Clofibrate improves myocardial ischemia-induced damage through regulation of renin-angiotensin system and favours a pro-vasodilator profile in left ventricle.

Myocardial ischemia initiates a chain of pathological conditions leading to cardiomyocyte death. Therefore, pharmacological treatment to stop ischemia-induced damage is necessary. Fibrates, have been reported to decrease inflammatory markers and to modulate the renin-angiotensin system (RAS). Our aim was to explore if clofibrate treatment, administered one week after myocardial event, decreases MI-induced cardiac damage. Wistar rats were assigned to: 1. Sham or 2. Coronary artery ligation (MI). Seven days after, rats were subdivided to receive vehicle (V) or clofibrate [100 mg/kg (C)] daily for 7 days. Blood samples and left ventricle were analyzed. RAS components [angiotensin II, angiotensin converting enzyme (ACE), and AT1-receptor] decreased in MI-C compared to MI-V, while [Ang-(1-7), bradykinin, ACE-2, and AT2-receptor] raised in response to clofibrate treatment. Oxidative stress markers increased in MI-V rats, a profile reverted in MI-C rats. Nitric oxide (NO) pathway (Akt, eNOS, and NO) exhibits a lower participation in MI-V, but clofibrate raised NO-pathway components and its production. MI-induced fibrosis and structural damage was also improved by clofibrate-treatment. In conclusion, clofibrate administration to 7 days MI-rats exerts an antioxidant, pro-vasodilator expression profile, and anti-fibrotic effect suggesting that PPARα activation can be considered a therapeutic target to improve cardiac condition posterior to ischemia.

Socio-demographic Moderators of Associations Between Psychological Factors and Latinas' Breast Cancer Screening Behaviors.

This study tested whether socio-demographic factors moderated associations between psychological factors and Latinas' breast cancer screening behaviors. 222 churchgoing Latinas (40-65 years) in San Diego, CA completed surveys assessing socio-demographics (e.g., income and acculturation), psychological factors (e.g., perceived barriers to screening), and cancer screening behaviors. Multilevel models examined associations of socio-demographic and psychological factors (and their interactions) with adherence to annual mammography or clinical breast exam (CBE) screening. Although no main effects were found, there were moderation effects. Acculturation moderated associations between perceived barriers to screening and both screening outcomes, with inverse associations only among the high-acculturation group. Education moderated the relationship between perceived barriers to screening and CBE screening, with an inverse association only among the low-education group. Marital status moderated the relationship between depressive symptoms and CBE screening, with an inverse association only among single/non-partnered participants. Interventions are needed targeting psychological barriers to breast cancer screening among Latinas.

Where and when adolescents are physically active: Neighborhood environment and psychosocial correlates and their interactions.

Female adolescents are less active than male peers in certain contexts including the neighborhood. Adolescents' physical activity can be explained by interactions between environmental and psychosocial factors, but few studies have tested such interactions in relation to context-specific behaviors. This study tested interactions between neighborhood environmental and psychosocial factors in relation to adolescents' context-specific physical activity. Data were collected in 2009-11 from 910 adolescents and a parent/guardian residing in the Baltimore/Seattle regions. Measures included adolescent-reported neighborhood leisure-time physical activity (LTPA) and non-neighborhood LTPA, accelerometer-based non-school moderate-to vigorous-physical activity (MVPA), psychosocial factors, and objective and parent-perceived neighborhood environmental factors. Gender-stratified mixed effects linear models tested associations of 6 environmental and 4 psychosocial factors and their interactions in relation to each physical activity outcome. The psychosocial factors had consistent associations with the physical activity outcomes but the environmental correlates were context-specific. Decisional balance (weighing of pros and cons of physical activity) moderated the association between recreation facility density and neighborhood LTPA among females, with a negative association only among those with high decisional balance (pros outweighed cons). Decisional balance also moderated associations of neighborhood walkability with non-school MVPA among females and non-neighborhood LTPA among males, with positive associations only among those with high decisional balance. Results support context-specific ecological models of physical activity. Targeting environmental factors that may promote opportunities for physical activity in specific contexts as well as adolescent decision-making may help promote their physical activity in those contexts, potentially leading to increased overall physical activity.

Promoting cancer screening among churchgoing Latinas: Fe en Acción/faith in action.

Cancer screening rates among Latinas are generally low, reducing the likelihood of early cancer detection in this population. This article examines the effects of a community intervention (Fe en Acción/Faith in Action) led by community health workers (promotoras) on promoting breast, cervical and colorectal cancer screening among churchgoing Latinas. Sixteen churches were randomly assigned to a cancer screening or a physical activity intervention. We examined cancer knowledge, barriers to screening and self-reported mammography, clinical breast exam, Pap test, fecal occult blood test and sigmoidoscopy or colonoscopy at baseline and 12 months follow-up. Participants were 436 adult Latinas, with 16 promotoras conducting a cancer screening intervention at 8 out of 16 churches. The cancer screening intervention had a significant positive impact on self-reported mammography (OR = 4.64, 95% CI: 2.00-10.75) and breast exams in the last year (OR= 2.82, 95% CI: 1.41-5.57) and corresponding reductions in perceived (87.6%) barriers to breast cancer screening (P < .008). Cervical and colorectal cancer screening did not improve with the intervention. These findings suggest Fe en Acción church-based promotoras had a significant impact on promoting breast cancer screening among Latinas. Colon cancer screening promotion, however, remains a challenge.

Interactions between individual and perceived environmental factors on Latinas' physical activity.

Background: Latinas have disproportionately low levels of physical activity (PA) and the ecological correlates of their PA remain unclear. This study aims to test interactions between individual and environmental factors on Latinas' PA. Methods: We analyzed baseline data from 436 Latinas participating in a PA randomized controlled trial in San Diego, CA [Fe en Acción/Faith in Action]. Measures included demographics, perceived environment, PA and anthropometrics. Mixed effects models examined interactions between individual and environmental factors on self-reported leisure-time and transportation, and accelerometer-assessed PA. Results: Significant positive associations were found between neighborhood aesthetics and leisure-time moderate-to-vigorous PA (MVPA) and between having destinations within walking distance from home and transportation PA (P < 0.05). We found significant interactions of income with aesthetics and sidewalk maintenance as well as between weight status and safety from crime. Favorable aesthetics was related to more leisure-time MVPA only among lower income women (odds ratio (OR) = 1.57; 95% confidence interval (CI): 1.18, 2.08); however, higher income women reporting better sidewalk maintenance reported more leisure-time MVPA (OR = 1.51; 95% CI: 1.06, 2.15). Higher perceived safety from crime was positively related to transportation PA only among overweight/obese women. Conclusions: Subgroup differences should be considered when developing interventions targeting the neighborhood environment to promote Latinas' PA.

Does the social environment moderate associations of the built environment with Latinas' objectively-measured neighborhood outdoor physical activity?

Favorable perceptions of the built and social neighborhood environment may promote outdoor physical activity (PA). However, little is known about their independent and interactive effects on neighborhood-specific outdoor PA. We examined associations of perceived built and social neighborhood environment factors, and their interactions, with objectively-measured neighborhood outdoor moderate-to-vigorous physical activity (MVPA) among a sample of Latina women in San Diego, CA. Analyses included baseline data collected in 2011-2013 from 86 Latinas with ≥ 2 days of combined accelerometer and global positioning system data and complete survey measures. We examined objective neighborhood outdoor MVPA within 500-meter home buffers. Generalized linear mixed models examined associations of 3 perceived built (e.g., sidewalk maintenance) and 3 social environmental (e.g., safety from crime) factors with engaging in any daily neighborhood outdoor MVPA. Models tested interactions between the built and social environmental factors. Although the perceived neighborhood environmental factors were not significantly related to daily neighborhood outdoor MVPA, we found 2 significant interactions: perceived sidewalk maintenance x safety from crime (p = 0.05) and neighborhood aesthetics x neighborhood social cohesion (p = 0.03). Sidewalk maintenance was positively related to daily neighborhood outdoor MVPA only among Latinas that reported low levels of safety from crime. Neighborhood aesthetics was positively related to daily neighborhood outdoor MVPA only among Latinas with high neighborhood social cohesion. Findings suggest several built and social environmental factors interact to influence Latinas' neighborhood outdoor MVPA. Interventions are needed targeting both built and social neighborhood environmental factors favorable to outdoor PA in the neighborhood.

[Clinical use of a quadruple biochemical marker in the prediction of obstetric complications]. / Utilidad clínica del cuádruple marcador bioquímico para predecir el desarrollo de complicaciones obstétricas.

The objective was to evaluate the association of an abnormal second trimester prenatal biochemical screening with the subsecuent development of pregnancy complications in women carrying chromosomally normal fetuses. A prospective study of 123 pregnant patients was performed. Specimens were assayed for alpha-fetoprotein, unconjugated estriol, free alpha hCG, and total hCG. The study included the evaluation of the mean and standard desviation as well as multiple of the median. Six women (4.6%) had positive results. The frequency of pregnancy complications in this group was 33.3%, while in the group with negative screening was 11.1%. As conclusion, positive four marker screening is associated with a adverse pregnancy evolution. However the usefulness of four marker screening for to predict pregnancy complications needs more investigations.

The entire SU subunit is required for the incorporation of the HIV-1 envelope glycoprotein complex into virions.

A modified envelope glycoprotein of the human immunodeficiency virus type 1 (HIV-1) containing an intact TM subunit, but lacking most of the gp120/SU subunit was transported and expressed on the membrane of COS-1 cells. However, this deleted glycoprotein, failed to be incorporated into the budding viral particles. This suggested that a particular domain(s) of the gp120/SU glycoprotein subunit could be required for envelope incorporation. To explore this possibilty, we constructed envelope genes containing specific domains of the SU protein in-frame with the TM subunit. Transient expression studies indicated that any envelope primary translation product containing one or more of the gp 20/SU variable domains and the entire gp41/TM protein was transported and stably expressed on the cell surface. However, efficient proteolytic processing of these Env precursors into gp41, was not observed. The addition of more than 90% of the SU sequences into the deleted Env product, including the five variable domains, were insufficient to promote incorporation of this glycoprotein precursor into virions. These results suggest that the native conformation of the SU subunit is an essential requirement for the efficient incorporation of the Env complex into virons. The C1 domain of the SU glycoprotein subunit constitutes an important determinant that makes the envelope complex assembly-competent, but, by itself, it is not sufficient to drive this process.

Coexpression of a nonsyncytium inducer HIV-1 glycoprotein inhibits syncytium formation by another HIV-1 Env protein.

The biosynthesis and biological properties of the envelope glycoprotein from a primary isolate of the human immunodeficiency virus type 1, HIV-1 YU2, and the Env product from the laboratory-adapted strain, HIV-1 LAI were compared in the absence of viral replication. We found that the level of expression and proteolytic processing into gp120/gp41 complexes of both glycoproteins was equivalent and independent of the cell type used. Although the two glycoproteins were detected on the surface of HeLa cells expressing high levels of CD4, only the HIV LAI Env product induced significant syncytium formation. Interestingly, when both glycoproteins were coexpressed in HeLa-CD4 cells, syncytium formation was greatly reduced. However, cell fusion could be restored by increasing amounts of the LAI envelope gene product. HeLa-CD4 cells expressing either glycoprotein fused with high efficiency to CEM-A cells, a hybrid of CEM and peripheral blood mononuclear cells, indicating that both glycoproteins were expressed in a biologically active form on the surface of these cells. These studies suggest that primary isolates and laboratory adapted stains may require, in addition to the CD4 receptor, independent accessory membrane components for the fusion activation step. Our results agree with the concept that virus entry requires the concerted activation of each glycoprotein subunit of the Env oligomeric complex.

The transmembrane glycoprotein of human immunodeficiency virus type 1 induces syncytium formation in the absence of the receptor binding glycoprotein.

To study the intracellular transport and biological properties of the human immunodeficiency virus type 1 (HIV-1) transmembrane glycoprotein (TM; gp41), we constructed a truncated envelope gene in which the majority of the coding sequences for the surface glycoprotein (SU; gp120) were deleted. Transient expression of this truncated env gene in primate cells resulted in the biosynthesis of two proteins with M(r)s of 52,000 and 41,000, respectively. Immunofluorescence studies with antibodies to the HIV-1 TM protein indicated that the intracellular and surface localization of these proteins were indistinguishable from those of the native HIV-1 gp120-gp41 complex. These results indicate that the oligosaccharide processing and cell surface transport of the HIV-1 TM protein were not dependent on the presence of the receptor binding subunit, gp120. Syncytium formation was readily detected upon expression of the deleted HIV-1 env gene into COS and CD4+ HeLa cell lines, suggesting that in the absence of gp120, the TM protein retained biological activity. This observation was confirmed by infection of primate and mouse cell lines with a recombinant vaccinia virus (vvgp41) expressing the truncated HIV-1 env gene. These results strongly suggest that (i) the two biological activities of the HIV-1 envelope glycoprotein can occur independently and (ii) the association of the two glycoprotein subunits may restrict the fusion activity of the transmembrane component to CD4+ cells.

Mutations within the proteolytic cleavage site of the Rous sarcoma virus glycoprotein define a requirement for dibasic residues for intracellular cleavage.

We investigated the amino acid sequence requirements for intracellular cleavage of the Rous sarcoma virus glycoprotein precursor by introducing mutations into the region encoding the cleavage recognition site (Arg-Arg-Lys-Arg). In addition to mutants G1 (Arg-Arg-Glu-Arg) and Dr1 (deletion of all four codons) that we have reported on previously (L. G. Perez and E. Hunter, J. Virol. 61:1609-1614, 1987), we constructed two additional mutants, AR1 (Arg-Arg-Arg-Arg), in which the highly conserved lysine is replaced by an arginine, and S19 (Ser-Arg-Glu-Arg), in which no dibasic pairs remain. The results of these studies demonstrate that when the cleavage sequence is deleted (Dr1) or modified to contain unpaired basic residues (S19), intracellular cleavage of the glycoprotein precursor is completely blocked. This demonstrates that the cellular endopeptidase responsible for cleavage has a stringent requirement for the presence of a pair of basic residues (Arg-Arg or Lys-Arg). Furthermore, it implies that the cleavage enzyme is not trypsinlike, since it is unable to recognize arginine residues that are sensitive to trypsin action. Substitution of the mutated genes into a replication-competent avian retrovirus genome showed that cleavage of the glycoprotein precursor was not required for incorporation into virions but was necessary for infectivity. Treatment of BH-RCAN-S19-transfected turkey cells with low levels of trypsin resulted in the release of infectious virus, demonstrating that exogenous cleavage could generate a biologically active glycoprotein molecule.

Mutants of the Rous sarcoma virus envelope glycoprotein that lack the transmembrane anchor and cytoplasmic domains: analysis of intracellular transport and assembly into virions.

The envelope glycoprotein complex of Rous sarcoma virus consists of a knoblike, receptor-binding gp85 polypeptide that is linked through disulfide bonds to a membrane-spanning gp37 spike. We used oligonucleotide-directed mutagenesis to assess the role of the hydrophobic transmembrane region and hydrophilic cytoplasmic domain of gp37 in intracellular transport and assembly into virions. Early termination codons were introduced on either side of the hydrophobic transmembrane region, and the mutated env genes were expressed from the late promoter of simian virus 40. This resulted in the synthesis of glycoprotein complexes composed of a normal gp85 and a truncated gp37 molecule that lacked the cytoplasmic domain alone or both the cytoplasmic and transmembrane domains. The biosynthesis and intracellular transport of the truncated proteins were not significantly different from those of the wild-type glycoproteins, suggesting that any protein signals for biosynthesis and intracellular transport of this viral glycoprotein complex must reside in its extracellular domain. The glycoprotein complex lacking the cytoplasmic domain of gp37 is stably expressed on the cell surface in a manner similar to that of the wild type. In contrast, the complex lacking both the transmembrane and cytoplasmic domains is secreted as a soluble molecule into the media. It can be concluded, therefore, that the transmembrane domain alone is essential for anchoring the RSV env complex in the cell membrane and that the cytoplasmic domain is not required for anchor function. Insertion of the mutated genes into an infectious proviral genome allowed us to assess the ability of the truncated gene products to be assembled into virions and to determine whether such virions were infectious. Viral genomes encoding the secreted glycoprotein were noninfectious, whereas those encoding a glycoprotein complex lacking only the cytoplasmic domain of gp37 were infectious. Virions produced from these mutant-infected cells contained normal levels of glycoprotein. The cytoplasmic tail of gp37 is thus not required for the assembly of envelope glycoproteins into virions. It is unlikely, therefore, that this region of gp37 interacts with viral core proteins during the selective incorporation of viral glycoproteins into the viral envelope.

Mutations within the proteolytic cleavage site of the Rous sarcoma virus glycoprotein that block processing to gp85 and gp37.

We have investigated the specificity of the proteolytic cleavage of the Rous sarcoma virus glycoprotein precursor by introducing two mutations into the putative cleavage region (Arg-Arg-Lys-Arg). We show that neither a deletion of the cleavage sequence nor a glutamic acid for lysine substitution altered intracellular transport or surface expression of the env gene products. However, both the four-amino-acid deletion and the glutamic acid substitution block processing of the env precursor. Susceptibility of the glutamic acid-substituted env precursor to proteases indicated that tertiary protein structure was unaffected. While inhibitor experiments suggested that more than one endopeptidase might be capable of mediating the proteolytic cleavage, the results presented here point to the presence in the Golgi apparatus of a novel endopeptidase, required for retroviral glycoprotein cleavage, that has a high specificity for lysine-containing peptides.

Purification and chemical and biological characterizations of seven toxins from the Mexican scorpion, Centruroides suffusus suffusus.

Seven polypeptides highly toxic to mice were isolated from the venom of the scorpion, Centruroides suffusus suffusus (Css), and their chemical and toxic properties were characterized. It was shown that the most active toxins by intracerebroventricular injection are less active when injected subcutaneously. The complete amino acid sequence (66 residues) of toxin II (Css II) has been determined. The C-terminal end is amidated as found for most other scorpion toxins. Css II is a beta-type toxin, previously used to define the binding site for activation of the sodium channel. Using rat brain synaptosomes, we demonstrated that all Css toxins compete with 125I-Css II to bind to site 4 and should be considered as beta-scorpion toxins. Specific binding parameters for Css VI, one of the most active toxins, were determined: KD = 100 pM; capacity in binding sites, 2.2 pmol of toxin/mg of synaptosomal protein. Css VI was shown to inhibit gamma-aminobutyric acid uptake by synaptosomes: K 0.5 = 100 pM, which agrees with its KD. Competition experiments between the seven Css toxins and 125I-Css II for antiserum raised against Css II demonstrated that all these toxins have common antigenic properties.