Tumour Necrosis Factor-α, Chemokines, and Leukocyte Infiltrate Are Biomarkers for Pathology in the Brains of Venezuelan Equine Encephalitis (VEEV)-Infected Mice.

Venezuelan equine encephalitis virus (VEEV) is a disease typically confined to South and Central America, whereby human disease is characterised by a transient systemic infection and occasionally severe encephalitis, which is associated with lethality. Using an established mouse model of VEEV infection, the encephalitic aspects of the disease were analysed to identify biomarkers associated with inflammation. Sequential sampling of lethally challenged mice (infected subcutaneously) confirmed a rapid onset systemic infection with subsequent spread to the brain within 24 h of the challenge. Changes in inflammatory biomarkers (TNF-α, CCL-2, and CCL-5) and CD45+ cell counts were found to correlate strongly to pathology (R>0.9) and present previously unproven biomarkers for disease severity in the model, more so than viral titre. The greatest level of pathology was observed within the olfactory bulb and midbrain/thalamus. The virus was distributed throughout the brain/encephalon, often in areas not associated with pathology. The principal component analysis identified five principal factors across two independent experiments, with the first two describing almost half of the data: (1) confirmation of a systemic Th1-biased inflammatory response to VEEV infection, and (2) a clear correlation between specific inflammation of the brain and clinical signs of disease. Targeting strongly associated biomarkers of deleterious inflammation may ameliorate or even eliminate the encephalitic syndrome of this disease.

Relation of Cardiometabolic Risk Factors between Parents and Children.

OBJECTIVES: To explore the relations of parent-child cardiometabolic risk factors and assess the influence of adiposity on these associations. STUDY DESIGN: Associations of adiposity, blood pressure (BP), lipids, fasting insulin and glucose, and a risk factor cluster score (CS) were evaluated in a cross-sectional study of 179 parents and their children (6-18 years, N = 255). Insulin resistance was assessed by euglycemic clamp in parents and children aged 10 years or older. Metabolic syndrome in parents was defined by National Cholesterol Education Program's Adult Treatment Panel III criteria. CSs of the risk factors were created based on age-specific z-scores. Analyses included Pearson correlation and linear regression, adjusted for parent and child age, sex, race, and body mass index (BMI), accounting for within-family correlation. RESULTS: We found positive parent-child correlations for measures of adiposity (BMI, BMI percentile, waist, subcutaneous fat, and visceral fat; r = 0.22-0.34, all P ≤ .003), systolic BP (r = 0.20, P = .002), total cholesterol (r = 0.39, P < .001), low-density lipoprotein cholesterol (r = 0.34, P < .001), high density lipoprotein cholesterol (r = 0.26, P < .001), triglycerides (r = 0.19, P = .01), and insulin sensitivity (r = 0.22, P = .02) as well as CSs (r = 0.15, P = .02). After adjustment for BMI all parent-child correlations, except systolic BP, remained significant. CONCLUSIONS: Although adiposity is strongly correlated between parents and children, many cardiometabolic risk factors correlate independent of parent and child BMI. Adverse parental cardiometabolic profiles may identify at-risk children independent of the child's adiposity status.

Increased NY-ESO-1 expression and reduced infiltrating CD3+ T cells in cutaneous melanoma.

NY-ESO-1 is a cancer-testis antigen aberrantly expressed in melanomas, which may serve as a robust and specific target in immunotherapy. NY-ESO-1 antigen expression, tumor features, and the immune profile of tumor infiltrating lymphocytes were assessed in primary cutaneous melanoma. NY-ESO-1 protein was detected in 20% of invasive melanomas (16/79), rarely in in situ melanoma (1/10) and not in benign nevi (0/20). Marked intratumoral heterogeneity of NY-ESO-1 protein expression was observed. NY-ESO-1 expression was associated with increased primary tumor thickness (P = 0.007) and inversely correlated with superficial spreading melanoma (P < 0.02). NY-ESO-1 expression was also associated with reduced numbers and density of CD3+ tumor infiltrating lymphocytes (P = 0.017). When NY-ESO-1 protein was expressed, CD3+ T cells were less diffusely infiltrating the tumor and were more often arranged in small clusters (P = 0.010) or as isolated cells (P = 0.002) than in large clusters of more than five lymphocytes. No correlation of NY-ESO-1 expression with gender, age, tumor site, ulceration, lymph node sentinel status, or survival was observed. NY-ESO-1 expression in melanoma was associated with tumor progression, including increased tumor thickness, and with reduced tumor infiltrating lymphocytes.

Implementation of lipid screening guidelines in children by primary pediatric providers.

OBJECTIVE: To assess the awareness and implementation of lipid guidelines among primary pediatric providers. STUDY DESIGN: An online survey was administered to primary pediatric providers (n = 1488): pediatricians, family medicine/general practitioners, and advanced practitioners (nurse practitioners/physician assistants) in Minnesota. The survey was conducted over 12 weeks in 2012-2013. A multiple-choice questionnaire was used to evaluate the participants' knowledge, screening, and management attitudes regarding pediatric lipid guidelines. RESULTS: The overall response rate was 39% (n = 548 of 1402 successful e-mails). Respondents were primarily pediatricians and family medicine practitioners (37% each), followed by general practitioners (11%) and advanced practitioners (nurse practitioners, 5.5%; physician assistants, 1.6%). Although 74% of providers reportedly believed that lipid screening and treatment would reduce future cardiovascular risk, 34% performed no screening, 50% screened selectively, and only 16% performed universal screening. Pediatricians were more likely to screen, with 30% performing universal screening and 41% performing selective screening. Among perceived barriers to screening, providers reported uneasiness addressing lipid disorders (43%), and unfamiliarity with screening guidelines (31%). The majority (83%) were uncomfortable managing lipid disorders, and 57% were opposed to the use of lipid-lowering medications in children. CONCLUSION: These findings underscore the need to further educate providers and supply easily accessible information on the screening and treatment of childhood lipid disorders.

Impact of pubertal development on endothelial function and arterial elasticity.

OBJECTIVES: Little is known about the relation of pubertal development on endothelial function and arterial elasticity in children and adolescents; therefore, we compared brachial artery flow-mediated dilation and carotid artery elasticity across Tanner (pubertal) stages in children and adolescents. STUDY DESIGN: Blood pressure, fasting lipids, glucose and insulin, body fat, insulin sensitivity adjusted for lean body mass, brachial flow-mediated dilation (percent dilation and area under the curve), endothelium-independent dilation (peak dilation and area under the curve), and carotid artery elasticity were evaluated across pubertal stages (Tanner I vs Tanner II-IV vs Tanner V) in 344 children and adolescents (184 males, 160 females; ages 6 to 21 years). RESULTS: One hundred twenty-four subjects (mean age 8.23 ± 0.15 years; 52 females) were Tanner stage I; 105 subjects (mean age 13.19 ± 0.17 years; 47 females) were Tanner stages II-IV; and 115 subjects (mean age 17.19 ± 0.16 years; 61 females) were Tanner stage V. There were no significant differences for any of the measures of vascular structure and function across pubertal stages. CONCLUSION: Results of the current study indicate that smooth-muscle and endothelial function, as well as carotid artery elasticity, do not differ throughout pubertal development and that accounting for pubertal stage when reporting vascular data in children and adolescents may be unnecessary.

A humanised murine monoclonal antibody protects mice from Venezuelan equine encephalitis virus, Everglades virus and Mucambo virus when administered up to 48 h after airborne challenge.

Currently there are no licensed antiviral treatments for the Alphaviruses Venezuelan equine encephalitis virus (VEEV), Everglades virus and Mucambo virus. We previously developed a humanised version of the mouse monoclonal antibody 1A3B-7 (Hu1A3B-7) which exhibited a wide range of reactivity in vitro and was able to protect mice from infection with VEEV. Continued work with the humanised antibody has now demonstrated that it has the potential to be a new human therapeutic. Hu1A3B-7 successfully protected mice from infection with multiple Alphaviruses. The effectiveness of the humanisation process was determined by assessing proliferation responses in human T-cells to peptides derived from the murine and humanised versions of the V(H) and V(L) domains. This analysis showed that the number of human T-cell epitopes within the humanised antibody had been substantially reduced, indicating that Hu1A3B-7 may have reduced immunogenicity in vivo.

Medidas antropométricas como preditoras de fatores de risco cardiovascular na população urbana do Irã / Anthropometric measures as predictors of cardiovascular disease risk factors in the urban population of Iran

FUNDAMENTO: O sobrepeso e a obesidade são um importante problema de saúde pública na sociedade pela sua associação com diversas doenças crônicas. OBJETIVO: O objetivo deste estudo é determinar a prevalência e a distribuição de sobrepeso e obesidade, usando diferentes medidas antropométricas, e identificar o melhor indicador antropométrico intimamente relacionado aos fatores de risco de Doenças Cardiovasculares (DCV) em população iraniana urbana. MÉTODOS: O presente estudo transversal foi realizado com 991 homens e 1.188 mulheres de 15 a 64 anos. Foram medidos Índice de Massa Corporal (IMC), Circunferência Abdominal (CA), Relação Cintura-Quadril (RCQ), Relação Cintura-Altura (RCA) e porcentagem de gordura corporal. Foi obtida amostra de sangue em jejum. Foram avaliados os fatores de risco cardiovascular, incluindo glicemia de jejum, triglicerídeos, colesterol total (col-T), colesterol de baixa densidade (LDL-colesterol) e colesterol da lipoproteína de alta densidade (HDL-colesterol). RESULTADOS: Em relação ao IMC, 49 por cento dos homens e 53 por cento das mulheres estavam acima do peso ou obesos, e 10,2 por cento dos homens e 18,6 por cento das mulheres encontravam-se obesos. Tanto nos homens quanto nas mulheres, a prevalência de sobrepeso esteve maior entre aqueles com 40-49 anos de idade, e a prevalência de obesidade esteve maior entre aqueles com 50 anos ou mais. Usando a análise de regressão múltipla, IMC, RCA e RCQ explicaram o maior percentual de variação de triglicerídeos, razão entre col-T e HDL-colesterol e LDL-colesterol em homens, respectivamente, ao passo que RCQ explicou o maior percentual de variação de triglicerídeos e CA explicou o maior percentual de variação da razão entre col-T e HDL-colesterol e LDL-colesterol em mulheres. CONCLUSÃO: Nossos dados indicam que RCQ e RCA foram os indicadores antropométricos que melhor previram fatores de risco cardiovascular em homens e RCQ e CA, em mulheres.

BACKGROUND: Overweight and obesity are an important public health problem in society, due to their association with various chronic diseases. OBJECTIVE: The purpose of this study is to determine the prevalence and distribution of overweight and obesity, using different anthropometric measurements and to identify the best anthropometric indicator which is most closely related to cardiovascular disease (CVD) risk factors in an Iranian urban population. METHODS: This cross-sectional study was conducted with 991 men and 1188 women aged 15 to 64 years. Body mass index (BMI), waist circumference (WC), waist-to-hip ratio (WHR), waist-to-height ratio (WHtR) and percentage of body fat were measured. A fasting blood specimen was obtained. CVD risk factors, including fasting blood glucose, triglycerides, total cholesterol (Tchol), low-density (LDL-C) and high-density-lipoprotein cholesterol (HDL-C) were assessed. RESULTS: Based on BMI, more than 49 percent of men and 53 percent of women were either overweight or obese with 10.2 percent of men and 18.6 percent of women being obese. In both men and women, the prevalence of overweight was greater among 40-49 year olds and the prevalence of obesity was greater among those 50+ years. Using the multiple regression analysis, BMI, WHtR and WHR explained the highest percentage of variation of triglycerides, Tchol/HDL-C ratio and LDL-C in men, respectively, whereas WHR explained the highest percentage of variation of triglycerides and WC explained the highest percentage of variation of Tchol/HDL-C ratio and LDL-C in women. CONCLUSION: Our data indicated that WHR and WHtR were the anthropometric indicators that best predicted CVD risk factors in men and WHR and WC in women.

Anthropometric measures as predictors of cardiovascular disease risk factors in the urban population of Iran.

BACKGROUND: Overweight and obesity are an important public health problem in society, due to their association with various chronic diseases. OBJECTIVE: The purpose of this study is to determine the prevalence and distribution of overweight and obesity, using different anthropometric measurements and to identify the best anthropometric indicator which is most closely related to cardiovascular disease (CVD) risk factors in an Iranian urban population. METHODS: This cross-sectional study was conducted with 991 men and 1188 women aged 15 to 64 years. Body mass index (BMI), waist circumference (WC), waist-to-hip ratio (WHR), waist-to-height ratio (WHtR) and percentage of body fat were measured. A fasting blood specimen was obtained. CVD risk factors, including fasting blood glucose, triglycerides, total cholesterol (Tchol), low-density (LDL-C) and high-density-lipoprotein cholesterol (HDL-C) were assessed. RESULTS: Based on BMI, more than 49% of men and 53% of women were either overweight or obese with 10.2% of men and 18.6% of women being obese. In both men and women, the prevalence of overweight was greater among 40-49 year olds and the prevalence of obesity was greater among those 50+ years. Using the multiple regression analysis, BMI, WHtR and WHR explained the highest percentage of variation of triglycerides, Tchol/HDL-C ratio and LDL-C in men, respectively, whereas WHR explained the highest percentage of variation of triglycerides and WC explained the highest percentage of variation of Tchol/HDL-C ratio and LDL-C in women. CONCLUSION: Our data indicated that WHR and WHtR were the anthropometric indicators that best predicted CVD risk factors in men and WHR and WC in women.

Cardiovascular risk and insulin resistance in childhood cancer survivors.

OBJECTIVE: Increased cardiovascular (CV) risk has been reported in adults who are childhood cancer survivors (CCS). We sought to determine the emergence of CV risk factors in CCS while still children. STUDY DESIGN: CCS in remission ≥5 years from cancer diagnosis (n=319, age=14.5 years) and their siblings (control subjects, n=208, age=13.6 years) participated in this cross-sectional study of CV risk, which included physiologic assessment of insulin sensitivity/resistance (hyperinsulinemic euglycemic clamp). Adjusted comparisons between CCS major diagnoses (leukemia [n=110], central nervous system tumors [n=82], solid tumors [n=127]) and control subjects were performed with linear regression for CV risk factors and insulin sensitivity. RESULTS: Despite no significant differences in weight and body mass index, CCS had greater adiposity (waist [73.1 versus 71.1 cm, P=.02]; percent fat [28.1 versus 25.9%, P=.007]), lower lean body mass (38.4 versus 39.9 kg, P=.01) than control subjects. After adjustment for adiposity, CCS had higher total cholesterol level (154.7 versus 148.3 mg/dL, P=.004), low-density lipoprotein cholesterol level (89.4 versus 83.7 mg/dL, P=.002), and triglyceride level (91.8 versus 84 mg/dL, P=.03) and were less insulin sensitive (insulin stimulated glucose uptake, measure of insulin resistance, adjusted for lean body mass 12.1 versus 13.4 mg/kg/min, P=.002) than control subjects. CONCLUSIONS: CCS have greater CV risk than healthy children. Because CV risk factors track from childhood to adulthood, early development of altered body composition and decreased insulin sensitivity in CCS may contribute significantly to their risk of early CV morbidity and mortality.

A humanised murine monoclonal antibody with broad serogroup specificity protects mice from challenge with Venezuelan equine encephalitis virus.

In murine models of Venezuelan equine encephalitis virus (VEEV) infection, the neutralising monoclonal antibody 1A3B-7 has been shown to be effective in passive protection from challenge by the aerosol route with serogroups I, II and Mucambo virus (formally VEE complex subtype IIIA). This antibody is able to bind to all serogroups of the VEEV complex when used in ELISA and therefore is an excellent candidate for protein engineering in order to derive a humanised molecule suitable for therapeutic use in humans. A Complementarity Determining Region (CDR) grafting approach using human germline IgG frameworks was used to produce a panel of humanised variants of 1A3B-7, from which a single candidate molecule with retained binding specificity was identified. Evaluation of humanised 1A3B-7 (Hu1A3B-7) in in vitro studies indicated that Hu1A3B-7 retained both broad specificity and neutralising activity. Furthermore, in vivo experiments showed that Hu1A3B-7 successfully protected mice against lethal subcutaneous and aerosol challenges with VEEV strain TrD (serogroup I). Hu1A3B-7 is therefore a promising candidate for the future development of a broad-spectrum antiviral therapy to treat VEEV disease in humans.

A recombinant humanized monoclonal antibody completely protects mice against lethal challenge with Venezuelan equine encephalitis virus.

A recombinant humanized antibody to Venezuelan equine encephalitis virus (VEEV) was constructed in a monocistronic adenoviral expression vector with a foot-and-mouth-disease virus-derived 2A self-cleavage oligopeptide inserted between the antibody heavy and light chains. After expression in mammalian cells, the heavy and light chains of the humanized antibody (hu1A4A1IgG1-2A) were completely cleaved and properly dimerized. The purified hu1A4A1IgG1-2A retained VEEV binding affinity and neutralizing activity similar to its parental murine antibody. The half-life of hu1A4A1IgG1-2A in mice was approximately 2 days. Passive immunization of hu1A4A1IgG1-2A in mice (50 microg/mouse) 24 h before or after virulent VEEV challenge provided complete protection, indicating that hu1A4A1IgG1-2A has potent prophylactic and therapeutic effects against VEEV infection.

Development of a novel monoclonal antibody with reactivity to a wide range of Venezuelan equine encephalitis virus strains.

BACKGROUND: There is currently a requirement for antiviral therapies capable of protecting against infection with Venezuelan equine encephalitis virus (VEEV), as a licensed vaccine is not available for general human use. Monoclonal antibodies are increasingly being developed as therapeutics and are potential treatments for VEEV as they have been shown to be protective in the mouse model of disease. However, to be truly effective, the antibody should recognise multiple strains of VEEV and broadly reactive monoclonal antibodies are rarely and only coincidentally isolated using classical hybridoma technology. RESULTS: In this work, methods were developed to reliably derive broadly reactive murine antibodies. A phage library was created that expressed single chain variable fragments (scFv) isolated from mice immunised with multiple strains of VEEV. A broadly reactive scFv was identified and incorporated into a murine IgG2a framework. This novel antibody retained the broad reactivity exhibited by the scFv but did not possess virus neutralising activity. However, the antibody was still able to protect mice against VEEV disease induced by strain TrD when administered 24 h prior to challenge. CONCLUSION: A monoclonal antibody possessing reactivity to a wide range of VEEV strains may be of benefit as a generic antiviral therapy. However, humanisation of the murine antibody will be required before it can be tested in humans.

Improved efficacy of a gene optimised adenovirus-based vaccine for venezuelan equine encephalitis virus.

BACKGROUND: Optimisation of genes has been shown to be beneficial for expression of proteins in a range of applications. Optimisation has increased protein expression levels through improved codon usage of the genes and an increase in levels of messenger RNA. We have applied this to an adenovirus (ad)-based vaccine encoding structural proteins (E3-E2-6K) of Venezuelan equine encephalitis virus (VEEV). RESULTS: Following administration of this vaccine to Balb/c mice, an approximately ten-fold increase in antibody response was elicited and increased protective efficacy compared to an ad-based vaccine containing non-optimised genes was observed after challenge. CONCLUSION: This study, in which the utility of optimising genes encoding the structural proteins of VEEV is demonstrated for the first time, informs us that including optimised genes in gene-based vaccines for VEEV is essential to obtain maximum immunogenicity and protective efficacy.

CpG used as an adjuvant for an adenovirus-based Venezuelan equine encephalitis virus vaccine increases the immune response to the vector, but not to the transgene product.

An adenovirus-based (ad-based) vaccine delivering antigens from the Alphavirus Venezuelan equine encephalitis virus (VEEV) is a strategy that offers clinical potential. A vaccine against VEEV is desirable because of the re-emerging nature of this virus, and also the potential that it may be used as a biological weapon. This study was designed to investigate whether the co-administration of CpG oligodeoxynucleotides (ODNs) with an ad-based VEEV vaccine could enhance the protective efficacy of the vaccine. We report that the co-administration of CpG ODN was unable to increase VEEV-specific antibody responses in mice, and was unable to increase the protective efficacy of the vaccine against aerosol challenge with virulent VEEV. However, it was noted that antibody responses directed against the adenovirus vaccine vector were increased, which may be detrimental, particularly in the context of homologous boosting.

Boosting with an adenovirus-based vaccine improves protective efficacy against Venezuelan equine encephalitis virus following DNA vaccination.

There is a requirement for a vaccine that protects against the alphavirus, Venezuelan equine encephalitis virus (VEEV). Previous work has shown that DNA vaccines encoding structural proteins of VEEV can elicit immune responses and protection against VEEV though this protection is incomplete against airborne VEEV. In this study, we demonstrate that particle-mediated epidermal delivery of a DNA vaccine encoding the E2 glycoprotein of VEEV can be boosted with a mucosally-delivered Ad-based vaccine encoding the same E2 glycoprotein. This results in an improved Th2-type IgG response, an increase in neutralising antibody and a significant increase in protection against airborne VEEV. This indicates that prime-boost may be a suitable immunisation regimen for providing protection against airborne VEEV.

Congenital melanocytic nevi: clinical and histopathologic features, risk of melanoma, and clinical management

Congenital melanocytic nevi occur a approximately 1% of newborns and are usually classified according to their size. Giant congenital melanocytic nevi are most simply defined as melanocytic nevi less that 1.5 cm in largest dimension. Congenital nevi can exhibit distinctive histologic features that can help in differentiating them from common acquired nevi. Giant congenital melanocytic nevi are associated with an increased risk of the development of melanoma. On the other hand, there is evidence of an increased melanoma risk in patientswith small congenital nevi. Nevertheless, the risk of malignant transformation in small congenital nevi and the lifetime melanoma risk in patients with congenital nevi remain controversial. In large part due to incosistency in the reported literature describing patients with congenital melanocytic nevi, the risk of melanoma in these patients remains unclear and consistent guidelines for clinical management for these patients at the Massachusetts General Hospital Pigmented Lesion Clinic