Development of Biotin-Prototrophic and -Hyperauxotrophic Corynebacterium glutamicum Strains

To develop the infrastructure for biotin production through naturally biotin-auxotrophic Corynebacterium glutamicum, we attempted to engineer the organism into a biotin prototroph and a biotin hyperauxotroph. To confer biotin prototrophy on the organism, the cotranscribed bioBF genes of Escherichia coli were introduced into the C. glutamicum genome, which originally lacked the bioF gene. The resulting strain still required biotin for growth, but it could be replaced by exogenous pimelic acid, a source of the biotin precursor pimelate thioester linked to either coenzyme A (CoA) or acyl carrier protein (ACP). To bridge the gap between the pimelate thioester and its dedicated precursor acyl-CoA (or -ACP), the bioI gene of Bacillus subtilis, which encoded a P450 protein that cleaves a carbon-carbon bond of an acyl-ACP to generate pimeloyl-ACP, was further expressed in the engineered strain by using a plasmid system. This resulted in a biotin prototroph that is capable of the de novo synthesis of biotin. On the other hand, the bioY gene responsible for biotin uptake was disrupted in wild-type C. glutamicum. Whereas the wildtype strain required approximately 1 mu g of biotin per liter for normal growth, the bioY disruptant (Delta bioY) required approximately 1 mg of biotin per liter, almost 3 orders of magnitude higher than the wild-type level. The Delta bioY strain showed a similar high requirement for the precursor dethiobiotin, a substrate for bioB-encoded biotin synthase. To eliminate the dependency on dethiobiotin, the bioB gene was further disrupted in both the wild-type strain and the Delta bioY strain. By selectively using the resulting two strains (Delta bioB and Delta bioBY) as indicator strains, we developed a practical biotin bioassay system that can quantify biotin in the seven-digit range, from approximately 0.1 mu g to 1 g per liter. This bioassay proved that the engineered biotin prototroph of C. glutamicum produced biotin directly from glucose, albeit at a marginally detectable level (approximately 0.3 mu g per liter).ArticleAPPLIED AND ENVIRONMENTAL MICROBIOLOGY. 79(15):4586-4594 (2013)journal articl

Index markers of chronic fatigue syndrome with dysfunction of TCA and urea cycles

Chronic fatigue syndrome (CFS) is a persistent and unexplained pathological state characterized by exertional and severely debilitating fatigue, with/without infectious or neuropsychiatric symptoms, lasting at least 6 consecutive months. Its pathogenesis remains incompletely understood. Here, we performed comprehensive metabolomic analyses of 133 plasma samples obtained from CFS patients and healthy controls to establish an objective diagnosis of CFS. CFS patients exhibited significant differences in intermediate metabolite concentrations in the tricarboxylic acid (TCA) and urea cycles. The combination of ornithine/citrulline and pyruvate/isocitrate ratios discriminated CFS patients from healthy controls, yielding area under the receiver operating characteristic curve values of 0.801 (95% confidential interval [CI]: 0.711–0.890, P < 0.0001) and 0.750 (95% CI: 0.584–0.916, P = 0.0069) for training (n = 93) and validation (n = 40) datasets, respectively. These findings provide compelling evidence that a clinical diagnostic tool could be developed for CFS based on the ratios of metabolites in plasma

Interplane Transport and Superfluid Density in Layered Superconductors

We report on generic trends in the behavior of the interlayer penetration depth $\lambda_c$ of several different classes of quasi two-dimensional superconductors including cuprates, Sr$_2$RuO$_4$, transition metal dichalcogenides and organic materials of the $(BEDT-TTF)_2X$-series. Analysis of these trends reveals two distinct patterns in the scaling between the values of $\lambda_c$ and the magnitude of the DC conductivity: one realized in the systems with a Fermi liquid (FL) ground state and the other seen in systems with a marked deviation from the FL response. The latter pattern is found primarily in under-doped cuprates and indicates a dramatic enhancement (factor $\simeq 10^2$) of the energy scale $\Omega_C$ associated with the formation of the condensate compared to the data for the FL materials. We discuss implications of these results for the understanding of pairing in high-$T_c$ cuprates.Comment: 4 pages, 2 figure

Weakened Bayesian Calibration for Tactile Temporal Order Judgment in Individuals with Higher Autistic Traits

Previous psychophysical studies reported a positive aftereffect in tactile temporal order judgments, which can be explained by the Bayesian estimation model (‘Bayesian calibration’). We investigated the relationship between Bayesian calibration and autistic traits in participants with typical development (TD) and autism spectrum disorder (ASD). Bayesian calibration was weakened in TD participants with high autistic traits, consistent with the ‘hypo-priors’ hypothesis for autistic perceptions. The results from the ASD group were generally observed as a continuation of those from the TD groups. Meanwhile, two ASD participants showed irregularly large positive or negative aftereffects. We discussed the mechanisms behind the general results among TD and ASD participants and two particular results among ASD participants based on the Bayesian estimation model. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10803-022-05442-0

Inhaled furosemide is not effective in severe asthma in children

The purpose of the present study was to determine whether or not inhaled furosemide has any therapeutic benefit in acute asthma in children. Aerosolized furosemide (20 mg) was administered four times per day together with conventional asthma therapy for 2 days in five children with severe asthma exacerbation. The oxygen saturation with a pulse oximeter (SpO2), and the peak expiratory flow (PEF) were measured before and 10 min after furosemide inhalation. Although a little improvement in SpO2 and PEF was observed after the furosemide inhalation, the results were inferior to those with an inhaled β2-agonist (procaterol hydrochloride). Long-term furosemide therapy was performed in two cases; however, amelioration of the SpO2 levels (>90%) and PEF (>80% of personal best) was not observed within 8 days of medication with furosemide. It is concluded that inhaled furosemide is not of additional benefit in children with severe exacerbation of asthma

Weakened Bayesian Calibration for Tactile Temporal Order Judgment in Individuals with Higher Autistic Traits

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