[Civil society promoting government accountability for health equity in the Caribbean: The Healthy Caribbean CoalitionPromoção, pela sociedade civil, da responsabilização do governo pela equidade em saúde no Caribe: a Coalizão do Caribe Saudável]. / Papel de la sociedad civil en la promoción de la rendición de cuentas de los gobiernos en materia de equidad en salud en el Caribe: la Coalición Caribe Saludable.

Achieving health equity and addressing the social determinants of health are critical to attaining the health and health-related targets of the 2030 Agenda for Sustainable Development and its Sustainable Development Goals. Frameworks for health, including the Sustainable Health Agenda for the Americas 2018 - 2030, emphasize reduction of health inequities and "leaving no one behind" in national sustainable development. Health equity includes advancing universal health and the primary health care approach, with equitable access for all people to timely, quality, comprehensive, people- and community-centered services that do not cause impoverishment. Equally important, and a hallmark of good governance, is accountability for such advances. Governments have primary responsibility for reducing health inequities and must be held accountable for their policies and performance. Civil society has been recognized as a key partner in advancing sustainable and equitable national development. Effective accountability mechanisms should include civic engagement. The Healthy Caribbean Coalition (HCC), the only Caribbean regional alliance of civil society organizations working to prevent and control noncommunicable diseases-a major health priority fueled by inequities-has played a significant role in holding governments accountable for advancing health equity. This case study examines factors contributing to the success of the HCC, highlighting work under its five strategic pillars- accountability, advocacy, capacity development, communication, and sustainability-as well as challenges, lessons learned, and considerations for greater effectiveness.

Conquistar a equidade em saúde e abordar os determinantes sociais da saúde são essenciais para atingir as metas de saúde e as relacionadas à saúde da Agenda 2030 para o Desenvolvimento Sustentável e seus Objetivos de Desenvolvimento Sustentável. As estruturas para a saúde, incluindo a Agenda de Saúde Sustentável para as Américas 2018-2030, enfatizam a redução das iniquidades em saúde "sem deixar ninguém para trás", quando se trata do desenvolvimento sustentável nacional. A equidade em saúde inclui impulsionar a saúde universal e a abordagem da atenção primária à saúde, habilitando o acesso equitativo por todas as pessoas a serviços oportunos, de qualidade, integrais, centrados no atendimento às pessoas e às comunidades de maneira a não causar o empobrecimento. A questão da responsabilidade por tais avanços é igualmente importante, e é um selo de distinção de boa gestão. Os governos são os principais responsáveis pela redução das iniquidades em saúde e precisam ser responsabilizados por suas políticas e por seu desempenho. Reconheceu-se que a sociedade civil desempenha um papel essencial na promoção do desenvolvimento nacional sustentável e equitativo. Para que sejam eficazes, os mecanismos de responsabilização devem incluir a participação cívica. A Coalizão do Caribe Saudável (HCC), a única aliança de organizações da sociedade civil que trabalha na prevenção e no controle de doenças não transmissíveis na região do Caribe ­ uma grande prioridade de saúde movida pelas iniquidades ­ tem desempenhado uma função significativa na responsabilização dos governos pelo avanço da equidade em saúde. Este estudo examina os fatores que contribuem para o sucesso da HCC e destaca o trabalho da perspectiva dos cinco pilares estratégicos ­ responsabilidade, promoção de causa, desenvolvimento das capacidades, comunicação e sustentabilidade ­, bem como os desafios, as lições aprendidas e as considerações para que se torne ainda mais eficaz.

Civil society promoting government accountability for health equity in the Caribbean: The Healthy Caribbean Coalition.

Achieving health equity and addressing the social determinants of health are critical to attaining the health and health-related targets of the 2030 Agenda for Sustainable Development and its Sustainable Development Goals. Frameworks for health, including the Sustainable Health Agenda for the Americas 2018 - 2030, emphasize reduction of health inequities and "leaving no one behind" in national sustainable development. Health equity includes advancing universal health and the primary health care approach, with equitable access for all people to timely, quality, comprehensive, people- and community-centered services that do not cause impoverishment. Equally important, and a hallmark of good governance, is accountability for such advances. Governments have primary responsibility for reducing health inequities and must be held accountable for their policies and performance. Civil society has been recognized as a key partner in advancing sustainable and equitable national development. Effective accountability mechanisms should include civic engagement. The Healthy Caribbean Coalition (HCC), the only Caribbean regional alliance of civil society organizations working to prevent and control noncommunicable diseases-a major health priority fueled by inequities-has played a significant role in holding governments accountable for advancing health equity. This case study examines factors contributing to the success of the HCC, highlighting work under its five strategic pillars-accountability, advocacy, capacity development, communication, and sustainability-as well as challenges, lessons learned, and considerations for greater effectiveness.

Alcanzar la equidad en salud y abordar los determinantes sociales de la salud son aspectos fundamentales para alcanzar las metas en materia de salud y relacionadas con la salud de la Agenda para el Desarrollo Sostenible 2030 y sus Objetivos de Desarrollo Sostenible. Los marcos de referencia para la salud, como la Agenda de Salud Sostenible para las Américas 2018-2030, hacen hincapié en la reducción de las desigualdades en salud y en "no dejar a nadie atrás" en el desarrollo sostenible a nivel nacional. La equidad en salud incluye la promoción de la salud universal y el enfoque de atención primaria de la salud, con un acceso equitativo de todas las personas a servicios de salud oportunos, de calidad, integrales y centrados en las personas y la comunidad que no ocasionen empobrecimiento. La rendición de cuentas por esos avances es igualmente importante, y un signo distintivo de una gobernanza adecuada. Los gobiernos tienen la responsabilidad primordial de reducir las desigualdades en salud y deben rendir cuentas de sus políticas y su desempeño. La sociedad civil es una parte interesada fundamental para promover un desarrollo nacional sostenible y equitativo, y debe formar parte de los mecanismos eficaces de rendición de cuentas.La Coalición Caribe Saludable ­la única alianza regional del Caribe de organizaciones de la sociedad civil dedicada a prevenir y controlar las enfermedades no transmisibles, una prioridad sanitaria importante acrecentada por las desigualdades­ ha desempeñado un papel importante en hacer que los gobiernos rindan cuentas de la promoción de la equidad en salud. En este estudio se examinan los factores que han contribuido al éxito de la Coalición Caribe Saludable, con énfasis en la labor realizada en el marco de sus cinco pilares estratégicos ­rendición de cuentas, promoción de la causa, desarrollo de capacidad, comunicación y sostenibilidad­ así como los retos, las enseñanzas extraídas y otras consideraciones para lograr una mayor eficacia.

Paroxysmal nocturnal haemoglobinuria testing in blood transfusion laboratories: do they go with the flow?

Paroxysmal nocturnal haemoglobinuria (PNH) is a rare stem cell disorder causing, in untreated patients, symptoms that include renal damage, thrombosis and increased mortality. When correctly diagnosed and treated, patients have reduced symptoms and normal life expectancies. Historically PNH testing resided within blood transfusion laboratories using techniques that were insensitive, for example, the Ham test. However, technology has evolved and flow cytometry is now regarded as the gold standard methodology. Given the clinical importance of diagnosing PNH correctly, we undertook a study to examine PNH testing procedures in blood transfusion laboratories within the UK and Ireland to determine implementation of best practices. An online survey was issued to 386 blood transfusion laboratories in the UK and Ireland requesting details of their current PNH testing practices and procedures. There were 143 responses, representing a 37% response rate. Of these, we identified seven laboratories undertaking PNH testing using obsolete methodologies. Furthermore, multiple centres did not refer samples for confirmatory testing by national PNH reference centres and inclusion on the national PNH disease registry. Staff handling requests for PNH testing should ensure that all samples are tested in accordance with current best practices using only flow cytometry.

Voluntary Surveillance Program for Equine Influenza Virus in the United States during 2008-2021.

A voluntary upper respiratory biosurveillance program in the USA received 9740 nasal swab submissions during the years 2008-2021 from 333 veterinarians and veterinary clinics. The nasal swabs were submitted for qPCR testing for six common upper respiratory pathogens:equine influenza virus (EIV), equine herpesvirus-1 (EHV-1), equine herpesvirus-4 (EHV-4), Streptococcus equi subspecies equi (S. equi), equine rhinitis A virus (ERAV), and equine rhinitis B virus (ERBV). Additional testing was performed for equine gamma herpesvirus-2 (EHV-2) and equine gamma herpesvirus-5 (EHV-5) and the results are reported. Basic frequency statistics and multivariate logistic regression models were utilized to determine the associations between risk factors and EIV positivity. The EIV qPCR-positivity rate was 9.9%. Equids less than 9 years of age with a recent history of travel and seasonal occurrence in winter and spring were the most common population that were qPCR positive for EIV. This ongoing biosurveillance program emphasizes the need for molecular testing for pathogen identification, which is critical for decisions associated with therapeutics and biosecurity intervention for health management and vaccine evaluations and development.

COVID-19 in Assisted Living: Protecting a Critical Long-Term Care Resource.

The COVID-19 pandemic had a big impact on assisted living (AL), a vital setting in long-term care (LTC). Understanding the strengths and opportunities for improvement through practice, policy, and research are essential for AL to be prepared for the next pandemic and other challenges. AL communities experienced the pandemic in unique ways, because of varying regulatory environments, differences in familiarity with using and procuring personal protective equipment not typically used in AL (such as N95 masks), loss of family involvement, the homelike environment, and lower levels of licensed clinical staff. Being state rather than federally regulated, much less national data are available about the COVID-19 experience in AL. This article reviews what is known about cases and deaths, infection control, and the impact on residents and staff. For each, we suggest actions that could be taken and link them to the Assisted Living Workgroup Report (ALW) recommendations. Using the Center for Excellence in Assisted Living (CEAL) 15-year ALW report, we also review which of these recommendations have and have not been implemented by states in the preceding decade and half, and how their presence or absence may have affected AL pandemic preparedness. Finally, we provide suggestions for policy, practice, and research moving forward, including improving state-level reporting, staff vaccine requirements, staff training and work-life, levels of research-provider partnerships, dissemination of research, and uptake of a holistic model of care for AL.

Pharmacokinetics of a FDA-labeled dose of diclazuril administered orally once weekly to adult horses.

Equine protozoal myeloencephalitis (EPM) has remained a devastating neurological disease of the Americas, especially in young performance horses. Prophylactic treatment strategies with diclazuril have shown to reduce seroprevalence and titer levels to Sarcocystis neurona in healthy horses continuously exposed to the apicomplexan parasite. The goal of this study was to determine if the FDA-labeled dose of 1 mg/kg of 1.56% diclazuril (ProtazilTM) given once weekly to healthy adult horses would achieve steady-state concentrations in plasma known to be inhibitory to S. neurona in cell culture. Five individual diclazuril doses were administered at weekly intervals to 8 adult horses. Blood was collected via venipuncture immediately before (trough concentration) and 10 hours after (peak concentration) each diclazuril administration. Following the fifth dose, additional blood samples were collected every 24 hours after the peak blood collection for 7 days. All plasma samples were analyzed by high-pressure liquid chromatography. The pharmacokinetic analysis was performed using a nonlinear mixed effects model. The mean population-derived peak concentration was 264 ng/mL and the mean terminal half-life was 3.6 days. Thus, the oral administration of an FDA-labeled dose of diclazuril to healthy horses once a week was able to produce steady-state plasma drug concentrations known to inhibit S. neurona in vitro.

Voluntary Biosurveillance of Streptococcus equi Subsp. equi in Nasal Secretions of 9409 Equids with Upper Airway Infection in the USA.

This study aimed to describe selected epidemiological aspects of horses with acute onset of fever and respiratory signs testing qPCR-positive for S. equi and to determine the effect of vaccination against S. equi on qPCR status. Horses with acute onset of fever and respiratory signs from all regions of the United States were included in a voluntary biosurveillance program from 2008 to 2020 and nasal secretions were tested via qPCR for S. equi and common respiratory viruses. A total of 715/9409 equids (7.6%) tested qPCR-positive for S. equi, with 226 horses showing coinfections with EIV, EHV-1, EHV-4, and ERBV. The median age for the S. equi qPCR-positive horses was 8 ± 4 years and there was significant difference when compared to the median age of the S. equi qPCR-negative horses (6 ± 2 years; p = 0.004). Quarter Horse, Warmblood, and Thoroughbred were the more frequent breed in this horse population, and these breeds were more likely to test qPCR-positive for S. equi compared to other breeds. There was not statistical difference for sex between S. equi qPCR-positive and qPCR-negative horses. Horses used for competition and ranch/farm use were more likely to test qPCR-positive for S. equi (p = 0.006). Horses that tested S. equi qPCR-positive were more likely to display nasal discharge, fever, lethargy, anorexia, and ocular discharge compared to horses that tested S. equi qPCR-negative (p = 0.001). Vaccination against S. equi was associated with a lower frequency of S. equi qPCR-positive status.

Human milk oligosaccharides, antimicrobial drugs, and the gut microbiota of term neonates: observations from the KOALA birth cohort study.

The infant gut microbiota affects childhood health. This pioneer microbiota may be vulnerable to antibiotic exposures, but could be supported by prebiotic oligosaccharides found in breast milk and some infant formulas. We sought to characterize the effects of several exposures on the neonatal gut microbiota, including human milk oligosaccharides (HMOs), galacto-oligosaccharides (GOS), and infant/maternal antimicrobial exposures. We profiled the stool microbiota of 1023 one-month-old infants from the KOALA Birth Cohort using 16S rRNA gene amplicon sequencing. We quantified 15 HMOs in breast milk from the mothers of 220 infants, using high-performance liquid chromatography-mass spectrometry. Both breastfeeding and antibiotic exposure decreased gut microbial diversity, but each was associated with contrasting shifts in microbiota composition. Other factors associated with microbiota composition included C-section, homebirth, siblings, and exposure to animals. Neither infant exposure to oral antifungals nor maternal exposure to antibiotics during pregnancy were associated with infant microbiota composition. Four distinct groups of breast milk HMO compositions were evident, corresponding to maternal Secretor status and Lewis group combinations defined by the presence/absence of certain fucosylated HMOs. However, we found the strongest evidence for microbiota associations between two non-fucosylated HMOs: 6'-sialyllactose (6'-SL) and lacto-N-hexaose (LNH), which were associated with lower and higher relative abundances of Bifidobacterium, respectively. Among 111 exclusively formula-fed infants, the GOS-supplemented formula was associated with a lower relative abundance of Clostridium perfringens. In conclusion, the gut microbiota is sensitive to some prebiotic and antibiotic exposures during early infancy and understanding their effects could inform future strategies for safeguarding a health-promoting infant gut microbiota.

Frequency of Detection and Prevalence Factors Associated with Common Respiratory Pathogens in Equids with Acute Onset of Fever and/or Respiratory Signs (2008-2021).

A voluntary biosurveillance program was established in 2008 in order to determine the shedding frequency and prevalence factors for common respiratory pathogens associated with acute onset of fever and/or respiratory signs in equids from the USA. Over a period of 13 years, a total of 10,296 equids were enrolled in the program and nasal secretions were analyzed for the qPCR detection of equine influenza virus (EIV), equine herpesvirus-1 (EHV-1), EHV-4, equine rhinitis A and B virus (ERVs), and Streptococcus equi subspecies equi (S. equi). Single infections with respiratory pathogens were detected in 21.1% of the submissions with EIV (6.8%) and EHV-4 (6.6%) as the two most prevalent viruses, followed by S. equi (4.7%), ERVs (2.3%), and EHV-1 (0.7%). Multiple pathogens were detected in 274 horses (2.7%) and no respiratory pathogens in 7836 horses (76.2%). Specific prevalence factors were determined for each of the six respiratory pathogen groups; most differences were associated with age, breed, and use of the horses, while the clinical signs were fairly consistent between viral and bacterial respiratory infections. Monitoring the frequency of detection of common respiratory pathogens is important in order to gain a better understanding of their epidemiology and to implement management practices aimed at controlling disease spread.

A Live-Attenuated Equine Influenza Vaccine Stimulates Innate Immunity in Equine Respiratory Epithelial Cell Cultures That Could Provide Protection From Equine Herpesvirus 1.

Equine herpesvirus 1 (EHV-1) ubiquitously infects horses worldwide and causes respiratory disease, abortion, and equine herpesvirus myeloencephalopathy. Protection against EHV-1 disease is elusive due to establishment of latency and immune-modulatory features of the virus. These include the modulation of interferons, cytokines, chemokines, antigen presentation, and cellular immunity. Because the modulation of immunity likely occurs at the site of first infection-the respiratory epithelium, we hypothesized that the mucosal influenza vaccine Flu Avert® I.N. (Flu Avert), which is known to stimulate strong antiviral responses, will enhance antiviral innate immunity, and that these responses would also provide protection from EHV-1 infection. To test our hypothesis, primary equine respiratory epithelial cells (ERECs) were treated with Flu Avert, and innate immunity was evaluated for 10 days following treatment. The timing of Flu Avert treatment was also evaluated for optimal effectiveness to reduce EHV-1 replication by modulating early immune responses to EHV-1. The induction of interferons, cytokine and chemokine mRNA expression, and protein secretion was evaluated by high-throughput qPCR and multiplex protein analysis. Intracellular and extracellular EHV-1 titers were determined by qPCR. Flu Avert treatment resulted in the modulation of IL-8, CCL2, and CXCL9 starting at days 5 and 6 post-treatment. Coinciding with the timing of optimal chemokine induction, our data also suggested the same timing for reduction of EHV-1 replication. In combination, our results suggest that Flu Avert may be effective at counteracting some of the immune-modulatory properties of EHV-1 at the airway epithelium and the peak for this response occurs 5-8 days post-Flu Avert treatment. Future in vivo studies are needed to investigate Flu Avert as a prophylactic in situations where EHV-1 exposure may occur.

Investigation of the Bi-Weekly Administration of Diclazuril on the Antibody Kinetics to Sarcocystis Neurona in Healthy Horses.

The aim of this study was to determine if bi-weekly administration of diclazuril at half the label dose would reduce seroprevalence and magnitude of titers to S. neurona in healthy horses naturally exposed to the apicomplexan protozoal parasite. 12 healthy adult horses were moved from a low-risk exposure to a farm with high exposure rate to S. neurona in their horse population. The horses were randomly assigned to either a treatment or a control group. Treatment consisted in the administration of half the label dose (0.5 mg/kg) of diclazuril (Protazil) pelleted top dress twice weekly (every 3-4 days) for 12 months. Prior to initiation of treatment and monthly thereafter, blood was collected for the detection of antibodies to S. neurona using a quantitative immunoassay. Further, trough plasma diclazuril levels were determined every 60 days. All 20 horses remained healthy during the entire study period. Seroprevalence to S. neurona decreased initially in the treatment group to 50% at 30 days post-treatment commencement. This was followed by a slow increase in seroprevalence in the treatment group before reaching 100% in both groups by 90 days post-treatment commencement. The seroprevalence remained 100% in both groups from 90 to 360 study days. While titer distribution between the two groups was similar at study commencement, treated horses had significantly lower titers throughout the treatment period (P < 0.05). All treated study horses had detectable plasma trough diclazuril levels at the 6 time points and the levels were above the concentration known to inhibit S. neurona in vitro (1.0 ng/mL). The administration of diclazuril pelleted top dress at half the label dose twice weekly was able to maintain low titers to S. neurona in healthy adult horses naturally exposed to the protozoal parasite. Further, trough diclazuril levels were in excess of the minimal concentration known to inhibit S. neurona.

Early prenatal androgenization results in diminished ovarian reserve in adult female rhesus monkeys.

BACKGROUND: Early prenatal androgenization (PA) accelerates follicle differentiation and impairs embryogenesis in adult female rhesus monkeys (Macaca mulatta) undergoing FSH therapy for IVF. To determine whether androgen excess in utero affects follicle development over time, this study examines whether PA exposure, beginning at gestational days 40-44 (early treated) or 100-115 (late treated), alters the decline in serum anti-Mullerian hormone (AMH) levels with age in adult female rhesus monkeys and perturbs their ovarian response to recombinant human FSH (rhFSH) therapy for IVF. METHODS: Thirteen normal (control), 11 early-treated and 6 late-treated PA adult female monkeys had serum AMH levels measured at random times of the menstrual cycle or anovulatory period. Using some of the same animals, basal serum AMH, gonadotrophins and steroids were also measured in six normal, five early-treated and three late-treated PA female monkeys undergoing FSH therapy for IVF during late-reproductive life (>17 years); serum AMH also was measured on day of HCG administration and at oocyte retrieval. RESULTS: Serum AMH levels in early-treated PA females declined with age to levels that were significantly lower than those of normal (P < or = 0.05) and late-treated PA females (P < or = 0.025) by late-reproductive life. Serum AMH levels positively predicted numbers of total/mature oocytes retrieved, with early-treated PA females having the lowest serum AMH levels, fewest oocytes retrieved and lowest percentage of females with fertilized oocytes that cleaved. CONCLUSIONS: Based on these animals, early PA appears to program an exaggerated decline in ovarian reserve with age, suggesting that epigenetically induced hormonal factors during fetal development may influence the cohort size of ovarian follicles after birth.

Chronologic and physiologic age affect replicative life-span of fibroblasts from diabetic, prediabetic, and normal donors.

Cultured skin fibroblasts from subjects with clinically apparent diabetes mellitus and from subjects genetically predisposed to diabetes have a replicative lifespan that is inversely related to donor age. Fibroblasts from carefully defined normal subjects not predisposed to diabetes fail to show this correlation. The data support the idea that physiologic status of the tissue donor is a more precise determinant of fibroblast replicative lifespan than chronologic age.

Therapeutic efficacy of undenatured type-II collagen (UC-II) in comparison to glucosamine and chondroitin in arthritic horses.

The present investigation evaluated arthritic pain in horses receiving daily placebo, undenatured type II collagen (UC-II) at 320, 480, or 640 mg (providing 80, 120, and 160 mg active UC-II, respectively), and glucosamine and chondroitin (5.4 and 1.8 g, respectively, bid for the first month, and thereafter once daily) for 150 days. Horses were evaluated for overall pain, pain upon limb manipulation, physical examination, and liver and kidney functions. Evaluation of overall pain was based upon a consistent observation of all subjects during a walk and a trot in the same pattern on the same surface. Pain upon limb manipulation was conducted after the walk and trot. It consisted of placing the affected joint in severe flexion for a period of 60 sec. The limb was then placed to the ground and the animal trotted off. The response to the flexion test was then noted with the first couple of strides the animal took. Flexion test was consistent with determining clinically the degree of osteoarthritis in a joint. Horses receiving placebo showed no change in arthritic condition, while those receiving 320 or 480 or 640 mg UC-II exhibited significant reduction in arthritic pain (P < 0.05). UC-II at 480 or 640 mg dose provided equal effects, and therefore, 480 mg dose was considered optimal. With this dose, reduction in overall pain was from 5.7 +/- 0.42 (100%) to 0.7 +/- 0.42 (12%); and in pain upon limb manipulation from 2.35 +/- 0.37 (100%) to 0.52 +/- 0.18 (22%). Although glucosamine and chondroitin treated group showed significant (P < 0.05) reduction in pain compared with pretreated values, the efficacy was less compared with that observed with UC-II. In fact, UC-II at 480 or 640 mg dose was found to be more effective than glucosamine and chondroitin in arthritic horses. Clinical condition (body weight, body temperature, respiration rate, and pulse rate), and liver (bilirubin, GGT, and ALP) and kidney (BUN and creatinine) functions remained unchanged, suggesting that these supplements were well tolerated.

99mTc-NC100692--a tracer for imaging vitronectin receptors associated with angiogenesis: a preclinical investigation.

INTRODUCTION: Technetium 99m (99mTc)-NC100692 is being developed as a marker of vitronectin receptor expression. The purpose of this study was to confirm the binding affinity [dissociation constant (Kd)] of 99mTc-NC100692 for a range of integrin receptors including alphavbeta3 and alphavbeta5 as well as to establish the biodistribution and metabolic stability of 99mTc-NC100692 in Wistar rats. METHODS: The Kd of 99mTc-NC100692 for a range of human integrin receptors was established in an in vitro saturation binding assay. The biodistribution and metabolic stability of 99mTc-NC100692 in normal Wistar rats was investigated. RESULTS: The Kd of 99mTc-NC100692 to alphavbeta3 and alphavbeta5 was less than 1 nM. It was not possible to saturate the binding of 99mTc-NC10092 towards alphaIIbbeta3, alpha5beta1, alpha3beta1 or alpha1beta1, and as a result, accurate Kd values could not be determined. The biodistribution of 99mTc-NC100692 in male and female Wistar rats showed that radioactivity was rapidly excreted, predominantly into the urine, with very little background tissue retention apart from the liver and kidneys. Kidney and liver retention was reduced in the presence of excess NC100692 ligand. In vivo, there was little systemic metabolism of 99mTc-NC100692. CONCLUSIONS: 99mTc-NC100692 has a high affinity for the vitronectin receptors that are associated with angiogenesis. 99mTc-NC100692 is metabolically stable in the systemic circulation of rats with a biodistribution that is favourable for imaging purposes. This evidence suggests that 99mTc-NC100692 might be a useful marker of vitronectin receptor expression in vivo.

5-lipoxygenase-dependent biosynthesis of novel 20:4 n-3 metabolites with anti-inflammatory activity.

5-lipoxygenase (5-LO) catalyzes the conversion of arachidonic acid (AA) into pro-inflammatory leukotrienes. N-3 PUFA like eicosapentaenoic acid are subject to a similar metabolism and are precursors of pro-resolving mediators. Stearidonic acid (18:4 n-3, SDA) is a plant source of n-3 PUFA that is elongated to 20:4 n-3, an analogue of AA. However, no 5-LO metabolites of 20:4 n-3 have been reported. In this study, control and 5-LO-expressing HEK293 cells were stimulated in the presence of 20:4 n-3. Metabolites were characterized by LC-MS/MS and their anti-inflammatory properties assessed using AA-induced autocrine neutrophil stimulation and leukotriene B4-mediated chemotaxis. 8­hydroxy­9,11,14,17-eicosatetraenoic acid (Δ17-8-HETE) and 8,15-dihydroxy-9,11,13,17-eicosatetraenoic acid (Δ17-8,15-diHETE) were identified as novel metabolites. Δ17-8,15-diHETE production was inhibited by the leukotriene A4 hydrolase inhibitor SC 57461A. Autocrine neutrophil leukotriene stimulation and neutrophil chemotaxis, both BLT1-dependent processes, were inhibited by Δ17-8,15-diHETE at low nM concentrations. These data support an anti-inflammatory role for Δ17-8,15-diHETE, a novel 5-LO product.

Diabetes mellitus and genetic prediabetes. Decreased replicative capacity of cultured skin fibroblasts.

The idea that the gene(s) that cause diabetes mellitus can be expressed in extrapancreatic cells has been examined by tissue culture techniques. Skin biopsies were obtained from 25 normal subjects (N), 26 overt diabetics (D), 16 of juvenile onset (JOD) and 9 of maturity onset (MOD), and 21 subjects genetically predisposed to diabetes (P) on the basis of maturity-onset diabetes in both parents. Each biopsy was subdivided, multiple skin fragments were explanted in vitro, and several parameters of cellular outgrowth were monitored in primary and secondary cultures until cell division ceased because of senescence. In general, the rank order of growth vigor was N greater than P greater than D although differences were often marginal and statistically significant between N and JOD and(or) MOD. Outgrowth of epithelial cells was more vigorous in N explants in early stages, but later, JOD and MOD cells grew better than those of N. Outgrowth of fibroblast cells from N explants was more vigorous both at early and later stages and required less time to achieve maximum percent outgrowth. In secondary cultures, N cells grew faster than the other three groups so that fewer days elapsed between subcultures but significant differences were only seen between N and one or two of the other groups over some of the first seven subcultures. The onset of cellular senescence occurred earlier in P and JOD cultures both in mean population doublings and calendar time. N cultures had a higher percent surviving clones after picking than MOD, and a shorter recloning time than clones of JOD. The replicative life-spans of cultures (mean population doublings +/- SE) were N = 52.54 +/- 2.24, P = 47.84 +/- 2.43, JOD = 47.12 +/- 2.99, and MOD = 46.40 +/- 4.04, but differences did not reach significance for N vs the other three groups. The data demonstrate that cellular growth is impaired in both JOD and MOD types of cultures and to a generally lesser extent in P cultures. This is consistent with intrinsic genetic defects but the possibility that persistent deleterious effects of in vivo pathophysiology contribute alone or in combination cannot be ruled out. Therefore, the diabetic defect(s) can be expressed in extrapancreatic cells of mesenchymal origin. This system should prove useful in exploring the interplay between genetic and environmental factors in diabetes, the mechanisms(s) of hyperglycemia and other metabolic derangements, and the propensity that affected individuals have to develop degenerative diseases.

99mTc-NC100668, an agent for imaging venous thromboembolism: The effect of anticoagulant or thrombolytic therapy on the uptake and retention of radioactivity in blood clots in vivo.

BACKGROUND: The purpose of this study was to evaluate the uptake of (99m)Tc-NC100668 into blood clots and elucidate the potential for medications commonly used to treat thromboembolism to interfere with the uptake and retention of (99m)Tc-NC100668. METHODS: (99m)Tc-NC100668 in vivo uptake and retention in a range of blood clot of various ages (up to 4 h. old) and in the presence of anticoagulants or thrombolytic therapies was measured in a rat model of deep vein thrombosis. RESULTS: (99m)Tc-NC100668 was rapidly absorbed into and retained by blood clots and was not significantly affected by the presence of unfractionated or low molecular weight heparin or thrombin inhibitor. Tissue plasminogen activator reduced the uptake of (99m)Tc-NC100668 into blood clot by a factor of 3 when adjusted to allow for changes in the weight of the blood clot. CONCLUSIONS: This study has demonstrated that the uptake and retention of (99m)Tc-NC100668 into blood clots in the rat model of deep vein thrombosis is rapid and maintained over at least a 4 h. post-injection period. It has been shown that (99m)Tc-NC100668 is retained in blood clots even in the presence of therapeutic doses of those anticoagulant and thrombolytic therapies typically used to treat pulmonary embolism and venous thrombosis.

Safety of an attenuated West Nile virus vaccine, live Flavivirus chimera in horses.

REASON FOR PERFORMING STUDY: West Nile virus (WNV) infection is endemic and able to cause disease in naive hosts. It is necessary therefore to evaluate the safety of new vaccines. OBJECTIVES: To establish: 1) the safety of a modified live Flavivirus/West Nile virus (WN-FV) chimera by administration of an overdose and testing for shed of vaccine virus and spread to uninoculated sentinel horses; 2) that this vaccine did not become pathogenic once passaged in horses; and 3) vaccine safety under field conditions. METHODS: There were 3 protocols: 1) In the overdose/shed and spread study, horses were vaccinated with a 100x immunogenicity overdose of WN-FV chimera vaccine and housed with sentinel horses. 2) A reversion to virulence study, where horses were vaccinated with a 20x immunogenicity overdose of WN-FV chimera vaccine. Horses in both studies were evaluated for abnormal health conditions and samples obtained to detect virus, seroconversion and dissemination into tissues. 3) In a field safety test 919 healthy horses of various ages, breeds and sex were used. RESULTS: Vaccination did not result in site or systemic reactions in either experimental or field-injected horses. There was no shed of vaccine virus, no detection of vaccine virus into tissue and no reversion to virulence with passage. CONCLUSIONS: WN-FV chimera vaccine is safe to use in horses with no evidence of ill effects from very high doses of vaccine. There was no evidence of reversion to virulence. In addition, administration of this vaccine to several hundred horses that may have been previously exposed to WNV or WNV vaccine resulted in no untoward reactions. POTENTIAL RELEVANCE: These studies establish that this live attenuated Flavivirus chimera is safe to use for immunoprophylaxis against WNV disease in horses.

Partial characterization of an allergenic glycoprotein from peanut (Arachis hypogaea L.).

A concanavalin A-reactive glycoprotein allergen has been isolated from peanut (Arachis hypogaea). The allergen was separated by affinity chromatography and purified by gel permeation and ion-exchange chromatography. The monomeric molecular weight is 65,000 and the pI is 4.6. The presence of one cysteine residue per molecule results in some dimer formation. Concanavalin A-reactive glycoprotein is a potent allergen for peanut-sensitive patients in both in vivo and in vitro tests. It is allergenically stable, on in vitro examination, at temperatures of up to 100 degrees C and over the pH range 2.8-10. Removal of the carbohydrate moiety failed to eliminate the allergenicity. Concanavalin A-reactive glycoprotein is identified in the crossed immunoelectrophoretic pattern as a major antigen of peanut protein extract but its structural characteristics indicate that it is probably not a component of the major storage-protein complex, arachin.