Oocyte in vitro maturation with eugenol improves the medium antioxidant capacity and total cell number per blastocyst.

This study investigates the impact of eugenol (EU) supplementation on bovine oocyte in vitro maturation (IVM) and antioxidant capacity, as well as in vitro embryo production and quality after conventional in vitro fertilization (IVF). A total of 1077 cumulus oocyte complexes were cultured in TCM-199+ without EU supplementation (control treatment) or supplemented with EU at the concentrations of 10 µM (EU-10), 20 µM (EU-20), or 40 µM (EU-40). After IVM, the oocytes were subjected to IVF and embryo culture. The addition of EU at 40 µM to the IVM medium improved (P < 0.05) the antioxidant capacity and cleavage rate when compared to the control treatment. Moreover, a positive correlation (r = 0.61, P < 0.03) was observed between cleavage rate and EU concentration. The addition of EU at concentrations of 10 and 20 µM decreased (P < 0.05) the calreticulin (CALR) levels in expanded blastocysts when compared to the control treatment and EU-40 treatment. However, the EU-10 and EU-20 treatments had a greater (P < 0.05) mean total cell number (TCN) per expanded blastocyst when compared to the control treatment and EU-40 treatment. In conclusion, the addition of EU to the enriched culture medium during IVM of bovine oocytes improved the antioxidant capacity of the spent medium, as well as the cleavage rate and embryonic quality (i.e., TCN/expanded blastocyst), and reduced the endoplasmic reticulum stress (i.e., CALR levels) in the embryos. Thus, we recommend enriching the IVM medium with 10 µM EU for in vitro bovine embryo production.

Energy consumption in a baffled membrane bioreactor (B-MBR): estimation based on long-term continuous operation.

We investigated the operating conditions of a baffled membrane bioreactor (B-MBR) under which long-term stable operation can be achieved through the continuous operation of a pilot-scale B-MBR. Under appropriate operating conditions, the B-MBR was capable of achieving excellent treated water quality in terms of biochemical oxygen demand and concentration of total nitrogen. Excellent removal of total phosphorus was also achieved. In addition, the degree of membrane fouling was acceptable, indicating that stable continuous operation of a B-MBR is possible under the operating conditions adopted in the present study. Estimation of the specific energy consumption in hypothetical full-scale B-MBRs operated under the conditions recommended by the findings was also performed in this study. The results suggest that energy consumption in full-scale B-MBRs would be in the range of 0.20-0.22 kWh/m3. These results strongly suggest that energy consumption in MBR operation can be significantly reduced by applying the concept of a B-MBR.

Effects of recirculation and separation times on nitrogen removal in baffled membrane bioreactor (B-MBR).

In this study, we investigated the effects of recirculation and separation times on removals of organic matter, nitrogen, and phosphorus in a baffled membrane bioreactor (B-MBR) treating real municipal wastewater. A pilot-scale B-MBR experimental apparatus was operated under two different sets of recirculation and separation times. The results revealed that, irrespective of operating conditions, the biochemical oxygen demand (BOD) and concentration of total nitrogen (T-N) in the treated water can be lowered to less than 3 and 5 mg/L, respectively. Although T-N was effectively removed in the two different operating conditions, increase in the fraction of recirculation time results in tiny deterioration of nitrogen removal efficiency in the B-MBR. Phosphorus removal efficiency was also slightly decreased as the fraction of recirculation time (ratio between recirculation and separation times) was increased. The results of the measurement of dissolved oxygen (DO) profiles at different points of the B-MBR apparatus indicate that the increase in DO concentration in the anoxic zone of the B-MBR becomes much more pronounced by increasing recirculation intensity. On the basis of the results obtained in this study, it can be concluded that efficient removal of BOD, T-N, and total phosphorus can be achieved by the B-MBR as long as appropriate recirculation intensity is selected.

Abnormal asymmetries in subcortical brain volume in schizophrenia.

Subcortical structures, which include the basal ganglia and parts of the limbic system, have key roles in learning, motor control and emotion, but also contribute to higher-order executive functions. Prior studies have reported volumetric alterations in subcortical regions in schizophrenia. Reported results have sometimes been heterogeneous, and few large-scale investigations have been conducted. Moreover, few large-scale studies have assessed asymmetries of subcortical volumes in schizophrenia. Here, as a work completely independent of a study performed by the ENIGMA consortium, we conducted a large-scale multisite study of subcortical volumetric differences between patients with schizophrenia and controls. We also explored the laterality of subcortical regions to identify characteristic similarities and differences between them. T1-weighted images from 1680 healthy individuals and 884 patients with schizophrenia, obtained with 15 imaging protocols at 11 sites, were processed with FreeSurfer. Group differences were calculated for each protocol and meta-analyzed. Compared with controls, patients with schizophrenia demonstrated smaller bilateral hippocampus, amygdala, thalamus and accumbens volumes as well as intracranial volume, but larger bilateral caudate, putamen, pallidum and lateral ventricle volumes. We replicated the rank order of effect sizes for subcortical volumetric changes in schizophrenia reported by the ENIGMA consortium. Further, we revealed leftward asymmetry for thalamus, lateral ventricle, caudate and putamen volumes, and rightward asymmetry for amygdala and hippocampal volumes in both controls and patients with schizophrenia. Also, we demonstrated a schizophrenia-specific leftward asymmetry for pallidum volume. These findings suggest the possibility of aberrant laterality in neural pathways and connectivity patterns related to the pallidum in schizophrenia.

Season's appreciations.

To follow the tradition set by the late Franz Halberg, highlights of research performed over the last year from his Minnesota Center are summarized. They illustrate the broad international cooperation enjoyed by his center and the diversity of applications of the discipline he founded. The results briefly summarized herein in the form of an annotated bibliography are a testimony that his legacy continues to live on and constitutes a tribute to his memory.

Toll-like receptor 4-dependent adjuvant activity of Kakkon-to extract exists in the high molecular weight polysaccharide fraction.

Kakkon-to, a traditional herbal medicine (Kampo formula), has been used historically in China and Japan for the treatment of infectious diseases such as influenza and the common cold. However, the biological mechanism of its therapeutic action has not yet been elucidated. In this study, we investigated the immunological function of Kakkon-to and found that the high molecular weight fraction of the extract activated macrophages in vitro. This fraction was found to be composed primarily of saccharides and in vitro intensively stimulated mouse peritoneal macrophages that produce Th1 inflammatory cytokines such as tumor necrosis factor α (TNFalpha), interleukin-1beta (IL-1beta), interferon-gamma (IFN-gamma), and interleukin-6 (IL-6). The fraction did not activate macrophages from C3H/HeJ lacking Toll-like receptor 4 (TLR4) or MyD88-deficient mice, indicating that macrophage activation by the fraction was mediated by TLR4. The route of administration of the fraction into mice regulated the kinetics of TNFalpha production in immune organs. Intravenous administration induced TNFalpha production in the four target organs of spleen, liver, lung, and Peyer’s patch; however, the most abundant production occurred in the liver and peaked at 30-60 min post administration. Peritoneal administration induced similar kinetics but the most abundant production occurred in the spleen. In contrast, oral administration induced TNFalpha production in the liver, lung, and Peyer’s patch, but not in the spleen. Although liver and lung are TNFalpha-abundant organs, production peaks in these organs occurred later than in Peyer’s patch. We also found that the fraction induced antibody production as an adjuvant against a specific antigen ovalbumin (OVA) when administered simultaneously and subcutaneously in a dose-dependent manner. Interestingly, the fraction induced IgG-class antibody in response to low doses of the antigen, which induced only IgM-class antibody when administered alone, suggesting that the fraction induces a class switch of immunoglobulin as an adjuvant in vivo. The high molecular weight fraction of Kakkon-to extract could be applicable as a potent immunostimulating drug and adjuvant.

In vitro evaluation of cashew nut shell liquid as a methane-inhibiting and propionate-enhancing agent for ruminants.

Cashew nut shell liquid (CNSL) containing antibacterial phenolic compounds was evaluated for its potency as a feed additive for ruminants. In experiment 1, ruminal responses to CNSL supplementation were assessed using a batch culture system. Rumen fluid from cattle was diluted with artificial saliva and incubated for 18h in a batch culture with a mixed diet containing a 30:70 hay:concentrate diet to which raw or heated CNSL was added at a final concentration of 500 µg/mL. In experiment 2, a Rusitec, using rumen fluid from the same cattle, was operated over a period of 7 d during which only raw CNSL was tested at concentrations of 0, 50, 100, or 200 µg/mL, and variations in fermentation and bacterial population were assessed. In experiment 3, a pure culture study was conducted using selected bacteria to determine their susceptibility to CNSL. In experiment 1, methane production was inhibited by raw CNSL (56.9% inhibition) but not by heated CNSL. Total volatile fatty acid concentration was not affected, whereas increased concentrations of propionate and decreased concentrations of acetate and butyrate were observed using either raw or heated CNSL. These changes were more obvious when raw CNSL was tested. In experiment 2, raw CNSL inhibited methanogenesis and increased propionate production in a dose-dependent manner, showing maximum methane inhibition (70.1%) and propionate enhancement (44.4%) at 200 µg/mL supplementation. Raw CNSL increased total volatile fatty acid concentration and dry matter digestibility. Raw CNSL also appeared to induce a dramatic shift in the population of rumen microbiota, based on decreased protozoa numbers and changes in quantitative PCR assay values for representative bacterial species. In experiment 3, using pure cultures, raw CNSL prevented the growth of hydrogen-, formate-, and butyrate-producing rumen bacteria, but not the growth of bacteria involved in propionate production. Based on these data, raw CNSL, rich in the antibacterial phenolic compound anacardic acid, is a potential candidate feed additive with selective activity against rumen microbes, leading to fermentation that results in decreased methane and enhanced propionate production.

Administration of dietary fish oil capsules in healthy middle-aged Japanese men with a high level of fish consumption.

The nutritional effects of fish oil, which is rich in the n-3 polyunsaturated fatty acids, have been reported. In this randomized, placebo-controlled, double-blind, crossover study, we evaluated the effects of dietary fish oil capsules on the hematological parameters of healthy middle-aged Japanese men with a high level of fish oil consumption. Over a 4-week period, subjects were administered five fish oil or olive oil (placebo) capsules with every meal (1,260 mg eicosapentaenoic acid and 540 mg docosahexaenoic acid/day). There was a 4-week washout period between the treatment phases. The results did not demonstrate a decrease in plasma triacylglycerol, cholesterol, low-density lipoprotein cholesterol, and whole-blood viscosity. Further, no changes in the fatty acid composition of plasma and erythrocyte phospholipids were noted. These results suggested that the supplementation of fish oil might be effective only for those subjects who have a lower level of fish oil consumption.

Variation of major neutral oligosaccharides levels in human colostrum.

OBJECTIVES: Human colostrum is known to be important for the protection of infants against infection by pathogenic microorganisms. This protection is thought to be due, partially, to various neutral and acidic oligosaccharides that are present in colostrum and milk. However, the concentrations of each of the oligosaccharide of human colostrum have not yet been determined. The aim of this present study was to determine the concentration of each of the major neutral oligosaccharide for three consecutive days from the start of lactation. METHOD: We analyzed the level of each neutral oligosaccharide in human colostrum, for three consecutive days from the start of lactation, obtained from 12 healthy Japanese women (ranging in age from 21 to 35 years; primipara 6 and multipara 6). The ABO blood groups of the donors were determined: A, three; B, three; O, five; AB, one. The determined human milk oligosaccharides were 2'-fucosyllactose (2'-FL), 3-fucosyllactose (3-FL), lactodifucotetraose (LDFT), lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), three lacto-N-fucopentaose (LNFP I, II and III) and two lacto-N-difucohexaose (LNFDH I and II) using high-performance liquid chromatography (HPLC) with two derivatization techniques. RESULTS: The concentrations of 2'-FL and LDFT in colostrum on day 1 were significantly higher than those on days 2 and 3 (P<0.05). An increase in LNT was observed on day 3 compared with day 1 (P<0.05). CONCLUSION: These changes in concentrations of 2'-FL, LDFT and LNT may reflect the requirements for prebiotics and anti-infection agents by human infants during early lactation.

Evaluation of the basic properties of the BANGkit gel dosimeter.

We evaluated the basic properties of a commercially available BANGkit gel dosimeter, which is a normoxic type of BANG gel. This gel-kit has the same composition as the BANG 3 gel, but is fully oxygenated. To exclude oxygen, oxygen scavenging ascorbic acid and copper sulfate as a catalyst are used. The properties that we examined are the effects of the concentrations of copper sulfate and ascorbic acid on the response, the reproducibility, the long-term stability, the temperature effect at irradiation and the dose-rate effect. In our results, the excellent linear fit of the R2-dose response in a dose range for clinical use and its reproducibility were observed. The precision of a linear fit was preserved for about 3 weeks. The temperature at irradiation showed a significant effect on the dose response. Although the dose-rate dependence in the high-dose range was observed, it was negligible for the clinical dose range up to 270 cGy. In conclusion, this gel dosimeter is thought to be utilizable in clinical practice, while we have to pay attention to the temperature during the entire measurement processes, and additionally there is room for improvement in the linearity and the dose-rate dependence in the high-dose range.

Mental fatigue-induced decrease in levels of several plasma amino acids.

To investigate the relation between plasma amino acid levels and mental fatigue, we measured the plasma concentrations of 20 amino acids in 9 healthy volunteers before and after a fatigue-inducing mental task session for 8 hr. As fatigue-inducing mental tasks, the subjects performed an advanced trail making test, a Japanese KANA pick up test, and a mirror drawing test. As a control, 8-hr relaxation session was performed in the same subjects at an interval of 4 weeks. Immediately after the fatigue session, the plasma levels of branched-chain amino acids, tyrosine, cysteine, methionine, lysine, and arginine were below those after a relaxation session. The values for other blood parameters including total protein, albumin, glucose, and total cholesterol did not show any differences between the 2 sessions. These results indicate that mental fatigue may be characterized by a decrease in the plasma level of these amino acids.

Hybrid membrane bioreactor for water recycling and phosphorus recovery.

This paper deals with the performance of hybrid membrane bioreactor (MBR) combining the precoagulation/sedimentation and membrane bioreactor. The hybrid MBR not only produces the treated water with excellent permeate quality but also shows much lower membrane fouling than the conventional MBR. It may come from its extremely low F/M ratio to maintain the low viscosity even in the high MLSS concentration range of about 20,000 mg/L. Some results of microbial community analysis in MBRs was conducted to demonstrate the other reason for its lower membrane fouling. Hybrid MBR has a high potential to be used for the recycling use of the municipal wastewater. Coagulated sludge produced in the hybrid MBR is a promising phosphorus resource. This paper also contains a recent progress of phosphorus recovery technology, which uses a new phosphoric acids absorbent, i.e. the hexagonal mesostructured zirconium sulfate (ZS). The ZS has the extremely high adsorption capacity of phosphoric acids through anion exchange. The adsorbed phosphoric acids are released from the ZS in a high pH range of about 13.

Development of an innovative vertical submerged membrane bioreactor (VSMBR) for simultaneous removal of organic matter and nutrients.

A novel vertical submerged membrane bioreactor (VSMBR) composed of anoxic and oxic zones in one reactor was developed in an attempt to reduce the problems concerning effective removal of pollutants from synthetic wastewater including glucose as a sole carbon source as well as membrane fouling. The optimal volume ratio of anoxic zone/oxic zone was found as 0.6. The desirable internal recycle rate and hydraulic retention time (HRT) for effective nutrient removal were 400% and 8h, respectively. Under these conditions, the average removal efficiencies of total nitrogen (T-N) and total phosphorus (T-P) were 75% and 71%, respectively, at the total chemical oxygen demand (T-COD)/T-N ratio of 10. In addition, the VSMBR showed high specific removal rates of nitrogen and phosphorus while the biomass growth yield from the reactor was about 20% of the conventional activated sludge process.

Effect of pre-coagulation on mitigating irreversible fouling during ultrafiltration of a surface water.

Membrane fouling can be divided into two types: reversible fouling and irreversible fouling. The former can be easily canceled by physical cleaning (e.g., backwashing) while the latter needs chemical cleaning to be mitigated. For more efficient use of membranes, the control of irreversible membrane fouling is of importance. In this study, the effectiveness of pre-coagulation/sedimentation on irreversible membrane fouling was investigated, based on the pilot-scale operation of the membrane unit installed at an existing water purification plant. The membrane employed was a low-pressure ultrafiltration (UF) membrane made of polysulfone and having a molecular weight cut-off of 750,000 daltons. Although pre-coagulation/ sedimentation significantly mitigated membrane fouling mainly through the reduction of reversible membrane fouling, the degree of irreversible fouling was not reduced by the pre-treatment. This was because the irreversible fouling observed during this study was mainly attributed to polysaccharides/protein like fractions of organic substances that cannot be efficiently removed by coagulation/sedimentation. Aluminium used as coagulant was thought to cause irreversible fouling to some extent but did not in the pilot operation, which could probable be explained by the fact that coagulation was conducted at relatively high pH (7.0) in this study.

Baffled membrane bioreactor (BMBR) for advanced wastewater treatment: easy modification of existing MBRs for efficient nutrient removal.

In this study, a novel membrane bioreactor (MBR) in which nitrification and denitrification simultaneously proceed in a single reaction chamber is proposed for advanced municipal wastewater treatment. Anoxic/aerobic environments are alternatively created in the proposed MBR by inserting baffles inside the membrane chamber. The performance of the proposed baffled membrane bioreactor (BMBR) was examined at an existing municipal wastewater treatment facility based on long-term operation. Although the procedure was simple, insertion of the baffles actually created the alternative anoxic/aerobic environments in the chamber at a constant interval and showed a great improvement in the nutrient removal. The insertion did not cause any adverse effect on membrane permeability. In this study, almost complete elimination of NH4+-N was observed while around 8 mg/L of NO(3-)-N was detected in the treated water. The modification proposed in this study can immediately be applied to most existing MBRs and is highly recommended for more efficient wastewater treatment.

Levofloxacin based triple therapy as a second-line treatment after failure of helicobacter pylori eradication with standard triple therapy.

BACKGROUND: Successful eradication of Helicobacter pylori infection after failure of standard triple therapy is difficult. The efficacy and safety of levofloxacin based triple therapy as a first-line therapy has-been studied. AIMS: The aim was to evaluate the efficacy and tolerability of levofloxacin based therapy after a failed standard triple therapy. PATIENTS: We conducted a prospective, uncontrolled study of a consecutive series of 33 patients who failed eradication with 1 week of lansoprazole-amoxicillin-clarithromycin triple therapy. METHODS: The subjects were retreated with 1 week of LA-LVFX triple therapy (lansoprazole, 30 mg twice daily; amoxicillin, 1000 mg twice daily: levofloxacin, 200 mg twice daily). Cure of infection was defined as negative results from culture, histology and a urea breath test 4 to 8 weeks after the second-line therapy. RESULTS: The eradication rate was 69.7% (23/33) by both intention-to-treat and per-protocol analyses (95% confidence interval=61-79%). Seven (21.2%) patients experienced mild side-effects, such as soft stools and taste disturbance. No patient stopped the medication on account of adverse effects. CONCLUSIONS: Levofloxacin based triple therapy is an effective second-line treatment after a failed standard triple therapy.

Differential activation in the medial temporal lobe during a sound-sequence discrimination task across age in human subjects.

To elucidate the brain mechanisms to encode sequential events, event-related potentials (ERPs) were recorded during a sound-sequence discrimination task using young and middle-aged adult subjects. In the task, a series of six or 12 kinds of natural sounds were sequentially presented; 70-80% of the stimuli were presented in a fixed order (Non-target), but the remaining stimuli, in a random order (Target). The subjects were instructed to detect the Targets and press a button at the end of each Target. In a control task, the same sounds were randomly presented (Control), and they were instructed to press the button at the end of each sound. Behavioral results indicated that the young subjects learned the task faster than did the middle-aged subjects. Positive ERP waves were evoked by Targets and Non-targets in the parieto-occipital area around 300-700 ms after stimulus onset. The mean amplitudes during this period in the young subjects were larger in Target than Control conditions, and those in Target condition were larger in the young than middle-aged subjects. Furthermore, the mean amplitudes in the Target condition were significantly correlated with behavioral performance. Equivalent dipoles for the ERPs evoked by Targets were estimated in the medial temporal lobe including the hippocampal formation and parahippocampal gyrus. The results suggest that the ERPs around 300-700 ms latency are involved in sound-sequence information processing. Furthermore, decrease in amplitudes of this positivity in the middle-aged subjects suggests that age-related memory decline is associated with deficits in encoding and retrieval of unfamiliar sequence.

Concentrations of sialyloligosaccharides in bovine colostrum and milk during the prepartum and early lactation.

Sialyloligosaccharides and sialylglycoconjugates in colostrum and milk are regarded to be important biological components with respect to be source of brain gangliosides in infant and to be antiinfectional components for the attack by the pathogenic bacteria and virus. Several acidic oligosaccharides have been characterised in both bovine and human milk or colostrum. The sialyloligosaccharide content of human colostrum and milk has been extensively studied, whereas that of cows milk and colostrum has received less attention. In this study, the concentrations of three sialyloligosaccharides of bovine colostrum and milk were determined at various stages during the prepartum and the first 7 d postpartum. The concentration of 3'SL (Neu5Ac(alpha2-3)Gal(beta1-4)Glc) reached a maximum value of 0.85 mg/ml immediately following parturition while the concentrations of 6'SL (Neu5Ac(alpha2-6)Gal(beta1-4)Glc) and 6'SLN (Neu5Ac(alpha2-6)Gal(beta1-4)GlcNAc) of 0.14 and 0.12 mg/ml, respectively, were much lower at this initial stage, although these concentration were maximum immediately following parturition. Bovine colostrum, especially that collected immediately after parturition, may be suitable as a source of 3'SL and other sialyloligosaccharides for use as additives by the food or pharmaceutical industries.

Pharmacokinetics of a novel anti-asthmatic, scoparone, in the rabbit serum assessed by a simple HPLC method.

The pharmacokinetics of scoparone (Sco), an anti-asthmatic coumarin derivative extracted from the traditional Chinese herb "Yin Chen", was determined in rabbit plasma by a high performance liquid chromatography (HPLC) method which was newly developed using a Nova-Pak Silica C(18) column and a mobile phase of acetonitrile-water (20:80, v/v). After intravenous administration of Sco at a dose of 2.0 or 3.6 mg/kg, the time course of the plasma concentration of Sco was found to fit an open two-compartment model. AUC values increased, dose-dependently. Mean T(1/2) values showed that half-life was dosage independent. Distribution and elimination of Sco in rabbit plasma were both relatively rapid. These results indicate that Sco could be easily administrated by spray inhalers and is expected to have few associated side effects.

Abnormal distributions of callosal commissural and corticothalamic neurons in the cerebral neocortex of Shaking Rat Kawasaki.

Shaking Rat Kawasaki (SRK) is an autosomal recessive mutant rat recognized by unstable gait and tremor and by early death around the time of weaning. We previously reported that corticospinal tract neurons are malpositioned in the motor cortex of the SRK rat [Ikeda and Terashima (1997) J. Comp. Neurol. 383, 370-380]. In the present study, we examined the distribution pattern of callosal commissural (CC) and corticothalamic (CT) neurons of SRK and normal rats with the injection of horseradish peroxidase (HRP) into the contralateral hemisphere or wheat germ agglutinin-conjugated HRP into the ventral lateral thalamic nucleus. The intracortical distribution pattern of retrogradely labeled CC and CT neurons in the motor cortex of SRK rat was abnormal: CC neurons were more deeply situated and CT neurons were more superficially situated in the SRK cortex than the corresponding components in the normal cortex. Most of labeled CC and CT neurons had abnormal dendritic configurations. Statistical analysis revealed that the difference of the mean intracortical position of CC and CT neurons of the SRK was significantly different from the normal counterparts (Student's t-test, P<0.01). Taken together with previous findings, our data demonstrate that the abnormal cytoarchitecture of SRK cortex resembles the reeler cortex.