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Aim: To determine if vitamin D3 treatment reduced the incidence of vaginal prolapse in pregnant sheep on a North Canterbury sheep breeding property. Methods: Pregnant ewes from a single farm were allocated to three treatment groups in May 2018. At this time, the first group (EarlyVitADE; n = 512) received an I/M 1 mL dose of 500,000 IU/mL vitamin D3, 60,000 IU/mL vitamin A, and 25 mg/mL vitamin E. This was repeated in July 2018, when the second group (LateVitADE; n = 695) also received the same treatment. The third group (n = 737) were untreated controls. All cases of vaginal prolapse on the property were recorded from pregnancy diagnosis in June 2018 until ewes were set-stocked in August 2018. The planned start of lambing was 10 August 2018. Results: During the period of observation, vaginal prolapses were recorded in 3/699 (0.4%) 2-year-old ewes, and the odds of vaginal prolapse were not associated with treatment group in these ewes (p > 0.3). Amongst ewes aged ≥3 years, during the same period, there were 6/333 (1.8%), 6/443 (1.4%) and 25/469 (5.3%) cases in the EarlyVitADE, LateVitADE and control groups, respectively. Compared to control ewes, the odds of vaginal prolapse were reduced in both the EarlyVitADE (OR = 0.37; 95% CI = 0.15–0.92) and LateVitADE (OR = 0.25; 95% CI = 0.10–0.62) treatment groups. Conclusions and clinical relevance: In this preliminary study, administration of injectable vitamins A, D3, and E to pregnant ewes reduced the incidence of vaginal prolapse during the period from pregnancy diagnosis to set-stocking on one North Canterbury hill-country farm. Due to the restricted data collection period, this investigation should be replicated to better quantify the repeatability of the observed treatment effect over the complete lambing period. [ABSTRACT FROM AUTHOR]

Pinhão is highly perishable due to its high water activity, being easily affected by fungi during storage and also susceptible to infestation by larvae. This seed is usually marketed in the pinhão cones itself, or bulk threshed, and packed in plastic bags, chilled or ground frozen. Pinhão conservation and industrialization techniques should be developed to promote its commercialization and consumption at other times of the year, besides the seasonal period, encouraging its sustainable production, extraction and commercialization, considering its essentially extractive character. The objective of this study was to evaluate the conservation of pinhão by the use of gamma radiation and refrigeration. The pinhões were irradiated with a cobalt–60 source at a dose rate of 1 kGy. A non-irradiated sample was used as a control. The pinhões were packed in high-density polyethylene bags and stored at ambient temperature and refrigerated at 4 ºC, during 90 days. Pinhões were evaluated for weight loss, acidity, reducing sugars, vitamin C, firmness, color, total phenolic compounds, antioxidant activity and microbiological analyzes. The isolated use of gamma radiation was not effective for the maintenance of the evaluated parameters. However, when used in conjunction with refrigerated storage, it reduced the growth of aerobic fungi, as well as mesophilic and psychrotrophic microorganisms. The isolated use of refrigeration showed a reduction in weight loss, reducing sugars and an increase in vitamin C content and antioxidant activity. Thus, to increase the benefits, we suggest evaluating higher doses of radiation as a function of the thick pinhão shell.

Apresentação do dossiê "Tecnologia e políticas culturais: o acesso à cultura digital no Brasil", que contempla os seguintes artigos: “Iniciativas brasileiras em torno da construção de uma Política Nacional para Acervos Digitais de Instituições de Memória: o desafio da memória em tempos de cultura digital”; “As Transformações da Era Digital e o Impacto na Economia da Cultura do Livro”; “Dialogando com e sobre o rural através das novas mídias: estratégias e desafios da comunicação” e “Softwares livres e políticas culturais no Brasil: o caso do Mapa Cultural”. Os artigos tratam das aplicações das tecnologias digitais às políticas culturais, localizando-as num contexto mais geral das aplicações das TICs e analisando casos setoriais.

Este trabalho tem como objeto a expansão do consumo cultural-digital das famílias brasileiras nos últimos dez anos. Tal expansão implicou diretamente a elevação de seus gastos orçamentários com cultura. De acordo com o IBGE, em 2009, as famílias brasileiras dispenderam em média 5% do seu orçamento mensal com bens culturais. Caso os serviços de telefonia e comunicação fossem adicionados à rubrica cultura, essa mesma média de gastos saltaria para 8,6%. Em face do aumento da produção, distribuição e consumo dos serviços culturais-digitais (séries, filmes, músicas, games, etc.) por meio dos dispositivos digitais móveis, notadamente os smartphones, sustentamos que a metodologia de cálculo e aferição do IBGE exige uma revisão. Para sustentar tal necessidade, demonstramos que os mercados culturais passam por uma profunda mudança: a digitalização do simbólico.

In the last decades, the themes of creative economy, creative industries and creative cities served as a technical and operational fundament for the formulation and execution of a series of economic development public policies. The main actors of this profusion and legitimation of power were the economists, managers and business schools theorists in close relation to national governments and UN agencies. This process was possible due to the emergence of post-industrial societies, the expansion of the knowledge economy and the digitalization of cultural markets. The combination of these three aspects produced a structural approximation between the symbolic-aesthetic domain and the economic-technological domain, resulting in the formation of native categories, such as creative economy, creative industries and creative cities. These categories do not have the analytical and descriptive potential which able to comprehend and explain the new relations between those domains, tough they are themselves a result of these new relations.

This work investigates the process of competition between some of the most important technology companies in the world, which created business models aimed at the culturaldigital services market - series, movies, games, music, books and animation. It is a process of global scope and great economic, technological and creative impact, which has been refashioning the cultural markets through the intense process of digitalization and corporate action. To analyze these aspects, two methodological and analytical strategies were adopted simultaneously along the text. The first one sought to use the concepts of digitalization of the symbolic and cultural-digital capitalism, both of which are woven into other works and research that combine a broader research agenda, whose efforts are engaged in the realization of an economic sociology of cultural markets. The second was to explore the latest economic decisions and commercial and creative strategies adopted by the Netflix, Disney and Apple companies, as well as to update data on the expansion of consumption of digital artistic-cultural content, especially in Brazil.

neste trabalho buscaremos reconstruir o início do mercado brasileiro da internet. Para isso partiremos de matérias publicadas em sites de naturezas distintas, no período dos 15 anos iniciais do mercado da internet no Brasil. Essas notícias são importantes porque trazem questões situadas no tempo sobre o uso da internet por parte dos brasileiros, bem como o esforço do Estado na criação e controle do mercado da internet. Ainda, traremos dilemas que preocuparam os sujeitos nesse período, seja do ponto de vista dos impactos da internet e da tecnologia na vida de seus usuários, ou dos dispositivos jurídicos necessários para a proteção dos sujeitos no cotidiano da relação com a internet. Esses dados serão mobilizados na operacionalização de diferentes teorias da rede que estão em disputa no campo de produção de conhecimento da sociologia, os representantes são Castells, Elias, Granovetter e Latour. Dessa forma, através dessas teorias, iremos desnaturalizar as preocupações relacionadas ao uso da internet no Brasil, bem como relembrar que as questões que nos afligem hoje, na verdade, são antigas.

To understand the relevance of the new media in the formation of the indignation nets, this text, of exploratory stamp, debates the digital activism in contemporary Brazil . Methodologically, we will make a discussion on cyberactivism, digital media, and national pressure groups starting from two examples, Movimento Brasil Livre (The Free Brazil Movement) and Vem pra Rua (Come to The Street movement) – these are key movements in the organisation of the big anti-government mobilisation that took place in 2015–2016 in Brazil. The theme is important because it embraces current and future challenges of the digital activism, once that this field faced significant changes in the last decades, with the development of interactive media and the technological convergence.

O Serviço Brasileiro de Apoio às Micro e Pequenas Empresas (SEBRAE) tem executado uma série de ações de estímulo ao empreendedorismo cultural. O SEBRAE é uma entidade privada sem fins lucrativos e de interesse público, com fortes vínculos jurídicos, financeiros e políticos junto ao Estado brasileiro. A atuação do SEBRAE no âmbito do empreendedorismo cultural o torna um agente estatal de mercado (AEM) bastante específico, cuja atuação o caracteriza como um agente das novas políticas culturais no Brasil, as políticas econômico-culturais. Por meio das suas modalidades de atuação, o SEBRAE corrobora diretamente para o processo de incorporação das disposições práticas que orientam a formulação de novos modelos de negócios e de novas racionalidades empresariais por parte das micro e pequenas empresas culturais.

Este artigo procura situar o advento e a operacionalização de uma formação discursiva responsável pela emergência de um novo universalismo global. Tal universalismo traz como imperativo ético-moral a defesa e a promoção de valores como a diversidade cultural. Para compreender as implicações políticas e culturais desse universalismo, este trabalho elege como eixo empírico a atuação da Organização das Nações Unidas Para Educação, a Ciência e a Cultura (UNESCO), destacando as ações dessa organização na coordenação e dinamização de um trânsito simbólico e discursivo transnacional envolvendo governos, organizações não-governamentais e grupos políticos.This article attempts to situate the emergence and development of a discursive formation responsible for the appearance of a new global universalism. The ethical and moral imperative of this universalism is the defense and promotion of values such as cultural diversity. To understand the political and cultural implications of this universalism, this article elects as its empirical axis the performance of the United Nations Educational, Scientific and Cultural Organization (UNESCO), highlighting the actions of the organization in coordinating and facilitating a symbolic and discursive transnational transit involving governments, nongovernmental organizations and political groups.

A discordance between sex hormone-binding globulin (SHBG) measurements by 2-site ELISAs was investigated using pairings of various "in house" SHBG antibodies together with a concordant control. The 2-site monoclonal ELISAs used the same base coat (11F11) and discordance was observed with one top coat monoclonal antibody (7H9) and also when a polyclonal SHBG antibody was paired with the basecoat antibody (11F11). Sialidase treatment of the discordant sample and purified SHBG revealed increased levels using 7H9 whereas there was no change in SHBG in the concordant sample. Conversely, following sialidase treatment, the discordant sample showed no change in SHBG measured using the other monoclonal antibody pairings whereas the SHBG levels in the concordant sample declined following sialidase using the same monoclonal antibody pairings. This implicated glycosylation as a factor in antibody recognition and synthetic peptides spanning the two N-linked and one O-linked glycosylation regions showed that SHBG recognition by monoclonal antibody 7H9 could be disrupted by a peptide spanning the O-linked glycosylation site. Hence rather than immunoassay discordance being attributed to heterophile antibodies or other circulating antibodies here it can be likely attributed to glycosylation affecting antibody recognition and hence the measurement of SHBG., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Objectives: Supplementation provides the best means of improving vitamin D status; however, individual responses vary partly owing to genetics. The aim of this study was to determine whether 28 single nucleotide polymorphisms (SNPs) in six key vitamin D pathway genes (GC, DHCR7, CYP2 R1, CYP24 A1, CYP27 B1, VDR) were associated with differences in response to supplementation., Methods: Participants (N = 313; n = 160 vitamin D, n = 153 placebo) were part of VIDARIS (Vitamin D and Acute Respiratory Infections Study), a double-blind, randomized controlled trial involving oral monthly supplementation of either vitamin D 3 (200 000 IU each for the first 2 mo, thereafter 100 000 IU monthly) or placebo for 18 mo. Circulating 25-hydroxyvitamin D (25[OH]D) concentrations at baseline and 2, 6, 12, and 18 mo, and vitamin D binding protein (Gc-globulin) and calculated free 25(OH)D concentrations at baseline and 2 mo were obtained. Multiple regression was used to model associations between genetic variants and 25(OH)D, Gc-globulin, and free 25(OH)D concentrations., Results: SNPs within GC, CYP2 R1, and CYP27 B1 were associated with 25(OH)D concentrations following supplementation. However, only two GC gene SNPs (rs2282679, rs1155563) were significant after adjustment for multiple testing. This effect disappeared after more than 2 mo of supplementation. None of the SNPs were significantly associated with Gc-globulin concentrations; however, there was a significant interaction with one SNP in DHCR7 (rs12785878), which was associated with reduced free 25(OH)D concentrations in the supplemented arm., Conclusion: Only variants of GC were associated with 25(OH)D concentrations after supplementation. This effect was modest and disappeared after >2 mo of supplementation, suggesting it may be time/dose-dependent and saturable., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Calprotectin, the major neutrophil protein, is a critical alarmin that modulates inflammation and plays a role in host immunity by strongly binding trace metals essential for bacterial growth. It has two cysteine residues favourably positioned to act as a redox switch. Whether their oxidation occurs in vivo and affects the function of calprotectin has received little attention. Here we show that in saliva from healthy adults, and in lavage fluid from the lungs of patients with respiratory diseases, a substantial proportion of calprotectin was cross-linked via disulfide bonds between the cysteine residues on its S100A8 and S100A9 subunits. Stimulated human neutrophils released calprotectin and subsequently cross-linked it by myeloperoxidase-dependent production of hypochlorous acid. The myeloperoxidase-derived oxidants hypochlorous acid, taurine chloramine, hypobromous acid, and hypothiocyanous acid, all at 10 μM, cross-linked calprotectin (5 μM) via reversible disulfide bonds. Hypochlorous acid generated A9-A9 and A8-A9 cross links. Hydrogen peroxide (10 μM) did not cross-link the protein. Purified neutrophil calprotectin existed as a non-covalent heterodimer of A8/A9 which was converted to a heterotetramer - (A8/A9) 2 - with excess calcium ions. Low level oxidation of calprotectin with hypochlorous acid produced substantial proportions of high order oligomers, whether oxidation occurred before or after addition of calcium ions. At high levels of oxidation the heterodimer could not form tetramers with calcium ions, but prior addition of calcium ions afforded some protection for the heterotetramer. Oxidation and formation of the A8-A9 disulfide cross link enhanced calprotectin's susceptibility to proteolysis by neutrophil proteases. We propose that reversible disulfide cross-linking of calprotectin occurs during inflammation and affects its structure and function. Its increased susceptibility to proteolysis will ultimately result in a loss of function., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)

The measurement of vitamin D-binding protein (VDBP) by immunoassay has been confounded by variable antibody recognition of the Gc1s, Gc1F and Gc2 phenotypes. This has led to spurious conclusions regarding vitamin D status in different ethnic groups. In order to overcome these problems there is a requirement for VDBP antibodies that are unaffected by phenotype status. Here we report the generation and testing of three monoclonal antibodies to VDBP which recognise linear epitopes and are unaffected by vast molar excesses of synthetic peptides spanning these phenotypic domains. These IgG1 kappa antibodies were purified and biotinylated to allow suitable pairings to develop a sandwich ELISA for circulating VDBP. The VDBP ELISA is unaffected by actin and confirms that VDBP levels are significantly reduced in sepsis patients and non-sepsis intensive care patients compared to normal healthy subjects. Levels of VDBP along with total 25OH vitamin D3 can be used to calculate free 25OH vitamin D3 levels and these compare well with consensus values determined independently. The VDBP ELISA meets acceptable performance criteria and as such can be used in conjunction with total 25OH vitamin D3 to determine the free 25OH vitamin D3 status in various cohorts., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The aim of this study was to determine if serum fibroblast growth factor (FGF23) concentrations were increased in dogs with chronic kidney disease (CKD). Serum samples submitted to a commercial laboratory were collected over a 15-month period, 14 samples were from dogs with a history of polyuria/polydipsia, azotaemia and low urine specific gravity, 20 samples were from non-azotaemic dogs. Serum FGF23, parathyroid hormone, total calcium and phosphorus, urea and creatinine were measured. Mann-Whitney test was used to determine differences between non-azotaemic and CKD groups; a one-way ANOVA with Tukey pairwise comparisons was used to determine any differences between International Renal Interest Society stages; and regression models were used to determine predictors of International Renal Interest Society stage, serum phosphorus and FGF23 concentrations. The median serum FGF23 concentration of dogs with CKD was 5194.6pg/mL, which was significantly greater (P<0.001) than the median serum FGF23 concentration of non-azotaemic dogs (259.2pg/mL). Log serum FGF23 and age were significantly associated with IRIS stage (P=0.027 and P=0.032 respectively), while log serum phosphorus concentration (P<0.001) was significantly associated with log serum FGF23 concentration. In summary, serum FGF23 concentration is increased in dogs with CKD, and is associated with serum phosphorus concentration. This phosphatonin pathway may be a useful target for the development of future treatments to control plasma phosphorus concentrations in chronic kidney disease., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Corticosteroid-binding globulin (CBG) binds most of the cortisol in circulation and is a non-functional member of the family of serine protease inhibitors (serpins) with an exposed elastase sensitive reactive centre loop (RCL). The RCL can be cleaved by human neutrophil elastase, released from activated neutrophils, and can also be cleaved at nearby site(s) by elastase released by Pseudomonas aeruginosa, and at two further sites, also within the RCL, by bovine chymotrypsin. Cleavage of the RCL results in a conformational change accompanied by a marked decrease in affinity for cortisol and hence its release at the site of proteolysis. These cleavages are irreversible and the similar half-lives of cleaved and intact CBG could mean that there may be some advantage in slowing the rate of CBG cleavage in acute inflammation thereby increasing the proportion of intact CBG in circulation. Here we show, for the first time, that pre-incubation of tethered human CBG with two monoclonal antibodies to the RCL of CBG protects against cleavage by all three enzymes. Furthermore, in plasma, pre-incubation with both RCL monoclonal antibodies delays neutrophil elastase cleavage of the RCL and one of these RCL monoclonal antibodies also delays bovine chymotrypsin cleavage of the RCL. These findings may provide a basis and rationale for the concept of the use of RCL antibodies as therapeutic agents to effectively increase the proportion of intact CBG in circulation which may be of benefit in acute inflammation., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Corticosteroid-binding globulin (CBG) is the predominant carrier of cortisol in circulation and is a non-inhibitory member of the serpin superfamily of serine protease inhibitors. In the stressed or "S" conformation, CBG possesses an intact exposed reactive centre loop (RCL) that can be irreversibly cleaved by elastase released from activated human neutrophils whereupon it adopts a relaxed or "R" conformation. The latter conformation has decreased affinity for cortisol, allowing the release of the majority of cortisol at sites of inflammation. Recently there has been speculation that mild increments in heat such as found in pyrexia (39-40°C) may also induce a reversible "flip-flop" of the RCL into the body of the protein structure, without cleavage, facilitating a reversible temperature-dependent release of cortisol. Here we raised a new monoclonal antibody to the RCL of human CBG and used this in concert with an existing RCL antibody and show by surface plasma resonance that, at temperatures up to 40°C, the RCL of purified CBG and the RCL of CBG in intact plasma is accessible to these two antibodies. Together, the epitopes of these antibodies span 11 consecutive amino acids (STGVTLNLTSK) of the 18 residues of the RCL. This adequate antibody cover of the RCL sequence leads to the conclusion that the proposed temperature-dependent "flip-flop" of the RCL of CBG is doubtful., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Corticosteroid-binding globulin (CBG) is a non-inhibitory member of the serpin superfamily of serine protease inhibitors and carries the majority of cortisol in circulation. It can be cleaved by neutrophil elastase at its exposed reactive centre loop which decreases its affinity for cortisol allowing the release of most of the cortisol at sites of inflammation. Intact and elastase cleaved CBG can be distinguished from each other and can coexist in circulation but with unknown half-lives. Here we treated a portion of purified human CBG with elastase, terminated the digestion and then combined this portion with intact human CBG and measured their respective half-lives in rabbits by ELISA. This investigation shows for the first time that the half-lives of intact and elastase cleaved CBG are identical (∼10h). This is an important finding as it implies that in conditions such as sepsis and septic shock where levels of intact CBG are low and the proportion of cleaved CBG is high that this is likely sustained which may affect the CBG mediated targeted delivery of cortisol to sites of inflammation. Furthermore the residual binding of cortisol to cleaved CBG may alter the overall buffering capacity of CBG for cortisol resetting the baseline concentration of free cortisol., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Background: Extraction followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis is the method of choice when it comes to the accurate quantification of 25-OH-vitamin D in blood samples. It is generally assumed that the addition of exogenous internal standard allows for the determination of the endogenous analyte concentration. In this study we investigated the extraction properties of endogenous and exogenous 25-OH-vitamin D., Methods: Eight samples were used for the evaluation of the extraction procedure and 59 patients' samples for a method comparison. The methanol-to-sample ratio (v/v) and the sample-to-hexane ratio (v/v) were varied and the LC-MS/MS signals of endogenous 25-OH-vitamin D3, spiked 25-OH-vitamin D2 and internal standard of the extracts recorded. The optimized 'in-house' LC-MS/MS assay was compared to two automated chemiluminescence immunoassays from DiaSorin and Abbott., Results: Mathematical analysis of the data revealed a differential extraction of endogenous 25-OH-vitamin D3, spiked 25-OH-vitamin D2 and non-equilibrated internal standard. Exogenous 25-OH-vitamin D can be measured accurately if a definite methanol-to-sample ratio is used. Endogenous 25-OH-vitamin D is affected by critical quantification issues due to a differential slope in the extraction profile. The actual 25-OH-vitamin D concentration can be one-third above the measured extractable concentration. Results confirm that the 'in-house' LC-MS/MS assay provides reproducible 25-OH-vitamin D results., Conclusions: Discordant concentrations of 25-OH-vitamin D from LC-MS/MS assays can be caused by selection of suboptimal extraction conditions. Furthermore, a different sample pretreatment or solvent extraction system may result in a different dissociation and extraction yield of endogenous 25-OH-vitamin D and therefore contribute to variations of LC-MS/MS results., (© The Author(s) 2014 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.)

Corticosteroid-binding globulin (CBG) binds more than 90% of circulating cortisol and is a non-inhibitory member of the family of serine protease inhibitors (SERPINS) with an exposed elastase sensitive reactive centre loop (RCL). At sites of inflammation neutrophil activation can release elastase which may cleave the RCL and result in cortisol release from CBG. The RCL sequence also has two theoretical chymotrypsin cleavage sites and we used a monoclonal antibody with specificity for the RCL to investigate chymotrypsin cleavage of CBG. Here we show, for the first time, rapid chymotrypsin cleavage of the RCL of CBG, resulting in undetectable levels of intact CBG, whereas total CBG levels were unchanged. Coincident with both chymotrypsin and elastase cleavage there was an increase in the free cortisol fraction of serum to levels similar to when CBG had been inactivated by heat indicating total cortisol release from CBG. These findings demonstrate a new mechanism for cortisol release from its binding globulin., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

The predominant carrier of cortisol in circulation is corticosteroid-binding globulin (CBG) which is a non-functional member of the family of serine protease inhibitors. Corticosteroid-binding globulin possesses an exposed elastase sensitive loop and upon cleavage it adopts a "relaxed" conformation promoting the delivery of cortisol to sites of inflammation. Recently we have developed monoclonal antibodies which recognise only the intact exposed elastase loop, including an N-glycosylation site, which, in concert with another monoclonal antibody to CBG, offered the potential for the determination of intact and total CBG which may both be present in circulation. Here we validate these parallel ELISAs and show that like total CBG there is little diurnal variation of intact plasma CBG. Furthermore in a normal reference population the majority of CBG is in the intact or active form but a significant level of apparently cleaved CBG is evident. In some subjects there is gross discordance between total CBG and intact CBG implying a predominance of apparently cleaved CBG in circulation and this significantly affects calculated free cortisol levels. Gross differences in total and intact CBG levels may not be due to differences in N-glycosylation affecting antibody binding as CBG levels are unaffected by PNGase F treatment., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Corticosteroid-binding globulin (CBG) is the principal carrier of cortisol in circulation and is a non-inhibitory member of the serpin family of serine proteinase inhibitors. It possesses an exposed elastase specific site which, when cleaved, allows a conformational change promoting the delivery of cortisol to sites of inflammation. Previously there was no ability to independently distinguish between the uncleaved, stressed, conformer of CBG and total CBG in circulation. Here we raised and characterized monoclonal antibodies generated against a synthetic peptide spanning the elastase cleavage site within the exposed reactive centre loop (RCL) and measured changes in CBG by ELISA following treatment with human neutrophil elastase. The antibodies recognized the synthetic peptide as well as intact CBG and the epitope (STGVTLNL) spanned the elastase cleavage site. Treatment of plasma with elastase resulted in a complete loss of CBG levels determined using these RCL antibodies whereas CBG levels measured with an unrelated CBG monoclonal antibody were unaffected. We also compared plasma levels of CBG measured by RCL antibodies and an unrelated CBG antibody and showed discordance in some samples. This study shows for the first time the ability to measure the intact, stressed conformer of CBG. We report discordance with total CBG in some samples implying the presence of cleaved CBG in circulation. This is an important finding as it has implications for free cortisol which hitherto have been determined from total cortisol and total CBG levels. This antibody could be used for determining the time course of intact CBG in various relevant patient cohorts and for structure/function studies on the biology of human CBG., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Circulating cortisol, corticosteroid-binding globulin, and sex hormone-binding globulin were measured retrospectively in plasma samples following the oral glucose tolerance test in 20 spinal cord-injured men and 20 able-bodied controls. Plasma-free cortisol responses attenuated more rapidly in the able-bodied men, compared to spinal cord-injured subjects, due to significant rise in circulating corticosteroid-binding globulin whereas changes in total plasma cortisol were similar in both groups. The changes in plasma-free cortisol in both groups paralleled changes in insulin and glucose and show that spinal cord-injured men had heightened exposure to free cortisol during this dynamic test. This raises the possibility that the mechanism of abdominal obesity and the propensity towards insulin resistance in spinal cord-injured men could be subtly mediated by perturbations in free cortisol. There were no significant changes in plasma sex hormone-binding globulin in either group., (© Georg Thieme Verlag KG Stuttgart · New York.)

Circulating sex hormone-binding globulin (SHBG), corticosteroid-binding globulin (CBG), and total and calculated free cortisol were measured in 206 overweight subjects to investigate whether or not they were markers of insulin resistance. Measurements were carried out on two occasions 36 months apart and subjects were grouped according to fasting plasma glucose. Fifty-one subjects, with a normal basal fasting glucose (<5.6 mmol/l) developed impaired fasting glucose 3 years later (> or = 5.6 mmol/l). Analysis either in toto or based on gender showed a highly significant increase in fasting insulin and insulin resistance, a modest increase in body mass index (BMI), but importantly no change in plasma SHBG, CBG, or cortisol concentrations. Subjects (n=101) with a normal fasting glucose both at baseline (<5.6 mmol/l) and at 36 months showed no significant change in fasting insulin, insulin resistance, SHBG, CBG, cortisol, or BMI. Cross-sectional analysis of the study population showed that plasma SHBG correlated negatively with insulin resistance both in men and women. Overall SHBG at baseline was not predictive of changes in fasting glucose. In females, plasma CBG correlated negatively with BMI. The major finding is that overweight subjects who developed impaired fasting glucose showed no significant change in plasma SHBG, CBG or cortisol, and therefore these indices are probably not early markers of insulin resistance in overweight subjects.

Objectives: This longitudinal study investigated whether or not plasma retinol-binding protein (RBP), recently referred to as RBP4, was a marker of insulin resistance in overweight subjects., Methods: We measured anthropometric markers as well as RBP, fasting glucose and insulin in 206 overweight subjects and repeated these measurements 36 months later. Subjects were grouped according to fasting plasma glucose concentration at baseline and 36 months., Results: Subjects (n=51) with a normal basal fasting glucose (<5.6 mmol/L) who developed impaired fasting glucose (IFG) 3 years later (>or=5.6 mmol/L) showed a highly significant increase in both fasting insulin and insulin resistance, but importantly no change in plasma RBP. This group had a significant increase in body mass index (BMI). Subjects (n=101) with a normal fasting glucose at both baseline (<5.6 mmol/L) and 36 months showed no significant change in fasting insulin, insulin resistance, RBP or BMI. The remaining subjects had impaired basal fasting glucose and were not analysed on a group-wise basis. Overall, RBP correlated significantly, but inversely, with anthropometric measures, but not with fasting glucose, insulin or insulin resistance., Conclusions: This is the first report of a long-term longitudinal study on RBP and the major finding is that subjects who developed insulin resistance showed no change in plasma RBP. On the basis of our results we consider that RBP cannot be construed as a marker of insulin resistance in overweight humans.

To assess whether plasma retinol-binding protein (RBP) is a marker of insulin resistance we measured RBP, insulin and glucose in 285 fasting subjects attending a Lipid Disorders Clinic as outpatients. They were grouped as either subjects without diabetes mellitus and with varying degrees of insulin resistance or subjects with diabetes mellitus according to the WHO criteria. We show that there was no association between plasma RBP and insulin-resistance, insulin, glucose, % body fat, waist circumference or BMI whether analysed together or in groups. We confirm, using the largest study cohort to date, that plasma RBP is unlikely to be a useful marker of insulin resistance.

Objectives: Plasma retinol-binding protein (RBP) has been linked to insulin resistance and cardiovascular risk, yet little is know of its natural variation in plasma. We examined this in normal subjects and compared plasma levels and variability in lean subjects and subjects with the metabolic syndrome., Methods: We established an "in house" ELISA for plasma RBP and measured levels in 20 normal subjects over daylight hours and 2 subject groups, either lean or classified with the metabolic syndrome., Results: Plasma RBP in normal subjects did not vary over the day with no differences between males and females. There was also no difference in plasma RBP levels and between the age- and sex-matched lean subjects compared to the metabolic syndrome group., Conclusion: The lack of variation in plasma RBP in normal subjects and the lack of difference between plasma RBP in normal and metabolic syndrome subjects suggest the link between plasma RBP and insulin resistance is tenuous. Investigating a large cohort over the diabetic non-diabetic spectrum may clarify this issue.

Sex hormone-binding globulin (SHBG) and corticosteroid-binding globulin (CBG) circulate in plasma and bind their cognate ligands with high affinity, offering a steroid delivery system to target tissues by a variety of mechanisms. Analysis of these steroid-binding proteins is gaining importance in the clinical setting, although more information is warranted on their diurnal and biological variation. This study shows that plasma SHBG (in normal subjects) exhibits little diurnal or biological variation over the 30 day period studied, in contrast to CBG, where plasma levels peak in the early afternoon. This leads to attenuation of the diurnal free cortisol level rhythm compared to total cortisol. We also show that plasma CBG is significantly lower in male subjects with the metabolic syndrome compared to age-matched lean counterparts, and may therefore act as a surrogate marker of insulin resistance. The consequence of lower levels of CBG in these obese male subjects is reflected by higher levels of circulating free cortisol, potentially offering a more favourable environment for adipogenesis.

Epidemiological evidence implicates dietary isoflavone intake as protective against prostate disease. A putative mechanism is attenuated circulating androgen levels in male populations consuming an isoflavone rich diet. We investigated this hypothesis by collecting plasma from 60 Japanese and 60 New Zealand males aged between 21 and 31 years each consuming their traditional diets. We measured plasma testosterone, dihydrotestosterone (DHT), androstenedione, dehydroepiandrosterone sulfate (DHEAS), the combined levels of androsterone sulfate and epiandrosterone sulfate (AoS/epiAoS), sex hormone-binding globulin, and cortisol and corticosteroid-binding globulin as well as the isoflavones genistein and equol. Plasma genistein and equol levels were several times higher in Japanese males as would be expected from an isoflavone rich diet. However, androstenedione, DHEAS, calculated free testosterone and paradoxically markers of 5alpha-reductase, DHT and AoS/epiAoS were all also significantly higher in Japanese rather than the New Zealand male counterparts. All other comparisons were not significant. Plasma DHT and DHEAS correlated positively with plasma equol and plasma AoS/epiAoS correlated positively with genistein levels. Taken together the results suggest that, rather than reduced levels of steroidogenesis, Japanese males may have increased 5alpha-reductase activity and possibly altered 17beta OH steroid dehydrogenase activity. Significantly the positive association between isoflavones levels and 5alpha-steroids is counter-intuitive to isoflavone intake offering prostate protection, unless this is postulated to occur through other mechanisms.