Aprendiendo de La Vida (Learning from Life): Development of a Radionovela to Promote Preventive Health Care Utilization among Indigenous Farmworkers from Mexico Living in California.

Mixtecs and Zapotecs, originating from the Oaxaca area in Mexico, are among the largest indigenous groups of workers in California. Many adults in this community only access the health care system when sick and as a last resort. This article describes the development of a radionovela to inform the community about the importance of preventive health care. It was developed following the Sabido Method. The methodology to develop a radionovela may be of interest to other public health practitioners who want to develop educational materials in an engaging format, especially for communities that rely on oral, not written information.

Peripheral blood T-cell receptor repertoire as a predictor of clinical outcomes in gastrointestinal cancer patients treated with PD-1 inhibitor.

BACKGROUND: Identifying valid biomarkers for patient selection impressively promotes the success of anti-PD-1 therapy. However, the unmet need for biomarkers in gastrointestinal (GI) cancers remains significant. We aimed to explore the predictive value of the circulating T-cell receptor (TCR) repertoire for clinical outcomes in GI cancers who received anti-PD-1 therapy. METHODS: 137 pre- and 79 post-treated peripheral blood samples were included. The TCR repertoire was evaluated by sequencing of complementarity-determining region 3 (CDR3) in the TRB gene. The Shannon index was used to measure the diversity of the TCR repertoire, and Morisita's overlap index was used to determine TCR repertoire similarities between pre- and post-treated samples. RESULTS: Among all enrolled patients, 76 received anti-PD-1 monotherapy and 61 received anti-PD-1 combination therapy. In the anti-PD-1 monotherapy cohort, patients with higher baseline TCR diversity exhibited a significantly higher disease control rate (77.8% vs. 47.2%; hazard ratio [HR] 3.92; 95% confidence interval [CI] 1.14-13.48; P = 0.030) and a longer progression-free survival (PFS) (median: 6.47 months vs. 2.77 months; HR 2.10; 95% CI 1.16-3.79; P = 0.014) and overall survival (OS) (median: NA vs. 8.97 months; HR 3.53; 95% CI 1.49-8.38; P = 0.004) than those with lower diversity. Moreover, patients with a higher TCR repertoire similarity still showed a superior PFS (4.43 months vs. 1.84 months; HR 13.98; 95% CI 4.37-44.68; P < 0.001) and OS (13.40 months vs. 6.12 months; HR 2.93; 95% CI 1.22-7.03; P = 0.016) even in the cohort with lower baseline diversity. However, neither biomarker showed predictive value in the anti-PD-1 combination therapy cohort. Interestingly, the combination of TCR diversity and PD-L1 expression can facilitate patient stratification in a pooled cohort. CONCLUSION: The circulating TCR repertoire can serve as a predictor of clinical outcomes in anti-PD-1 therapy in GI cancers.

Dietary Supplementation with Ferula Improves Productive Performance, Serum Levels of Reproductive Hormones, and Reproductive Gene Expression in Aged Laying Hens

This study investigated the effects of dietary Ferula supplementation on productive performance, egg quality, follicular development, serum levels of reproductive hormones, and reproductive gene expression in aged laying hens. A total of 300 Dawu pink laying hens (65 weeks old) were assigned to four treatments with six replicates per treatment and 10 hens per replicate. The birds were individually housed in wire cages and fed a corn-soybean meal-based diet with added Ferula at doses of 0, 50, 100, and 200 mg/kg for 12 weeks. The results showed that the laying rate in the 100 mg/kg Ferula-supplemented group was higher than in birds of the control group during weeks 1 to 12 (p<0.05). The average egg weight in the 100 mg/kg Ferula-supplemented group was significantly higher than in the other groups (p<0.01), while the feed-to-egg ratio was significantly lower than in other groups (p<0.01). The numbers of small yellow follicle, middle white follicle, and small white follicle were higher in the 100 mg/kg Ferula-supplemented birds than in the other groups (p<0.01). On weeks 69, the serum levels of estradiol, follicle-stimulating hormone, and luteinizing hormone were significantly higher in the 100 mg/kg Ferula-supplemented group than in the other groups (p<0.05). Additionally, expressions of ERα, FSHR, and LHR in the ovarian tissue were up-regulated by Ferula supplementation, especially in the 100 mg/kg group (p<0.01). These results indicate that the Ferula supplementation can significantly improve productive performance, egg quality, reproduction of hormonal profile, and reproductive gene expression of aged laying hens.(AU)

Dietary Supplementation with Ferula Improves Productive Performance, Serum Levels of Reproductive Hormones, and Reproductive Gene Expression in Aged Laying Hens

This study investigated the effects of dietary Ferula supplementation on productive performance, egg quality, follicular development, serum levels of reproductive hormones, and reproductive gene expression in aged laying hens. A total of 300 Dawu pink laying hens (65 weeks old) were assigned to four treatments with six replicates per treatment and 10 hens per replicate. The birds were individually housed in wire cages and fed a corn-soybean meal-based diet with added Ferula at doses of 0, 50, 100, and 200 mg/kg for 12 weeks. The results showed that the laying rate in the 100 mg/kg Ferula-supplemented group was higher than in birds of the control group during weeks 1 to 12 (p<0.05). The average egg weight in the 100 mg/kg Ferula-supplemented group was significantly higher than in the other groups (p<0.01), while the feed-to-egg ratio was significantly lower than in other groups (p<0.01). The numbers of small yellow follicle, middle white follicle, and small white follicle were higher in the 100 mg/kg Ferula-supplemented birds than in the other groups (p<0.01). On weeks 69, the serum levels of estradiol, follicle-stimulating hormone, and luteinizing hormone were significantly higher in the 100 mg/kg Ferula-supplemented group than in the other groups (p<0.05). Additionally, expressions of ERα, FSHR, and LHR in the ovarian tissue were up-regulated by Ferula supplementation, especially in the 100 mg/kg group (p<0.01). These results indicate that the Ferula supplementation can significantly improve productive performance, egg quality, reproduction of hormonal profile, and reproductive gene expression of aged laying hens.

Clinician offering is a key factor associated with HPV vaccine uptake among Mexican mothers in the USA and Mexico: a cross-sectional study.

OBJECTIVE: To compare the knowledge, beliefs, and practices regarding HPV vaccination among mothers of vaccine-eligible girls in Mexico and the USA. METHODS: Similar samples of Mexican mothers with vaccine-eligible daughters were surveyed at two clinics in Cuernavaca, Morelos, from July to October 2012 (n = 200) and at two clinics in Oxnard, California, from August to November 2013 (n = 200). RESULTS: Although mothers in the USA had less knowledge and more negative attitudes toward the vaccine than their counterparts in Mexico, vaccine uptake rates were higher in the USA (49% vs. 40%). US mothers were more likely to have discussed and been offered the HPV vaccine by a clinician than mothers in Mexico. In multivariate analyses, having been offered the HPV vaccine was the most important predictor of vaccine uptake. CONCLUSIONS: Our results suggest that healthcare access or other system, clinic, or provider factors are the main drivers of vaccine receipt in this binational sample of Mexican mothers. Interventions and programs that encourage clinicians to offer the HPV vaccine should be developed to increase vaccine uptake in both countries.

miR-26a-5p suppresses tumor metastasis by regulating EMT and is associated with prognosis in HCC.

OBJECTIVE: To determine the role of miR-26a-5p in tumor invasion and metastasis in hepatocellular carcinoma (HCC). METHODS: We evaluated miR-26a-5p expression in HCC tissues by quantitative PCR and then analyzed its clinical significance using a Cox regression model. Transwell and nude mouse models were used to examine tumor metastasis in vitro and in vivo, respectively. The relationship between miR-26a-5p and epithelial-mesenchymal transition was also investigated by q-PCR and western blot. RESULTS: Strong downregulation of miR-26a-5p was observed in tumor tissues compared to paired adjacent normal tissues. Moreover, patients with low miR-26a-5p expression had a significantly poorer prognosis than those with high expression. The multivariate analysis indicated that miR-26a-5p expression was an independent prognostic indicator. The experimental transwell model and athymic mouse model revealed that miR-26a-5p depressed tumor metastasis in vitro and in vivo, respectively. In addition, the decreased miR-26a-5p level observed in HCC was associated with reduced E-cadherin expression and upregulation of vimentin, which affects the molecular mechanism of EMT. CONCLUSION: Downregulation of miR-26a-5p promotes tumor metastasis by targeting EMT and influences the prognosis of HCC patients. Therefore, miR-26a-5p has potential as a new biomarker and therapeutic target.

Cardiac damage and dysfunction in diabetic cardiomyopathy are ameliorated by Grx1.

Glutaredoxin 1 (Grx1) has been found to be an important endogenous antioxidant enzyme closely related to the pathogenesis of diabetes and cardiovascular diseases caused by oxidative stress. In this study, the functional changes of the Grx1 redox system in blood of hyperglycemic patients were examined. Furthermore, using a rat model of streptozotocin (STZ)- and high-fat-diet-induced type 2 diabetes, we explored the correlation between functional changes of the Grx1 redox system in the left ventricular tissue and blood of the diabetic rats. Moreover, we studied the protective effect of Grx1 against cardiac toxicity caused by the high-glucose-induced expression of cardiac matrix metalloproteinases (MMPs) in primary cultured cardiac fibroblasts. Finally, we investigated the protective effects and signaling regulatory mechanism of Grx1 against diabetic cardiomyopathy (DCM) in terms of oxidative stress and NF-kB-mediated fibrosis-associated signaling pathways. In the serum of hyperglycemic patients, Grx1 levels were elevated, total/protein thiol or sulfhydryl (Total-SH/P-SH) levels were decreased, glutathione was downregulated, and oxidized glutathione was upregulated. In addition, in the left ventricular myocardium and blood of the diabetic rats, Grx1 levels were significantly increased and glutathione reductase and P-SH levels were decreased. Moreover, endogenous Grx1 was highly expressed in cardiac fibroblasts during high-glucose treatment, and exogenous Grx1 can prevent DCM by controlling oxidative damage and MMP expression. These findings are suggestive of changes in the Grx1 redox system, and Grx1-regulated protein oxidative modifications may serve as molecular markers for diabetes caused by high-glucose-induced oxidative stress.

Identification of an SCAR marker related to female phenotype in Idesia polycarpa Maxim.

Idesia polycarpa Maxim. is a dioecious species. Because of the lack of morphological and cytological methods available for identifying its sex during the long juvenile stage, the application of molecular markers in sex identification may facilitate sex determination in the seedling stage. The objective of this study was to use sequence-related amplified polymorphism to identify sex-linked markers in I. polycarpa and convert these markers into sequence-characterized amplified region markers, which are much easier to identify. A total of 342 primer combinations were screened and 2770 bands were examined. Only me14/em8 could amplify a specific fragment (210 base pairs) in all female but none in male plants. We analyzed this fragment using GenBank and found that the sequence similarity was 80% to the Populus trichocarpa clone POP006-H09 (sequence ID: gb|AC212923.1|) and that of the deduced amino acid sequence was 73% to the integrase of Mendicago truncatula (sequence ID: gb|ABD28291.1|) and 71% to the predicted retrotransposon integrase-like protein 1-like in Cicer arietinum (sequence ID: ref|XP 004515460.1|) (NCBI database through December 17, 2013). This fragment was converted into a stable and simple sequence-characterized amplified region marker approximately 200 base pairs in length. This marker can be utilized for early sexual identification in I. polycarpa, which will facilitate future breeding programs.

CYP4F2 gene single nucleotide polymorphism is associated with ischemic stroke.

The objective of this study was to explore the relationship between CYP4F2 gene polymorphism and ischemic stroke (IS) in the Han Chinese population. We performed a case-control study to genotype four single nucleotide polymorphisms (SNPs) (rs2108622, rs3093100, rs3093105, rs3093135) in the CYF4F2 gene. The genotype and haplotype distributions were compared between the case and control groups. We found that the GG genotype of rs2108622 in the CYP4F2 gene was associated with risk of IS (P = 0.023). Haplotype analysis indicated that the GGGT haplotype comprising rs2108622-rs3093100-rs3093105-rs3093135 was associated with IS, which suggests that the GGGT haplotype may be a risk factor for IS (P = 0.012). CYP4F2 gene polymorphism might increase the risk of IS in the Chinese population.

Association of the ApoE gene polymorphism and dietary factors with cerebral infarction and circulating lipid concentrations.

The aim of this study was to explore the relationship between the ApoE gene polymorphism and dietary factors with stroke and circulating lipid levels in the Chinese population. We selected 580 patients with stroke and 580 age- and gender-matched healthy controls, and examined their ApoE polymorphism genotype using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. We also analyzed the relationship between the ApoE gene polymorphism and dietary factors as well as plasma lipid concentrations in this cohort. We detected six ApoE genotypes in the study populations, and determined that the E4 allele was positively associated with cerebral infarction (CI), whereas allele E2 was negatively associated with total cholesterol (TC) and low-density lipoprotein-cholesterol levels. The dietary habits of the subjects with descending order of average total TC and triglyceride levels were: subjects addicted to oily food > subjects addicted to sweets > subjects addicted to smoking > subjects addicted to alcohol > subjects following a vegetarian diet (P < 0.05). Our results demonstrated that the ApoE gene polymorphism was associated with a risk for CI in a Chinese population.

Isolation and characterization of the papaya MADS-box E-class genes, CpMADS1 and CpMADS3, and a TM6 lineage gene CpMADS2.

Papaya (Carica papaya L.) plants are polygamous, with female, male, and hermaphroditic flowers. To understand the roles of MADS-box genes in flower development and sex determination, we cloned cDNAs of E-class genes CpMADS1 and CpMADS3 and a TM6 lineage of the B-class gene CpMADS2 from young flower buds of papaya. Reverse transcription-polymerase chain reaction (RT-PCR) and real-time PCR analyses revealed that CpMADS1 and CpMADS3 were preferentially expressed in the carpel and also in petals and stamens. CpMADS2 was expressed in both petals and stamens early during floral development. Comparison of 10 papaya genotypes of 5 different sex phenotypes - hermaphrodite, male, female, progeny-all-hermaphrodite, and progeny-all-male - by Southern blot analysis of genomic DNAs with probes of the 3 genes revealed similar restriction patterns and copy number, suggesting a low relationship of the 3 CpMADS genes with sex expression of papaya plants at the genomic level.

Relationship between progeny growth performance and molecular marker-based genetic distances in Eucommia ulmoides parental genotypes.

Progeny performances, variations and combining abilities for growth traits were evaluated in a factorial mating design of Eucommia ulmoides. Three marker systems, sequence-related amplified polymorphism, amplified fragment length polymorphism, and inter-simple sequence repeat, were used to determine genetic distances between parents. Correlations of genetic distances with progeny performances, within-family coefficients of variation and specific-combining abilities were established for height and basal diameter traits. Significant positive correlations were found between progeny performances of growth traits and genetic distances of parents based on sequence-related amplified polymorphism markers or a combination of all 3 marker systems. This revealed that crosses between genetically distant parents produced progenies with excellent growth performances. The lack of correlations between parental genetic distances and within-family coefficients of variation or specific-combining abilities suggested that these characteristics were unpredictable. The results of this study represent a potential criterion to predict progeny performances and choose parents in the breeding program.

Significant association between lower pulse pressure and increasing levels of a novel type of phospholipid.

The aim of this study was to analyze the association between pulse pressure and a novel type of phospholipid with solubility similar to that of lysophosphatidic acid (LPA), designated as AP, which was reported to be elevated during ischemia. In this cross-sectional study, 416 hypertensive patients and 252 controls aged between 35 and 70 years were enrolled consecutively. Fasting blood samples were extracted for assays of phospholipids and other biomarkers. Compared to controls, the hypertensive patients had higher levels of both LPA [odds ratio (OR) = 3.83] and AP (OR = 4.30). Changes in blood pressure did not affect the levels of LPA or AP. However AP, but not LPA, levels were significantly higher in patients with lower or higher pulse pressure (Pearson χ(2) = 11.239, P = 0.001). For patients whose pulse pressure was ≤60 mmHg, plasma levels of AP were significantly negatively correlated with pulse pressure. However, this was not observed for LPA and nine other biomarkers, including lipoproteins. Plasma levels of AP increased in hypertensive patients with higher or lower pulse pressure. Thus, attention should be paid to the possibility of cerebral ischemia in hypertensive patients when they have abnormal pulse pressure, especially for those with relatively low pulse pressure.

Demographic predictors of leukotriene antagonist monotherapy among children with persistent asthma.

OBJECTIVE: To describe the children with persistent asthma receiving non-preferred controller therapy in the form of leukotriene receptor antagonist monotherapy (LTRAM). STUDY DESIGN: In this cross-sectional study, we analyzed 2007-2009 South Carolina Medicaid data of children aged 2- to 18 years with persistent asthma, defined by Healthcare Effectiveness Data and Information Set (HEDIS). Those without either LTRAM or inhaled corticosteroids (ICS) were excluded. With multivariable logistic regression modeling, we compared the outcome of LTRAM with the primary predictor of age and adjusted for covariates of race, sex, HEDIS class, rurality, and disease severity. We also used negative binomial regression to compare outcomes of albuterol and oral steroid claims, outpatient and emergency department visits, and hospitalizations with predictors of LTRAM vs ICS therapy. RESULTS: A total of 19,512 patients with asthma aged 2- to 18-years were studied: 2658 (13.6%) without controllers were excluded, 2508 (12.9%) received LTRAM, and 14 346 (73.5%) received ICS. Age, race, rurality, and HEDIS classification were all significantly associated with LTRAM (all P < .01): 5- to 13-year-olds relative to children <5 years old (OR 1.46, 95% CI 1.30-1.64), Caucasians relative to African Americans (OR 1.40, 95% CI 1.27-1.53), and rural children relative to urban (OR 1.18, 95% CI 1.08-1.3) were all more likely to receive LTRAM. Albuterol, oral steroid, and outpatient visits were lower in LTRAM (P < .01). No difference was detected in emergency department visits or admissions. CONCLUSIONS: Children 5- to 13-years of age, rural children, and Caucasian children were more likely to receive LTRAM. Uncovering provider rationale and practices as well as patient influences on this prescribing pattern may be helpful in optimizing asthma controller therapy.

Review article: the treatment of functional abdominal bloating and distension.

BACKGROUND: Abdominal bloating and distension are common symptoms in patients with functional gastrointestinal disorders (FGIDs), however, relatively little is known about their treatment. AIM: To review the treatment trials for abdominal bloating and distension. METHODS: A literature review in Medline for English-language publications through February 2010 of randomised, controlled treatment trials in adults. Study quality was assessed according to Jadad's score. RESULTS: Of the 89 studies reviewed, 18% evaluated patients with functional dyspepsia, 61% with irritable bowel syndrome (IBS), 10% with chronic constipation and 10% with other FGIDs. No studies were conducted in patients diagnosed with functional abdominal bloating. The majority of trials investigated the efficacy of prokinetics or probiotics, although studies are heterogeneous with respect to diagnostic criteria and outcome measures. In general, bloating and/or distension were evaluated as secondary endpoints or as individual symptoms as part of a composite score rather than as primary endpoints. A greater proportion of IBS patients with constipation reported improvement in bloating with tegaserod vs. placebo (51% vs. 40%, P<0.0001) and lubiprostone (P<0.001). A greater proportion of nonconstipating IBS patients reported adequate relief of bloating with rifaximin vs. placebo (40% vs. 30%, P<0.001). Bloating was significantly reduced with the probiotics, Bifidobacterium infantis 35624 (1×10(8) dose vs. placebo: -0.71 vs. -0.44, P<0.05) and B. animalis (live vs. heat-killed: -0.56±1.01 vs. -0.31±0.87, P=0.03). CONCLUSIONS: Prokinetics, lubiprostone, antibiotics and probiotics demonstrate efficacy for the treatment of bloating and/or distension in certain FGIDs, but other agents have either not been studied adequately or have shown conflicting results.

Interdisciplinary communication in the intensive care unit at the University Hospital of the West Indies.

OBJECTIVE: To assess the perceptions of physicians and nurses working full-time in the Intensive Care Unit (ICU) at the University Hospital of the West Indies (UHWI) regarding interdisciplinary communication. METHOD: A cross-sectional survey of all medical personnel working full-time in the ICU was conducted in January 2008 using a self-administered, validated questionnaire. Data on perceived communication, teamwork and leadership, comprehension of patient care goals, perceived effectiveness and satisfaction were collected and analysed using the SPSS Version 14. Internal reliability was tested using Cronbach's alpha score and differences and correlations were assessed using Pearson's Chi-square and correlation analysis. RESULTS: Ninety-five per cent (105/111) of questionnaires were completed. More doctors than nurses experienced open communication with other staff members (73% vs 32%; p < 0.01), with less openness occurring with increasing seniority. More doctors (53%) than nurses (32%) reported receiving inaccurate information from doctors (p < 0.05), with 67% and 51% respectively receiving incorrect information from nurses (p < 0.05). Communication across shifts was felt to be better amongst doctors than nurses (73% vs 63%). Only 50% of doctors compared to 88% of nurses felt they received relevant information quickly (p < 0.05). More nurses than doctors (86% vs 63%; p < 0.01) felt that they had a good understanding of patient care goals. Negative perceptions of the leadership characteristics of consultants (62% amongst doctors and 74% of nurses) and sisters (79% and 73%, respectively) were high. CONCLUSIONS: Communication within the ICU, UHWI, is unsatisfactory with an overall poor perception of senior leadership. Improvement in staff morale and leadership training may create a working environment where team members can communicate openly without fear of chastisement.

Stem cell factor protects against neuronal apoptosis by activating AKT/ERK in diabetic mice

Neuronal apoptosis occurs in the diabetic brain due to insulin deficiency or insulin resistance, both of which reduce the expression of stem cell factor (SCF). We investigated the possible involvement of the activation of the MAPK/ERK and/or AKT pathways in neuroprotection by SCF in diabetes. Male C57/B6 mice (20-25 g) were randomly divided into four groups of 10 animals each. The morphology of the diabetic brain in mice treated or not with insulin or SCF was evaluated by H&E staining and TUNEL. SCF, ERK1/2 and AKT were measured by Western blotting. In diabetic mice treated with insulin or SCF, there was fewer structural change and apoptosis in the cortex compared to untreated mice. The apoptosis rate of the normal group, the diabetic group receiving vehicle, the diabetic group treated with insulin, and the diabetic group treated with SCF was 0.54 ± 0.077 percent, 2.83 ± 0.156 percent, 1.86 ± 0.094 percent, and 1.78 ± 0.095 percent (mean ± SEM), respectively. SCF expression was lower in the diabetic cortex than in the normal cortex; however, insulin increased the expression of SCF in the diabetic cortex. Furthermore, expression of phosphorylated ERK1/2 and AKT was decreased in the diabetic cortex compared to the normal cortex. However, insulin or SCF could activate the phosphorylation of ERK1/2 and AKT in the diabetic cortex. The results suggest that SCF may protect the brain from apoptosis in diabetes and that the mechanism of this protection may, at least in part, involve activation of the ERK1/2 and AKT pathways. These results provide insight into the mechanisms by which SCF and insulin exert their neuroprotective effects in the diabetic brain.

Stem cell factor protects against neuronal apoptosis by activating AKT/ERK in diabetic mice.

Neuronal apoptosis occurs in the diabetic brain due to insulin deficiency or insulin resistance, both of which reduce the expression of stem cell factor (SCF). We investigated the possible involvement of the activation of the MAPK/ERK and/or AKT pathways in neuroprotection by SCF in diabetes. Male C57/B6 mice (20-25 g) were randomly divided into four groups of 10 animals each. The morphology of the diabetic brain in mice treated or not with insulin or SCF was evaluated by H&E staining and TUNEL. SCF, ERK1/2 and AKT were measured by Western blotting. In diabetic mice treated with insulin or SCF, there was fewer structural change and apoptosis in the cortex compared to untreated mice. The apoptosis rate of the normal group, the diabetic group receiving vehicle, the diabetic group treated with insulin, and the diabetic group treated with SCF was 0.54 +/- 0.077%, 2.83 +/- 0.156%, 1.86 +/- 0.094%, and 1.78 +/- 0.095% (mean +/- SEM), respectively. SCF expression was lower in the diabetic cortex than in the normal cortex; however, insulin increased the expression of SCF in the diabetic cortex. Furthermore, expression of phosphorylated ERK1/2 and AKT was decreased in the diabetic cortex compared to the normal cortex. However, insulin or SCF could activate the phosphorylation of ERK1/2 and AKT in the diabetic cortex. The results suggest that SCF may protect the brain from apoptosis in diabetes and that the mechanism of this protection may, at least in part, involve activation of the ERK1/2 and AKT pathways. These results provide insight into the mechanisms by which SCF and insulin exert their neuroprotective effects in the diabetic brain.

High expression of the circadian gene mPer2 diminishes the radiosensitivity f NIH 3T3 cells

Period2 is a core circadian gene, which not only maintains the circadian rhythm of cells but also regulates some organic functions. We investigated the effects of mPeriod2 (mPer2) expression on radiosensitivity in normal mouse cells exposed to 60Co-γ-rays. NIH 3T3 cells were treated with 12-O-tetradecanoylphorbol-13-acetate (TPA) to induce endogenous mPer2 expression or transfected with pcDNA3.1(+)-mPer2 and irradiated with 60Co-γ-rays, and then analyzed by several methods such as flow cytometry, colony formation assay, RT-PCR, and immunohistochemistry. Flow cytometry and colony formation assay revealed that irradiated NIH 3T3 cells expressing high levels of mPer2 showed a lower death rate (TPA: 24 h 4.3 percent vs 12 h 6.8 percent and control 9.4 percent; transfection: pcDNA3.1-mPer2 3.7 percent vs pcDNA3.1 11.3 percent and control 8.2 percent), more proliferation and clonogenic survival (TPA: 121.7 ± 6.51 vs 66.0 ± 3.51 and 67.7 ± 7.37; transfection: 121.7 ± 6.50 vs 65.3 ± 3.51 and 69.0 ± 4.58) both when treated with TPA and transfected with mPer2. RT-PCR analysis showed an increased expression of bax, bcl-2, p53, c-myc, mre11, and nbs1, and an increased proportionality of bcl-2/bax in the irradiated cells at peak mPer2 expression compared with cells at trough mPer2 expression and control cells. However, no significant difference in rad50 expression was observed among the three groups of cells. Immunohistochemistry also showed increased protein levels of P53, BAX and proliferating cell nuclear antigen in irradiated cells with peak mPer2 levels. Thus, high expression of the circadian gene mPer2 may reduce the radiosensitivity of NIH 3T3 cells. For this effect, mPer2 may directly or indirectly regulate the expressions of cell proliferation- and apoptosis-related genes and DNA repair-related genes.

High expression of the circadian gene mPer2 diminishes the radiosensitivity of NIH 3T3 cells.

Period2 is a core circadian gene, which not only maintains the circadian rhythm of cells but also regulates some organic functions. We investigated the effects of mPeriod2 (mPer2) expression on radiosensitivity in normal mouse cells exposed to 60Co-gamma-rays. NIH 3T3 cells were treated with 12-O-tetradecanoylphorbol-13-acetate (TPA) to induce endogenous mPer2 expression or transfected with pcDNA3.1(+)-mPer2 and irradiated with 60Co-gamma-rays, and then analyzed by several methods such as flow cytometry, colony formation assay, RT-PCR, and immunohistochemistry. Flow cytometry and colony formation assay revealed that irradiated NIH 3T3 cells expressing high levels of mPer2 showed a lower death rate (TPA: 24 h 4.3% vs 12 h 6.8% and control 9.4%; transfection: pcDNA3.1-mPer2 3.7% vs pcDNA3.1 11.3% and control 8.2%), more proliferation and clonogenic survival (TPA: 121.7 +/- 6.51 vs 66.0 +/- 3.51 and 67.7 +/- 7.37; transfection: 121.7 +/- 6.50 vs 65.3 +/- 3.51 and 69.0 +/- 4.58) both when treated with TPA and transfected with mPer2. RT-PCR analysis showed an increased expression of bax, bcl-2, p53, c-myc, mre11, and nbs1, and an increased proportionality of bcl-2/bax in the irradiated cells at peak mPer2 expression compared with cells at trough mPer2 expression and control cells. However, no significant difference in rad50 expression was observed among the three groups of cells. Immunohistochemistry also showed increased protein levels of P53, BAX and proliferating cell nuclear antigen in irradiated cells with peak mPer2 levels. Thus, high expression of the circadian gene mPer2 may reduce the radiosensitivity of NIH 3T3 cells. For this effect, mPer2 may directly or indirectly regulate the expressions of cell proliferation- and apoptosis-related genes and DNA repair-related genes.