Evidence-Based Guidelines for Cardiovascular Disease Prevention in Women

Significant advances in our knowledge about interventions to prevent cardiovascular disease (CVD) have occurred since publication of the first female-specific recommendations for preventive cardiology in 1999.1 Despite research-based gains in the treatment of CVD, it remains the leading killer of women in the United States and in most developed areas of the world.2–3 In the United States alone, more than one half million women die of CVD each year, exceeding the number of deaths in men and the next 7 causes of death in women combined. This translates into approximately 1 death every minute.2 Coronary heart disease (CHD) accounts for the majority of CVD deaths in women, disproportionately afflicts racial and ethnic minorities, and is a prime target for prevention.1–2 Because CHD is often fatal, and because nearly two thirds of women who die suddenly have no previously recognized symptoms, it is essential to prevent CHD.2 Other forms of atherosclerotic/thrombotic CVD, such as cerebrovascular disease and peripheral arterial disease, are critically important in women. Strategies known to reduce the burden of CHD may have substantial benefits for the prevention of noncoronary atherosclerosis, although they have been studied less extensively in some of these settings

<i>Giardia duodenalis</i> Infection Reduces Granulocyte Infiltration in an <i>In Vivo</i> Model of Bacterial Toxin-Induced Colitis and Attenuates Inflammation in Human Intestinal Tissue

<div><p><i>Giardia duodenalis (syn. G. intestinalis, G. lamblia)</i> is a predominant cause of waterborne diarrheal disease that may lead to post-infectious functional gastrointestinal disorders. Although <i>Giardia</i>-infected individuals could carry as much as 10⁶ trophozoites per centimetre of gut, their intestinal mucosa is devoid of overt signs of inflammation. Recent studies have shown that in endemic countries where bacterial infectious diseases are common, <i>Giardia</i> infections can protect against the development of diarrheal disease and fever. Conversely, separate observations have indicated <i>Giardia</i> infections may enhance the severity of diarrheal disease from a co-infecting pathogen. Polymorphonuclear leukocytes or neutrophils (PMNs) are granulocytic, innate immune cells characteristic of acute intestinal inflammatory responses against bacterial pathogens that contribute to the development of diarrheal disease following recruitment into intestinal tissues. <i>Giardia</i> cathepsin B cysteine proteases have been shown to attenuate PMN chemotaxis towards IL-8/CXCL8, suggesting <i>Giardia</i> targets PMN accumulation. However, the ability of <i>Giardia</i> infections to attenuate PMN accumulation <i>in vivo</i> and how in turn this effect may alter the host inflammatory response in the intestine has yet to be demonstrated. Herein, we report that <i>Giardia</i> infection attenuates granulocyte tissue infiltration induced by intra-rectal instillation of <i>Clostridium difficile</i> toxin A and B in an isolate-dependent manner. This attenuation of granulocyte infiltration into colonic tissues paralled decreased expression of several cytokines associated with the recruitment of PMNs. <i>Giardia</i> trophozoite isolates that attenuated granulocyte infiltration <i>in vivo</i> also decreased protein expression of cytokines released from inflamed mucosal biopsy tissues collected from patients with active Crohn’s disease, including several cytokines associated with PMN recruitment. These results demonstrate for the first time that certain <i>Giardia</i> infections may attenuate PMN accumulation by decreasing the expression of the mediators responsible for their recruitment.</p></div

Streptococcus pneumoniae colonization is required to alter the nasal microbiota in cigarette smokeexposed mice

Smokers have nasal microbiota dysbiosis, with an increased frequency of colonizing bacterial pathogens. It is possible that cigarette smoke increases pathogen acquisition by perturbing the microbiota and decreasing colonization resistance. However, it is difficult to disentangle microbiota dysbiosis due to cigarette smoke exposure from microbiota changes caused by increased pathogen acquisition in human smokers. Using an experimental mouse model, we investigated the impact of cigarette smoke on the nasal microbiota in the absence and presence of nasal pneumococcal colonization. We observed that cigarette smoke exposure alone did not alter the nasal microbiota composition. The microbiota composition was also unchanged at 12 h following low-dose nasal pneumococcal inoculation, suggesting that the ability of the microbiota to resist initial nasal pneumococcal acquisition was not impaired in smoke-exposed mice. However, nasal microbiota dysbiosis occurred as a consequence of established high-dose nasal pneumococcal colonization at day 3 in smoke-exposed mice. Similar to clinical reports on human smokers, an enrichment of potentially pathogenic bacterial genera such as Fusobacterium, Gemella, and Neisseria was observed. Our findings suggest that cigarette smoke exposure predisposes to pneumococcal colonization independent of changes to the nasal microbiota and that microbiota dysbiosis observed in smokers may occur as a consequence of established pathogen colonization.</p

<i>Giardia</i> NF trophozoites attenuate several PMN-associated mediators released from human colonic mucosal biopsies.

<p>Human descending colon mucosal biopsy tissues obtained from areas of active inflammation from patients with active Crohn’s disease (CD) were incubated with 5.0×10⁶<i>Giardia</i> NF trophozoites for 6 hours. Supernatant levels of (A) IL-8/CXCL8, (B) GRO, (C) CCL3, (D) IL-17A, (E) G-CSF, and (F) GM-CSF were determined via a bead-based cytokine assay. All data are representative of three independent experiments (n = 4–6/group) and represented as median with the range. * p<0.05.</p

<i>Giardia</i> NF infections attenuate granulocyte recruitment in a model of <i>C. difficile</i> TcdAB-induced colitis.

<p>Male C57BL/6 mice aged 4 to 6 weeks were infected with <i>Giardia</i> NF or GS/M trophozoites. 7 days post-infection, animals were given 100 µg of TcdAB or PBS i.r. and, after 3 hours, animals were euthanized. (A) Trophozoites counts within the upper 3 cm of the jejunum were determined. (B) Colonic myeloperoxidase (MPO) activity was determined between uninfected controls and <i>Giardia</i> NF-infected animals given 100 µg TcdAB or PBS. (C) Colonic MPO activity was determined between uninfected controls and <i>Giardia</i> GS/M-infected animals given 100 µg TcdAB or PBS. (D) Values were determined via bead-based cytokine assay. All data are representative of two independent experiments (n = 5–11/group) and represented as mean ± SEM. * p<0.05.</p

<i>Giardia</i> NF trophozoites attenuate the release of several chemokines from colonic mucosal biopsies.

<p>Human descending colon mucosal biopsy tissues obtained from areas of active inflammation from patients with active Crohn’s disease (CD) were incubated with 5.0×10⁶<i>Giardia</i> NF trophozoites for 6 hours. Supernatant levels of (A) CCL2, (B) CCL4, (C) CCL5, (D) CCL7, (E) CCL11, (F) CCL22, (G) CXCL10, and (H) CX₃CL1 were determined via a bead-based cytokine assay. All data are representative of three independent experiments (n = 4–6/group) and represented as median with the range. * p<0.05.</p

<i>Giardia</i> NF trophozoites attenuate the expression of certain pro-inflammatory mediators assessed in colonic mucosal biopsy tissue homogenates.

<p>Human descending colon mucosal biopsy tissues obtained from areas of active inflammation from patients with active Crohn’s disease (CD) were incubated with 5.0×10⁶<i>Giardia</i> NF trophozoites for 6 hours. Tissue homogenate levels of (A) IL-1α, (B) IL-6, (C) IL-7, (D) IL-10, (E) IL-12p70, (F) IFNα, (G) TNFα, (H) sCD40L, and (I) VEGF were determined via a bead-based cytokine assay. All data are representative of three independent experiments (n = 4–6/group) and represented as median with the range. * p<0.05.</p

<i>Giardia</i> NF trophozoites attenuate the release of several pro-inflammatory mediators from colonic mucosal biopsies.

<p>Human descending colon mucosal biopsy tissues obtained from areas of active inflammation from patients with active Crohn’s disease (CD) were incubated with 5.0×10⁶<i>Giardia</i> NF trophozoites for 6 hours. Supernatant levels of (A) IL-1α, (B) IL-6, (C) IL-7, (D) IL-10, (E) IL-12p70, (F) IFNα, (G) TNFα, (H) sCD40L, and (I) VEGF were determined via a bead-based cytokine assay. All data are representative of three independent experiments (n = 4–6/group) and represented as median with the range. * p<0.05.</p

<i>Giardia</i> NF infections attenuate colonic expression levels of several PMN-associated mediators following administration of <i>C. difficile</i> TcdAB.

<p>Male C57BL/6 mice, aged 4 to 6 weeks, were infected with <i>Giardia</i> NF or GS/M trophozoites. 7 days post-infection, animals were given 100 µg of TcdAB or PBS i.r. and, after 3 hours, animals were euthanized. Levels of (A) CXCL1, (B) CXCL2, (C) IL-17, and (D) G-CSF were compared between <i>Giardia</i> NF-infected animals and uninfected animals given i.r. 100 µg of TcdAB or PBS. Levels of (E) CXCL1, (F) CXCL2, (G) IL-17, and (H) G-CSF were compared between <i>Giardia</i> GS/M-infected animals and uninfected animals given i.r. 100 µg of TcdAB or PBS. Values were determined via bead-based cytokine assay. All data are representative of two independent experiments (n = 5–11/group) and represented as mean ± SEM. n.s. = not significant * p<0.05.</p