Globular adiponectin induces esophageal adenocarcinoma cell pyroptosis via the miR-378a-3p/UHRF1 axis.

BACKGROUND: Antiapoptosis is a major factor in the resistance of tumor cells to chemotherapy and radiotherapy. Thus, activation of cell pyroptosis may be an effective option to deal with antiapoptotic cancers such as esophageal adenocarcinoma (EAC). METHODS: Differential expression of ubiquitin-like versus PHD and ring finger structural domain 1 (UHRF1) in EAC and near normal tissues was analyzed, as well as the prognostic impact on survival in EAC. Also, the same study was done for globular adiponectin (gAD). Simultaneously, the mRNA expression of UHRF1 was observed in different EAC cell lines. Real time cellular analysis (RTCA) was used to detect cell proliferation, and flow cytometry and inverted fluorescence microscopy were used to detect pyroptosis. Biocredit analysis was conducted to observe the correlation between UHRF1 and key pyroptosis proteins. OD values and CCK8 assay were used to determine the effect of miR-378a-3p on EAC cells. Quantitative real-time polymerase chain reaction and Western blot were used to detect the correlation between UHRF1, gAD, and miR-378a-3p in EAC cells. Moreover, in vivo and in vitro experiments were performed to detect the relevant effects on tumor migration and invasion after inhibiting UHRF1 expression. RESULTS: UHRF1 was negatively correlated with the survival of patients with EAC, while miR-378a-3p showed the opposite effect. Additionally, gAD promoted EAC cell pyroptosis, upregulated miR-378a-3p, and significantly inhibited the proliferation of EAC cells. gAD directly reduced UHRF1 expression in EAC cells by upregulating miR-378a-3p. In cell migration and invasion assays, inhibition of UHRF1 expression significantly suppressed EAC cell metastasis. In animal experiments, we again demonstrated that gAD induced pyroptosis in EAC cells by inhibiting the expression of UHRF1. CONCLUSION: gAD-induced upregulation of miR-378a-3p significantly inhibited the proliferation of EAC by targeting UHRF1. Therefore, gAD may serve as an alternative therapy for chemotherapy- and radiation-refractory EAC or other cancers with the same mechanism of pyroptosis action.

Bioinformatics identify the role of chordin-like 1 in thyroid cancer.

The abnormal expression of chordin-like 1 (CHRDL1) is identified in many cancers, while the effect of CHRDL1 in thyroid cancer (THCA) remains unclear. The University of California Santa Cruz, Gene Expression Profiling Interactive Analysis, University of Alabama at Birmingham Cancer, and Gene Expression Omnibus database (GSE33570, GSE33630, and GSE60542) were used for determining the mRNA and methylation expression of CHRDL1 in tumor and normal tissues. Human Protein Atlas was used for exploring the protein expression level of CHRDL1. The genes correlated to CHRDL1 were assessed by cBioPortal database. The prognostic value of CHRDL1 was evaluated through Kaplan-Meier method, cox regression, and nomogram analysis. Kyoto Encyclopedia of Genes and Genomes, Gene Ontology, and gene set enrichment analysis were used for predicting potential function of CHRDL1. The relationship between CHRDL1 and immune cell infiltration was determined by Pearson method. The downregulated mRNA and protein expressions of CHRDL1 were identified in THCA through the analysis of data from The Cancer Genome Atlas, Gene Expression Omnibus, and Human Protein Atlas database. The survival analysis showed that the CHRDL1 expression significantly affected disease-free interval (DFI) and progression-free interval, and CHRDL1 was an independent predictor of DFI. Besides, we found that C-C motif chemokine ligand 21 could significantly affect DFI time when it was co-expressed with CHRDL1. Additionally, the function of CHRDL1 was enriched in cell migration, apoptosis, and immune cell receptor. The downregulated expression of CHRDL1 was observed in THCA and caused poor prognosis. CHRDL1 may be involved in signal pathway related to cancer development and immune response, which suggested it could be a potential biomarker.

Circular RNA NF1-419 Inhibits Proliferation and Induces Apoptosis by Regulating Lipid Metabolism in Astroglioma Cells.

BACKGROUND: Astroglioma is the most common primary tumor of the central nervous system. Currently, there is no effective treatment for astroglioma. In the present study, the extract (L3) from Ganoderma Lucidum (G. lucidum) was found to inhibit the growth of astroglioma U87 cells and change the expression of circular RNAs (circRNAs). One of these, including the circular NF1-419 (circNF1-419), was of interest because NF1 gene is a classic tumor suppressor gene. OBJECTIVES: The functional role of circ-NF1-419 in the inhibition of astroglioma cells remains unknown. This study focuses on the role of circNF1-419 in functional abnormalities of U87 astroglioma cells and aims to elaborate on its regulatory mechanism. METHODS: The circNF1-419 overexpressing U87 (U87-NF1-419) cells were constructed. We generated U87-NF1-419 to evaluate the role of circNF1-419 on cell cycle, apoptosis, proliferation, tumor growth and metabolic regulation. Finally, we used docking screening to identify compounds in G. lucidum extracts that target circ-419. RESULTS: U87-NF1-419 can promote cell apoptosis and regulate lipid metabolism through glycerophospholipid metabolism and retrograde endocannabinoid signaling. Further examinations revealed that the expression of metabolic regulators, such as L-type voltage-operated calcium channels (L-VOCC), phospholipase C-ß3 (PLCß3), Mucin1, cationic amino acid transporter 4 (CAT4), cationic amino acid transporter 1 (CAT1) and a kinase (PRKA) anchor protein 4 (AKAP4) was inhibited, while phosphatidylserine synthase 1 (PTDSS1) was enhanced in U87-NF1-419 cells. In vivo experiments showed that circNF1-419 inhibits tumor growth in BALB/C nude mice, and enhanced AKAP4 and PTDSS1 in tumor tissues. The virtual docking screening results supported that ganosporeric acid A, ganodermatriol, ganoderic acid B and α-D-Arabinofuranosyladenine in L3 could activate circNF1-419 in astroglioma treatment. CONCLUSION: This study indicated that circNF1-419 could be a therapeutic target for the clinical treatment of astroglioma. L3 from Ganoderma Lucidum (G. lucidum) could inhibit astroglioma growth by activating circNF1-419.

Do Statins Have a Positive Impact on Patients with Coronary Microvascular Dysfunction on Long-Term Clinical Outcome? A Large Retrospective Cohort Study.

OBJECTIVES: To investigate the influence of statins on major adverse cardiovascular events (MACE) in patients with coronary microvascular dysfunction (CMVD). PARTICIPANTS: 23,494 patients who received coronary angiography (CAG) were included. Thrombolysis in Myocardial Infarction, Myocardial Perfusion Grading (TMPG), a useful angiographic method, was used to evaluate CMVD. RESULTS: Using multivariate analysis, NYHA III/IV (HR, 1.44; 95% CI, 1.03-2.01; P=0.031), PCI history (HR, 3.69; 95% CI, 2.57-5.31; P<0.001), TG (HR, 1.15; 95% CI, 1.06-1.26; P=0.001), creatinine (HR, 1.00; 95% CI, 1.00-1.01; P<0.001), cTnT (HR, 0.98; 95% CI, 0.96-0.99; P<0.001), heart rate (HR, 0.98; 95% CI, 0.97-0.99; P=0.001), ß-blocker (HR, 0.68; 95% CI, 0.51-0.91; P=0.008), aspirin (HR, 0.38; 95% CI, 0.24-0.61; P<0.001), and statins (HR, 0.33; 95% CI, 0.19-0.60; P<0.001) significantly correlated with reduced MACE in CMVD patients. In subgroups analysis, statins decreased MACE overall (HR, 0.33; 95% CI, 0.19-0.59; P<0.001) and in CMVD patients with smoking history (HR, 0.64; 95% CI, 0.43-0.93; P=0.014), diabetes (HR,0.27; 95% CI,0.12-0.61; P=0.002), hypertension (HR, 0.10; 95% CI, 0.03-0.36; P=0.001), and hypertension and diabetes (HR, 0.09; 95% CI, 0.014-0.53; P=0.008). CONCLUSION: Statins could reduce MACE in patients with CMVD.

One-pot synthesis of nickel-cobalt hydroxyfluorides nanowires with ultrahigh energy density for an asymmetric supercapacitor.

A novel and unique nickel-cobalt hydroxyfluorides (NiCo-HF) nanowires material is fabricated by one-pot solvothermal synthesis method for asymmetric supercapacitor. The synthesis mechanism and factors that influence the formation of the NiCo-HF nanowires have been further discussed. The as-prepared NiCo-HF electrode exhibits a high specific capacitance of 3,372.6 F g-1, and the capacitance retention of 94.3% can be achieved at a high current density of 20 A g-1 after 10,000 cycles. The outstanding electrochemical performance of the electrode can be attributed to the synergistic effect of the nanowires morphology and complicated redox process of active material. Furthermore, an asymmetric supercapacitor assembled with NiCo-HF nanowires as positive electrode and activated carbon as the negative electrode shows an ultrahigh energy density of 83.6 Wh kg-1 at a power density of 379.4 W kg-1 and an excellent cycling stability with 86.3% capacitance retention after 10,000 cycles, indicating that this novel material has great promise for potential application in energy storage device.

Associations Between Women's Perception of Their Husbands'/Partners' Social Support and Pap Screening in Pacific Islander Communities.

Pacific Islanders experience high rates of cervical cancer incidence and mortality. This cross-sectional study examined the extent to which Samoan, Chamorro, and Tongan women's perceived receipt of social support from their husbands or male partners was associated with rates of routine cancer screening- specifically Pap testing. A total of 585 Pacific Islander women who live in the United States completed a self-report survey. Women who reported having a Pap test within the past 3 years had significantly higher scores on support from their husbands/male partners. Furthermore, the relationship of emotional support and informational support with increased Pap testing was significantly stronger for Tongan women. The findings suggest that men play an important role in promoting women's cancer prevention behaviors in Pacific Islander and potentially other collectivistic populations. Incorporating social support messages into interventions may be a simple yet effective strategy to increase women's Pap testing.

Vaccination with a potent DNA vaccine targeting B-cell epitopes of hGRP induces prophylactic and therapeutic antitumor activity in vivo.

Gastrin-releasing peptide (GRP), a bombesin-like peptide, is an autocrine or paracrine growth factor that can stimulate the growth of various cancer cells, making it an ideal target antigen to develop vaccines against cancer. In this study, we developed a novel DNA vaccine that encodes six tandem repeats of B-cell epitope GRP(18-27) (GRP6) flanked by HSP65 as carrier and four tandem repeats of mycobacterial HSP70(407-426) (M4) as helper T-cell epitopes for enhancement of immunogenicity. When intramuscularly immunized to mice, this anti-GRP DNA vaccine-induced GRP-specific antibody (Ab) responses that were at least 10-fold higher in magnitude compared with HSP65-GRP6 protein vaccine. Both prophylactic and therapeutic antitumor immunities induced by vaccination significantly suppressed the growth of GRP-dependent prostate carcinoma RM-1 in vivo and prolonged the survival of tumor-inoculated mice. Out results also showed that the immune sera with high titer of GRP-specific Abs effectively inhibited the growth of tumor in mice and dose dependently inhibited proliferation of cultured RM-1 cells in vitro, suggesting that the GRP neutralizing Ab is responsible for the protective and therapeutic antitumor activity of vaccination. These findings may be of great importance in the further exploration of the applications of growth factors identified in human in cancer therapy.

Is image everything? The role of self-image in the relationship between family functioning and substance use among Hispanic adolescents.

This cross-sectional self-report study examined (1) whether family functioning (Family Functioning in Adolescence Questionnaire, FFAQ) and self-image (Piers-Harris Children's Self-Concept Scale) have independent associations with smoking and alcohol use and (2) whether self-image mediates the relationship between family functioning, smoking, and alcohol use among 1,598 Hispanic males and females in grades 7 through 12 in Los Angeles County, California, in 2001. The findings supported the main effects and, to some degree, the mediational effects that were hypothesized. Limitations and implications for this study are discussed, and directions for future research are suggested. This study was funded by the California Tobacco Related Disease Research Program.

Association between psychological factors and adolescent smoking in seven cities in China.

BACKGROUND: Cigarette smoking is a culturally accepted behavior among adult males in China. However, there is limited information on smoking among Chinese adolescents, particularly the information on the relationship between psychological well-being and smoking behavior among this population. PURPOSE: This study explored associations between three psychological factors--anxiety, hostility, and depressive symptoms--and smoking behavior among Chinese adolescents. METHODS: The data presented in this study are a cross-sectional slice from a longitudinal investigation of tobacco use and lifestyle carried out in China. The study population consisted of 4,724 7th and 11th grade students from seven large cities in China. RESULTS: Odds ratios showed that anxiety, hostility, and depressive symptoms were significantly associated with a higher risk of lifetime smoking for both boys and girls. CONCLUSION: Much like their Western counterparts, Chinese adolescents face developmental stress. The resulting distress may increase their risk for substance use and other health-risk behaviors.

A novel vaccine targeting gastrin-releasing peptide: efficient inhibition of breast cancer growth in vivo.

Gastrin-releasing peptide (GRP), a bombesin-like peptide, is an autocrine growth factor that can stimulate the growth of various cancer cells. We developed a novel protein vaccine HSP65-(GRP-10)(6) (HG6) that consists of six copies of a 10-amino acid residue epitope of GRP C-terminal fragment carried by mycobacterial 65 kDa HSP65 and then immunized mice via subcutaneous injection. Strong humoral and cell-mediated immune responses were induced. High titer of anti-GRP antibodies was detected in immunized mice sera by ELISA and verified by Western blot analysis. Activity of CD4+T lymphocytes, especially high levels of interferon (INF)-gamma, were developed in mice immunized with HG6 when compared with HSP65 or PBS. We found that immunogene tumor therapy with a vaccine based on GRP was effective at both protective and therapeutic antitumor immunity in breast tumor models in mice. The purified GRP monoclonal antibody (McAb) was proved to be potential in inhibiting EMT-6 tumor cell proliferation in vitro. The attenuation induced by active immune responses on tumor-induced angiogenesis was observed with an intradermal tumor model in mice. Taken together, we demonstrate for the first time that immune responses that are elicited by a novel chimeric protein vaccine targeting GRP can suppress the proliferation of breast tumor cell EMT-6 in mice, and it may be of importance in the further exploration of the applications of other autocrine growth factor identified in human and other animal in cancer therapy.

Ethnic variation in the association between weight concern and adolescent smoking.

This study examined differences in associations between weight concern and smoking initiation among culturally diverse adolescents. Statistical analyses were conducted on responses from 3,515 students in the 8th and 9th grades from three school districts in Los Angeles County. The restrained eating scale, adapted from the Dutch Eating Behavior Questionnaire, was used to measure students' weight concerns. Our results indicated that weight concern was significantly associated with increased risk for smoking. Those who scored higher on weight concern were approximately 40% more likely to report having tried smoking and smoked in the past 30 days. Compared to White students who reported weight concerns, Asian-American and African-American students were significantly less likely to report having tried smoking whereas Hispanic students were more likely to report having tried smoking. Health educators may wish to design smoking prevention programs which advocate for alternative methods of weight reduction rather than using smoking as a means of weight control.

Longitudinal effects of pro-tobacco and anti-tobacco messages on adolescent smoking susceptibility.

We examined the longitudinal impact of self-reported exposure to pro- and anti-tobacco media on adolescents' susceptibility to smoking, using in-school surveys from a culturally diverse sample. Ethnicity and acculturation also were examined as potential moderators. Middle-school students (N = 2,292) completed self-report questionnaires during the 6th, 7th, and 8th grades. Chi-square analyses were conducted to determine whether reported exposure to pro- and anti-tobacco media varied according to ethnicity, acculturation, and immigration status. Logistic regression models were used to examine whether pro- and anti-tobacco media exposure in 6th grade was associated with susceptibility to smoking by later grades. Recall of people smoking in television programs and pro-tobacco advertisements in stores was associated with adolescent smoking susceptibility. Exposure to anti-tobacco advertisements on television protected against susceptibility. No significant interaction effects between pro- and anti-tobacco media exposure on smoking susceptibility were found. Ethnicity and acculturation did not moderate these associations. Our longitudinal study provides evidence that pro-tobacco media and advertising increases susceptibility to smoking over time. More important, anti-tobacco advertisements are not sufficient to reduce the harmful effects of adolescent exposure to pro-tobacco media. Policy-level interventions such as restrictions in tobacco advertising may be necessary to prevent adolescent smoking.

Ten tandem repeats of beta-hCG 109-118 enhance immunogenicity and anti-tumor effects of beta-hCG C-terminal peptide carried by mycobacterial heat-shock protein HSP65.

The beta-subunit of human chorionic gonadotropin (beta-hCG) is secreted by many kinds of tumors and it has been used as an ideal target antigen to develop vaccines against tumors. In view of the low immunogenicity of this self-peptide,we designed a method based on isocaudamer technique to repeat tandemly the 10-residue sequence X of beta-hCG (109-118), then 10 tandemly repeated copies of the 10-residue sequence combined with beta-hCG C-terminal 37 peptides were fused to mycobacterial heat-shock protein 65 to construct a fusion protein HSP65-X10-betahCGCTP37 as an immunogen. In this study, we examined the effect of the tandem repeats of this 10-residue sequence in eliciting an immune by comparing the immunogenicity and anti-tumor effects of the two immunogens, HSP65-X10-betahCGCTP37 and HSP65-betahCGCTP37 (without the 10 tandem repeats). Immunization of mice with the fusion protein HSP65-X10-betahCGCTP37 elicited much higher levels of specific anti-beta-hCG antibodies and more effectively inhibited the growth of Lewis lung carcinoma (LLC) in vivo than with HSP65-betahCGCTP37, which should suggest that HSP65-X10-betahCGCTP37 may be an effective protein vaccine for the treatment of beta-hCG-dependent tumors and multiple tandem repeats of a certain epitope are an efficient method to overcome the low immunogenicity of self-peptide antigens.

Effects of acculturation and social norms on adolescent smoking among Asian-American subgroups.

This study provides new information about how acculturation and perceived social norms affect adolescents smoking among four Asian-American subgroups. Results showed differences in smoking prevalence rates across subgroups, with Koreans having the highest rates of smoking, while Chinese have the lowest rates. In contrast to the large gender disparity in the ancestral countries, smoking rates were equal for Asian-American boys and girls. Acculturation was significantly associated with an increased risk for lifetime smoking for Asian-American girls, but not for boys. Perceived social norms regarding peer smoking were significantly associated with smoking behaviors for both genders and for all subgroups.

Smoking among adolescents in China: an analysis based upon the meanings of smoking theory.

PURPOSE: This study utilized the Meanings of Behavior theory to examine whether meanings of smoking differ among adolescents in China who were never smokers, ever smokers, and past-30-day smokers. The Meanings of Behavior theory argues that affect takes precedence over cognitive constructs in motivating behavior This study also examined whether the associations among meanings of smoking and smoking behavior vary by age and gender DESIGN: This study was a cross-sectional study using survey data. SETTING: Middle and high schools in seven cities in China. SUBJECTS: A random sample of 4724 students comprised this study. MEASURE: A self-administered questionnaire asked about smoking behavior and incorporated the meanings of smoking scale. RESULTS: Overall prevalence rates of ever smokers and past-30-day smokers in this sample were 24.3% and 9.0%. Smoking was much more prevalent in boys than in girls. Students in the 11th grade were more likely than those in the 7th grade to have tried smoking at some time and to have smoked within the past 30 days. Odds ratios confirmed that meanings of smoking were significantly associated with smoking behaviors. CONCLUSIONS: Meanings of smoking are associated with smoking behavior among Chinese adolescents in that smoking may connote autonomy, control, or social relatedness. Prevention programs in China require new strategies to incorporate meanings of smoking in order to meet adolescent psychological needs for autonomy, competence, and social connections.

Improved efficacy of DNA vaccination against breast cancer by boosting with the repeat beta-hCG C-terminal peptide carried by mycobacterial heat-shock protein HSP65.

Studies have demonstrated that active-specific immunotherapy has potential for controlling mammary tumor progression. Human chorionic gonadotropin (hCG) is expressed and extremely sensitive, easily detectable and highly correlated with breast cancer. We developed a gene vaccine using a plasmid vector to deliver the six copies of 10-amino acid residues of beta-hCG 109-118 and beta hCG C-terminal 37-amino acid (CTP37). BALB/c female mice were immunized with a combination of pCR-HBc-X6-betahCGCTP37 DNA vaccine and HSP-X6-betahCGCTP37 protein vaccine. pCR-HBc-X6-betahCGCTP37 DNA vaccine were injected intramuscularly three times, on days -46,-25 and -11 and HSP-X6-betahCGCTP37 protein were applied two times, 21 and 14 days before tumor cell challenge. We assessed a combined DNA and protein vaccine for its effect of against murine EMT6 mammary tumor cells. In this study, animals vaccinated DNA vaccination boosting with the repeat beta-hCG C-terminal peptide carried by mycobacterial heat-shock protein HSP65 induced higher avidity antibodies and effectively inhibited the growth of tumor, compared with treatment using DNA alone or BCG priming HSP-X6-betahCGCTP37 protein boosting. The data presented demonstrate that improve immunogenicity of DNA vaccination by boosting with the repeat beta-hCG C-terminal peptide carried by mycobacterial heat-shock protein HSP65, which should prove useful in the development of new DNA vaccine against growth factors for cancer immunotherapy.

A synthetic gonadotropin-releasing hormone (GnRH) vaccine for control of fertility and hormone dependent diseases without any adjuvant.

Active immunization against self-peptides have gained widespread acceptance inspite of their low immunogenicity. Recent applications involving multiple copies of self-peptides in linear alignment and conjugation with carrier proteins appear to increase the immune response against self-peptides. As with most vaccines, however, immunogens require supplementation with adjuvants to elicit an optimum immune response. In the present study, we prepared a double-chain mini-protein with each chain containing three linear repeats of the self-peptide gonadotropin-releasing hormone (GnRH3), the hinge region of human IgG1 (hinge), and a T-helper epitope from the measles virus protein (MVP). The GnRH3-hinge-MVP mini-protein was conjugated to purified recombinant heat shock protein 65 (Hsp 65) of Mycobacterium bovis and used to immunize rats primed with subcutaneous injections of Bacillus Calmette-Guerin (BCG) in the absence of adjuvants. The GnRH3-hinge-MVP-Hsp 65 stimulated the production of specific anti-GnRH antibodies in the absence of adjuvants and the antibody titer was comparable to that produced in rats immunized with the dimeric mini-protein in the presence of Freund's adjuvant. Moreover, immunization with the adjuvant-free GnRH3-hinge-MVP-Hsp 65 induced degeneration of the reproductive organs in both male and female rats unlike those immunized in the absence of Hsp 65 or in control animals inoculated with the vehicle only. Histological examination of the affected organs showed atrophy of the seminiferous tubules with diminished spermatogenesis in the testes of male rats. In female rats, the uteri were much smaller in size and the ovaries exhibited reduced follicular development. These findings demonstrated that GnRH3-hinge-MVP-Hsp 65 mounted a strong immune response in the absence of conventional adjuvants, and could prove useful in control of fertility and the treatment of conditions/diseases where GnRH ablation is required.

Asparaginase display of polypeptides in the periplasm of Escherichia coli: potential rapid pepscan technique for antigen epitope mapping.

Asparaginase of Escherichia coli, a tetramer of identical subunits, was tested as a vector to display linear peptides on the surface of each enzyme subunit. A recombinant gene encoding a chimeric protein composed of asparaginase, a tetanus toxin peptide (TTP) spacer (831-854 fragment), and the foreign cholesteryl ester transfer protein C-terminal fragment (CETPC) was expressed and targeted to the periplasm of E. coli. The purified chimeric enzyme exhibited approximately 83% activity of the native enzyme, allowing the rapid screening of recombinant clones. In contrast, an asparaginase-CETPC fusion protein without the TTP spacer produced only about 23% activity of the native enzyme. Rats immunized with bacterial cells containing the chimeric enzymes induced CETP-specific immunoresponse. In contrast, rats inoculated with the cells expressing asparaginase only did not generate specific anti-CETP antibodies. Our study showed that asparaginase of E. coli was an effective carrier for displaying foreign peptides or epitopes. Moreover, the use of the TTP spacer appeared to play a critical role in maintaining the catalytic activity of the chimeric enzymes by redirecting the foreign CETPC peptide to the surface of the enzyme. The chimeric enzyme constructs fusing asparaginase with foreign peptides via a TTP spacer could be utilized as a rapid pepscan technique for antigen epitope mapping. The fusion protein of asparaginase-TTP-CETPC could also be useful for the development of a vaccine against atherosclerosis.

Hostility, depressive symptoms, and smoking in early adolescence.

Using logistic and multiple regression, we examined the association between hostility, level of depressive symptoms, and smoking in a sample of 1699 ethnically diverse students in California. Self-reports were collected twice from each student, at the beginning of the 6th and 7th grade years. Among 6th graders who had not smoked, depressive symptoms and hostility were associated with smoking initiation by the 7th grade. Among those students who had already tried smoking, increases in depressive symptoms and hostility were associated with more frequent smoking. The association between hostility and smoking was stronger for students reporting higher levels of depressive symptoms.

Bioeffects of low-frequency ultrasonic gene delivery and safety on cell membrane permeability control.

OBJECTIVE: To develop a novel method of ultrasonic naked gene delivery (UNGD); to examine the relationship between optimal parameters of ultrasound exposure and cell membrane permeability, enzymes, and free radicals; and to find optimal control parameters that were realizable, reliable, and noncytotoxic for use in gene therapy. METHODS: Suspensions of chicken, rabbit, and rat red blood cells and S180 cells were exposed to a calibrated ultrasonic field with different parameters in both the still and flowing states to obtain optimal parameters for UNGD. The optimal parameters then were used to implement UNGD. We examined morphologic characteristics, membrane permeability, enzymes, free radicals, naked gene expression efficiency, cell damage threshold, and cell viability by laser scanning confocal microscopy, fluorescent microscopy, flow cytometry, and spectrophotometry. RESULTS: Green fluorescent protein (GFP) as a reporter was delivered into S180 cells under the optimal parameters without cell damage or cytotoxicity. The transfection rate (mean +/- SD) was approximately 35.83% +/- 2.53% (n = 6) in viable cells, and cell viability was 90.17% +/- 1.47% (n = 6). The intensity of GFP expression with UNGD showed a higher fluorescent peak over both an adeno-associated virus vector-GFP group and a control group (P < .001). Additionally, malondialdehyde, hydroxyl free radicals, alkaline phosphatase, and acid phosphatase displayed an S-shaped growth model (r = 0.98 +/- 0.01) in response to permeability and morphologic alteration. CONCLUSIONS: Under optimal conditions, low-frequency ultrasound can safely deliver naked genes into cells without causing cell damage. The analytical results indicate that, except for subcavitation, free radical products are responsible for bioeffects in gene delivery. The constant E of energy deposition at 90% cell viability is the optimal control factor, and 80% viability represents the damage threshold. Optimal gene uptake by cells and safety depend on E. Constant E can be applied to control the gene delivery effect in combination with other parameters.