Caring for Sexual and Gender Minority Adults with Cardiovascular Disease.

This article summarizes existing evidence on cardiovascular disease (CVD) risk and CVD diagnoses among sexual and gender minority adults and provides recommendations for providing nursing care to sexual and gender minority adults with CVD. More research is needed to develop evidence-based strategies to care for sexual and gender minority adults with CVD.

Differences in Ideal Cardiovascular Health Between Sexual Minority and Heterosexual Adults.

Importance: Research on the cardiovascular health (CVH) of sexual minority adults has primarily examined differences in the prevalence of individual CVH metrics rather than comprehensive measures, which has limited development of behavioral interventions. Objective: To investigate sexual identity differences in CVH, measured using the American Heart Association's revised measure of ideal CVH, among adults in the US. Design, Setting, and Participants: This cross-sectional study analyzed population-based data from the National Health and Nutrition Examination Survey (NHANES; 2007-2016) in June 2022. Participants included noninstitutional adults aged 18 to 59 years. We excluded individuals who were pregnant at the time of their interview and those with a history of atherosclerotic cardiovascular disease or heart failure. Exposures: Self-identified sexual identity categorized as heterosexual, gay/lesbian, bisexual, or something else. Main Outcomes and Measures: The main outcome was ideal CVH (assessed using questionnaire, dietary, and physical examination data). Participants received a score from 0 to 100 for each CVH metric, with higher scores indicating a more favorable CVH profile. An unweighted average was calculated to determine cumulative CVH (range, 0-100), which was recoded as low, moderate, or high. Sex-stratified regression models were performed to examine sexual identity differences in CVH metrics, disease awareness, and medication use. Results: The sample included 12 180 participants (mean [SD] age, 39.6 [11.7] years; 6147 male individuals [50.5%]). Lesbian (B = -17.21; 95% CI, -31.98 to -2.44) and bisexual (B = -13.76; 95% CI, -20.54 to -6.99) female individuals had less favorable nicotine scores than heterosexual female individuals. Bisexual female individuals had less favorable body mass index scores (B = -7.47; 95% CI, -12.89 to -1.97) and lower cumulative ideal CVH scores (B = -2.59; 95% CI, -4.84 to -0.33) than heterosexual female individuals. Compared with heterosexual male individuals, gay male individuals had less favorable nicotine scores (B = -11.43; 95% CI, -21.87 to -0.99) but more favorable diet (B = 9.65; 95% CI, 2.38-16.92), body mass index (B = 9.75; 95% CI, 1.25-18.25), and glycemic status scores (B = 5.28; 95% CI, 0.59-9.97). Bisexual male individuals were twice as likely as heterosexual male individuals to report a diagnosis of hypertension (adjusted odds ratio [aOR], 1.98; 95% CI, 1.10-3.56) and use of antihypertensive medication (aOR, 2.20; 95% CI, 1.12-4.32). No differences in CVH were found between participants who reported their sexual identity as something else and heterosexual participants. Conclusion and Relevance: Results of this cross-sectional study suggest that bisexual female individuals had worse cumulative CVH scores than heterosexual female individuals, whereas gay male individuals generally had better CVH than heterosexual male individuals. There is a need for tailored interventions to improve the CVH of sexual minority adults, particularly bisexual female individuals. Future longitudinal research is needed to examine factors that might contribute to CVH disparities among bisexual female individuals.

Examination of Sexual Identity Differences in the Prevalence of Hypertension and Antihypertensive Medication Use Among US Adults: Findings From the Behavioral Risk Factor Surveillance System.

BACKGROUND: Recent evidence suggests that sexual minority (eg, gay/lesbian, bisexual) adults might be at increased risk of hypertension compared with heterosexual adults. However, disparities by sexual identity in antihypertensive medication use among adults with hypertension have not been comprehensively examined. METHODS: We analyzed data from the Behavioral Risk Factor Surveillance System (2015-2019), to examine sexual identity differences in the prevalence of hypertension and antihypertensive medication use among adults. We ran sex-stratified logistic regression models to estimate the odds ratios of diagnosis of hypertension and antihypertensive medication use among sexual minority (ie, gay/lesbian, bisexual, and other) and heterosexual adults (reference group). RESULTS: The sample included 420 340 participants with a mean age of 49.7 (±17.0) years, of which 66.7% were Non-Hispanic White. Compared with heterosexual participants of the same sex, bisexual women (adjusted odds ratio, 1.19 [95% CI, 1.03-1.37]) and gay men (adjusted odds ratio, 1.18 [95% CI, 1.03-1.35]) were more likely to report having been diagnosed with hypertension. Among women with diagnosed hypertension, bisexual women had lower odds of current antihypertensive medication use (adjusted odds ratio, 0.71 [95% CI, 0.56-0.90]). Among men with diagnosed hypertension, gay men were more likely than heterosexual men to report current antihypertensive medication use (adjusted odds ratio, 1.39 [95% CI, 1.10-1.78]). Compared with heterosexual participants of the same sex, there were no differences in hypertension or antihypertensive medication use among lesbian women, bisexual men, and participants who reported their sexual identity as other. CONCLUSIONS: Clinical and public health interventions are needed to reduce the risk of hypertension among bisexual women and gay men. Bisexual women were at higher risk of untreated hypertension, which may be attributed to lower health care utilization due to fear of discrimination from health care providers and socioeconomic disadvantage. Future research is needed to better understand factors that may contribute to untreated hypertension among bisexual women with hypertension.

Diet, Food Insecurity, and CVD Risk in Sexual and Gender Minority Adults.

PURPOSE OF REVIEW: Sexual and gender minority (SGM) adults experience significant cardiovascular health disparities, yet little is known about diet and food insecurity in this population. This review summarizes recent literature on diet and food insecurity in SGM adults and their contribution to cardiovascular disease (CVD) risk in this population. RECENT FINDINGS: Existing evidence on diet and food insecurity disparities among SGM adults is inconclusive and research examining their link with CVD risk in SGM adults is limited. The majority of existing studies lack standardized and validated assessments of diet and food insecurity. Correlates of unhealthy diet and food insecurity among SGM adults are poorly understood. Research examining the associations between diet and food insecurity with CVD risk in SGM adults is limited. Longitudinal studies are needed to investigate whether diet and food insecurity contribute to the cardiovascular health disparities observed in SGM adults.

Lifetime Trauma and Cardiovascular Health in Latinas.

BACKGROUND: Potentially traumatic experiences throughout the life course are associated with poor cardiovascular health among women. However, research on the associations of trauma with cardiovascular health among Latino populations is limited. Understanding the impact of trauma on cardiovascular health within marginalized populations may provide guidance on developing interventions with a particular focus on preventative care. OBJECTIVE: The purpose of this descriptive cross-sectional study was to examine the associations of lifetime trauma with cardiovascular health among middle-aged and older Latina women. METHODS: Participants were recruited from an existing study in New York City. All participants completed a structured questionnaire to assess lifetime trauma, demographic characteristics (such as age and education), financial resource strain, and emotional support. The Trauma History Questionnaire was used to assess lifetime exposure to potentially traumatic experiences (range 0-24). Cardiovascular health was measured with a validated measure of cardiovascular health from the American Heart Association (Life's Simple 7). We used self-reported and objective data to calculate cardiovascular health scores (range 0-14). Multiple linear regression was used to examine the associations of lifetime trauma with cardiovascular health, adjusted for age, education, financial resource strain, and emotional support. RESULTS: The sample included 50 Latina women with a mean age of 63.1 years, 88% were Dominican, and only 6% had completed a college degree. Women reported an average of 4.8 traumatic experiences. Mean cardiovascular health score was 6.5 (SD = 1.6, range 3-10). Linear regression models found that, after adjusting for age, education, financial resource strain, and emotional support, a higher count of lifetime trauma was associated with worse cardiovascular health. However, this association did not reach statistical significance. DISCUSSION: Women with a higher count of lifetime trauma had worse cardiovascular health scores; this association was not statistically significant. Future studies should investigate associations of lifetime trauma and cardiovascular health in larger and more diverse samples of Latinas. Nurses and other clinicians should incorporate trauma-informed approaches to cardiovascular disease risk reduction to improve the cardiovascular health of Latina women who are survivors of trauma.

Exploring the binding of peptidic West Nile virus NS2B-NS3 protease inhibitors by NMR.

West Nile virus (WNV) NS2B-NS3 protease is an important drug target since it is an essential protein for the replication of the virus. In order to determine the minimum pharmacophore for protease inhibition, a series of dipeptide aldehydes were synthesized. The 50% inhibitory concentration (IC(50)) measurements revealed that a simple acetyl-KR-aldehyde was only threefold less active than 4-phenyl-phenylacetyl-KKR-aldehyde (1) (Stoermer et al., 2008) that was used as the reference compound. The ligand efficiency of 0.40 kcal/mol/HA (HA=heavy atom) for acetyl-KR-aldehyde is much improved compared to the reference compound 1 (0.23 kcal/mol/HA). The binding of the inhibitors was examined using (1)H-(15)N-HSQC experiments and differential chemical shifts were used to map the ligand binding sites. The biophysical studies show that the conformational mobility of WNV protease has a major impact on the design of novel inhibitors, since the protein conformation changes profoundly depending on the structure of the bound ligand.

Transplantation of a hydrogen bonding network from West Nile virus protease onto Dengue-2 protease improves catalytic efficiency and sheds light on substrate specificity.

The two-component serine protease of flaviviruses such as Dengue virus (DENV) and West Nile virus (WNV) are attractive targets for inhibitor/therapeutic design. Peptide aldehyde inhibitors that bind to the covalently tethered two-component WNV protease (WNVpro) with 50% inhibitory concentration (IC(50)) at sub-micromolar concentrations, bind the equivalent DENV-2 protease (DEN2pro) with IC(50) of micromolar concentrations at best. Conversely, the protease inhibitor aprotinin binds DEN2pro ∼1000-fold more tightly than WNVpro. To investigate the residues that are crucial for binding specificity differences, a binding-site network of hydrogen bonds was transplanted from WNVpro onto DEN2pro. The transplantations were a combination of single, double and triple mutations involving S79D, S83N and S85Q. The mutant DENV proteases, except those involving S85Q, proved to be more efficient enzymes, as measured by their kinetic parameters. The binding affinities of the mutants to peptide inhibitors however showed only marginal improvement. Protein structure modeling suggests that the negatively charged residue cluster, Glu89-Glu92, of the NS2B cofactor may play an important role in determining substrate/inhibitor-binding specificity. These same residues may also explain why aprotinin binds more tightly to DEN2pro than WNVpro. Our results suggest that structure-based inhibitor design experiments need to explicitly consider/include this C-terminal region whose negative charge is conserved across the four DENV serotypes and also among the flavivirus family of proteases.

Tripeptide inhibitors of dengue and West Nile virus NS2B-NS3 protease.

A series of tripeptide aldehyde inhibitors were synthesized and their inhibitory effect against dengue virus type 2 (DENV2) and West Nile virus (WNV) NS3 protease was evaluated side by side with the aim to discover potent flaviviral protease inhibitors and to examine differences in specificity of the two proteases. The synthesized inhibitors feature a varied N-terminal cap group and side chain modifications of a P2-lysine residue. In general a much stronger inhibitory effect of the tripeptide inhibitors was observed toward WNV protease. The inhibitory concentrations against DENV2 protease were in the micromolar range while they were submicromolar against WNV. The data suggest that a P2-arginine shifts the specificity toward DENV2 protease while WNV protease favors a lysine in the P2 position. Peptides with an extended P2-lysine failed to inhibit DENV2 protease suggesting a size-constrained S2 pocket. Our results generally encourage the investigation of di- and tripeptide aldehydes as inhibitors of DENV and WNV protease.

Structure-guided fragment-based in silico drug design of dengue protease inhibitors.

An in silico fragment-based drug design approach was devised and applied towards the identification of small molecule inhibitors of the dengue virus (DENV) NS2B-NS3 protease. Currently, no DENV protease co-crystal structure with bound inhibitor and fully formed substrate binding site is available. Therefore a homology model of DENV NS2B-NS3 protease was generated employing a multiple template spatial restraints method and used for structure-based design. A library of molecular fragments was derived from the ZINC screening database with help of the retrosynthetic combinatorial analysis procedure (RECAP). 150,000 molecular fragments were docked to the DENV protease homology model and the docking poses were rescored using a target-specific scoring function. High scoring fragments were assembled to small molecule candidates by an implicit linking cascade. The cascade included substructure searching and structural filters focusing on interactions with the S1 and S2 pockets of the protease. The chemical space adjacent to the promising candidates was further explored by neighborhood searching. A total of 23 compounds were tested experimentally and two compounds were discovered to inhibit dengue protease (IC(50) = 7.7 µM and 37.9 µM, respectively) and the related West Nile virus protease (IC(50) = 6.3 µM and 39.0 µM, respectively). This study demonstrates the successful application of a structure-guided fragment-based in silico drug design approach for dengue protease inhibitors providing straightforward hit generation using a combination of homology modeling, fragment docking, chemical similarity and structural filters.

High affinity human antibody fragments to dengue virus non-structural protein 3.

BACKGROUND: The enzyme activities catalysed by flavivirus non-structural protein 3 (NS3) are essential for virus replication. They are distributed between the N-terminal protease domain in the first one-third and the C-terminal ATPase/helicase and nucleoside 5' triphosphatase domain which forms the remainder of the 618-aa long protein. METHODOLOGY/PRINCIPAL FINDINGS: In this study, dengue full-length NS3 protein with residues 49 to 66 of NS2B covalently attached via a flexible linker, was used as bait in biopanning with a naïve human Fab phage-display library. Using a range of truncated constructs spanning the NS2B cofactor region and the full-length NS3, 10 unique Fab were identified and characterized. Of these, monoclonal Fab 3F8 was shown to bind α3″ (residues 526 through 531) within subdomain III of the helicase domain. The antibody inhibits the ATPase and helicase activites of NS3 in biochemical assays and reduces DENV replication in HEK293 cells that were previously transfected with Fab 3F8 compared with mock transfected cells. CONCLUSIONS/SIGNIFICANCE: Antibodies such as 3F8 are valuable tools for studying the molecular mechanisms of flaviviral replication and for the monospecific detection of replicating dengue virus in vivo.

Flexibility between the protease and helicase domains of the dengue virus NS3 protein conferred by the linker region and its functional implications.

The dengue virus (DENV) NS3 protein is essential for viral polyprotein processing and RNA replication. It contains an N-terminal serine protease region (residues 1-168) joined to an RNA helicase (residues 180-618) by an 11-amino acid linker (169-179). The structure at 3.15 A of the soluble NS3 protein from DENV4 covalently attached to 18 residues of the NS2B cofactor region (NS2B(18)NS3) revealed an elongated molecule with the protease domain abutting subdomains I and II of the helicase (Luo, D., Xu, T., Hunke, C., Grüber, G., Vasudevan, S. G., and Lescar, J. (2008) J. Virol. 82, 173-183). Unexpectedly, using similar crystal growth conditions, we observed an alternative conformation where the protease domain has rotated by approximately 161 degrees with respect to the helicase domain. We report this new crystal structure bound to ADP-Mn(2+) refined to a resolution of 2.2 A. The biological significance for interdomain flexibility conferred by the linker region was probed by either inserting a Gly residue between Glu(173) and Pro(174) or replacing Pro(174) with a Gly residue. Both mutations resulted in significantly lower ATPase and helicase activities. We next increased flexibility in the linker by introducing a Pro(176) to Gly mutation in a DENV2 replicon system. A 70% reduction in luciferase reporter signal and a similar reduction in the level of viral RNA synthesis were observed. Our results indicate that the linker region has evolved to an optimum length to confer flexibility to the NS3 protein that is required both for polyprotein processing and RNA replication.

The gpQ portal protein of bacteriophage P2 forms dodecameric connectors in crystals.

Double-stranded bacteriophages code for a protein called a connector or portal protein that serves as the entry and exit portal for DNA during genome packaging and ejection, as well as the connection point between heads and tails, and possibly as a nucleator for capsid assembly. The gpQ connector protein from bacteriophage P2 has been overexpressed in Escherichia coli and purified by sucrose gradient centrifugation. Negative stain electron microscopy and image analysis revealed a 135 A diameter dodecameric ring structure with a central 25 A hole. The connector showed a strong propensity to aggregate at low ionic strength and would form microcrystalline structures in solution. Consequently, the connectors were crystallized by hanging-drop vapor diffusion against low ionic strength buffer. Two crystal forms were observed: a P4(1)22 form with unit cell parameters a=b=96.33 A and c=454.42 A that diffracted X-rays to 4.5 A resolution and an I222 crystal form with a=168.86 A, b=171.88 A and c=168.68 A that diffracted to 4.1A resolution. Self-rotation functions confirmed the presence of 12-fold symmetry in the crystals.

Three-dimensional reconstruction of hibiscus chlorotic ringspot virus.

Hibiscus chlorotic ringspot virus (HCRSV) is a positive-sense, single-stranded RNA virus, which belongs to the Tombusviridae family and infects plants of the Hibiscus genus, including kenaf, a woody plant of agricultural importance. These icosahedral viruses have a capsid consisting of 180 copies of coat protein (CP) arranged with T=3 symmetry. The CP consists of an internal RNA-binding domain, a shell-forming domain and a protruding domain. The HCRSV virion was reconstructed to about 12A resolution from cryo-EM images using the program EMAN. The structure had the arrangement of 90 dimers of protruding domains characteristic of the Tombusviridae. Reconstructions were also made from negatively stained samples, and showed essentially the same features. In addition, a particle of a different, "smooth" appearance was also identified in the negatively stained samples. These particles were slightly smaller and lacked protruding domains. Biochemical analysis confirmed the presence of two protein products: a 37 kDa protein identified as HCRSV CP and a 54 kDa protein that appeared to be of non-HCRSV origin.

Structure of the nucleocapsid protein of porcine reproductive and respiratory syndrome virus.

Porcine reproductive and respiratory syndrome virus (PRRSV) is an enveloped RNA virus of the Arteriviridae family, genomically related to the coronaviruses. PRRSV is the causative agent of both severe and persistent respiratory disease and reproductive failure in pigs worldwide. The PRRSV virion contains a core made of the 123 amino acid nucleocapsid (N) protein, a product of the ORF7 gene. We have determined the crystal structure of the capsid-forming domain of N. The structure was solved to 2.6 A resolution by SAD methods using the anomalous signal from sulfur. The N protein exists in the crystal as a tight dimer forming a four-stranded beta sheet floor superposed by two long alpha helices and flanked by two N- and two C-terminal alpha helices. The structure of N represents a new class of viral capsid-forming domains, distinctly different from those of other known enveloped viruses, but reminiscent of the coat protein of bacteriophage MS2.

Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of the structural domain of the nucleocapsid N protein from porcine reproductive and respiratory syndrome virus (PRRSV).

The structural domain of the PRRSV nucleocapsid N protein was overexpressed in Escherichia coli and purified to homogeneity. Crystals of the expressed protein, designated His-Ndelta(57), were obtained by hanging-drop vapour diffusion using PEG 3350 as precipitant at pH 6.5. A native data set from a frozen crystal was collected to 2.7 A resolution using synchrotron radiation. The crystals belong to space group P3(1)21 or P3(2)21, with unit-cell parameters a = 44.41, c = 125.05, and contain a dimer in the asymmetric unit.