ABSTRACT Tigecycline In-vitro Surveillance in Taiwan (TIST), initiated in 2006, is a nationwide surveillance programme designed to monitor longitudinally the in-vitro activity of tigecycline against ...commonly encountered resistant bacteria. This study compared the in-vitro activity of tigecycline against clinical isolates of resistant Gram-negative bacteria determined by the broth microdilution and Etest methods. A total of 622 isolates were collected from patients treated at 20 teaching hospitals. Tigecycline had excellent in-vitro activity against extended-spectrum β-lactamase (ESBL)-producing Escherichia coli ( N = 275) with MIC90 0.5 μg/mL and a 99.6% susceptibility rate, and also against ESBL-producing Klebsiella pneumoniae ( N = 324) with MIC90 2 μg/mL and a 98.5% susceptibility rate. For ESBL-producing Proteus mirabilis ( N = 15) the MIC90 was 4 μg/mL with a 73.3% susceptibility rate. For ESBL-producing Klebsiella oxytoca ( N = 8) the MIC50 and MIC90 were 0.5 and 1 μg/mL, respectively, with a 100% susceptibility rate. Limited agreement (<80%) was found between the broth microdilution and the Etest methods when determining the in-vitro activity of tigecycline against ESBL- producing K. pneumoniae and K. oxytoca.
We examined the influences of patients' background characteristics on the frequency of performing five diabetes self-care behaviours that 185 Taiwanese outpatients reported. All patients had type 2 ...diabetes diagnosed for more than a year and attended an outpatient clinic at a large university hospital where they had received at least one dietitian- led individual nutrition education session and one nurse-led diabetes education session during the course of their care. Seventy nine percent of the patients regularly (defined as responses often or always on the questionnaire) took their medications and over half followed recommended meal plans and exercised, but fewer performed foot care (38%) or checked their blood glucose levels (20%) regularly. The associations between patients' demographics and disease-related characteristics and their performance of self-care behaviours were assessed with logistic regression. Although checking blood glucose levels and performing diabetes foot care were unrelated to any clinical outcome examined, patients who took their diabetes medications had lower hemoglobin A1c levels and fewer chronic complications than those who did not. Furthermore, patients who followed a diabetes meal plan also had lower hemoglobin A1c levels, and those who exercised regularly had healthier body mass indices (BMI) than those who did not.
The effects of patient characteristics on reported adherence to dietary self-care behaviours in 184 Taiwanese outpatients 40 years or older with type 2 diabetes was assessed. Patient characteristics ...included the presence of predisposing factors affecting diabetes adherence (knowledge and attitudes about the disease, self-efficacy, and the absence of psychological problems), enabling factors (understanding of diabetes and environmental factors affecting it), and reinforcing factors (presence of medical and social support) which were evaluated using a 72 item self-administered questionnaire with 8 subscales. Adherence was assessed by patients' reports of carrying out 7 self-care behaviours (following a diabetic meal plan, following the diabetes exchange system, eating meals providing the same amount of carbohydrate every day, counting carbohydrates, reducing dietary fat, consuming high fiber foods, and keeping a daily food record).
Reported adherence ranged from 17% to 74%. No single predisposing, enabling, or reinforcing factor predicted adherence to all of the dietary self-care behaviours. However, more self-efficacy, better understanding, and a better attitude toward diabetes were associated with performing five or more of the dietary self-care behaviours examined. With respect to specific self-care behaviours, women were more likely than men to count carbohydrates (OR=5.75) and reduce fat in their diets (OR=2.57). Patients who attended more nutrition education sessions were more likely to follow diabetes meal plans (OR=2.11) and the diabetes exchange system (OR=3.07). Efforts are needed to encourage providers to teach diabetes self-care behaviours to patients and to capitalize upon demographic and psychosocial characteristics that can enhance patient adherence.
Background 3T3-L1 cells are widely used to study adipogenesis and insulin response. Their adipogenic potential decreases with time in the culture. Expressing exogenous genes in 3T3-L1 cells can be ...challenging. This work tries to establish and characterize an alternative model of cultured adipocytes that is easier to work with than the 3T3-L1 cells. Methodology/Principal Findings Induced cells were identified as adipocytes based on the following three characteristics: (1) Accumulation of triglyceride droplets as demonstrated by oil red O stain. (2) Transport rate of 2-deoxyglucose increased after insulin stimulation. (3) Expression of fat specific genes such as Fabp4 (aP2), Slc2a4 (Glut4) and Pparg (PPARgamma). Among the cell lines induced under different conditions in this study, only NIH/3T3 cells differentiated into adipocytes after prolonged incubation in 3T3-L1 induction medium containing 20% instead of 10% fetal bovine serum. Rosiglitazone added to the induction medium shortened the incubation period from 14 to 7 days. The PI3K/AKT pathway showed similar changes upon insulin stimulation in these two adipocytes. C/EBPalpha mRNA was barely detectable in NIH/3T3 adipocytes. NIH/3T3 adipocytes induced in the presence of rosiglitazone showed higher 2-deoxyglucose transport rate after insulin stimulation, expressed less Agt (angiotensinogen) and more PPARgamma. Knockdown of C/EBPalpha using shRNA blocked 3T3-L1 but not NIH/3T3 cell differentiation. Mouse adipose tissues from various anatomical locations showed comparable levels of C/EBPalpha mRNA. Conclusions/Significance NIH/3T3 cells were capable of differentiating into adipocytes without genetic engineering. They were an adipocyte model that did not require the reciprocal activation between C/EBPalpha and PPARgamma to differentiate. Future studies in the C/EBPalpha independent pathways leading to insulin responsiveness may reveal new targets to diabetes treatment.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Abstract
Esophageal cancer is the 5th leading cause of cancer death among male population in Eastern Asia. The esophageal squamous cell carcinoma (ESCC) is the major type of esophageal cancer in ...Taiwan. Due to the poor overall 5-year survival rate, effective treatment strategies are required to improve the long-term survival rate. miRNAs are important gene regulators that may be dysregulated in many cancer types. The expression level of miR-338-5p was found to be downregulated in esophageal tumor tissues compared to the adjacent nontumor tissues. However, the functions and the potential target genes of miR-338-5p in ESCC are still unclear. In this study, we transfected miR-338-5p mimic into the ESCC cell line, CE-81T/VGH, which expresses low levels of miR-338-5p. The results showed that miR-338-5p mimic can inhibit cell proliferation, migration, and colony formation. In order to identify the target genes of miR-338-5p, microarray analysis was performed to select significantly downregulated genes in the miR-338-5p mimic transfected CE-81T/VGH cells. Moreover, miRSystem was used to predict the miR-338-5p putative target genes. By cross-comparing the results from the microarray and miRSystem analysis, FERMT2 was identified as the most likely candidate gene, and it was validated by real-time PCR. The function of FERMT2 was further studied in the CE-81T/VG cell. Silencing of FERMT2 can inhibit cell proliferation, migration, and colony formation in the ESCC cells. The luciferase activity assays results showed the direct binding between miR-338-5p and 3'UTR region of FERMT2. To further confirm that miR-338-5p can go through FERMT2 to regulate cell functions, the “rescue” experiments were performed. As the results, overexpression of FERMT2 after miR-338-5p mimic transfection could restore cell migration and cell proliferation. Moreover, it has been reported that FERMT2 can increase chemoresistance in glioma cells. Therefore, the effects of miR-338-5p and FERMT2 on drug resistance were studied, and the results showed that miR-338-5p mimic transfection or FERMT2 silencing can increase cisplatin-induced cytotoxicity in the CE-81T/VGH cells. These data indicated that miR-338-5p can inhibit cell proliferation, migration, colony formation, and enhance cisplatin sensitivity in the ESCC cells by targeting FERMT2 expression. Therefore, miR-338-5p plays an important role in the ESCC cells via repressing FERMT2 and it could be used as a new research target for cancer treatment.
Citation Format: Wen-chun Lin, Yao-chin Hsieh, Li-han Chen, Pei-wen Yang, Chi-Cheng Huang, Liang-chuan Lai, Jang-ming Lee, Eric Yao-yu Chuang, Mong-hsun Tsai. miR-338-5p modulates growth and chemoresistance of esophageal cancer cell via targeting FERRMT2 abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4432.
In this ongoing longitudinal study of adolescent twins/sibling pairs and their parents in Taipei, we aimed to investigate the genetic and environmental influences on adolescent behavioral ...development, metabolic risk factors, and the associations between the two. Special focus is on anxious depression, metabolic profile, and hormonal factors such as cortisol and leptin. The first wave of assessment was completed during the period of 2002 to 2005 and included 192 twin pairs, 6 triplets, 56 sibling pairs and their first-degree relatives (484 parents and 142 siblings). We are currently in the process of a second wave assessment as follow-up. Dimensional psychological assessments using self-report questionnaires, as well as categorical assessments based on semistructured clinical interviews, were performed. All subjects received a 2-hour oral glucose tolerance test at the examination site. The metabolic phenotypes including body mass index, blood pressure, levels of glucose, insulin, and lipid profile as well as related hormonal levels were measured. Zygosity was determined using DNA, except for a few twins whose DNA was not available. Such a combination of detailed psychological assessments and metabolic function tests is expected to help shed light on the interrelation of psychological well-being and metabolic functioning.
Diabetic kidney disease(DKD) is one of the most common diabetic complications, as well as the leading cause of chronic kidney disease and end-stage renal disease around the world. To prevent the ...dreadful consequence, development of new assays for diagnostic of DKD has always been the priority in the research field of diabetic complications. At present, urinary albumin-to-creatinine ratio and estimated glomerular filtration rate(eG FR) are the standard methods for assessing glomerular damage and renal function changes in clinical practice. However, due to diverse tissue involvement in different individuals, the so-called "non-albuminuric renal impairment" is not uncommon, especially in patients with type 2 diabetes. On the other hand, the precision of creatinine-based GFR estimates is limited in hyperfiltration status. These facts make albuminuria and eG FR less reliable indicators for early-stage DKD. In recent years, considerable progress has been made in the understanding of the pathogenesis of DKD, along with the elucidation of its genetic profiles and phenotypic expression of different molecules. With the help of ever-evolving technologies, it has gradually become plausible to apply the thriving information in clinical practice. The strength and weakness of several novel biomarkers, genomic, proteomic and metabolomic signatures in assisting the early diagnosis of DKD will be discussed in this article.
Abstract 3984
The rates of treatment-related mortality (TRM) and relapse are unacceptably high in adults undergoing antileukemia treatments for acute lymphoblastic leukemia (ALL). So far has no ...better therapy with low side effects to improve long-term survival in these patients. Indirubin, a Chinese translational anti-chronic myelogenous leukemia (CML) agent, is able to induce cellular apoptosis. However, until now the functional action of IO on ALL remains still unknown. Therefore, here we investigated and compared the cytotoxic efficacy and action of indirubin-3'-monoxime (IO) in JM1 (ALL cell) and K562 (CML cell). ALL and CML cells were treated with a series of IO dose for 24 and 48h, and cell survival was determined by WST-1 assay. ALL and CML cells were shown to be similar susceptible to IO cytotoxicity. In order to clear in which way of cell death induced by IO, we performed analyses for apoptosis, necrosis and autophagy, respectively. After IO treatment, both ALL and CML cells were arrested in the G2/M cell cycle phase. In addition, an increase of sub-G1 proportion was caused. We found also increasing of caspase-3 activation and formation of cleaved PARP in a dose dependent manner. These were associated with the form of apoptosis. However, the caspase inhibitor Z-VAD-FMK only could partially prevent cell death in ALL and CML cells. When further analyzing the necrotic phenomenon through measuring LDH release, the result clearly showed that LDH release was not remarkable after cell treatment with high dose of IO. Besides, we observed surprisingly in Western blot the increasing expression of microtubule-associated protein light chain 3-II (LC3-II), which generally correlates to formation of autophagosomes. Because better antileukemic drug should not induce toxicity in normal blood cells as much as possible, so the cytotoxic effect of IO in CD34+ hematopoietic stem cells, lymphocytes and granulocytes was analyzed. Excitingly, results showed that IO could not affect cell viability of granulocytes, and IO cytotoxicity in lymphocytes was only marginal. If CD34+ hematopoietic stem cells were treated with IO, the rate of cell survival and their ability of differentiation were almost identical in contrast with non-treated control. Apparently, these data indicated that IO possesses the capability to induce apoptosis and autophagy in both CML and ALL cells. The most important is that the IO hardly influences the cell survival and the differentiation of CD34+ hematopoietic stem cells, the cytotoxic effect in granulocytes and lymphocytes is only limited. In conclusion, IO can be considered as a potential agent for clinical anti- ALL treatment.
No relevant conflicts of interest to declare.
Abstract 4106
Indirubin-3-monoxime (IO) is the active ingredient of Danggui Longhui Wan, a mixture of plants that is used in traditional Chinese medicine. It is a potent inhibitor of cyclin-dependent ...kinases (CDKs), especially CDK2. In clinical studies demonstrated that indirubin did not cause major side effects in patients with chronic myelogenous leukemia. However, the functional action of indirubin on acute lymphoblastic leukemia (ALL) is still unclear. In the present study, we investigated that cytotoxic effect and mechanism study of IO in human acute lymphocytic leukemia cell line JM1 and human chronic myeloid leukemia cell line K562. We also analyzed the viability influence of IO in normal human granulocytes and lymphocytes.
After the treatment of IO, cell viability of JM1 and K562 cells was determined by WST-1 assay. The influence of IO on cell-cycle distribution was analyzed by FACScan. To identify which cell death types were induced during IO treatment we measured the caspase-3 activity for analysis the induction of apoptosis; evaluated the LDH release in the necrotic analysis and the expression level of LC3-II was determined by Western blotting analysis for autophagic cell death. Furthermore, we also analyzed the toxicity of IO in normal human granulocytes and lymphocytes by the treatment dose of IO with value of IC50.
IO significantly affected the cell viability on JM1 but also on K562 cells in a dose dependent manner. The G2/M phase of cell cycle was arrested and sub-G1 proportion was relative increased. In addition, the finding showed that IO initiated caspase-3-dependent apoptosis in JM1 and K562 cells. The expression of autophagosome-incorporated LC3-II protein was also clearly increased once cells treated with IO. However, the necrotic phenomenon through measuring LDH release from K562 and JM1 cells could not be observed. Excitingly, by the treatment dose of IO with value of IC50, the cell viability of lymphocytes was marginally affected, whereas the cell cytotoxitity of granulocytes was not induced.
IO has the ability to induce potent cytotoxic effect on K562 and JM1 cells. The cell death after IO treatment is mainly caused by apoptosis and autophagy, not by necrosis. This effect is also associated with interrupted cell cycle. Importantly, IO has few or little cytotoxic effect on human lymphocytes and granulocytes. The results suggest that IO might be useful for clinical anti- ALL treatment.
No relevant conflicts of interest to declare.
Pancreatic beta-cells are particularly susceptible to fatty-acid-induced endoplasmic reticulum (ER) stress and apoptosis. To understand how beta-cells sense fatty acid stimuli and translate into a ...long-term adaptive response, we investigated whether palmitic acid (PA) regulates early growth response-1 (Egr-1), an immediate-early transcription factor, which is induced by many environmental stimuli and implicated in cell proliferation, differentiation, and apoptosis. We found that Egr-1 was rapidly and transiently induced by PA in MIN6 insulinoma cells, which was accompanied by calcium influx and ERK1/2 phosphorylation. Calcium chelation and MEK1/2 inhibition blocked PA-induced Egr-1 upregulation, suggesting that PA induces Egr-1 expression through a calcium influx-MEK1/2-ERK1/2 cascade. Knockdown of Egr-1 increased PA-induced caspase-3 activation and ER stress markers and decreased PA-induced Akt phosphorylation and insulin secretion and signaling. Akt replenishment and insulin supplementation rescued PA-induced apoptosis in Egr-1 knockdown cells. These results suggest that the absence of Egr-1 loses its ability to couple the short-term insulin/Akt pathway to long-term survival adaptation. Finally, Egr-1-deficient mouse islets are more susceptible to ex vivo stimuli of apoptosis. In human pancreatic tissues, EGR1 expression correlated with expression of ER stress markers and anti-apoptotic gene. In conclusion, Egr-1 is induced by PA and further attempts to rescue beta-cells from ER stress and apoptosis through improving insulin/Akt signaling. Our study underscores Egr-1 as a critical early sensor in pancreatic beta-cells to translate fatty acid stimuli into a cellular adaptation mechanism. PA stimulates Egr-1 expression via a calcium influx-MEK1/2-ERK1/2-Elk-1 cascade. Egr-1 attenuates PA-induced ER stress and apoptosis. Egr-1 maintains Akt survival pathway to protect beta-cells from PA-induced apoptosis. Egr-1-deficient islets are prone to ex vivo stimuli of apoptosis. Human EGR1 expression correlates with genes for ER stress and anti-apoptosis.
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