Epigallocatechin-3-gallate potentiates curcumin's ability to suppress uterine leiomyosarcoma cell growth and induce apoptosis.

BACKGROUND: Uterine leiomyosarcoma (LMS) has an unfavorable response to standard chemotherapeutic regimens. Two natural occurring compounds, curcumin and epigallocatechin gallate (EGCG), are reported to have anti-cancer activity. We previously reported that curcumin reduced uterine LMS cell proliferation by targeting the AKT-mTOR pathway. However, challenges remain in overcoming curcumin's low bioavailability. METHODS: The human LMS cell line SKN was used. The effect of EGCG, curcumin or their combination on cell growth was detected by MTS assay. Their effect on AKT, mTOR, and S6 was detected by Western blotting. The induction of apoptosis was determined by Western blotting using cleaved-PARP specific antibody, caspase-3 activity and TUNEL assay. Intracellular curcumin level was determined by a spectrophotometric method. Antibody against EGCG cell surface receptor, 67-kDa laminin receptor (67LR), was used to investigate the role of the receptor in curcumin's increased potency by EGCG. RESULTS: In this study, we showed that the combination of EGCG and curcumin significantly reduced SKN cell proliferation more than either drug alone. The combination inhibited AKT, mTOR, and S6 phosphorylation, and induced apoptosis at a much lower curcumin concentration than previously reported. EGCG enhanced the incorporation of curcumin. 67LR antibody partially rescued cell proliferation suppression by the combination treatment, but was not involved in the EGCG-enhanced intracellular incorporation of curcumin. CONCLUSIONS: EGCG significantly lowered the concentration of curcumin required to inhibit the AKT-mTOR pathway, reduce cell proliferation and induce apoptosis in uterine LMS cells by enhancing intracellular incorporation of curcumin, but the process was independent of 67LR.

The antidiabetic drug metformin inhibits uterine leiomyoma cell proliferation via an AMP-activated protein kinase signaling pathway.

Uterine leiomyomas are the most common gynecological benign tumors and greatly affect reproductive health and wellbeing. Metformin is the most widely used antidiabetic drug in the world, and there is increasing evidence of a potential efficacy of this agent as an anticancer drug. In order to understand metformin's anti-tumorigenic potential better, in this study, we investigated the inhibitory effect of metformin and expression of key targets of metformin cell signaling in leiomyoma cells. Cell proliferation was assessed after exposure to metformin. Apoptosis was assessed by western blotting for cleaved-PARP and TUNEL staining. The expressions of phosphorylated AMPK and phosphorylated S6 were determined by western blotting. Metformin potently inhibited ELT-3 cell proliferation in a dose-dependent manner. Western blotting analysis demonstrated that metformin induced phosphorylation of AMPK and the inhibitory effect was attenuated with AMPK inhibitor, compound C. In parallel, treatment with metformin decreased phosphorylation of S6 protein. These experimental findings show that metformin is a potent inhibitor of cell proliferation in leiomyoma cells. This effect is mediated by AMPK activation and subsequent inhibition of the mTOR pathway. Thus, this study provides a possible mechanism of the action of metformin in the inhibition of leiomyoma cell growth.

Relationship between premenstrual symptoms and dysmenorrhea in Japanese high school students.

To determine the relationship between premenstrual symptoms and dysmenorrhea among Japanese adolescent girls, a total of 1,431 high school students were assessed. Of them, 11.3% were classified with "moderate to severe premenstrual syndrome (PMS)" and 3.2% with "premenstrual dysphoric disorder (PMDD)." Eighty-five percent of the girls had dysmenorrhea. The rates of prevalence of PMDD and moderate to severe PMS were increased according to the severity of dysmenorrhea (rs = 0.479), showing a correlation between the severity of PMS/PMDD and dysmenorrhea in adolescents.

Impact of Masticatory Behaviors Measured With Wearable Device on Metabolic Syndrome: Cross-sectional Study.

BACKGROUND: It has been widely recognized that mastication behaviors are related to the health of the whole body and to lifestyle-related diseases. However, many studies were based on subjective questionnaires or were limited to small-scale research in the laboratory due to the lack of a device for measuring mastication behaviors during the daily meal objectively. Recently, a small wearable masticatory counter device, called bitescan (Sharp Co), for measuring masticatory behavior was developed. This wearable device is designed to assess objective masticatory behavior by being worn on the ear in daily life. OBJECTIVE: This study aimed to investigate the relation between mastication behaviors in the laboratory and in daily meals and to clarify the difference in mastication behaviors between those with metabolic syndrome (MetS) and those without (non-MetS) measured using a wearable device. METHODS: A total of 99 healthy volunteers (50 men and 49 women, mean age 36.4 [SD 11.7] years) participated in this study. The mastication behaviors (ie, number of chews and bites, number of chews per bite, and chewing rate) were measured using a wearable ear-hung device. Mastication behaviors while eating a rice ball (100 g) in the laboratory and during usual meals for an entire day were monitored, and the daily energy intake was calculated. Participants' abdominal circumference, fasting glucose concentration, blood pressure, and serum lipids were also measured. Mastication behaviors in the laboratory and during meals for 1 entire day were compared. The participants were divided into 2 groups using the Japanese criteria for MetS (positive/negative for MetS or each MetS component), and mastication behaviors were compared. RESULTS: Mastication behaviors in the laboratory and during daily meals were significantly correlated (number of chews r=0.36; P<.001; number of bites r=0.49; P<.001; number of chews per bite r=0.33; P=.001; and chewing rate r=0.51; P<.001). Although a positive correlation was observed between the number of chews during the 1-day meals and energy intake (r=0.26, P=.009), the number of chews per calorie ingested was negatively correlated with energy intake (r=-0.32, P=.002). Of the 99 participants, 8 fit the criteria for MetS and 14 for pre-MetS. The number of chews and bites for a rice ball in the pre-MetS(+) group was significantly lower than the pre-MetS(-) group (P=.02 and P=.04, respectively). Additionally, scores for the positive abdominal circumference and hypertension subgroups were also less than the counterpart groups (P=.004 and P=.01 for chews, P=.006 and P=.02 for bites, respectively). The number of chews and bites for an entire day in the hypertension subgroup were significantly lower than in the other groups (P=.02 and P=.006). Furthermore, the positive abdominal circumference and hypertension subgroups showed lower numbers of chews per calorie ingested for 1-day meals (P=.03 and P=.02, respectively). CONCLUSIONS: These results suggest a relationship between masticatory behaviors in the laboratory and those during daily meals and that masticatory behaviors are associated with MetS and MetS components. TRIAL REGISTRATION: University Hospital Medical Information Network Clinical Trials Registry R000034453; https://tinyurl.com/mwzrhrua.

Curcumin disrupts uterine leiomyosarcoma cells through AKT-mTOR pathway inhibition.

OBJECTIVE: Uterine leiomyosarcoma generally has an unfavorable response to standard chemotherapy. The loss of PTEN which results in constitutive AKT-mTOR activation causes an increase in leiomyosarcoma formation in mice. The active ingredient derived from the herb Curcuma longa, curcumin, shows antitumor properties in a variety of cancer cell lines by altering a number of oncogenic pathways. To explore the possibility of curcumin as an alternative to standard chemotherapy, we decided to investigate curcumin's antitumor effect on uterine leiomyosarcoma cells. METHODS: Human leiomyosarcoma cell lines, SKN and SK-UT-1, were cultured for in vitro experiments. Rapamycin or curcumin was added in different doses and their effect on cell growth was detected by MTS assay. The influence of rapamycin or curcumin on AKT, mTOR, p70S6 and S6 phosphorylation and protein expression was detected by Western Blotting. The ability of rapamycin or curcumin to induce apoptosis was determined by Western blotting using cleaved-PARP specific antibody, Caspase-3 activity assay and TUNEL assay. RESULTS: Both rapamycin and curcumin significantly reduced SKN cell proliferation. Curcumin inhibited mTOR, p70S6 and S6 phosphorylation similar with rapamycin. Cleaved PARP, caspase-3 activity and DNA fragmentation increased proportional with curcumin concentration. At a high concentration, curcumin significantly induced apoptosis in SKN cells, but not rapamycin. CONCLUSIONS: Curcumin inhibited uterine leiomyosarcoma cells' growth by targeting the AKT-mTOR pathway for inhibition. However, rapamycin, a specific mTOR inhibitor, did not induce apoptosis in SKN cells unlike curcumin that also has a pro-apoptotic potential in SKN cells.

Perioperative infusion of dexmedetomidine at a high dose reduces postoperative analgesic requirements: a randomized control trial.

PURPOSE: We hypothesized that a high dose of dexmedetomidine (1 µg/kg/h) could reduce postoperative analgesic requirements of patients. METHODS: This was a prospective, randomized, double-blind, placebo-controlled study carried out in Tohoku University Hospital. Thirty-two patients who underwent open gynecological abdominal surgery were randomly divided into a control (group C) and a dexmedetomidine group (group D). In both groups of patients, an epidural catheter was put in position prior to the induction of anesthesia, and continuous epidural infusion was started using a patient-controlled epidural analgesia (PCEA) pump. During the induction of anesthesia, group D patients received a loading dose of dexmedetomidine (1 µg/kg over 10 min), followed by a continuous infusion at a rate of 1 µg/kg/h. The patients in group C received a volume-matched infusion of normal saline as placebo. Consumption of PCEA bolus (local anesthetics) during the first postoperative 24 h, postoperative pain scores, and side effects related to the use of dexmedetomidine were recorded. RESULTS: Dexmedetomidine (1 µg/kg/h) significantly reduced PCEA bolus consumption [15.9 ± 6.5 (group C) vs. 5.3 ± 5.0 ml (group D); P = 0.0001] and postoperative pain scores. The infusion of dexmedetomidine produced no serious side effects, such as hemodynamic changes. CONCLUSIONS: Among this small patient cohort, perioperative infusion of dexmedetomidine (1 µg/kg/h) resulted in antinociception without severe side effects. These results suggest that this method could be of interest with respect to improving postoperative pain status.

Primary sellar lymphoma: intravascular large B-cell lymphoma diagnosed as a double cancer and improved with chemotherapy, and literature review of primary parasellar lymphoma.

Lymphoma is one of the causative factors of hypothalamus-pituitary dysfunction, and intravascular large B-cell lymphoma (IVLBCL) is a subtype of primary extranodal neoplasm. A 69-year-old woman visited our hospital because of general fatigue. We diagnosed her with presumable non-functional primary pituitary adenoma and subsequent dysfunction. Eight months after, the patient revisited our hospital because of dyspnea. Though we conducted systemic investigations including chest and abdomen enhanced computer tomography, transbronchial lung biopsy, and bone marrow biopsy, the diagnosis was not confirmed. Inadvertently, a breast cancer was found, and the surgical specimen proved that the patient had double cancer-adenocarcinoma and IVLBCL. Rituximab, cyclophosphamide, adriamycin, vincristine, and prednisolone regimen was initiated, and complete remission was achieved. Notably, the sellar mass returned to normal size with improved function. We reviewed 32 patients with primary parasellar lymphoma. In affected sites, both sellar and pituitary stalk (6.7%), both hypothalamus and pituitary stalk (6.7%), only sellar (63.3%), only pituitary stalk (6.7%), only hypothalamus (13.3%), and only clivus (3.3%) were observed. In hypothalamus-pituitary dysfunction, both anterior and posterior dysfunction (20.7%), only anterior dysfunction (58.6%), only posterior dysfunction (3.4%), and no dysfunction (17.2%) were observed. It seemed that hypothalamic lesion is related to both anterior and posterior dysfunction, while sellar lesion is related to mainly anterior dysfunction. In cranial nerve dysfunction, 2nd nerve dysfunction (45.2%) and 6th nerve dysfunction (35.5%) were frequently observed. It seemed that sellar lesion is related to both 2nd and 6th nerve dysfunction, while hypothalamic lesion is related to mainly 2nd nerve dysfunction.

New approach to diagnose arytenoid dislocation and subluxation using three-dimensional computed tomography.

Understanding the complex three-dimensional (3D) arrangement of the arytenoid cartilage is necessary for diagnosing arytenoid dislocation (AD) and arytenoid subluxation (AS). We examined the 3D arrangements of AD and AS (AD/AS) cases by region and considered their new diagnoses. This retrospective study included 2 patients with AD, 10 with AS, and 23 with unilateral vocal fold paralysis (UVFP) for comparison. The etiologies were intubation-induced and idiopathic. We classified the AD/AS position into four joint regions: mediocaudal, laterocaudal, mediocranial, and laterocranial. We generated 3D computed tomography (3DCT) images during rest and phonation to analyze functional movements. We attempted to compare the endoscopic findings and 3DCT images of patients with UVFP and AD/AS. To examine the joint status, we especially focused on the position and movements of the muscular process (MP) on the joint because the arytenoid facet is mainly located on the back of the MP. We were able to obtain endoscopic and 3DCT findings characteristic of each AD/AS region. The dislocated MPs were localized to the mediocaudal, mediocranial, and laterocranial regions. Two AD cases were diagnosed due to complete separation of the joint surfaces during rest and phonation. The finding of MPs displacing partially outside the cricoid facet is common to both severe UVFP and AS. The most important differentiation point was that the MP in UVFP cases was located on both the medial and lateral side regions of the joint, but that of AS was on one side region only. Furthermore, no cases of passive gliding movements characteristic of UVFP that have been described previously by us were observed in AD/AS cases. AD can be diagnosed by findings of complete joint separation. AS can be diagnosed based on positions and movements distinct from those of UVFP.

Dimethylarsinous acid disturbs cytokinesis.

Dimethylarsinous acid (DMAIII) induced tetraploidy and multinucleation in V79 cells. While the highest yield of tetraploids and multinucleated cells was at 1.25 microM of DMAIII, the mitotic index was highest at DMAIII 2.5 microM due to mitotic arrest. Mitosis was not observed at 5 microM. We observed V79 cells treated with DMAIII and visualized with rhodamine-phalloidin. Abnormal actin location was observed in the dividing cells which were treated with DMAIII and visualized with rhodamine-phalloidin. These findings suggested strongly that DMAIII inhibits not only formation of the normal mitotic spindle but cytokinesis and induces the formation of multinuclear cells.

IL-12 and IL-18 induction and subsequent NKT activation effects of the Japanese botanical medicine Juzentaihoto.

In this study, we first measured some cytokine concentrations in the serum of patients treated with Juzentaihoto (JTT). Of the cytokines measured interleukin (IL) -18 was the most prominently up-regulated cytokine in the serum of patients under long term JTT administration. We next evaluated the effects of JTT in mice, focusing especially on natural killer T (NKT) cell induction. Mice fed JTT were compared to control group ones. After sacrifice, the liver was fixed, embedded and stained. Transmission electron microscope (TEM) observations were performed. Although the mice receiving the herbal medicine had same appearance, their livers were infiltrated with massive mononuclear cells, some of which were aggregated to form clusters. Immunohistochemical staining revealed that there was abundant cytokine expression of IL-12 and IL-18 in the liver of JTT treated mice. To clarify what the key molecules that induce immunological restoration with JTT might be, we next examined in vitro lymphocyte cultures. Mononuclear cells isolated and prepared from healthy volunteers were cultured with and without JTT. Within 24 hours, JTT induced the IL-12 and IL-18 production and later (72 hours) induction of interferon (IFN)-gamma. Oral administration of JTT may induce the expression of IL-12 in the early stage, and IL-18 in the chronic stage, followed by NKT induction. Their activation, following immunological restoration could contribute to anti-tumor effects.

Estrogen formulations and beauty care practices in Japanese women.

PURPOSE: Traditionally, oral estrogens have been used for hormone replacement therapy. However, in Japan, additional estrogen formulations have been used, including transdermal patches and transdermal gels. The latter have a unique commonality with cosmetics because both of them are applied to the skin. Beauty care is one of the most important lifestyle factors for women, and it has been reported that the amount of attention paid to beauty care has an effect in determining whether or not women will choose to undergo HRT during menopause. Therefore, our study focused on estrogen formulations and beauty care practices. PATIENTS AND METHODS: Fifty women who use hormone replacement therapy were recruited from the outpatient clinic of Tohoku University Hospital. They were treated with oral conjugated estrogen (n = 11), transdermal 17ß-estradiol patch (n = 11), and transdermal 17ß-estradiol gel (n = 28). They completed a questionnaire to assess their lifestyle (beauty care practices and exercise habits) and their compliance. The transdermal gel users were further interviewed about their subjective impressions regarding "smell", "sticky feeling", "spreadability", and "irritation" on the skin using a five-grade scale. RESULTS: There were no differences in the usability of medicines and patient compliance among the estrogen formulations. We observed a positive tendency between the level of beauty care and transdermal gel use (P = 0.0645, ordinary logistic regression analysis). The gel users placed top priority on a lack of "sticky feeling" but the subjective impression regarding "sticky feeling" was worst among the four factors (P < 0.01, Steel-Dwass test). Correspondence analysis showed that the subjective impressions of transdermal gel corresponding to usability in the range of "moderate" to "very good" and "sticky feeling" greatly affected the usability of the formulation. CONCLUSION: These results suggest that the level of attention to beauty care plays some role in the choice of estrogen formulations.

Severe duodenal hemorrhage induced by Lugol's solution administered for thyroid crisis treatment.

Lugol's solution is an iodinated agent used for treating thyroid crisis. It is primarily used in diagnostic tests for esophageal diseases. However, Lugol's solution can cause local mucosal injury and hemorrhage. We report, for the first time, a case of 34-year-old man who exhibited severe duodenal hemorrhage induced by Lugol's solution that was used to treat thyroid crisis. The quantity of Lugol's solution used for treating thyroid crisis is much higher than that used for mucosal disease investigation. Clinical practitioners should be aware of gastrointestinal hemorrhage when using Lugol's solution for the treatment of thyroid crisis.