Cytosolic UDP-L-arabinose synthesis by bifunctional UDP-glucose 4-epimerases in Arabidopsis.

L-Arabinose (L-Ara) is a plant-specific sugar found in cell wall polysaccharides, proteoglycans, glycoproteins, and small glycoconjugates, which play physiologically important roles in cell proliferation and other essential cellular processes. L-Ara is synthesized as UDP-L-arabinose (UDP-L-Ara) from UDP-xylose (UDP-Xyl) by UDP-Xyl 4-epimerases (UXEs), a type of de novo synthesis of L-Ara unique to plants. In Arabidopsis, the Golgi-localized UXE AtMUR4 is the main contributor to UDP-L-Ara synthesis. However, cytosolic bifunctional UDP-glucose 4-epimerases (UGEs) with UXE activity, AtUGE1, and AtUGE3 also catalyze this reaction. For the present study, we first examined the physiological importance of bifunctional UGEs in Arabidopsis. The uge1 and uge3 mutants enhanced the dwarf phenotype of mur4 and further reduced the L-Ara content in cell walls, suggesting that bifunctional UGEs contribute to UDP-L-Ara synthesis. Through the introduction of point mutations exchanging corresponding amino acid residues between AtUGE1 with high UXE activity and AtUGE2 with low UXE activity, two mutations that increase relative UXE activity of AtUGE2 were identified. The crystal structures of AtUGE2 in complex forms with NAD+ and NAD+/UDP revealed that the UDP-binding domain of AtUGE2 has a more closed conformation and smaller sugar-binding site than bacterial and mammalian UGEs, suggesting that plant UGEs have the appropriate size and shape for binding UDP-Xyl and UDP-L-Ara to exhibit UXE activity. The presented results suggest that the capacity for cytosolic synthesis of UDP-L-Ara was acquired by the small sugar-binding site and several mutations of UGEs, enabling diversified utilization of L-Ara in seed plants.

Higher prefrontal activity based on short-term neurofeedback training can prevent working memory decline in acute stroke.

This study aimed to clarify whether short-term neurofeedback training during the acute stroke phase led to prefrontal activity self-regulation, providing positive efficacy to working memory. A total of 30 patients with acute stroke performed functional near-infrared spectroscopy-based neurofeedback training for a day to increase their prefrontal activity. A randomized, Sham-controlled, double-blind study protocol was used comparing working memory ability before and after neurofeedback training. Working memory was evaluated using a target-searching task requiring spatial information retention. A decline in spatial working memory performance post-intervention was prevented in patients who displayed a higher task-related right prefrontal activity during neurofeedback training compared with the baseline. Neurofeedback training efficacy was not associated with the patient's clinical background such as Fugl-Meyer Assessment score and time since stroke. These findings demonstrated that even short-term neurofeedback training can strengthen prefrontal activity and help maintain cognitive ability in acute stroke patients, at least immediately after training. However, further studies investigating the influence of individual patient clinical background, especially cognitive impairment, on neurofeedback training is needed. Current findings provide an encouraging option for clinicians to design neurorehabilitation programs, including neurofeedback protocols, for acute stroke patients.

Expression of factor XIII originating from synovial fibroblasts and macrophages induced by interleukin-6 signaling.

BACKGROUND: Blood coagulation factor XIII (FXIII) promotes cross-linking between fibrin molecules at the final stage of the blood coagulation cascade. However, its expression in cells or tissues and function, particularly factor XIII subunit B (FXIII-B), remains controversial. Hemorrhagic FXIII deficiency following anti-interleukin-6 (IL-6) receptor antibody treatment has been reported in patients with rheumatoid arthritis (RA). Patients receiving this biologics have reduced FXIII activity when compared to the activity in those treated with other biologics. The relationship between pro-inflammatory cytokines and FXIII expression remains unknown. METHODS: To investigate the expression pattern of FXIII in synovial tissues, immunohistochemistry, RT-qPCR, and western blotting were performed. FXIII-A expressed monocyte-derived macrophages were treated with recombinant IL-6 and anti-IL-6 receptor antibody. RNA sequencing of FXIII-B-overexpressing cells was performed to clarify the function of FXIII-B. RESULTS: The immunohistochemical analysis of synovial tissues revealed that factor XIII subunit A (FXIII-A) was expressed in M2 macrophages, and FXIII-B was expressed in fibroblast-like synoviocytes. IL-6 stimulation upregulated FXIII-A expression in IL-4-induced monocyte-derived macrophages, and the anti-IL-6 receptor antibody suppressed FXIII-A expression. FXIII-B was more abundantly secreted in the supernatant of fibroblast-like synoviocytes compared with that of other cells. RNA sequencing showed that FXIII-B elevated the expression of genes associated with anti-apoptotic molecules and chemokines. CONCLUSIONS: Our findings highlight that synovial tissue is one of the sources of FXIII production. We also have demonstrated IL-6-dependent FXIII-A expression and the novel potential functions of FXIII-B.

Individual Sensory Modality Dominance as an Influential Factor in the Prefrontal Neurofeedback Training for Spatial Processing: A Functional Near-Infrared Spectroscopy Study.

Neurofeedback is a neuromodulation technique used to improve brain function by self-regulating brain activity. However, the efficacy of neurofeedback training varies widely between individuals, and some participants fail to self-regulate brain activity. To overcome intersubject variation in neurofeedback training efficacy, it is critical to identify the factors that influence this type of neuromodulation. In this study, we considered that individual differences in cognitive ability may influence neurofeedback training efficacy and aimed to clarify the effect of individual working memory (WM) abilities, as characterized by sensory modality dominance, on neurofeedback training efficacy in healthy young adults. In particular, we focused on the abilities of individuals to retain internal (tactile or somatosensory) or external (visual) body information in their WM. Forty participants performed functional near-infrared spectroscopy-based neurofeedback training aimed at producing efficient and lower-level activity in the bilateral dorsolateral prefrontal cortex and frontopolar cortex. We carried out a randomized, sham-controlled, double-blind study that compared WM ability before and after neurofeedback training. Individual WM ability was quantified using a target searching task that required the participants to retain spatial information presented as vibrotactile or visual stimuli. Participants who received feedback information based on their own prefrontal activity showed gradually decreasing activity in the right prefrontal area during the neurofeedback training and demonstrated superior WM ability during the target searching task with vibrotactile stimuli compared with the participants who performed dummy neurofeedback training. In comparison, left prefrontal activity was not influenced by the neurofeedback training. Furthermore, the efficacy of neurofeedback training (i.e., lower right prefrontal activity and better searching task performance) was higher in participants who exhibited tactile dominance rather than visual dominance in their WM. These findings indicate that sensory modality dominance in WM may be an influential neurophysiological factor in determining the efficacy of neurofeedback training. These results may be useful in the development of neurofeedback training protocols tailored to individual needs.

A versatile cis-prenyltransferase from Methanosarcina mazei catalyzes both C- and O-prenylations.

Polyprenyl groups, products of isoprenoid metabolism, are utilized in peptidoglycan biosynthesis, protein N-glycosylation, and other processes. These groups are formed by cis-prenyltransferases, which use allylic prenyl pyrophosphates as prenyl-donors to catalyze the C-prenylation of the general acceptor substrate, isopentenyl pyrophosphate. Repetition of this reaction forms (Z,E-mixed)-polyprenyl pyrophosphates, which are converted later into glycosyl carrier lipids, such as undecaprenyl phosphate and dolichyl phosphate. MM_0014 from the methanogenic archaeon Methanosarcina mazei is known as a versatile cis-prenyltransferase that accepts both isopentenyl pyrophosphate and dimethylallyl pyrophosphate as acceptor substrates. To learn more about this enzyme's catalytic activity, we determined the X-ray crystal structures of MM_0014 in the presence or absence of these substrates. Surprisingly, one structure revealed a complex with O-prenylglycerol, suggesting that the enzyme catalyzed the prenylation of glycerol contained in the crystallization buffer. Further analyses confirmed that the enzyme could catalyze the O-prenylation of small alcohols, such as 2-propanol, expanding our understanding of the catalytic ability of cis-prenyltransferases.

Distinct bilateral prefrontal activity patterns associated with the qualitative aspect of working memory characterized by individual sensory modality dominance.

In addition to quantitative individual differences in working memory (WM) capacity, qualitative aspects, such as enhanced sensory modality (modality dominance), can characterize individual WM ability. This study aimed to examine the neurological basis underlying the individual modality dominance component of WM using functional near-infrared spectroscopy (fNIRS). To quantify the degree of individual WM modality dominance, 24 participants were required to find seven hidden targets and hold their spatial location and appearance order with vibrotactile or visual stimuli aids. In this searching task, eight participants demonstrated higher performance with the tactile condition (tactile-dominant) whereas sixteen demonstrated visual dominance. We then measured prefrontal activity by fNIRS during memorization of visual stimulus numbers while finger tapping as a cognitive-motor dual-task. Individual modality dominance significantly correlated with bilateral frontopolar and dorsolateral prefrontal activity changes over repeated fNIRS sessions. In particular, individuals with stronger visual dominance showed marked decreases in prefrontal area activity. These results suggest that distinct processing patterns in the prefrontal cortex reflect an individual's qualitative WM characteristics. Considering the individual modality dominance underlying the prefrontal areas could enhance cognitive or motor performance, possibly by optimizing cognitive resources.

Orthogonal Folding of Amphiphilic/Fluorous Random Block Copolymers for Double and Multicompartment Micelles in Water.

Here, we report orthogonal folding and self-assembly systems of amphiphilic/fluorous random block copolymers for double core and multicompartment micelles in water. For this, we developed the precision folding techniques of polymer chains via the selective self-assembly of the pendant groups. Typically, A/C-B/C random block copolymers were designed: Hydrophobic dodecyl groups (A) and fluorous fluorinated octyl groups (B) were introduced into the respective blocks, while hydrophilic poly(ethylene glycol) chains (C) were randomly incorporated into all the segments. By controlling the chain length and composition of the respective blocks, the copolymers induce orthogonal single-chain folding in water to form double-compartment micelles comprising hydrophobic and fluorous cores. The copolymers were site-selectively folded in a fluoroalcohol to result in tadpole unimer micelles comprising a hydrophobic A/C unimer micelle and an unfolded fluorous B/C chain. Additionally, asymmetric A/C-B/C random block copolymers with short and highly hydrophobic or fluorous segments were effective for multicompartment micelles in water.

Compartmentalization Technologies via Self-Assembly and Cross-Linking of Amphiphilic Random Block Copolymers in Water.

Orthogonal self-assembly and intramolecular cross-linking of amphiphilic random block copolymers in water afforded an approach to tailor-make well-defined compartments and domains in single polymer chains and nanoaggregates. For a double compartment single-chain polymer, an amphiphilic random block copolymer bearing hydrophilic poly(ethylene glycol) (PEG) and hydrophobic dodecyl, benzyl, and olefin pendants was synthesized by living radical polymerization (LRP) and postfunctionalization; the dodecyl and benzyl units were incorporated into the different block segments, whereas PEG pendants were statistically attached along a chain. The copolymer self-folded via the orthogonal self-assembly of hydrophobic dodecyl and benzyl pendants in water, followed by intramolecular cross-linking, to form a single-chain polymer carrying double yet distinct hydrophobic nanocompartments. A single-chain cross-linked polymer with a chlorine terminal served as a globular macroinitiator for LRP to provide an amphiphilic tadpole macromolecule comprising a hydrophilic nanoparticle and a hydrophobic polymer tail; the tadpole thus self-assembled into multicompartment aggregates in water.

Evaluation of the effects of a combination of Japanese honey and hydrocolloid dressing on cutaneous wound healing in male mice.

The aim of this study was to evaluate the effect of the combined use of Japanese honey and hydrocolloid dressing (HCD) on cutaneous wound healing. Mice were divided into four groups: the Acacia (Japan) + HCD, Manuka (New Zealand) + HCD, Chinese milk vetch (Japan) + HCD, and HCD (control) groups. The mice received two full-thickness wounds. The wounds of the HCD group were covered with HCD, whereas those of the other groups were treated with 0.1 mL of the relevant type of honey, before being covered with HCD. Wound area was significantly smaller in the HCD group than in the Acacia + HCD and Manuka + HCD groups on day 13 and days 8-14, respectively. Moreover, compared with the HCD group, reepithelialization was delayed in the Acacia + HCD group and reepithelialization and collagen deposition were delayed in the Chinese milk vetch + HCD and Manuka + HCD groups. These results indicate that the combined use of Japanese honey and HCD does not promote cutaneous wound healing compared with the use of HCD alone. Thus, this method is probably not useful for promoting healing.

[Adenocarcinoma of the rete testis. A case report].

Adenocarcinoma of the rete testis is a rare malignant tumor with a poor prognosis. About 60 cases of this adenocarcinoma have been reported in the literature. The diagnosis is often difficult and made incidentally. Herein, we report a case of adenocarcinoma of the rete testis and review the literature. Our patient was an 80-year-old man who presented with painless scrotal swelling for 2 years. Physical examination revealed an enlarged, hard mass of the left scrotum. The serum markers alpha-fetoprotein (AFP), beta-human chorionic gonadotropin (beta-HCG), and carcinoembryonic antigen (CEA) were negative. Magnetic resonance imaging (MRI) showed a left hydrocele with central necrosis of the testis. After 4 months, the patient presented with appetite loss, general fatigue, and pain in the left scrotum. Positron emission tomography (PET) was performed in another hospital, and the patient was referred for a left testicular tumor, multiple lung metastases, and para-aorta lymph node metastasis. The patient underwent left high inguinal orchiectomy. Pathological examination revealed a hard whitish mass around the testis involving the epididymis and tunica vaginalis and spreading under the subcutaneous tissue. Histological examination revealed adenocarcinoma in the hilum of the testis, which extended to the subcutaneous tissue but not to the surface of the scrotum. The tunica albuginea was intact, and no invasion of carcinoma in the testis was seen. After the histological diagnosis of adenocarcinoma of the rete testis was confirmed, computed tomography (CT) was performed and showed multiple pulmonary nodules and para-aortica lymph node swelling of 3 cm diameter. Because the patient did not wish to receive chemotherapy or other aggressive treatment, he has been followed-up with palliative care since his diagnosis. Although local recurrence has occurred 4 months later, he is still alive for 8 months since his diagnosis.

Polycomb group protein Ezh1 represses Nodal and maintains the left-right axis.

The left-right (LR) axis is essential for the proper function of internal organs. In mammals and fish, left-sided Nodal expression governs LR patterning. Here, we show that the Polycomb group protein Ezh1, which is highly conserved from fish to human, participates in LR patterning. Knockdown of olezh1, a medaka homologue of Ezh1, led to LR reversal of internal organs. It was shown that OLEZH1 acts in silencing the expression of Spaw (a medaka homolog of Nodal) via a previously unknown pathway. Furthermore, coimmunoprecipitation showed physical interaction of Ezh1 with FoxH1, a Nodal regulator. This represents a novel mechanism for LR patterning and implies that Ezh1 has developmental importance.

Effects of estrogen on age-related changes in muscarinic responsiveness of the urinary bladder and lumbosacral dorsal root ganglion cells in female rats.

The aim of this study was to investigate estrogen's effect on age-related changes in bladder function. Female Wistar rats were divided into three groups that included young rats (3-month-old) (YR), old rats (13-month-old) (OR), and old rats given subcutaneous treatments of estradiol for 6 weeks (OR + E). The groups were evaluated for (i) micturition behavior, (ii) changes of detrusor contractility and frequency of detrusor contraction in response to muscarinic stimulation in cystometrograms, (iii) messenger RNA (mRNA) expression of the muscarinic receptor subtype in the detrusor muscle, as measured by real-time polymerase chain reaction, and (iv) the immunoreactivity of P2X(3), CGRP, and substance P in the lumbosacral dorsal root ganglia. There were no significant changes for the average micturition volumes or micturition frequencies seen among the three groups. Moreover, there were no significant differences in the proportion of the immunoreactivities of P2X(3), CGRP, and substance P in afferent neurons among these three groups. However, an intravenous administration of muscarine significantly increased the frequency during continuous cystometrograms in the OR and OR + E groups. In these groups, there was a significant increase in the expression of the M(2) receptor mRNAs as compared to YR. It appears that the up-regulation of the M(2) receptor may lead to a decreased intercontraction interval by muscarinic stimuli. OR + E rats showed a significant increase in bladder weight as compared to the OR group. The muscarine-stimulated contractility of the detrusor in the cystometrogram also exhibited a significant increase in the OR + E group as compared to the OR group, which resulted from estrogen-induced functional hypertrophy of the detrusor muscle. These findings suggest that as little as 6 weeks of estrogen treatment is capable of improving the detrusor contractility, although the treatment contributes little to the storage phase of the micturition cycle.

Effects of long-term estradiol treatment on the contractile response to muscarine and muscarinic receptor subtypes in the bladder of aged female rats.

In this study, we tried to elucidate the effect of estrogen treatment on the detrusor contractile response to muscarine and muscarine receptor subtypes of the bladder in 13-month-old female Wistar rats. The rats were divided into two groups, controls and rats treated with estradiol for 12 weeks. After the treatment phase, we monitored micturition behavior in addition to performing cystometrograms after the administration of muscarine, and real-time polymerase chain reaction for mRNA expression of the muscarinic receptor subtypes in the detrusor muscle. Our data indicated that there was a significant increase in the maximum micturition volume in the estradioltreated rats. The urodynamic results indicated significant changes in the maximum detrusor pressure following the administration of muscarine in the estradiol-treated rats, in contrast to the controls for which no significant changes were observed. Furthermore, M(3) receptor mRNAs in the detrusor muscle were significantly decreased in the estradiol-treated rats as compared to the control rats, while there were no differences noted for the M2 receptor mRNAs. Our data demonstrates that long-term estradiol treatment might be capable of increasing the potential detrusor contractility, and thus, estradiol might be a therapeutic agent that can be used to target the M3 receptors during the treatment of detrusor instability.

Lymphovascular invasion independently predicts increased disease specific survival in patients with transitional cell carcinoma of the upper urinary tract.

PURPOSE: We investigated the prognostic impact of lymphovascular invasion (LVI) and traditional prognostic factors for survival in a large series of patients treated surgically for upper tract transitional cell carcinoma (TCC). We also developed a prognostic factors-based model for risk stratification of upper tract TCC. MATERIALS AND METHODS: We identified a study population of 173 consecutive patients treated surgically for upper tract TCC at our institution between 1980 and 2002. We compared LVI with other pathological features and determined the disease specific survival rate. RESULTS: LVI was found in 52 patients (30.1%). As tumor grade and pathological stage increased, the incidence of LVI increased significantly. LVI was found in 12 of 133 patients (9.0%) without lymph node metastasis compared with 40 of 40 patients (100%) with lymph node metastasis. Five and 10-year disease specific survival rates were 84.9% and 80.4% in the absence of LVI, and 40.2% and 21.1% in the presence of LVI, respectively (p <0.001). In multivariate analysis LVI, pathological T stage and tumor grade were independent predictors for disease specific survival. The relative risk of death could be expressed with the formula, exp(0.729 x tumor grade + 1.659 x pathological T stage + 1.160 x LVI). Using this equation the patients were stratified into low risk (grade 1 or 2, LVI negative, stage pT2 or lower), high risk (any tumor grade, LVI positive, stage pT3 or greater) and intermediate risk (all others) groups with significant differences in survival. Five and 10-year disease specific survival rates were 93.0% and 89.4% in the low risk group (82 patients), 66.8% and 62.9% in the intermediate risk group (53 patients), and 25.6% and 0% in the high risk group (38 patients), respectively. CONCLUSIONS: In addition to pathological stage and tumor grade, LVI is an independent prognostic factor for disease specific survival in upper tract TCC. Patients in the high and/or intermediate risk groups may benefit from integrated therapies with surgery and postoperative systemic chemotherapy.

Modulation of bcl-2 family proteins in MAPK independent apoptosis induced by a cdc25 phosphatase inhibitor Cpd 5 in renal cancer cells.

Protein phosphatases have been classified into two basic types, namely protein serine/threonine phosphatase (PP), and protein tyrosine phosphatase (PTP). Cpd 5 is a selective inhibitor of cdc25 phosphatases, which belong to members of PTPs and regulate cell proliferation by controlling cyclin-dependent kinases (cdks). The present study was undertaken to investigate the potential utility of Cpd 5 as an anti-neoplastic agent for renal cell carcinomas (RCCs). Three renal cancer cell lines, 769P, Sw839, and A498 were used. The effects of Cpd 5 on the viability of renal cancer cell lines was analyzed using an Alamar Blue assay. Apoptosis was determined by flow cytometric TUNEL analysis. Changes in the expression of cdc25 phosphatases, mitogen-activated protein kinases (MAPKs), and bcl-2 family proteins were detected using Western blot analysis. The apoptosis-inducing effect of Cpd 5 on human RCC tissue was analyzed through TUNEL staining of organ cultures from RCCs. Cpd 5 showed a strong cytotoxicity against all renal cancer cell lines with an apoptosis-inducing effect. All cell lines treated with Cpd 5 resulted in a down-regulation of cdc25A, cdc25B, and cdc25C, however, the MAPK pathways were not affected. In addition, the up-regulation of bax, and the down-regulation of bcl-2 and bcl-xL, was observed. In organ cultures from RCCs, TUNEL-positive apoptotic nuclei were observed when treated with Cpd 5. Cpd 5 was thus found to effectively inhibit the proliferation of human renal cancer cells while also inducing apoptosis by inhibiting cdc25 phosphatases and modulating bcl-2 family proteins. The administration of Cpd 5 may thus be an effective therapeutic approach for RCCs.