Metabolic syndrome — a new definition and management guidelines

The joint position paper by Polish Society of Hypertension, Polish Society for the Treatment of Obesity, Polish Lipid Association, Polish Association for Study of Liver, Polish Society of Family Medicine, Polish Society of Lifestyle Medicine, Division of Prevention and Epidemiology Polish Cardiac Society, “Club 30” Polish Cardiac Society, and Division of Metabolic and Bariatric Surgery Society of Polish Surgeons Reviewers: Agnieszka Olszanecka, Krzysztof J. Filipia

Tannic Acid Modified Silver Nanoparticles Show Antiviral Activity in Herpes Simplex Virus Type 2 Infection

The interaction between silver nanoparticles and herpesviruses is attracting great interest due to their antiviral activity and possibility to use as microbicides for oral and anogenital herpes. In this work, we demonstrate that tannic acid modified silver nanoparticles sized 13 nm, 33 nm and 46 nm are capable of reducing HSV-2 infectivity both in vitro and in vivo. The antiviral activity of tannic acid modified silver nanoparticles was size-related, required direct interaction and blocked virus attachment, penetration and further spread. All tested tannic acid modified silver nanoparticles reduced both infection and inflammatory reaction in the mouse model of HSV-2 infection when used at infection or for a post-infection treatment. Smaller-sized nanoparticles induced production of cytokines and chemokines important for anti-viral response. The corresponding control buffers with tannic acid showed inferior antiviral effects in vitro and were ineffective in blocking in vivo infection. Our results show that tannic acid modified silver nanoparticles are good candidates for microbicides used in treatment of herpesvirus infections.This work was supported by the Polish National Science Centre grant No. 2011/03/B/NZ6/04878 (for MK) and Centre for Preclinical Research and Technology (CePT) Project No. POIG.02.02.00-14-024/08-0 (for MG and MD). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscrip

Psychometric properties and cultural adaptation of the polish version of the healthy lifestyle and personal control questionnaire (Hlpcq)

Background: Chronic non-communicable diseases (NCDs), sometimes referred to as lifestyle diseases, are the most common cause of death and disability worldwide. Thus, healthcare professionals should be equipped with tools, knowledge, skills, and competencies in the newly distinguished field of lifestyle medicine. The purpose of this study was to test the psychometric properties of the Polish version of the Healthy Lifestyle and Personal Control Questionnaire (HLPCQ). The Polish version of the HLPCQ would further provide Polish healthcare professionals with a useful and convenient tool for routine lifestyle assessment while giving HLPCQ novel use and potential for further research. Methods: Before testing its psychometric properties, the HLPCQ was translated and adapted from the original Greek version into Polish. Subsequently, we tested the instrument’s psychometric properties on a sample of 2433 participants. In addition, we tested the factorial validity of the HLPCQ using confirmatory and exploratory factor analysis. Results: There were more female than male participants (91.78%). Most of them were middle-aged (30.40 ± 7.71), single (39.62%), and living with family (70.65%). In terms of residence, 1122 (46.12%) participants lived in cities with a population of over 500,000. In terms of reliability, the internal consistency of the Polish version and its domains is excellent. Cronbach’s alpha for each of the domains of the scale ranged between 0.6 and 0.9. Conclusions: The Polish version of the Healthy Lifestyle and Personal Control Questionnaire (HLPCQ) has good characteristics of factorial validity and can be used in clinical practice and research

Decreased recruitment of NK, CD4+ and CD8+ T cells in HSV-2 infected Fas and FasL-deficient mice.

<p>Vaginal tissue samples from Fas <i>(−/−)</i>, FasL <i>(−/−)</i> and WT (C57BL/6) mice (n = 5 mice per strain) isolated at 3 and 7 day of HSV-2 infection were pooled and used to prepare single cell suspensions and further analyzed for the total numbers of CD4+ T cells (A), CD8+ T cells (B) and NK cells (C). Uninfected mice were used as controls. Each bar represents the mean total cell numbers from 5 separate experiments (N = 5) ± SEM. * represent significant differences with <i>p</i>≤0.05, **<i>p</i>≤0.01.</p

HSV-2 infection of mouse monocytes leads to early induction of apoptosis in comparison to keratinocytes.

<p>(A) HSV-2 infected monocyte cell culture showed significantly less infected cells at 24 and 48 h p.i. (hours post infection) in comparison to HSV-2 infected keratinocyte cell cultures. (B) Kinetics of total percentage of M30-positive (apoptotic) cells in monocyte RAW 264.7 and keratinocyte 03C cell line cultures during 24 hours after infection with HSV-2. (C) Representative dot plots showing M30 and/or HSV-2 positive cells detected by flow cytometry at 6 and 18 h p.i. in monocyte and keratinocyte cultures and in control uninfected cultures. (D) Kinetics of HSV-2 infected cells undergoing apoptosis in monocyte RAW 264.7 and keratinocyte 03C cell line cultures during 24 hours after infection with HSV-2 (R3 gate from panel C). (E) Kinetics of uninfected cells undergoing apoptosis in monocyte RAW 264.7 and keratinocyte 03C cell line cultures during 24 hours after infection with HSV-2 (R2 gate from panel C). Each bar represents the mean of 5 independent experiments (N = 5) ± SEM. * represents significant differences with <i>p</i>≤0.05, while ** means <i>p</i>≤0.001. *k means significant differences for keratinocyte cultures, while *m means significant differences for monocyte cultures.</p

HSV-2 infected monocytes up-regulate FasL, but are susceptible to Fas-induced apoptosis.

<p>The cells in the HSV-2 infected RAW 264.7 cultures were analysed for Fas and FasL expression using flow cytometry and all event from gate R2 were divided into infected and uninfected cells expressing Fas or FasL. Fas expression is shown in the form of representative dot plots (A) or as the mean fluorescence intensity (MFI) for infected (white bars) and uninfected cells (grey bars) (B). FasL expression is presented as the representative dot plots (A) or as the percentage of FasL - positive cells (C). (D) Susceptibility to Fas-induced apoptosis was tested in RAW 264.6 cultures at 6 and 18 hours after infection with MOI = 1 in the presence of cytotoxic anti-Fas antibody (10 µg/ml) able to induce apoptosis through binding Fas receptor or FasL-blocking antibody (10 µg/ml). Apoptosis was detected using intracellular M30 staining. (E) Apoptotic monocytes (annexinV+/CD11b+ cells) after 12 h of co-culture with murine 291.O3C keratinocytes uninfected or HSV-2 infected at 18 h p.i. Each bar represents the mean from 3 experiments (N = 3) ± SEM. * represents significant differences with <i>p</i>≤0.05, while ** means <i>p</i>≤0.001.</p

Monocytes during in vivo HSV-2 infection are susceptible to Fas-induced apoptosis.

<p>(A) CD11b+ cells (green) and HSV-2 antigen localisation (red) in the vaginal tissue isolated from control, uninfected mice C57BL/6 mice (left) and at 3 days of HSV-2 infection (right). The nuclei were counterstained with Hoechst 33342 (blue). (B) Gating strategy for identification of monocytes in the vaginal tissue (CD11b+/Ly6G−/CD11c-). Histograms (C) and bars (D) showing total percentage of Fas and FasL-expressing monocytes (CD11b+/Ly6G-) in the cell suspensions prepared from HSV-2 infected C57BL/6 mice at 3 day of infection and from control uninfected mice. (E) Total counts of monocytes (CD11b+/Ly6G−/CD11c-) in cell suspensions prepared from the vaginal tissues isolated from Fas <i>(−/−)</i>, FasL <i>(−/−)</i> and WT (C57BL/6) mice at 3 and 7 day of HSV-2- infection and from control uninfected mice. (F) Total counts of annexin V-positive monocytes in the vaginal tissue cell suspensions prepared from HSV-2-infected Fas <i>(−/−)</i>, FasL <i>(−/−)</i> and WT (C57BL/6) mice at 3 and 7 day of infection. The bars represent mean from 5 separate experiments ± SEM. * represents significant differences with <i>p</i>≤0.05, while ** means <i>p</i>≤0.01.</p

Fas is involved in production of TNF-α, CXCL9 and CXCL10 during HSV-2 infection.

<p>Vaginal tissues and vaginal lavages of Fas <i>(−/−)</i>, FasL <i>(−/−)</i> and WT (C57BL/6) mice (n = 5 mice per strain) infected with HSV-2 were collected at 3 and 7 day of infection. Uninfected mice were used as controls. The pooled vaginal tissues were used to prepare single cell suspensions and further stained for monocytes and intracellular expression of TNF-α, CXCL9 and CXCL10. Vaginal lavages from single mouse were used for further determination of TNF-α (A), CXCL9 (B) and CXCL10 (C). Percentages of monocytes with TNF-α (D), CXCL9 (E) and CXCL10 (F) expression in the vaginal tissues. (G) TNF-α, IL-1β, CXCL9 and CXCL10 levels in culture supernatants at 18 h p.i. of RAW 264.7 cell cultures infected with HSV-2 and exposed or not, to anti-Fas cytotoxic antibody (10 µg/ml). The bars represent the mean from 3 separate experiments (N = 3) ± SEM. * represents significant differences with <i>p</i>≤0.05, **<i>p</i>≤0.01. n.d. – not detected.</p

Monocytes do not up-regulate Bcl-2 expression during HSV-2 infection and show significant early caspase-9 activation.

<p>(A) Representative dot plots showing cells positive for HSV-2 and/or active form of caspase-9 detected by flow cytometry at 6 and 18 h p.i. and in control uninfected cultures of monocytes and keratinocytes. (B) Percentage of HSV-2-positive cells with active caspase-9 form in HSV-2 infected monocyte RAW 264.7 and keratinocyte 291.03C cell cultures (R2 gate from panel A). (C) Percentage of uninfected cells with active caspase-9 form in HSV-2 infected monocyte and keratinocyte cell cultures (R3 gate from panel A). (D), (E) Mean fluorescence intensity (MFI) showed as histograms (left panels) and bars (right panels) for Bcl-2 (D) and Bax (E) proteins expression in the monocyte (white bars) and keratinocyte (black bars) cell cultures infected with HSV-2 at 6 and 18 hp.i., detected by intracellular staining and flow cytometry. Controls are uninfected cultures (0 h p.i.). Each bar represents the mean from 5 experiments (N = 5) ± SEM. * represents significant differences with <i>p</i>≤0.05, while ** means <i>p</i>≤0.001.</p