miRNA-21 promotes gastric cancer growth by adjusting prostaglandin E2.

OBJECTIVE: It is to study the stimulation and possible active mechanism of miRNA-21 on AGS proliferation of gastric cancer. MATERIALS AND METHODS: AGS gastric cancer cells were cultivated in vitro and then divided into the blank control group, the PGE2 (prostaglandin E2) group, the anti-miRNA-21 group and the PGE2 + anti-miRNA-21 group and the MTT and the flow cytometry methods were adopted to test the effect of PGE2 or/and anti-miRNA-21 intervention on AGS cell proliferation and apoptosis and the differences to miRNA-21 expression. In addition, the cells were also divided into the blank control group, the PGE2 group, the PGE2 + Perifosine group, the PGE2 + anti-miRNA-21 group and the PGE2 + anti-miRNA-21 + Perifosine group and the MTT and flow cytometry methods were adopted to test the effect of Perifosine intervention on AGS cell proliferation and apoptosis and on PTEN and p-AktmRNA and protein expressions. RESULTS: Compared with the control group, AGS cell proliferation activity increased significantly, the apoptosis rate decreased and the miRNA-21tmRNA and protein expression increased in the PGE2 group (p < 0.05); compared with the PGE2 group, the AGS cell proliferation rate decreased, the apoptosis rate increased and the miRNA-21mRNA and protein expressions decreased (p < 0.05) in the anti-miRNA-21 group and the PGE2 + anti-miRNA-21 group. In addition, after intervention of Perifosine, the AGS cell proliferation rate decreased, the apoptosis rate increased, the PTEN mRNA and protein expressions increased and the pAktmRNA and protein expressions decreased (p < 0.05). CONCLUSIONS: miRNA-21 may promote the growth of gastric cancer cells by adjusting and controlling PTEN/Akt signal passage mediated PEG2.

Protective effects and mechanisms of Ndfipl on SH-SY5Y cell apoptosis in an in vitro Parkinson's disease model.

The aim of the current study was to examine the protective effects and mechanisms of Ndfipl on neurocytes in an experimental in vitro Parkinson's disease model induced by MPP+. The cell model was developed with dominant negative expression and suppressed expression of Ndfipl by means of transient transfection of Ndfipl-dominant negative and -inhibitory vectors. In total, four different Ndfipl cell models were established. Different methods were used to analyze the cells. The MTT method was used to detect the effect of Ndfipl on the survival rate and apoptosis of the cells induced by MPP(+). We further studied the roles of Ndfipl in inhibiting MPP(+)-induced SH-SY5Y apoptosis, protection, and ubiquitination of SH-SY5Y cells. Our results showed that Ndfipl reduced apoptosis and improved cell survival rate, indicating that Ndfipl has a neuroprotective effect. Furthermore, we found that Ndfipl binds to Nedd4-1, and that increased expression of Ndfipl significantly reduced Itch expression. We also found that increased ubiquitination played a role in Ndfipl-mediated processes, and that Ndfipl and α-synuclein interact. Additionally, the expression of Ndfipl reduced expression of α-synuclein. In conclusion, Ndfipl plays a significant role in protecting SH-SY5Y cells in in vitro Parkinson's disease models.

Accuracy of enhanced and unenhanced MRI in diagnosing scaphoid proximal pole avascular necrosis and predicting surgical outcome.

PURPOSE: Determine the sensitivity, specificity and accuracy of unenhanced and enhanced MRI in diagnosing scaphoid proximal pole (PP) avascular necrosis (AVN) and correlate whether MRI can help guide the selection of a vascularized or nonvascularized bone graft. METHODS: The study was approved by the IRB. Two MSK radiologists independently performed a retrospective review of unenhanced and enhanced MRIs from 18 patients (16 males, 2 females; median age, 17.5 years) with scaphoid nonunions and surgery performed within 65 days of the MRI. AVN was diagnosed on the unenhanced MRI when a diffusely decreased T1-W signal was present in the PP and on the enhanced MRI when PP enhancement was less than distal pole enhancement. Surgical absence of PP bleeding was diagnostic of PP AVN. Postoperative osseous union (OU) was assessed with computed tomography and/or radiographs. RESULTS: Sensitivity, specificity and accuracy for PP AVN were 71, 82 and 78% for unenhanced and 43, 82 and 67% for enhanced MRI. Patients with PP AVN on unenhanced MRI had 86% (6/7) OU; 100% (5/5) OU with vascularized bone grafts and 50% (1/2) OU with nonvascularized grafts. Patients with PP AVN on enhanced MRI had 80% (4/5) OU; 100% (3/3) OU with vascularized bone grafts and 50% (1/2) OU with nonvascularized grafts. Patients with viable PP on unenhanced and enhanced MRI had 91% (10/11) and 92% (12/13) OU, respectively, all but one with nonvascularized graft. CONCLUSIONS: When PP AVN is evident on MRI, OU is best achieved with vascularized grafts. If PP AVN is absent, OU is successful with nonvascularized grafts.

DEEP BIOSPHERE. Exploring deep microbial life in coal-bearing sediment down to ~2.5 km below the ocean floor.

Microbial life inhabits deeply buried marine sediments, but the extent of this vast ecosystem remains poorly constrained. Here we provide evidence for the existence of microbial communities in ~40° to 60°C sediment associated with lignite coal beds at ~1.5 to 2.5 km below the seafloor in the Pacific Ocean off Japan. Microbial methanogenesis was indicated by the isotopic compositions of methane and carbon dioxide, biomarkers, cultivation data, and gas compositions. Concentrations of indigenous microbial cells below 1.5 km ranged from <10 to ~10(4) cells cm(-3). Peak concentrations occurred in lignite layers, where communities differed markedly from shallower subseafloor communities and instead resembled organotrophic communities in forest soils. This suggests that terrigenous sediments retain indigenous community members tens of millions of years after burial in the seabed.

First evidence for the contribution of the genetic variations of BRCA1-interacting protein 1 (BRIP1) to the genetic susceptibility of cervical cancer.

BRIP1 (BRCA1-interacting protein 1), a DNA-dependent ATPase and a DNA helicase, is critical for BRCA-associated DNA damage repair functions, and may be involved in the development of cervical cancer. Genetic markers in different regions of the BRIP1 gene have a plausible role in modulating the risk of cervical cancer. In this study, we evaluate the association between the BRIP1 variations and the risk of cervix cancer. We examined the potential association between cervical cancer and eighteen single nucleotide polymorphisms (SNPs, rs2048718, rs16945692, rs4968451, rs6504074, rs4988344, rs8077088, rs10515211, rs9897121, rs9906313, rs2159450, rs4986764, rs11871785, rs4986763, rs11079454, rs7213430, rs34289250, rs4988345 and rs12937080) of the BRIP1 gene using the MassARRAY system. The participants enrolled in this study included 298 patients with cervical cancer and 286 healthy women as the healthy controls from a Chinese Han population. The results showed that rs16945692 (intron 1), rs4968451 (intron 4), rs4986764 (exon 18) and rs7213430 (3'UTR) were significantly associated with cervical cancer (P<0.05). Furthermore, strong linkage disequilibrium (LD) was observed in three blocks (D'>0.9), and significantly more T-A-C-A haplotypes (block 1) (P=0.001) were found in the patients with cervical cancer. Significantly higher frequencies of C-A-T haplotypes (block 2) (P=0.018) and A-A haplotypes (block 3) (P=0.009) were detected in the healthy controls than in the patients with cervical cancer, suggesting that they may show protective effects against cervical cancer. These findings point to a role for the BRIP1 gene polymorphisms in cervical cancer in a Chinese Han population, and may be informative for future genetic or biological studies on cervical cancer.

BRIP1 variations analysis reveals their relative importance as genetic susceptibility factor for cervical cancer.

To evaluate the association between gene variations in BRIP1 (BRCA1-interacting protein 1) and the risk of cervical cancer, we examined eight single nucleotide polymorphisms (SNPs: rs2048718, rs12937080, rs4988344, rs6504074, rs4988345, rs4986764, rs4986763, and rs11079454) in the BRIP1 gene in cervical tissue from a Chinese population using the MassARRAY system. The participants enrolled included 454 cervical cancer patients and 562 healthy controls. Quantitative real-time reverse transcription PCR (qRT-PCR) was performed to examine the potential correlation between functional BRIP1 SNP genotypes and mRNA levels in cervical cancer tissues. Our results first showed that rs4986764, located in exon 18 in the BRIP1 gene, was significantly associated with cervical cancer (χ(2)=11.191, P=0.001, odds ratio (OR)=1.384, 95% confidence interval (CI)=1.144-1.675). Another significant association was observed for rs4986763 located in exon 20 in BRIP1 (χ(2)=4.988, P=0.026, OR=1.241, 95% CI=1.027-1.500). Strong linkage disequilibrium was observed in the rs11079454-rs4986763-rs4986764 SNP block (D'>0.9). The frequencies of haplotype T-T-T are higher in controls than in these patients (P=2.01E-5). Moreover, cervical cancer tissues with a homozygous C/C genotype for rs4986764 had the lowest level of BRIP1, which was 2.8 and 2.9-fold lower than the C/T heterozygote and the T/T homozygote, respectively. These findings indicate a role for BRIP1 gene variations in cervical cancer and may be informative for future genetic or biological studies on cervical cancer.

A miniaturized pressure sensor with inherent biofouling protection designed for in vivo applications.

The design, fabrication, and measurement results for a diaphragm-based single crystal silicon sensor element of size 820 µm × 820 µm × 500 µm are presented. The sensor element is designed for in vivo applications with respect to size and measurement range. Moreover, it is optimized for longtime operation in the human body through a built-in protection preventing biofouling on the piezoresistors. The sensitivity is about 20 mV/V for a change from 500 to 1500 mbar absolute pressure. This result is comparable to conventional sized micromachined pressure sensors. The output signal is not found to be influenced by exposure to 60 °C for three hours, a normal temperature load for a typical sterilization process for medical devices (Ethylene Oxide Sterilization). The hysteresis is low; < 0.25% of full scale output signal. The sensor element withstands an overload pressure of 3000 mbar absolute pressure. Observed decrease in the output signal with temperatures and observed nonlinearity can easily be handled by traditional electronic compensation techniques.

Measurement of parenting self-efficacy and outcome expectancy related to discussions about sex.

The purpose of this study was to evaluate the psychometric properties of two scales--one to measure the self-efficacy of parents to discuss sexual health issues with their adolescents and the other to measure parents' outcome expectancy associated with such discussions. Understanding how parents feel about their confidence in talking with their children about important sexual health issues and the outcomes they expect as a result of such discussions can be useful in guiding both the development and refinement of educational programs to promote parent-child discussions. The responses of 491 mothers who participated in an HIV prevention intervention with their adolescents were used for the present analysis. Mothers ranged in age from 25 to 68 years with a mean of 37.9 years (SD = 6.9). Of mother participants, 33% were married, 96.7% were African American, and 89.2% had completed high school. Their adolescents ranged in age from 11 to 14 years, and 61.5% were male. Assessment of reliability for both scales showed that internal consistency reliability was acceptable for the total scales as well as three of the five subscales. With the exception of one item on the outcome expectancy scale, the inter-item correlations, the mean inter-item correlations, and the item-to-total correlations meet the standard criteria for scale development for both scales. Factor analysis was used to identify the underlying structure of the scales, and hypothesis testing was used to assess construct validity. The results of these analyses provide support for the construct validity of the scales.

High-level expression and single-step purification of human tryptophanyl-tRNA synthetase.

Human trpS gene was cloned into the expression vector pET-24a(+) to yield pET-24a(+)-HTrpRS, which could direct the synthesis of a mammalian derived protein in Escherichia coli BL21-CodonPlus(DE3)-RIL. The vector allows overproduction and single-step purification of His(6)-tagged human tryptophanyl-tRNA synthetase by the facilitation of metal (Ni(2+)) chelate affinity chromatography. The expression level of human TrpRS was about 40% of total cell proteins after isopropyl beta-D-thiogalactoside induction. The overproduced human TrpRS-His(6) could be purified to homogeneity within 2 h and about 24 mg purified enzyme could be obtained from 400 ml cell culture. The His(6) tag at C terminus had little effect on the binding ability of its substrates.

GO GIRLS!: results from a nutrition and physical activity program for low-income, overweight African American adolescent females.

This article describes the development, implementation, and results of an intervention designed for inner-city, overweight African American adolescent women. Fifty-seven participants were recruited from four public housing developments. Participants were administered physiologic, dietary, and cognitive assessments at baseline and immediately postintervention. Each session comprised three elements: (1) an educational/behavioral activity, (2) 30 to 60 minutes of physical activity, and (3) preparation and tasting of low-fat meals. In the absence of a control group, results were compared for high and low attenders, the former defined as attending at least 50% of the sessions. High attenders (n = 26) showed more favorable 6-month posttest values for most outcomes compared with low attenders (n = 31). These effects achieved statistical significance for nutrition knowledge, low-fat practices, perceived changes in low-fat practices, and social support.

Social cognitive factors associated with mother-adolescent communication about sex.

To better understand why some mothers talk to their children about sex and others do not, we examined the role of two social cognitive variables--self-efficacy and outcome expectancies--in explaining sex-based communication. The present study was part of a larger study to test the efficacy of two HIV prevention programs for mothers and their adolescents. Mothers and their adolescents were recruited from a large community organization that serves youth who live in disadvantaged circumstances. The sample for the present study included 486 mothers who averaged 38.4 years of age (SD = 6.73). The majority were African American (97.7%), not married (66.7%), and had a high school degree (89.5%). Their adolescents ranged in age from 11 through 14 years of age and most were male (61.3%). The results of the analysis revealed that mothers who expressed higher levels of self-efficacy and more favorable outcomes associated with talking to their children about sex were more likely to do so. In a regression analysis, we learned that the mother's degree of efficacy beliefs, along with her expected outcomes associated with talking about sex, the importance of religious beliefs to her, and the age and sex of her adolescents were important factors associated with talking with them about sex.

Gimme 5 fruit and vegetables for fun and health: process evaluation.

Gimme 5 (Georgia) was a school-based nutrition education effectiveness trial to help fourth- and fifth-grade students eat more fruit, 100% juice, and vegetables (FJV). Process evaluation assessed fidelity of implementation, reach, and use of intervention materials and environmental mediators: teacher training, curriculum delivery, participation in family activities, attendance at evening point-of-purchase grocery store activities, and availability and accessibility of FJV at home. Approximately half of the curriculum activities were implemented in fourth and fifth grades. The lowest proportion completed were those most pertinent to behavior change. Eighty-seven percent of parents reported participating in homework activities with their fourth grader, 66% with fifth graders. Sixty-five percent of parents reported viewing a video with their child in both grades. Ten percent attended evening point-of-purchase grocery store activities. The low level of implementation and modest level of participation in family activities suggest that higher levels of behavior change may have occurred if exposure to the intervention had been higher.

Gimme 5 fruit, juice, and vegetables for fun and health: outcome evaluation.

A theory-based multicomponent intervention (Gimme 5) was designed and implemented to impact fourth- and fifth-grade children's fruit, juice, and vegetable (FJV) consumption and related psychosocial variables. Gimme 5 was a randomized controlled intervention trial with school (n = 16 elementary) as unit of random assignment and analysis. Participants included the cohort of students who were in the third grade in the winter of 1994 and students who joined them in the fourth and fifth grades. The intervention included a curriculum, newsletters, videotapes, and point-of-purchase education. Evaluation included 7-day food records and psychosocial measures from students, telephone interviews with parents, and observational assessments. Favorable results were observed for consumption of FJV combined, FJV consumed at weekday lunch, eating FJV self-efficacy, social norms, asking behaviors, and knowledge. A theory-based school nutrition education program can help change children's FJV consumption and impact factors at home that predispose to FJV consumption, but changes were small, and their persistence is unknown.

Evaluation of developmentally appropriate programs for adolescent tobacco cessation.

Despite introduction of numerous smoking prevention programs in schools, tobacco use has not declined among adolescents. Schools face the dilemma of what to do with students who smoke and are not ready to quit. This study evaluated two programs based on the stages of change model. The educational program, the Tobacco Education Group (TEG), was designed for adolescents not yet thinking about quitting. The cessation program, the Tobacco Awareness Program (TAP), was intended for adolescents who want to quit. Evaluation was completed on 351 students at six public high schools. Compared to a control group of adolescent smokers not assigned to programs, both intervention groups significantly decreased tobacco use. Self-reported use was validated biochemically. Self-efficacy for quitting increased in both programs. Posttest use was predicted by posttest self-efficacy, peer support, and parental support, after controlling for initial use and initial self-efficacy.

Determination of diquat and paraquat in water by liquid chromatography-(electrospray ionization) mass spectrometry.

A method for the determination of the herbicides diquat and paraquat in water was developed using liquid chromatography-(electrospray ionization) mass spectrometry [LC-(ESI)MS]. The analytes were isolated on an ENVI-8 DSK solid phase extraction (SPE) disk and eluted with 5-M trifluoroacetic acid (TFA). The eluate was evaporated to dryness and the analytes were redissolved in the mobile phase (7% methanol/93% water/25-mM TFA). The extract was analyzed by liquid chromatography (C1 column) with postcolumn addition of propionic acid/methanol followed by (ESI)MS. Diquat was detected using the [M(2+)-H+] ion (M2+ = dication) at m/z 183, whereas paraquat was detected using the mono-trifluoroacetate ion pair [M2 +/- OOCCF3] at m/z 299. Quantitation was done by isotope dilution mass spectrometry using d4-diquat and d8-paraquat and the corresponding ions [M(2+)-D+] and [M2 +/- OOCCF3] at m/z 186 and m/z 307, respectively. Detection limits of 0.1 and 0.2 microgram/L, respectively (based on the dications), were adequate to meet the Ontario Drinking Water Objectives of 70 and 10 micrograms/L, respectively, and the Ontario Provincial Water Quality Objective for diquat of 0.5 microgram/L. Precision and accuracy were 14% and 6% for diquat and 12% and 3% for paraquat.

How best to measure implementation of school health curricula: a comparison of three measures.

The impact of school health education programs is often attenuated by inadequate teacher implementation. Using data from a school-based nutrition education program delivered in a sample of fifth graders, this study examines the discriminant and predictive validity of three measures of curriculum implementation: class-room observation of fidelity, and two measures of completeness, teacher self-report questionnaire and post-implementation interview. A fourth measure, obtained during teacher observations, that assessed student and teacher interaction and student receptivity to the curriculum (labeled Rapport) was also obtained. Predictive validity was determined by examining the association of implementation measures with three study outcomes; health knowledge, asking behaviors related to fruit and vegetables, and fruit and vegetable intake, assessed by 7-day diary. Of the 37 teachers observed, 21 were observed for two sessions and 16 were observed once. Implementation measures were moderately correlated, an indication of discriminant validity. Predictive validity analyses indicated that the observed fidelity, Rapport and interview measures were significantly correlated with post-test student knowledge. The association between health knowledge and observed fidelity (based on dual observation only), Rapport and interview measures remained significant after adjustment for pre-test knowledge values. None of the implementation variables were significantly associated with student fruit and vegetable intake or asking behaviors controlling for pre-test values. These results indicate that the teacher self-report questionnaire was not a valid measure of implementation completeness in this study. Post-implementation completeness interviews and dual observations of fidelity and Rapport appear to be more valid, and largely independent methods of implementation assessment.

Results of the TeachWell worksite wellness program.

OBJECTIVE: This study examined whether providing a school-based teacher wellness program enhances the impact of a health curriculum on student outcomes and improves cognitive, behavioral, and physiological outcomes among participating teachers. METHODS: Thirty-two elementary schools were randomly assigned to experimental or comparison conditions. Comparison group schools received the Gimme-5 program, a curriculum designed to increase fourth and fifty graders' consumption of fruits and vegetables. Experimental schools received Gimme-5 and the teacher wellness program, which included 54 workshops over 2 years, along with several schoolwide health activities. Physiological, behavioral, and cognitive outcomes were assessed in teachers and students. RESULTS: There was no evidence that the intervention favorably modified any student or teacher end points; nor did intervention teachers deliver the Gimme-5 program with greater fidelity than comparison teachers. CONCLUSION: Confidence in the null results is bolstered by the randomized design, baseline sample equivalence, appropriate mixed-model analyses, and lack of selective or differential attrition. Insufficient participation in the wellness program appears a likely explanation for the lack of teacher and student effects. Factors specific to the school setting and intervention may have diminished participation and, thus, intervention effects.