Differences in smoking attitudes of adolescents and young adults

This study employed the Theory of Planned Behavior to examine the differences between adolescent (n = 182) and young adults (n = 209) in their intention to smoke and examined possible differences. Analysis showed that young adults had more positive self-reported attitudes toward smoking than adolescents, had higher intentions to smoke, lower perceived behavioral control over smoking and perceived they were more informed about smoking. The Theory of Planned Behavior provided good prediction of intention for both young adults (R-2 = .70, attitudes, information, and past behavior significant) and adolescents (R-2 = .68, attitudes, past behavior significant). For both samples attitudes were the strongest predictor of intentions to smoke. Implications for understanding intention toward smoking between adolescents and young adults are discussed

Isolation and phenotypic characterization of large vein endothelial cells.

<p>Cells isolated from surgical specimens were characterized <i>in-vitro</i> by morphological assessment (<b>A</b>) and multiparametric flow cytometry analysis (<b>B</b>). In <b>A</b>, cell morphology was examined by using phase-contrast microscopy. VEC cultures were characterized by regular polygonal shape and dimensions and uniform monolayer. On the other hand, non-VEC cultures appeared with a fibroblast-like morphology characterized by elongated shapes and growing in an uneven manner. Representative images of VEC and non-VEC cultures, at two different <i>in vitro</i> passages (p = 0 and p = 3), are shown. Left panels: 10X, original magnification; right panels: 20X original magnifications. In <b>B</b>, multiparametric flow cytometry analyses were performed with a specific panel of endothelial cells defining antibodies. VEC were defined as CD146⁺/CD144⁺/CD31⁺/CD105⁺/CD34⁺/CD45⁻/CD14⁻ while non-VEC displayed a more variable and random pattern of antigens expression. Two representative multiparametric flow cytometry analysis panels of a non-VEC and pure VEC cultures are shown as two-colors dot plots.</p

Baseline characteristics of subjects.

<p>Baseline characteristics of subjects.</p

Differential phenotype between control and pathological VEC.

<p>Surface expression of CD31 and CD146 was evaluated by flow cytometry in either pathological and control-VEC. In <b>A</b>, colored histograms represent cells stained with monoclonal antibodies specific for the indicated antigens and white histograms represent background fluorescence obtained by staining the same cells with isotype-matched control antibodies. Representative panels for control, C2- and C3-VEC are shown. In <b>B</b>, the expression levels of the indicated antigens were determined for all VEC samples (8 C2-VEC, 13 C3-VEC and 5 control-VEC) by flow cytometry analysis and expressed as mean fluorescence intensity (MFI). Horizontal bars are median, upper and lower edges of box are 75th and 25th percentiles, lines extending from box are 10th and 90th percentiles. *<i>P</i><0.05 compared to control VEC.</p

Analysis of ICAM-1 and VCAM expression in control and pathological VEC.

<p>In <b>A–C</b>, surface expression of ICAM-1 antigen was evaluated by flow cytometry in either pathological and control-VEC. In <b>A</b>, colored histograms represent cells stained with monoclonal antibodies specific for the indicated antigens and white histograms represent background fluorescence obtained by staining the same cells with isotype-matched control antibodies. A representative panel for control-, C2- and C3-VEC is shown. In <b>B</b>, the expression levels of ICAM-1 were determined for all VEC samples (8 C2-VEC, 13 C3-VEC and 5 control VEC) by flow cytometry analysis and expressed as mean fluorescence intensity (MFI). *<i>P</i><0.05 compared to control VEC. In <b>C</b>, comparative analysis of ICAM-1 surface expression, reported as MFI, at different VEC passages. In <b>D–E</b>, pathological and control-VEC cultures were exposed to TNF-α for 18 hours before ICAM-1 and VCAM surface expression analysis by flow cytometry (<b>D</b>), and mRNA levels analysis by quantitative RT-PCR (<b>E</b>). In <b>D</b>, two representative panels are shown: dotted histograms represent background fluorescence obtained by staining the same cells with isotype-matched control antibodies. The expression levels of ICAM-1 and VCAM, determined for all VEC samples by flow cytometry analysis upon TNF-α stimulation, are expressed as mean fluorescence intensity (MFI). In <b>E</b>, mRNA expression levels of ICAM-1 and VCAM were determined both in unstimulated and TNF-α-stimulated VEC cultures. Results from amplifications, done in duplicate, are expressed as arbitrary units, after normalization for the housekeeping gene. Horizontal bars are median, upper and lower edges of box are 75th and 25th percentiles, lines extending from box are 10th and 90th percentiles.</p

Analysis of the NF-kB, OPG and VEGF levels in VEC culture supernatants and serum samples.

<p>In <b>A</b>, NF-kB-p65 DNA binding activity was assessed in duplicate using the TransAM assay. Results are reported as absorbance values (O.D.) per 20 µg of cell lysate protein. In <b>B</b>, OPG and VEGF levels were determined by ELISA in VEC culture supernatants. In <b>C</b>, OPG and VEGF levels were determined by ELISA in sera of C2 and C3 patients as well as in sex and age-matched normal controls. In <b>A–C</b>, horizontal bars are median, upper, and lower edges of box are 75th and 25th percentiles; lines extending from box are 10th and 90th percentiles. *, <i>P</i><0.05 compared to either control VEC (<b>A–B</b>) or control sera (<b>C</b>).</p

An Attenuated Herpes Simplex Virus Type 1 (HSV1) Encoding the HIV-1 Tat Protein Protects Mice from a Deadly Mucosal HSV1 Challenge

<div><p>Herpes simplex virus types 1 and 2 (HSV1 and HSV2) are common infectious agents in both industrialized and developing countries. They cause recurrent asymptomatic and/or symptomatic infections, and life-threatening diseases and death in newborns and immunocompromised patients. Current treatment for HSV relies on antiviral medications, which can halt the symptomatic diseases but cannot prevent the shedding that occurs in asymptomatic patients or, consequently, the spread of the viruses. Therefore, prevention rather than treatment of HSV infections has long been an area of intense research, but thus far effective anti-HSV vaccines still remain elusive. One of the key hurdles to overcome in anti-HSV vaccine development is the identification and effective use of strategies that promote the emergence of Th1-type immune responses against a wide range of epitopes involved in the control of viral replication. Since the HIV1 Tat protein has several immunomodulatory activities and increases CTL recognition of dominant and subdominant epitopes of heterologous antigens, we generated and assayed a recombinant attenuated replication-competent HSV1 vector containing the <i>tat</i> gene (HSV1-Tat). In this proof-of-concept study we show that immunization with this vector conferred protection in 100% of mice challenged intravaginally with a lethal dose of wild-type HSV1. We demonstrate that the presence of Tat within the recombinant virus increased and broadened Th1-like and CTL responses against HSV-derived T-cell epitopes and elicited in most immunized mice detectable IgG responses. In sharp contrast, a similarly attenuated HSV1 recombinant vector without Tat (HSV1-LacZ), induced low and different T cell responses, no measurable antibody responses and did not protect mice against the wild-type HSV1 challenge. These findings strongly suggest that recombinant HSV1 vectors expressing Tat merit further investigation for their potential to prevent and/or contain HSV1 infection and dissemination.</p></div

Determination of HSV1-Tat and HSV1-LacZ viral attenuation in Balb/c mice.

<p>Groups of mice (n = 10) were infected by the intravaginal route with a single dose (ranging from 10⁴ to 10⁸ pfu/mouse) of HSV1-Tat or HSV1-LacZ at a single time point. Mice were observed daily for survival up to 20 days post-infection, for survival. The results of the group inoculated with 10⁸ pfu/mouse are shown. The results of one representative experiment (out of three) are shown. The Kaplan-Meier test was used to estimate the probability of clinical manifestation.</p

Disease severity and survival of BALB/c (A and B) or C57BL/6 (C and D) mice inoculated intravaginally with HSV1-LacZ or HSV1-Tat after challenge with a lethal dose of wild-type HSV1.

<p>Ten mice/group were inoculated intravaginally with 10³ pfu/mouse of HSV1-LacZ, HSV1-Tat or control buffer, and, 28 days later, challenged intravaginally with a lethal dose of wild-type HSV1 (2×10⁶ pfu/mouse for BALB/c mice and 2×10⁸ pfu/mouse for C57BL/6 mice). Mice were observed daily for appearance of signs of HSV1 disease and death. Disease severity scores (A for BALB/c and C for C57BL/6) and survival (B for BALB/c and D for C57BL/6) were assessed. Mean disease scores (± SD) for each group is shown. The results of one representative experiment (out of three) are shown. Data were analysed statistically using the two-tailed Mann Whitney test (panels A and C) and the Kaplan-Meier test. **P<0.01 (panels B and D).</p

Proteolytic activity of proteasomes isolated from CB1 cells, uninfected or infected with 5 MOI of HSV1-Tat or HSV1-LacZ. Trypsin-like (A) and chymotrypsin-like (B) proteasome activities measured at 6, 12, and 24 h after infection are shown.

<p>Proteasome activity is expressed as arbitrary fluorescence units (AFU). The means (± SD) of three independent experiments are shown. The two-tailed Wilcoxon signed-rank test was used for statistical analysis, *p<0.05.</p