper presents a 4 kW 400 kHz voltage source inverter for induction heating applications, which is characterized by power control based on PDM (pulse density modulation). The pulse density modulated inverter makes an induction heating system simple and compact, thus leading to higher efficiency. The modulation strategy is proposed to realize an induction heating system capable of operation at the frequency and power level of interest. Some experimental results are shown to verify the validity of the concept. >

We examined the heart rate (HR) of subjects with spastic cerebral palsy (CP) in order to estimate exercise intensity while walking. The subjects were 17 subjects with CP (14.0 +/- 3.7 years of age) containing 7 subjects rated as level 1, 4 subjects rated as level 2, and 6 subjects rated as level 3 by the Gross Motor Function Classification System, and 7 normal subjects (12.4 +/- 2.8 years of age) were used as a controls. Even in subjects whose gross motor function was excellent (rated as level 1), the HR significantly increased while walking when compared to normal subjects (p0.05), although the walking speed between the groups was not different. According to the HR, the exercise intensity while walking was adapted from weakly to moderately and thought to be appropriate for exercise. On the other hand, walking speed was significantly reduced in the subjects rated as level 2 and 3 (p0.05), and the HR increased significantly (p0.05). Seven of the ten subjects rated as either level 2 or 3 showed a high HR of over 150 beats/min while walking. The HR while walking of the two subjects rated as level 3 continued to increase although the walking speed was kept constant. The walking exercise would be too strong and become detrimental to such subjects.

Reactive oxygen species play a critical role in the onset of apoptosis induced by various extracellular stimuli, including ionizing radiation. Therefore active regulation of reactive oxygen species-metabolizing enzymes may be one response to an apoptotic stimulus. In this regard, HP100 cells, H(2)O(2)-resistant variants derived from human leukemia HL60 cells, display an interesting phenotype in which the activity of catalase is constitutively high, whereas its mRNA is reduced after X-ray irradiation. In the present study, we investigated the molecular mechanisms underlying this phenomenon. By combining analyses from nuclear run-on, reporter gene transient transfection, genomic footprinting, site-directed mutagenesis, electrophoretic mobility shift analysis, and Western blotting experiments, we found that constitutively elevated catalase expression is strongly regulated at the transcriptional level by both Sp1 and CCAAT-recognizing factors and that much higher levels of nuclear Sp1 and NF-Y are present in HP100 nuclei as compared with HL60 nuclei. In addition, we demonstrated an X-ray-inducible association of a WT1/Egr-related factor with an overlapping Sp1/Egr-1 recognition sequence located within the core promoter of the catalase gene. This association may lead to inactivation of the promoter by disturbing or competing with the transactivating ability of Sp1.

Fully grown immature oocytes acquire the ability to be fertilized with sperm after meiotic maturation, which is finally accomplished by the formation and activation of the maturation-promoting factor (MPF). MPF is the complex of Cdc2 and cyclin B, and its function in promoting metaphase is common among species. The Mos/mitogen-activated protein kinase (MAPK) pathway is also commonly activated during vertebrate oocyte maturation, but its function seems to be different among species. We investigated the function of the Mos/MAPK pathway during oocyte maturation of the frog Rana japonica. Although MAPK was activated in accordance with MPF activation during oocyte maturation, MPF activation and germinal vesicle breakdown (GVBD) was not initiated when the Mos/MAPK pathway was activated in immature oocytes by the injection of c-mos mRNA. Inhibition of Mos synthesis by c-mos antisense RNA and inactivation of MAPK by CL100 phosphatase did not prevent progesterone-induced MPF activation and GVBD. However, continuous MAPK activation and MAPK inhibition through oocyte maturation accelerated and delayed MPF activation, respectively. Furthermore, Mos induced a low level of cyclin B protein synthesis in immature oocytes without the aid of MAPK. These results suggest that the general function of the Mos/MAPK pathway, which is not essential for MPF activation and GVBD in Rana oocytes, is to enhance cyclin B translation by Mos itself and to stabilize cyclin B protein by MAPK during oocyte maturation.

The aim of the present study was to elucidate the muscle contractile properties of preadolescent boys using the mechanomyogram (MMG) and electromyogram (EMG). In 8 preadolescent boys and 10 male young adults, the EMG and MMG were recorded from quadriceps muscles during isometric knee extension contractions at levels of 10 to 80% MVC. The relationship of EMG and MMG to absolute force was approximated by a regression line (r0.8). The regression line of the EMG in preadolescent was located above that of the adults, whereas the MMG in both subject groups was fitted by virtually the same regression line. Thus, the MMG seems to be indicative of absolute force in preadolescents as well as young adults. Both groups showed a non-linear increase in the EMG and MMG with relative force (%MVC). The EMG and %MVC relationship was not significantly different between the two groups, whereas the MMG in preadolescents was smaller than that of adults over the range of force studied. The MMG increment with %MVC was more remarkable above 30 or 40% MVC in both groups; however, the rate of the increment in preadolescents was smaller than that in adults. The MMG and %MVC relationship suggests that the muscle contractile properties in preadolescent boys are characterized by immaturity of the fast twitch fibers.

Protamine is an arginine-rich basic protein found in the sperm nuclei of many vertebrates, but its actual roles in spermatozoa remain to be elucidated. In this study, we investigated the physiological roles of protamine by examining protamine-less spermatozoa produced in vitro in the presence of the transcriptional inhibitor actinomycin D. Even under inhibited transcription, medaka spermatocytes underwent meiosis and differentiated into spermatozoa with a condensed nucleus and an elongated flagellum. Using a newly produced anti-medaka protamine antibody, we confirmed the absence of protamine protein in the spermatozoa differentiated in the presence of actinomycin D. These findings clearly indicate that sperm nuclear condensation in medaka is independent of protamine. Since medaka spermatozoa are shed into water upon natural fertilization, we also investigated the roles of protamine by comparing the differences between the nuclear morphology of protamine-equipped and protamine-less spermatozoa immersed in water. The nuclei without protamine more rapidly swelled than did those with protamine and completely broke down within 10 min, whereas more than 80% of the sperm nuclei with protamine resisted the disruption under similar conditions. These findings strongly suggest that a physiological role of protamine in medaka spermatozoa is to protect the ejaculated spermatozoa against the disruption by low osmotic pressure until arrival at the eggs for successful fertilization.

Stimulated by maturation-inducing hormone secreted from follicle cells surrounding the oocytes, fully-grown oocytes mature and become fertilisable. During maturation, immature oocytes resume meiosis arrested at the first prophase and proceed to the first or second metaphase at which they are naturally inseminated. Paying special attention to general and species-specific aspects, we summarise the mechanisms regulating the initial phase of oocyte maturation, from the reception of hormonal signals on the oocyte surface to activation of the maturation-promoting factor in the cytoplasm, in amphibians, fishes, mammals and marine invertebrates.