We have recently developed a Corynebacterium glutamicum strain that generates NADPH via the glycolytic pathway by replacing endogenous NAD-dependent glyceraldehyde 3-phosphate dehydrogenase (GapA) with a nonphosphorylating NADP-dependent glyceraldehyde 3-phosphate dehydrogenase (GapN) from Streptococcus mutans. Strain RE2, a suppressor mutant spontaneously isolated for its improved growth on glucose from the engineered strain, was proven to be a high-potential host for L-lysine production (Takeno et al., 2010). In this study, the suppressor mutation was identified to be a point mutation in rho encoding the transcription termination factor Rho. Strain RE2 still showed retarded growth despite the mutation rho696. Our strategy for reconciling improved growth with a high level of L-lysine production was to use GapA together with GapN only in the early growth phase, and subsequently shift this combination-type glycolysis to one that depends only on GapN in the rest of the growth phase. To achieve this, we expressed gapA under the 2 myo-inositol-inducible promoter of iolT1 encoding a myo-inositol transporter in strain RE2. The resulting strain RE2A iol was engineered into an L-lysine producer by introduction of a plasmid carrying the desensitized lysC, followed by examination for culture conditions with myo-inositol supplementation. We found that as a higher concentration of myo-inositol was added to the seed culture, the following fermentation period became shorter while maintaining a high level of L-lysine production. This finally reached a fermentation period comparable to that of the control GapA strain, and yielded a 1.5-fold higher production rate compared with strain RE2. The transcript level of gapA, as well as the GapA activity, in the early growth phase increased in proportion to the myo-inositol concentration and then fell to low levels in the subsequent growth phase, indicating that improved growth was a result of increased GapA activity, especially in the early growth phase. Moreover, blockade of the pentose phosphate pathway through a defect in glucose 6-phosphate dehydrogenase did not significantly affect L-lysine production in the engineered GapN strains, while a drastic decrease in L-lysine production was observed for the control GapA strain. Determination of the intracellular NADPH/NADP + ratios revealed that the ratios in the engineered strains were significantly higher than the ratio of the control GapA strain irrespective of the pentose phosphate pathway. These results demonstrate that our strain engineering strategy allows efficient L-lysine production independent of the oxidative pentose phosphate pathway.
Purpose:The aim was to evaluate the value of multifocal electroretinogram (mfERG) and optical coherence tomography (OCT) in predicting post-operative visual acuity (post-VA) after removal of an idiopathic epiretinal membrane (ERM). Methods: Fifty-one eyes of 51 patients with an idiopathic macular ERM were examined by OCT and mfERG before three-port pars plana vitrectomy with removal of the ERM. The parameters of the OCT images and mfERGs, pre-operative visual acuity (pre-VA) and the duration of symptoms (DS) were compared with the post-VA. Results: The pre-VA (r = 0.570, p < 0.001) and the duration of the symptoms (r = 0.389, p < 0.035; n = 35) were significantly correlated with the post-VA. The central retinal thickness (CRT) was significantly correlated with the pre-VA (r = 0.379, p < 0.01) and the post-VA (r = 0.318, p < 0.03). The amplitudes of macular electroretinogram (amERGs) were not significantly correlated with the pre-VA or post-VA. The CRT to amERG ratio (CRT/amERG) was correlated with the post-VA (r = 0.429, p < 0.002) but not with the pre-VA (p > 0.1). Comparative analyses among groups of cases based on CRT/amERG showed significant differences in the post-VA but not in the pre-VA and the duration of symptoms. Conclusion: As with the large CRT/amERG, the pre-operative morphologically thickened and electroretinographically reduced retinas have a greater likelihood of being affected by the irreversible photoreceptor damages that could have been masked or modified by other factors before the removal of the ERM. Because the CRT/amERG values are obtained objectively, they should be valuable in predicting the post-operative visual function along with the pre-VA and the duration of symptoms.
Objective The primary outcome measure used in mechanical thrombectomy (MT) trials is the modified Rankin Scale (mRS). However, the accuracy of mRS might be limited. On the other hand, the functional independence measure (FIM) is a widely used tool to quantify the extent to which patients require assistance during their activities of daily living. The current study aimed to reveal different clinical backgrounds that affect the efficacy of MT measured either by mRS or FIM. Methods Patients who underwent MT at our institution from January 2019 to July 2022 were included and divided into groups based on mRS scores of 0–2 and ≥ 3. Patients were also divided into two groups based on a cut-off value of FIM of ≥ 108, as patients with FIM ≥ 108 are capable of living an independent life. Results The mRS score was 0–2 in 33% of the patients, while the FIM score was ≥ 108 in only 15% of the patients. In the mRS groups, there were significant differences in terms of duration of hospitalization, National Institutes of Health Stroke Scale (NIHSS) scores, achievement of thrombolysis in cerebral infarction (TICI) reperfusion grade of 2b or 3, and postoperative bleeding. Multivariate logistic regression analysis revealed that NIHSS score and achievement of TICI 2b or 3 were significant factors related to mRS 0–2 at discharge. The FIM groups differed significantly in terms of age and, duration of hospitalization, NIHSS score, although multivariate logistic regression analysis revealed that only the NIHSS score was significantly associated with an FIM score of ≥ 108. Conclusion The study showed that the percentage of independent patients is significantly reduced when we evaluated the patients by the FIM. In addition, there are some differences in the clinical background that led to a good outcome between that evaluated by mRS and FIM.
The authors have developed a new technique for removing large and hard lens fragments or crystalline lenses with hard nuclei dislocated into the vitreous cavity using an optic fiber-free intravitreal surgical system (OFFISS). The OFFISS visualizes the fundus without a fiberoptic and facilitates the bimanual technique. The lens materials are stabilized with one hand and the fragmatome to emulsify the lens fragments is controlled with the other. The lens fragments or subluxated crystalline lenses can be removed through 20-gauge incisions. This technique was used in 14 cases and was found to be safe and not invasive.
Objective: To elucidate the relationship between sideto-side asymmetry and sit-to-stand and stand-to-sit duration in patients with right or left hemiplegia by three-dimensional motion analysis of the two movements. Methods: Forty-fi ve patients with hemiplegia (right hemiplegia in 21, left hemiplegia in 24) and 20 normal healthy adults were studied. Using three-dimensional motion analysis, an asymmetry index (AI) was calculated from the trajectory of the left-right component at the midpoint between two acromion markers as a function of time. Normal range of the sitto-stand and stand-to-sit duration was calculated as the mean ± SD obtained from normal subjects. Patients were divided into two groups according to duration: normal duration (within the mean ± 2SD of normal subjects) and abnormal duration (outside the mean ± 2SD of normal subjects). Motor function of the affected lower extremity and other parameters were compared between the normal and abnormal duration groups. Results: Sit-to-stand and stand-to-sit duration was signifi cantly prolonged in both right hemiplegic and left hemiplegic patients compared with normal subjects. In left hemiplegic patients, AI was signifi cantly higher in those with abnormal duration for both sit-to-stand and stand-to-sit movements. In left hemiplegic patients, the lower extremity motor function was signifi cantly poorer in those with abnormal duration for sit-to-stand movement. Conclusion:In patients with left hemiplegia, AI deviates toward the unaffected side, and impaired lower extremity motor function is associated with prolonged sit-to-stand duration.
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