The discovery of azetidine-piperazine di-amides as potent, selective and reversible monoacylglycerol lipase (MAGL) inhibitors.

Monoacylglycerol lipase (MAGL) is the enzyme that is primarily responsible for hydrolyzing the endocannabinoid 2-arachidononylglycerol (2-AG) to arachidonic acid (AA). It has emerged in recent years as a potential drug target for a number of diseases. Herein, we report the discovery of compound 6g from a series of azetidine-piperazine di-amide compounds as a potent, selective, and reversible inhibitor of MAGL. Oral administration of compound 6g increased 2-AG levels in rat brain and produced full efficacy in the rat complete Freund's adjuvant (CFA) model of inflammatory pain.

Effects of Educational Intervention on State Anxiety and Pain in People Undergoing Spinal Surgery: A Randomized Controlled Trial.

Preoperative educational intervention for anxiety and pain affects patients undergoing spinal surgery. The effects, however, have never been examined using randomized controlled designs. To investigate the effects of education on anxiety and pain for patients undergoing spinal surgery, a randomized trial with block design was used. Patients were recruited from a medical center in central Taiwan. We invited 90 patients to participate in this study. Inclusion criteria were (a) age ≥20 years, (b) voluntary participation, (c) able to understand Taiwanese Mandarin Chinese or Taiwanese, and (4) no hearing or vision impairments after using aids. Patients (n = 86) undergoing lumbar spinal surgery were randomized into either an Intervention group (using educational intervention; n = 43) or a Control group (n = 43); four patients voluntarily dropped out after surgery (one in Intervention group; three in Control group). Patients had their anxiety (using the State-Trait Anxiety Inventory; STAI) and pain (using a visual analog scale) measured the day before surgery, 30 minutes before surgery, and the day after surgery. After controlling for demographics, the adjusted anxiety and pain levels were significantly lower for the Intervention group: mean STAI scores were 52.67 at baseline and 47.54 at 30 minutes before surgery (p < .001); mean pain scores were 6.07 at baseline and 5.28 on day after surgery (p < .001). Preoperative educational intervention is effective in informing patients undergoing spinal surgery that can lead to a reduction in pain, anxiety, and fear postoperatively.

Sirolimus induces apoptosis and reverses multidrug resistance in human osteosarcoma cells in vitro via increasing microRNA-34b expression.

AIM: Multi-drug resistance poses a critical bottleneck in chemotherapy. Given the up-regulation of mTOR pathway in many chemoresistant cancers, we examined whether sirolimus (rapamycin), a first generation mTOR inhibitor, might induce human osteosarcoma (OS) cell apoptosis and increase the sensitivity of OS cells to anticancer drugs in vitro. METHODS: Human OS cell line MG63/ADM was treated with sirolimus alone or in combination with doxorubicin (ADM), gemcitabine (GEM) or methotrexate (MTX). Cell proliferation and apoptosis were detected using CCK-8 assay and flow cytometry, respectively. MiRNAs in the cells were analyzed with miRNA microarray. The targets of miR-34b were determined based on TargetScan analysis and luciferase reporter assays. The expression of relevant mRNA and proteins was measured using qRT-PCR and Western blotting. MiR-34, PAK1 and ABCB1 levels in 40 tissue samples of OS patients were analyzed using qRT-PCR and in situ hybridization assays. RESULTS: Sirolimus (1-100 nmol/L) dose-dependently suppressed the cell proliferation (IC50=23.97 nmol/L) and induced apoptosis. Sirolimus (10 nmol/L) significantly sensitized the cells to anticancer drugs, leading to decreased IC50 values of ADM, GEM and MTX (from 25.48, 621.41 and 21.72 µmol/L to 4.93, 73.92 and 6.77 µmol/L, respectively). Treatment of with sirolimus increased miR-34b levels by a factor of 7.5 in the cells. Upregulation of miR-34b also induced apoptosis and increased the sensitivity of the cells to the anticancer drugs, whereas transfection with miR-34b-AMO, an inhibitor of miR-34b, reversed the anti-proliferation effect of sirolimus. Two key regulators of cell cycle, apoptosis and multiple drug resistance, PAK1 and ABCB1, were demonstrated to be the direct targets of miR-34b. In 40 tissue samples of OS patients, significantly higher miR-34 ISH score and lower PAK5 and ABCB1 scores were detected in the chemo-sensitive group. CONCLUSION: Sirolimus increases the sensitivity of human OS cells to anticancer drugs in vitro by up-regulating miR-34b interacting with PAK1 and ABCB1. A low miR-34 level is an indicator of poor prognosis in OS patients.

7-(4-Alkylidenylpiperidinyl)-quinolone bacterial topoisomerase inhibitors.

Novel antibacterial fluoroquinolone agents bearing a 4-alkylidenylpiperidine 7-position substituent are active against quinolone-susceptible and quinolone-resistant gram-positive bacteria, including Streptococcus pneumoniae and MRSA. Analogs 22b, 23c, and 24 demonstrated superior in vitro and in vivo efficacy to ciprofloxacin against these cocci.

The Influence of a Celebrity Chef on Customer Repurchase Behavior: Empirical Study of Taiwan's F&B Industry During COVID-19 Pandemic.

The spread of COVID-19 pandemic has inflicted severe blows on the global hospitality industry. In Taiwan, revenue from the food and beverage (F&B) department has decreased by more than 90%. This study aims to understand whether celebrity chefs can effectively help and enhance their corporates' business performance under COVID-19's severe impacts via leveraging their personal brand value, explores the influence of a celebrity chef on customer repurchase behavior during the epidemic and examines whether such a chef has a mediation effect on the relationship between corporate brand and customer satisfaction. The primary data were collected from the respondents through online questionnaire in Taiwan to get 245 respondents as a sample size of the research from Nov. 10 to Nov. 25 in 2021, and through validity and reliability analysis that processed by statistical software using factor analysis and structural equation modeling to see if celebrity chefs' personality branding could influence customer repurchase behavior, and also examine the relationship between corporate brand and celebrity chef. The findings show that corporate brand enhances both a celebrity chef's personal brand and customer satisfaction, and that a celebrity chef has a positive effect on both customer satisfaction and loyalty, which can partially mediate the effect of corporate brand; furthermore, a celebrity chef has a positive effect on customer repurchase behavior. In Taiwan relative studies into aspects of a celebrity chef's effect on consumer behavior are limited, and so this research offers new insights into the celebrity chef phenomenon there as well as elsewhere.

Obesity Affects Short-Term Renal Function After Renal Transplantation.

BACKGROUND: The literature has shown a significant association between body mass index (BMI) and patient and graft outcomes after renal transplantation. The purpose of this study was to reveal the effect of obesity on graft function in a Taiwanese kidney transplant cohort. METHODS: Two hundred consecutive patients who received kidney transplantation were enrolled in our study. Eight pediatric cases were excluded due to differing definitions of BMI among children. According to the national obesity criteria, these patients were divided into underweight, normal, overweight, and obese groups. Their estimated glomerular filtration rate (eGFR) was compared accordingly using t tests. Cumulative graft and patient survivals were calculated using Kaplan-Meier analysis. A P value of ≤ .05 was considered significant. RESULTS: The mean age of our cohort (105 men and 87 women) was 45.3 years. There was no significant difference comparing biopsy-proven acute rejection, acute tubular necrosis, and delayed graft function between the obese and nonobese groups (P values: .293, .787, and .304, respectively). Short-term eGFR was inferior in the overweight group, but this effect was insignificant beyond 1 month. The 1-month and 3-month eGFR were found to be correlated with BMI groups (P = .012 and P = .008, respectively) but not significant after 6 months post-kidney transplantation. CONCLUSIONS: Our study found that short-term renal function was affected by obesity and being overweight, possibly due to the higher prevalence of diabetes and dyslipidemia in obese patients and the increased surgical difficulty.

Clonality and prognostic implications of p53 and epidermal growth factor receptor somatic aberrations in multiple primary lung cancers.

PURPOSE: For treatment decision and prognostic applications, we evaluated p53/epidermal growth factor receptor (EGFR) somatic aberrations in multiple primary lung cancers to differentiate multifocal tumors from intrapulmonary metastasis. EXPERIMENTAL DESIGN: Fifty-eight multiple primary lung cancers of 1,037 patients in a 10-year period were identified to investigate somatic mutations and altered expression of p53 and EGFR for clonality assessment. Genomic DNA was extracted from microdissected cells of paraffin-embedded multiple primary lung cancer tissues. Overexpression and somatic mutations in exons of p53 (exons 5-8) and tyrosine kinase domain of EGFR (exons 18-22) were examined by immunohistochemical staining and DNA sequencing, respectively. RESULTS: High frequency of somatic mutations in p53 (33 of 58, 56.9%) and/or EGFR (44 of 58, 75.9%) resulted in high discrimination rate of tumor clonality (50 of 58, 86.2%) of multiple primary lung cancers. Twenty-two cases (37.9%) were assessed as having the same clonality and 28 cases (48.3%) were determined as having different clonality, which further supported the carcinogenic theory of field cancerization. Notably, the occurrence of lymph node metastasis was more commonly observed in tumors with the same clonality (P = 0.045) and was associated with poor patient 5-year survival rate (P = 0.001). However, no correlation was found between tumor clonality and patient survival (P = 0.630). The EGFR somatic aberrations in 58 multiple primary lung cancers, including vascular invasion associated with EGFR overexpression (P = 0.012) and mutation (P = 0.025), further suggested the potential benefits of target therapy of inoperable multiple primary lung cancers. CONCLUSIONS: Our results suggest that analysis of somatic alterations in p53 and EGFR can significantly improve the clonality assessment and impact management of multiple primary lung cancer patients.

Luteolin inhibited proliferation and induced apoptosis of prostate cancer cells through miR-301.

Luteolin is a falvonoid compound derived from Lonicera japonica Thunb. Numerous reports have demonstrated that luteolin has anticancer effects on many kinds of tumors. This study investigated the effects of luteolin on prostate cancer (PCa), assessing the PC3 and LNCaP cells. The cell viability and apoptosis were assessed by performing Cell Counting Kit-8 assay and Annexin V-fluorescein isothiocyanate/propidium iodide double staining. Luteolin was found to inhibit androgen-sensitive and androgen-independent PCa cell lines' growth and induced apoptosis. To uncover the exact mechanisms and molecular targets, microRNA (miR) array analysis was performed. miR-301 was found to be markedly downregulated. Then, the expression of miR-301 was retrospectively analyzed in the primary PCa tissues by quantitative reverse transcription polymerase chain reaction and in situ hybridization methods. According to the quantitative reverse transcription polymerase chain reaction results of miR-301, the 54 PCa patients were divided into two groups: high and low miR-301 groups. The division indicator is a relative expression ≥5. Compared to the low-expression group, high miR-301 expression was associated with a significantly shorter overall survival (P=0.029). The proapoptotic gene, DEDD2, was predicted to be the direct target of miR-301. It was clarified in accordance with bioinformatics and luciferase activity analyses. The overexpression of miR-301 by plasmid decreased the luteolin effect. Taken together, these results suggest that luteolin inhibits PCa cell proliferation through miR-301, the poor predictive factor of PCa.

Glutamate-gated chloride channels inhibit juvenile hormone biosynthesis in the cockroach, Diploptera punctata.

Juvenile hormone (JH) synthesized and released from endocrine gland corpus allatum (CA) plays an important role in insect metamorphosis, vitellogenesis and reproduction. Glutamate is a major neurotransmitter in the nervous system and its activated receptors possess excitatory and inhibitory forms in muscle fibers of invertebrates. Previously, we have shown that the rise of intracellular calcium through excitatory glutamate receptors, N-methyl-d-aspartate (NMDA) and non-NMDA-type channels stimulates JH synthesis in the cockroach, Diploptera punctata. Here, we demonstrate the occurrence of inhibitory chloride permeable glutamate (GluCl) receptors on CA cell membranes. Application of the GluCl channel activators, ibotenic acid (Ibo) and ivermectin, but not gamma-aminobutyric acid caused a decline in JH synthesis in glands of either high or low activity during the gonadotrophic cycle. Also, while recording the membrane potential of the isolated whole CA glands intracellularly, Ibo induced a hyperpolarizated response. Both changes in the membrane potential and inhibition of JH synthesis could be abolished by the application of the chloride channel blocker picrotoxin. Finally, we found both excitatory and inhibitory glutamate receptors cause antagonistic effects on rates of JH synthesis. These results indicate a novel function of GluCl channels in the inhibition of JH synthesis that could be a potential pathway for developing a new generation of insecticides.

The Prognostic Value of Forkhead Box P3 Expression in Operable Breast Cancer: A Large-Scale Meta-Analysis.

BACKGROUND: Recent studies have shown that the forkhead box P3 (FOXP3) protein has a prognostic role in breast cancer. However, these results are controversial. Therefore, the aim of this meta-analysis was to clarify the prognostic role of FOXP3 expression in operable breast cancer cases. METHODS: Eligible studies describing the use of FOXP3 as a prognostic factor for operable breast cancer cases were identified. Clinicopathological features, disease-free survival (DFS), and overall survival (OS) data were collected from these studies and were analyzed using Stata software. RESULTS: A total of 16 articles containing data from 13,217 breast cancer patients met the inclusion criteria established for this study. The subsequent meta-analysis that was performed showed that high levels of FOXP3 are not significantly associated with DFS and OS with significant heterogeneity. An additional subgroup analysis demonstrated that intratumoral FOXP3+ regulatory T cells (Tregs) were positively correlated with adverse clinicopathological parameters, yet they did not show an association with DFS or OS. For tumor cells, the pooled results revealed that FOXP3 is significantly associated with DFS (HR: 2.55, 95% CI: 1.23-5.30) but is not associated with clinicopathological parameters or OS. We also observed a significant correlation between FOXP3 expression and survival in the estrogen receptor-positive (ER)+ subgroup (HR: 1.83, 95% CI: 1.36-2.47 for DFS, HR: 1.87, 95% CI 1.28-2.73 for OS), in the Asian region (HR: 1.98, 95% CI: 1.56-2.50 for DFS, HR: 1.93, 95% CI: 1.12-3.35 for OS) and using the median as the FOXP3-positive cut-off value (HR: 1.94, 95% CI: 1.57-2.39 for DFS, HR: 2.06; 95% CI: 1.36-3.11 for OS). CONCLUSION: This meta-analysis indicates that a prognostic role for FOXP3 expression in operable breast cancer cases depends on the FOXP3-positive region, ER status, geographic region and the FOXP3-positive cut-off value.

Ionotropic glutamate receptors mediate juvenile hormone synthesis in the cockroach, Diploptera punctata.

By monitoring changes in the cytosolic [Ca2+](i) and rates of juvenile hormone (JH) synthesis in response to L-glutamate agonists and antagonists, we identified and characterized glutamate receptor subtypes in corpus allatum (CA) cells of the cockroach, Diploptera punctata. During the first ovarian cycle, corpora allata exhibited a cycle of changes in sensitivity to L-glutamate correlated to cyclic changes in rates of JH synthesis. When exposed to 60 microM L-glutamate in vitro, the active corpora allata of day-4 mated females produced 60% more JH, while inactive corpora allata at other ages showed 10-20% stimulatory response. Pharmacological characterization using various L-glutamate receptor agonists and antagonists indicated that several ionotropic subtypes of L-glutamate receptors were present in the CA. The CA showed an increase in rates of JH synthesis in response to NMDA, kainate, and quisqualate, but not to AMPA in both L-15 medium and minimum incubation medium. In contrast, applications of the metabotropic receptor-specific agonist trans-ACPD failed to elicit a change in the cytosolic [Ca2+](i) and JH production. An elevation of cytosolic calcium concentration, followed by 20-30% rise in JH production, was observed when active CA cells were exposed to 10-40 microM kainate. Kainate had no stimulatory effect on JH synthesis in calcium-free medium. The kainate-induced JH synthesis was blocked by 20 microM CNQX but was not affected by 20 microM NBQX. Kainate-stimulated JH production was not suppressed by MK-801 (a specific blocker of NMDA-receptor channel), nor was NMDA-stimulated JH production affected by CNQX (a specific antagonist of kainate receptor). These data suggest that active CA cells are stimulated to synthesize more JH by a glutamate-induced calcium rise via NMDA-, kainate- and/or quisqualate-sensitive subtypes of ionotropic L-glutamate receptors. The metabotropic-subtype and ionotropic AMPA-subtype L-glutamate receptors are unlikely to be present on active CA cells.

Health related quality of life of long-term kidney transplantation recipients.

BACKGROUND: Health related quality of life (HRQOL) is an important issue for long-term kidney transplantation (KT) patients. Nevertheless, few studies have focused on long-term HRQOL in KT recipients with a functional graft. Thus, the aim of this study is to describe the long-term (10-year) HRQOL of KT recipients. METHODS: This is a cross-sectional and correlational design. The Medical Outcome Survey (MOS SF-36) questionnaire was used to collect data on HRQOL. The data were collected from November 2009 to September 2010 at a medical center in Northern Taiwan. RESULTS: A total of 88 patients were interviewed. The mean years after transplantation was 14.48 (SD = 3.9). The mean score of each of the HRQOL subscales ranged from 59.4 to 82.5. The mean scores on the bodily pain (BP) subscale were the highest and, on the general health (GH) subscale, the lowest. Compared to the general population, with the exception of the BP subscale, long-term KT patients had a lower mean score (poorer HRQOL) on all subscales. Age, gender, serum creatinine level, and employment status were significantly related to HRQOL. CONCLUSION: HRQOL of long-term KT patients was, overall, poorer than that of the general population. When comparing the HRQOL of KT patients with that of the general population, one should take into account age and gender. Finally, the physical, psychological, and social adjustment domains of HRQOL of KT patients warrant further attention.

Identification of oncogenic point mutations and hyperphosphorylation of anaplastic lymphoma kinase in lung cancer.

The oncogenic property of anaplastic lymphoma kinase (ALK) plays an essential role in the pathogenesis of various cancers and serves as an important therapeutic target. In this study, we identified frequent intragenic loss of heterozygosity and six novel driver mutations within ALK in lung adenocarcinomas. Overexpression of H694R or E1384K mutant ALK leads to hyperphosphorylation of ALK, and activation of its downstream mediators STAT3, AKT, and ERK resulted in enhanced cell proliferation, colony formation, cell migration, and tumor growth in xenograft models. Furthermore, the activated phospho-Y1604 ALK was increasingly detected in 13 human lung cancer cell lines and 263 lung cancer specimens regardless of tumor stages and types. Treatment of two different ALK inhibitors, WHI-P154 and NVP-TAE684, resulted in the down-regulation of aberrant ALK signaling, shrinkage of tumor, and suppression of metastasis and significantly improved survival of ALK mutant-bearing mice. Together, we identified that novel ALK point mutations possessed tumorigenic effects mainly through hyperphosphorylation of Y1604 and activation of downstream oncogenic signaling. The upregulated phospho-Y1604 ALK could serve as a diagnostic biomarker for lung cancer. Furthermore, targeting oncogenic mutant ALKs with inhibitors could be a promising strategy to improve the therapeutic efficacy of fatal lung cancers.

Assessment of xerostomia and its impact on quality of life in head and neck cancer patients undergoing radiotherapy, and validation of the Taiwanese version of the xerostomia questionnaire.

The purposes of this study were to (a) explore the impact of xerostomia and saliva flow on quality of life and (b) validate the Taiwanese version of the Xerostomia Questionnaire (XQ) for patients undergoing radiotherapy (RT) for head and neck cancer in Taiwan. This was a prospective longitudinal study. Instruments consisted of the Xerostomia Questionnaire-Taiwan version (XQ-T) and the Medical Outcomes Study Short Form-36 Taiwan Version. Salivary output was measured by collecting unstimulated whole saliva. The questionnaires and measurements of salivary output were completed before RT was initiated and at two, four, six, and eight weeks after RT had started. Changes in xerostomia scores, quality of life, saliva flow, and predictors of quality of life over time were examined by using general estimating equations. The XQ-T is the first xerostomia measurement instrument developed for use with Taiwanese cancer patients and demonstrated excellent reliability and validity. Saliva flow was significantly correlated with XQ-T scores at two, four, six, and eight weeks after RT had started, but not before RT had begun. Saliva flow and quality-of-life scores significantly diminished and xerostomia scores significantly increased over the eight-week period. Saliva flow and XQ-T scores significantly predicted quality of life, after adjusting for the maturation effect. The results of this study show that the XQ-T is the first xerostomia measurement instrument to be developed for Taiwanese cancer patients and demonstrates excellent reliability and validity.

A new tumor suppressor DnaJ-like heat shock protein, HLJ1, and survival of patients with non-small-cell lung carcinoma.

BACKGROUND: We previously identified DnaJ-like heat shock protein (HLJ1) as a gene associated with tumor invasion. Here, we investigated the clinical significance of HLJ1 expression in non-small-cell lung cancer (NSCLC) patients and its role in cancer progression. METHODS: We induced HLJ1 overexpression or knockdown in human lung adenocarcinoma CL1-5 cells and analyzed cell proliferation, anchorage-independent growth, in vivo tumorigenesis, cell motility, invasion, and cell cycle progression. Expression of genes that act downstream of HLJ1 was examined by DNA microarray analysis, pathway analysis, and western blotting. We measured HLJ1 expression in tumors and adjacent normal tissues of 71 NSCLC patients by quantitative reverse transcription-polymerase chain reaction. Associations between HLJ1 expression and disease-free and overall survival were determined using the log-rank test and multivariable Cox proportional hazards regression analysis. Validation was performed in an independent cohort of 56 NSCLC patients. Loss of heterozygosity (LOH) mapping of the HLJ1 locus was analyzed in 48 paired microdissected NSCLC tumors. All statistical tests were two-sided. RESULTS: HLJ1 expression inhibited lung cancer cell proliferation, anchorage-independent growth, tumorigenesis, cell motility, and invasion, and slowed cell cycle progression through a novel STAT1/P21(WAF1) pathway that is independent of P53 and interferon. HLJ1 expression was lower in tumors than in adjacent normal tissue in 55 of 71 patients studied. NSCLC patients with high HLJI expressing tumors had reduced cancer recurrence (hazard ratio [HR] = 0.47; 95% confidence interval [CI] = 0.23 to 0.93; P = .03) and longer overall survival (HR = 0.38; 95% CI = 0.16 to 0.89; P = .03) than those with low-expressing tumors. Validation in the independent patient cohort confirmed the association between HLJ1 expression and patient outcome. LOH mapping revealed high frequencies (66.7% and 70.8%) of allelic loss and microsatellite instability (87.5% and 95.2%) of the HLJ1 locus at chromosome 1p31.1. CONCLUSIONS: HLJ1 is a novel tumor suppressor in NSCLC, and high HLJ1 expression is associated with reduced cancer recurrence and prolonged survival of NSCLC patients.