Wnt/ß­catenin signaling is a novel therapeutic target for tumor suppressor CYLD­silenced glioblastoma cells.

Tumor suppressor cylindromatosis (CYLD) dysfunction by its downregulation is significantly associated with poor prognosis in patients with glioblastoma (GBM), the most aggressive and malignant type of glioma. However, no effective treatment is currently available for patients with CYLD­downregulated GBM. The aim of the present study was to identify the crucial cell signaling pathways and novel therapeutic targets for CYLD downregulation in GBM cells. CYLD knockdown in GBM cells induced GBM malignant characteristics, such as proliferation, metastasis, and GBM stem­like cell (GSC) formation. Comprehensive proteomic analysis and RNA sequencing data from the tissues of patients with GBM revealed that Wnt/ß­catenin signaling was significantly activated by CYLD knockdown in patients with GBM. Furthermore, a Wnt/ß­catenin signaling inhibitor suppressed all CYLD knockdown­induced malignant characteristics of GBM. Taken together, the results of the present study revealed that Wnt/ß­catenin signaling is responsible for CYLD silencing­induced GBM malignancy; therefore, targeting Wnt/ß­catenin may be effective for the treatment of CYLD­negative patients with GBM with poor prognosis.

Changes in homocysteine and non-mercaptoalbumin levels after acute exercise: a crossover study.

BACKGROUND: Acute exercise is one factor that increases blood homocysteine levels, and elevated homocysteine levels cause oxidative stress. Albumin, which is abundant in blood, is an antioxidant, and the redox state of albumin is used as an index of oxidative stress in blood. This study aimed to assess the effect of acute exercise on plasma homocysteine levels and the blood non-mercaptoalbumin/mercaptoalbumin ratio as an oxidative stress marker. METHODS: This study used a crossover design with exercise and control conditions. Under exercise conditions, a bicycle ergometer was used to perform 40 min of transient constant-load exercise at 65% heart rate reserve. Under control conditions, participants rested for 40 min. Blood was collected before, 30 min after, and 90 min after exercise, and at the same time points under control conditions. Samples were analyzed for the homocysteine concentration and non-mercaptoalbumin/mercaptoalbumin ratio. RESULTS: The results revealed that a 65% heart rate reserve and 40 min of acute exercise increased plasma homocysteine concentration and non-mercaptoalbumin ratio. In the intra-condition comparison, the plasma Hcy concentration was significantly increased at Post 30 min (+ 0.83 ± 0.70 µmol/L, P = 0.003) compared with that at Pre in the exercise condition. Furthermore, 90 min after exercise, the blood non-mercaptoalbumin ratio was significantly increased (+ 0.35 ± 0.71%, P = 0.030) compared to Pre. CONCLUSION: These results indicate that the plasma Hcy concentration first increased, and then the non-mercaptoalbumin/mercaptoalbumin ratio increased as the elevated state was maintained. This study revealed that 65% heart rate reserve, 40 min of acute exercise increased plasma Hcy concentration and non-mercaptoalbumin ratio.

Effect of Acute Dietary Nitrate Supplementation on the Venous Vascular Response to Static Exercise in Healthy Young Adults.

The purpose of this study was to test the hypothesis that acute intake of inorganic nitrate (NO3−) via supplementation would attenuate the venoconstriction and pressor response to exercise. Sixteen healthy young adults were assigned in a randomized crossover design to receive beetroot juice (BRJ) or an NO3−-depleted control beverage (prune juice: CON). Two hours after consuming the allocated beverage, participants rested in the supine position. Following the baseline period of 4 min, static handgrip exercise of the left hand was performed at 30% of the maximal voluntary contraction for 2 min. Mean arterial pressure (MAP) and heart rate (HR) were measured. Changes in venous volume in the right forearm and right calf were also measured using venous occlusion plethysmography while cuffs on the upper arm and thigh were inflated constantly to 30−40 mmHg. The plasma NO3− concentration was elevated with BRJ intake (p < 0.05). Exercise increased MAP and HR and decreased venous volume in the forearm and calf, but there were no differences between CON and BRJ. Thus, these findings suggest that acute BRJ intake does not alter the sympathetic venoconstriction in the non-exercising limbs and MAP response to exercise in healthy young adults, despite the enhanced activity of nitric oxide.

Acute dietary nitrate supplementation does not change venous volume and compliance in healthy young adults.

In this crossover study, we investigated the influence of inorganic nitrate ([Formula: see text]) supplementation on venous volume and compliance in the resting forearm and calf. Twenty healthy young adults were assigned to receive an [Formula: see text]-rich beverage [beetroot juice (BRJ): 140 mL; ∼8 mmol [Formula: see text]] or an [Formula: see text]-depleted control beverage [prune juice (CON): 166 mL; < 0.01 mmol [Formula: see text]). Two hours after consuming the allocated beverage, each participant rested in the supine position for 20 min. Cuffs were then placed around the right upper arm and right thigh, inflated to 60 mmHg for 8 min, and then decreased to 0 mmHg at a rate of 1 mmHg/s. During inflation and deflation of cuff pressure, changes in venous volume in the forearm and calf were measured by venous occlusion plethysmography. Venous compliance was calculated as the numerical derivative of the cuff pressure-venous volume curve in the limbs. The plasma [Formula: see text] concentration was elevated by intake of BRJ (before, 15.5 ± 5.8 µM; after, 572.0 ± 116.1 µM, P < 0.05) but not by CON (before, 14.8 ± 7.2 µM; after, 15.3 ± 7.4 µM, P > 0.05). On the other hand, there was no significant difference in venous volume or compliance in the forearm or calf between BRJ and CON. These findings suggest that although acute inorganic NO3- supplementation may enhance the activity of nitric oxide (NO) via nitrite → NO pathway, it does not influence venous volume or compliance in the limbs in healthy young adults.

Tocotrienol-rich fraction from annatto ameliorates expression of lysyl oxidase in human osteoblastic MG-63 cells.

Lysyl oxidase (LOX) is required for the formation of bone collagen cross-links. Inactivation of the LOX gene in osteoblasts by DNA methylation and JAK signaling has been reported to cause loss of cross-links and an increased risk of fractures. Tocotrienols (T3s) have proven benefits on bone strength, but their potential effects on LOX remain largely unknown. Thus, the present study investigates the in vitro effects of T3s on LOX expression in human osteoblastic MG-63 cells. Results indicated that Tocotrienol-Rich Fraction (TRF), the δ-T3 rich oil extracted from Annatto was the most effective and significantly increased LOX expression. TRF treatment decreased de-novo methyltransferases (DNMTs), DNMT3A and DNMT3B levels. In addition, TRF significantly inhibited JAK2 activation and decreased expression of Fli1, a transcription factor of DNMTs. We conclude that TRF induced an increase in LOX expression via inhibition of de-novo methylation and reduction of Fli1 expression by the inactivation of JAK2.Abbreviations: CpG: cytosine-guanine dinucleotide; DNMT: DNA methyltransferase; Fli1: friend leukemia virus integration 1; JAK: janus kinase; LOX: lysyl oxidase; PCR: polymerase chain reaction; STAT: signal transducers and activators of transcription; T3s: tocotrienols; TPs: tocopherols; TRF: Tocotrienol-Rich Fraction.

Salt Restriction Affects the Excretions of Minerals (Na, K, Ca, Mg, P and Zn) in the Second Voided Fasting Early Morning Urine.

The plasma concentrations of mineral (sodium (Na), potassium (K), calcium (Ca), magnesium (Mg), phosphorus (P), and zinc (Zn)) are kept within narrow ranges to maintain homeostasis; hence, it is difficult to use them as indicators of nutritional status. We selected the excretion of these minerals in the second voided fasting early morning urine (EMU) as potential indicators of nutritional status. We previously reported that Na restriction caused a negative balance of Ca and Mg. Therefore, Na restriction can cause changes in EMU-minerals. This study aimed to examine the relationship between dietary Na restriction and urinary mineral excretion. The study lasted for 21 d, including 16 d of balance period and 3 d of recovery period. The participants (11 healthy young women) were divided into the Na restriction group (n=5) (NaCl: 6 g/d) and control group (n=6) (NaCl: 12 g/d). The Na restriction group changed to the control diet (NaCl: 12 g/d) during only the recovery period. The EMU-Na, Ca, Mg, P and Zn in the Na restriction group significantly decreased compared with that of the control group. The EMU-Na, K, Ca, Mg, and Zn in the group with NaCl intake of 6 g/d significantly decreased compared with that of the group with NaCl intake of 12 g/d (in the Na restriction group). We conclude that the decrease in excretion of Na, Ca, Mg and Zn in the EMU can lead to Na restriction. This result can serve as basis when considering EMU as an indicator of mineral status.

Association of Frequency of Milk or Dairy Product Consumption with Subjective Sleep Quality during Training Periods in Japanese Elite Athletes: A Cross-Sectional Study.

The purpose of the study was to examine the association of the frequencies of milk and dairy product consumption with subjective sleep quality during the training period in Japanese elite athletes. In this cross-sectional study, 682 Japanese elite athletes who were candidates for the 2016 Rio Olympic Games underwent medical evaluations at the medical center of The Japan Institute of Sports Sciences. Self-reported questionnaires were used to collect information on demographics and lifestyle (age, height, weight, sports, presence of milk allergy, smoking and drinking habits), subjective sleep quality (good, normal, or poor), bedtime, waking time, sleep duration, and frequencies of milk and dairy product consumption. Data from 679 athletes (379 men, 300 women) without milk allergy, were analyzed. Based on the frequencies of both milk and dairy product consumption, the athletes were divided into three groups: low (0-2 d/wk), middle (3-5 d/wk), and high (6-7 d/wk). Multiple logistic regression models showed that in comparison with the low milk consumption group, the middle [OR (95% CI): 0.48 (0.26-0.91)] and high groups [0.38 (0.21-0.71)] were significantly associated with a lower risk of decrease in subjective sleep quality (0: good, 1: normal or poor) only in women, after adjusting for possible confounders, such as smoking, drinking habits, and sleep duration. Accordingly, the present study elucidated that a greater frequency of milk consumption was significantly associated with a lower risk of decrease in subjective sleep quality, during training periods in women.

How elevated CO2 affects our nutrition in rice, and how we can deal with it.

Increased concentrations of atmospheric CO2 are predicted to reduce the content of essential elements such as protein, zinc, and iron in C3 grains and legumes, threatening the nutrition of billions of people in the next 50 years. However, this prediction has mostly been limited to grain crops, and moreover, we have little information about either the underlying mechanism or an effective intervention to mitigate these reductions. Here, we present a broader picture of the reductions in elemental content among crops grown under elevated CO2 concentration. By using a new approach, flow analysis of elements, we show that lower absorption and/or translocation to grains is a key factor underlying such elemental changes. On the basis of these findings, we propose two effective interventions-namely, growing C4 instead of C3 crops, and genetic improvements-to minimize the elemental changes in crops, and thereby avoid an impairment of human nutrition under conditions of elevated CO2.

Association between Adherence to the Japanese Food Guide Spinning Top and Sleep Quality in College Students.

This study aimed to elucidate the association between adherence to the Japanese Food Guide Spinning Top (Food Guide score) and sleep quality in Japanese college students. We conducted a cross-sectional study of 175 Japanese college students aged 19⁻22 years in the eastern part of Gunma Prefecture to examine the association between the Food Guide score and sleep quality. A self-administered diet history questionnaire and the Pittsburgh Sleep Quality Index were used to assess habitual dietary intake and sleep quality, respectively. In the fully adjusted model, the odds ratios for poor sleep quality in the middle and highest tertile categories of the Food Guide score were 0.50 (95% confidence interval, 0.18⁻1.37) and 0.30 (95% confidence interval, 0.11⁻0.84), respectively, compared with those in the lowest tertile category (p for trend = 0.033). A well-balanced diet may be associated with good sleep quality in Japanese college students.

Association between Skin Condition and Sleep Efficiency in Japanese Young Adults.

There has been little epidemiological evidence that has comprehensively clarified whether alterations in lifestyle, such as sleep quality and dietary intake, explain changes in the skin condition of healthy young adults. Therefore, the aim of our study was to elucidate the association between skin condition and lifestyle behaviors such as diet and sleep, after statistically controlling confounding factors. The subjects were 54 participants who were aged 20-32 y, and who attended college in the Kanto region (in Japan). Transepidermal water loss (TEWL) was obtained by putting a probe on the skin surface based on the European Society of Contact Dermatitis guidelines for TEWL measurements. Self-administered questionnaires on demographic characteristics, dietary habits, and health status, such as sleep condition, depression, and fatigue, were handed to participants on the day of measurement. We found that lower sleep efficiency was significantly associated with higher TEWL (p=0.023), while other demographic factors, dietary intakes, and eating behaviors were not (p>0.05). This significant association remained unchanged after controlling for confounding variables, such as sex, BMI, and dietary intake. Our findings may have important implications in the development of valuable health strategies that may suggest behavior modifications for young to middle-aged men and women.

A redox-silent analogue of tocotrienol inhibits cobalt(II) chloride-induced VEGF expression via Yes signaling in mesothelioma cells.

Vascular endothelial growth factor (VEGF) plays a crucial role in tumor angiogenesis and represents an attractive anticancer target. We have previously demonstrated that a redox-silent analogue of α-tocotrienol, 6-O-carboxypropyl-α-tocotrienol (T3E) exhibits potent anti-carcinogenic property in human malignant mesothelioma (MM) cells. However, inhibition of tumor growth by targeting VEGF pathway remains undetermined. In this study, we explored the inhibitory effect of T3E on the paracrine secretion of VEGF in MM cells under mimicked hypoxia by cobalt chloride (CoCl2). In this study we examine whether T3E can suppress the secretion of VEGF in MM cells exposed to mimic hypoxia by cobalt chloride (CoCl2). We found that CoCl2-induced hypoxia treatment leads to increased up-regulated hypoxia-inducible factor-2α (HIF-2α) and subsequently induced the secretion of VEGF in MM cells. This up-regulation activation mainly depended on the activation of Yes, a member of the Src family of kinases. Treatment of hypoxic MM cells with T3E effectively inhibited the secretion of VEGF, On the other hand, T3E inhibited CoCl2-induced gene expression of VEGF due to the inactivation of Yes/HIF-2α signaling. These data suggest that Yes/HIF2-α/VEGF could be a promising therapeutic target of T3E in MM cells.

Induction of the connexin 32 gene by epigallocatechin-3-gallate potentiates vinblastine-induced cytotoxicity in human renal carcinoma cells.

BACKGROUND/AIM: Enforced expression of the connexin (Cx) 32 gene, a member of the gap junction gene family and a tumor suppressor gene in human renal cell carcinoma (RCC), enhanced vinblastine (VBL)-induced cytotoxicity in RCC cells due to suppression of multidrug resistance 1 (MDR1) expression. Furthermore, in RCC the Cx32 gene is silenced by hypermethylation of CpG islands in a promoter region of the Cx gene. In this study, we investigated if the green tea polyphenol epigallocatechin-3-gallate (EGCG) could enhance susceptibility of RCC cells (Caki-1, a human metastatic RCC cell) to VBL. METHODS: The effects of EGCG on Caki-1 cells were estimated by WST-1 (cell viability), real-time RT-PCR (mRNA level) and immunoblotting (protein level). We estimated the methylation status in the promoter region of the Cx32 gene in RCC cells by methylation-specific PCR. Each protein function was inhibited by small interfering RNA (siRNA) and specific inhibitors. RESULTS: The EGCG treatment elicited significant upregulation of Cx32 in Caki-1 cells, and the induction of the Cx led to the suppression of MDR1 mRNA expression through inactivation of Src and subsequent activation of c-Jun NH2-terminal kinase (JNK). Chemical sensitivity to VBL in Caki-1 cells was increased by EGCG pretreatment, and this effect was abrogated by siRNA-mediated knockdown of Cx32. CONCLUSION: This study suggests that the restoration of Cx32 by EGCG pretreatment improves chemical tolerance on VBL in Caki-1 cells via the inactivation of Src and the activation of JNK.

Redox-inactive analogue of tocotrienol as a potential anti-cancer agent.

Vitamins are prominent among natural or endogenous compounds that are considered to be beneficial for both prevention and therapy of various human ailments. The vitamin E group of compounds composed of tocopherol and tocotrienol isoforms, has been subsequently proven to have health benefits including antioxidant and related protective properties. However, individual isoforms exhibit a wide-range of antioxidant potencies. Tocotrienol (T3) displays powerful anticancer activity that is often not exhibited by tocopherols, by modulating multiple intracellular signaling pathways associated with tumor cell proliferation and survival. The anticancer effect of T3 remains not fully understood but generally is mediated independently of its antioxidant activity. Further we have synthesized a new redox-inactive analogue of T3, 6-O-carboxypropyl-α-tocotrienol (T3E) showing considerable promise for stronger anticancer potency than its mother compound. In this mini-review, we particularly focus upon the anticancer action of the above active components of vitamin E and describe current research on the anticancer effects of T3 irrespective of antioxidant activity.

Yes is a central mediator of cell growth in malignant mesothelioma cells.

The constitutive activation of the Src family kinases (SFKs) has been established as a poor prognostic factor in malignant mesothelioma (MM), however, the family member(s) which contribute to the malignancy have not been defined. This study aimed to identify the SFK member(s) contributing to cell growth using RNA interference in various MM cell lines. Silencing of Yes but not of c-Src or Fyn in MM cells leads to cell growth suppression. This suppressive effect caused by Yes silencing mainly depends on G1 cell cycle arrest and partly the induction of apoptosis. Also, the knockout of Yes induces the inactivation of ß-catenin signaling and subsequently decreases the levels of cyclin D necessary for G1-S transition in the cell cycle. In addition, Yes knockout has less effect on cell growth suppression in ß-catenin-deficient H28 MM cells compared to other MM cells which express the catenin. Overall, we conclude that Yes is a central mediator for MM cell growth that is not shared with other SFKs such as c-Src.

Bowman-Birk protease inhibitor from soybeans enhances cisplatin-induced cytotoxicity in human mesothelioma cells.

Malignant mesothelioma (MM) is an aggressive cancer with no effective treatment options. Enforced expression of the gap junction (GJ) component connexin 43 (Cx43) increases the sensitivity of MM cells to cisplatin. Bowman-Birk protease inhibitor (BBI) induces the restoration of Cx43 in several types of tumor cells. In this study, we examined the capability of BBI to enhance the cytotoxic effect of cisplatin in MM cells via the induction of Cx43. Human MM H28 cells were used. Cell viability was evaluated by a WST-1 assay and proteasomal activity was determined by fluorometric analysis. Protein and mRNA levels were determined by immunoblot analysis and real-time RT-PCR, respectively. GJ function mediated by Cx43 was evaluated using the scrape-loading method. BBI effectively inhibited H28 cell growth in a dose-dependent manner (200-400 µg/ml). In parallel with the growth inhibition, Cx43 levels (mRNA and protein) and GJ function were elevated by BBI treatment. Knockdown of BBI-induced Cx43 by an antisense nucleotide treatment almost cancelled the growth inhibition. BBI enhanced cisplatin-induced cytotoxicity in H28 cells, and down-regulation of Cx43 by the antisense nucleotide treatment abrogated the enhancing effect of BBI. The induction of Cx43 by BBI contributed to Src inactivation and subsequent induction of Bax. Furthermore, an Src inhibitor (SU6656) also enhanced cisplatin-induced cytotoxicity in H28 cells. These results suggest that BBI improves the cytotoxic efficacy of cisplatin in H28 cells via the inhibition of Src signaling.

Long-term survival of a patient with Marshall-Smith syndrome.

A 7-year-old girl presented with classic signs of typical Marshall-Smith syndrome, a rare early overgrowth syndrome. Her long-term survival, which is extremely uncommon in such patients, can be attributed to the successful securing and maintenance of an airway. Continued follow-up is extremely important in finding out how to ensure long-term survival of such patients.

An Unusual Case of Multiple Gastric Carcinoids Associated with Diffuse Endocrine Cell Hyperplasia and Parietal Cell Hypertrophy.

We describe a case of multiple gastric carcinoid tumors in a 47 year-old Japanese man. The patient had markedly elevated serum gastrin levels (>800 pg/mL), which were suppressed by secretin-pancreozymin administration. In the partial gastrectomy specimen, a total of 19 carcinoids arose from diffuse linear and micronodular hyperplasia of the oxyntic mucosal endocrine cells. The carcinoid cells were chromogranin A-positive. Except for a very small number of serotonin-positive cells in several carcinoids, none of the 19 reacted with a battery of antibodies to other bioactive neuroendocrine substances. The most prominent findings in this case were peculiar fundic glands that were distended with a proteinaceous substance and lined with large hypertrophic parietal cells. At the ultra-structural level, these cells showed poorly developed intracytoplasmic canaliculi and vesicotubular profiles, yet their large cytoplasm had numerous mitochondria. On the basis of our histological and ultrastructural findings we suggest that an intrinsic HCI secretion abnormality of the parietal cells may be responsible for the patient's hypergastranemia. Since there have been no reports on similar parietal cells changes, it is possible that our case may represent a pathological entity not previously described.