Application of Ga(68) -DOTA-exendin-4 PET/CT to localise an occult insulinoma

A 49 year old female presented to our Neuroendocrine Tumour (NET) centre with recurrent severe and disabling hypoglycaemia. She had previously been extensively investigated with a clinical and biochemical diagnosis of endogenous hyperinsulinemic hypoglycaemia although the source of hormonal hypersecretion could not be localised with MRI, EUS and (111) In-Octreotide scans. After extensive discussion the patient opted for blind surgical resection undergoing a pylorus-preserving pancreaticoduodenectomy in December 2010. Histological examination of the resected operative specimen demonstrated a normal pancreas with no evidence of neuroendocrine tumour. Consistent with this, post-surgery her hypoglycaemic symptoms persisted with fasting capillary blood glucose of 2.1-6.0 mmol/l with increasing hypoglycaemia unawareness. Consequently she sought alternative clinical opinions from two European Neuroendocrine Tumour Society (ENETS) Centres of Excellence who investigated her collaboratively. This article is protected by copyright. All rights reserved.

Integrated mRNA and miRNA expression profiling in blood reveals candidate biomarkers associated with endurance exercise in the horse.

The adaptive response to extreme endurance exercise might involve transcriptional and translational regulation by microRNAs (miRNAs). Therefore, the objective of the present study was to perform an integrated analysis of the blood transcriptome and miRNome (using microarrays) in the horse before and after a 160 km endurance competition. A total of 2,453 differentially expressed genes and 167 differentially expressed microRNAs were identified when comparing pre- and post-ride samples. We used a hypergeometric test and its generalization to gain a better understanding of the biological functions regulated by the differentially expressed microRNA. In particular, 44 differentially expressed microRNAs putatively regulated a total of 351 depleted differentially expressed genes involved variously in glucose metabolism, fatty acid oxidation, mitochondrion biogenesis, and immune response pathways. In an independent validation set of animals, graphical Gaussian models confirmed that miR-21-5p, miR-181b-5p and miR-505-5p are candidate regulatory molecules for the adaptation to endurance exercise in the horse. To the best of our knowledge, the present study is the first to provide a comprehensive, integrated overview of the microRNA-mRNA co-regulation networks that may have a key role in controlling post-transcriptomic regulation during endurance exercise.

The role of cardiorespiratory fitness and weight status on 24-hr ambulatory blood pressure among adolescents

BACKGROUND: This observational research study investigated the association of cardiorespiratory fitness and weight status with repeated measures of 24-hr ambulatory blood pressure (24-hr ABP). Little is known about these associations and few data exist examining the interaction between cardiorespiratory fitness and weight status and the contributions of each on 24-hr ABP in youth. ^ METHODS: This research study used secondary analysis data from the "Adolescent Blood Pressure and Anger: Ethnic Differences" study. This current study sample included 374 African-American, Anglo-American, and Mexican-American adolescents 11-16 years of age. Mixed-effects models were used for testing the relationship between weight status and cardiorespiratory fitness and repeated measures of ambulatory blood pressure over 24 hours (24-hr ABP). Weight status was categorized into "normal weight" (BMI<85th percentile), "overweight" (85th≤BMI<95th), and "obese" (BMI≥95th). Cardiorespiratory fitness, determined by heart rate recovery (HRR), was defined as the difference between heart rate at peak exercise and heart rate at two minutes post-exercise, as measured by a height-adjusted step test and stratified into two groups: low and high fitness, using a median split. Ambulatory blood pressure (ABP) was monitored for a 24-hr period on a school day using the Spacelabs ambulatory monitor (Model 90207). Blood pressure and heart rate were recorded at 30 minute intervals throughout the day of recording and at 60 minute intervals during sleep. ^ RESULTS: No significant associations were found between weight status and mean 24-hr systolic blood pressure (SBP) or mean arterial pressure (MAP). A significant and inverse association between weight status and mean 24-hr diastolic blood pressure (DBP) was revealed. Cardiorespiratory fitness was significantly and inversely associated with mean 24-hr ABP. High fitness adolescents had significantly lower mean 24-hr SPB, DBP, and MAP measurements than low fitness adolescents. Compared to low fitness adolescents, high fitness adolescents had 1.90 mmHg, 1.16 mmHg, and 1.68 mmHg lower mean 24-hr SBP, DBP, and MAP, respectively. Additionally, high fitness appeared to afford protection from higher mean 24-hr SBP and MAP, irrespective of weight status. Among normal weight adolescents, low fitness resulted in higher mean 24-hr SBP and MAP, compared to their fit counterparts. Among adolescents categorized as high fitness, increasing weight status did not appear to result in higher mean 24-hr SBP or MAP. Cardiorespiratory fitness, rather than weight status, appeared to be a more dominant predictor of mean 24-hr SBP and MAP. ^ CONCLUSIONS: To our knowledge, this research is the first study to investigate the independent and combined contributions of cardiorespiratory fitness and weight status on 24-hr ABP, all objectively measured. The results of this study may potentially guide and inform future research. It appears that early cardiovascular disease (CVD) prevention should focus on improving cardiorespiratory fitness levels among all adolescents, particularly those adolescents least fit, regardless of their weight status, while obesity prevention efforts continue.^

Bicarbonato ingestion has no ergogenic effect on consecutive all out sprint test in MWX elite cyclists

The aim of the present study was to examine the effect of sodium bicarbonate ingestion on consecutive "all out" sprint tests, analyzing the acid-base status and its influence on performance and perceived effort. Ten elite bicycle motocross (BMX) riders (20.7 ± 1.4 years, training experience 8-12 years) participated in this study which consisted of two trials. Each trial consisted of three consecutive Wingate tests (WTs) separated by 15 min recovery. Ninety minutes prior to exercise subjects ingested either NaHCO(3) (-) (0.3 g kg(-1) body weight) or placebo. Blood samples were collected for the assessment of blood acid-base status: bicarbonate concentration ([HCO(3) (-)]), pH, base excess (BE) and blood lactate concentration ([La(-)]). Performance variables of peak power (PP), mean power (MP), time to peak power and fatigue index were calculated for each sprint. Significant differences (p < 0.05) were observed in acid-base variables [pH before WT1: 7.47 ± 0.05 vs. 7.41 ± 0.03; [HCO(3) (-)] before WT1: 29.08 ± 2.27 vs. 22.85 ± 0.24 mmol L(-1) (bicarbonate vs. placebo conditions, respectively)], but there were not significant differences in performance variables between trials [PP WT1: 1,610 ± 373 vs. 1,599 ± 370 W; PP WT2: 1,548 ± 460 vs. 1,570 ± 428 W; PP WT3: 1,463 ± 361 vs. 1,519 ± 364 W. MP WT1: 809 ± 113 vs. 812 ± 108 W; MP WT2: 799 ± 135 vs. 799 ± 124 W; MP WT3: 762 ± 165 vs. 782 ± 118 W (bicarbonate vs. placebo conditions, respectively)]. Rating of perceived effort (RPE) was not influenced nor ratings of perceived readiness. Sodium bicarbonate ingestion modified significantly the blood acid-base balance, although the induced alkalosis did not improve the Wingate test performance, RPE and perceived readiness across three consecutive WTs in elite BMX cyclists.

Crowdsourcing malaria parasite quantification: an online game for analyzing images of infected thick blood smears

Background: There are 600,000 new malaria cases daily worldwide. The gold standard for estimating the parasite burden and the corresponding severity of the disease consists in manually counting the number of parasites in blood smears through a microscope, a process that can take more than 20 minutes of an expert microscopist’s time. Objective: This research tests the feasibility of a crowdsourced approach to malaria image analysis. In particular, we investigated whether anonymous volunteers with no prior experience would be able to count malaria parasites in digitized images of thick blood smears by playing a Web-based game. Methods: The experimental system consisted of a Web-based game where online volunteers were tasked with detecting parasites in digitized blood sample images coupled with a decision algorithm that combined the analyses from several players to produce an improved collective detection outcome. Data were collected through the MalariaSpot website. Random images of thick blood films containing Plasmodium falciparum at medium to low parasitemias, acquired by conventional optical microscopy, were presented to players. In the game, players had to find and tag as many parasites as possible in 1 minute. In the event that players found all the parasites present in the image, they were presented with a new image. In order to combine the choices of different players into a single crowd decision, we implemented an image processing pipeline and a quorum algorithm that judged a parasite tagged when a group of players agreed on its position. Results: Over 1 month, anonymous players from 95 countries played more than 12,000 games and generated a database of more than 270,000 clicks on the test images. Results revealed that combining 22 games from nonexpert players achieved a parasite counting accuracy higher than 99%. This performance could be obtained also by combining 13 games from players trained for 1 minute. Exhaustive computations measured the parasite counting accuracy for all players as a function of the number of games considered and the experience of the players. In addition, we propose a mathematical equation that accurately models the collective parasite counting performance. Conclusions: This research validates the online gaming approach for crowdsourced counting of malaria parasites in images of thick blood films. The findings support the conclusion that nonexperts are able to rapidly learn how to identify the typical features of malaria parasites in digitized thick blood samples and that combining the analyses of several users provides similar parasite counting accuracy rates as those of expert microscopists. This experiment illustrates the potential of the crowdsourced gaming approach for performing routine malaria parasite quantification, and more generally for solving biomedical image analysis problems, with future potential for telediagnosis related to global health challenges.

A common mutation in the methylenetetrahydrofolate reductase gene is associated with an accumulation of formylated tetrahydrofolates in red blood cells

A common mutation (C677T) in the gene encoding for methylenetetrahydrofolate reductase (MTHFR) (5-methyltetrahydrofolate:(acceptor) oxidoreductase, EC 1.7.99.5), a key regulatory enzyme in one-carbon metabolism, results in a thermolabile variant of the MTHFR enzyme with reduced activity in vitro. In the present study we used a chromatographic method for folate analysis to test the hypothesis that this mutation would be associated with altered distribution of red blood cell (RBC) folates. An alteration was found as manifested by the presence of formylated tetrahydrofolate polyglutamates in addition to methylated derivatives in the RBCs from homozygous mutant individuals. 5-Methyltetrahydrofolate polyglutamates were the only folate form found in RBCs from individuals with the wild-type genotype. Existence of formylated folates in RBCs only from individuals with the thermolabile MTHFR is consistent with the hypothesis that there is in vivo impairment in the activity of the thermolabile variant of MTHFR and that this impairment results in an altered distribution of RBC folates.

The effect of 4,4′-diisothiocyanato-stilbene-2,2′-disulfonate on CO2 permeability of the red blood cell membrane

It has long been assumed that the red cell membrane is highly permeable to gases because the molecules of gases are small, uncharged, and soluble in lipids, such as those of a bilayer. The disappearance of 12C18O16O from a red cell suspension as the 18O exchanges between labeled CO2 + HCO3− and unlabeled HOH provides a measure of the carbonic anhydrase (CA) activity (acceleration, or A) inside the cell and of the membrane self-exchange permeability to HCO3− (Pm,HCO−3). To test this technique, we added sufficient 4,4′-diisothiocyanato-stilbene-2,2′-disulfonate (DIDS) to inhibit all the HCO3−/Cl− transport protein (Band III or capnophorin) in a red cell suspension. We found that DIDS reduced Pm,HCO−3 as expected, but also appeared to reduce intracellular A, although separate experiments showed it has no effect on CA activity in homogenous solution. A decrease in Pm,CO2 would explain this finding. With a more advanced computational model, which solves for CA activity and membrane permeabilities to both CO2 and HCO3−, we found that DIDS inhibited both Pm,HCO−3 and Pm,CO2, whereas intracellular CA activity remained unchanged. The mechanism by which DIDS reduces CO2 permeability may not be through an action on the lipid bilayer itself, but rather on a membrane transport protein, implying that this is a normal route for at least part of red cell CO2 exchange.

High-sensitivity array analysis of gene expression for the early detection of disseminated breast tumor cells in peripheral blood

Early detection is an effective means of reducing cancer mortality. Here, we describe a highly sensitive high-throughput screen that can identify panels of markers for the early detection of solid tumor cells disseminated in peripheral blood. The method is a two-step combination of differential display and high-sensitivity cDNA arrays. In a primary screen, differential display identified 170 candidate marker genes differentially expressed between breast tumor cells and normal breast epithelial cells. In a secondary screen, high-sensitivity arrays assessed expression levels of these genes in 48 blood samples, 22 from healthy volunteers and 26 from breast cancer patients. Cluster analysis identified a group of 12 genes that were elevated in the blood of cancer patients. Permutation analysis of individual genes defined five core genes (P ≤ 0.05, permax test). As a group, the 12 genes generally distinguished accurately between healthy volunteers and patients with breast cancer. Mean expression levels of the 12 genes were elevated in 77% (10 of 13) untreated invasive cancer patients, whereas cluster analysis correctly classified volunteers and patients (P = 0.0022, Fisher's exact test). Quantitative real-time PCR confirmed array results and indicated that the sensitivity of the assay (1:2 × 108 transcripts) was sufficient to detect disseminated solid tumor cells in blood. Expression-based blood assays developed with the screening approach described here have the potential to detect and classify solid tumor cells originating from virtually any primary site in the body.

Tissue factor promotes melanoma metastasis by a pathway independent of blood coagulation.

Several studies have established a link between blood coagulation and cancer, and more specifically between tissue factor (TF), a transmembrane protein involved in initiating blood coagulation, and tumor metastasis. In the study reported here, a murine model of human melanoma metastasis was used for two experiments. (i) The first experiment was designed to test the effect of varying the level of TF expression in human melanoma cells on their metastatic potential. Two matched sets of cloned human melanoma lines, one expressing a high level and the other a low level of the normal human TF molecule, were generated by retroviral-mediated transfections of a nonmetastatic parental line. The metastatic potential of the two sets of transfected lines was compared by injecting the tumor cells into the tail vein of severe combined immunodeficiency (SCID) mice and later examining the lungs and other tissues for tumor development. Metastatic tumors were detected in 86% of the mice injected with the high-TF lines and in 5% of the mice injected with the low-TF lines, indicating that a high TF level promotes metastasis of human melanoma in the SCID mouse model. This TF effect on metastasis occurs with i.v.-injected melanoma cells but does not occur with primary tumors formed from s.c.-injected melanoma cells, suggesting that TF acts at a late stage of metastasis, after tumor cells have escaped from the primary site and entered the blood. (ii) The second experiment was designed to analyze the mechanism by which TF promotes melanoma metastasis. The procedure involved testing the effect on metastasis of mutations in either the extracellular or cytoplasmic domains of the transmembrane TF molecule. The extracellular mutations introduced two amino acid substitutions that inhibited initiation by TF of the blood-coagulation cascade; the cytoplasmic mutation deleted most of the cytoplasmic domain without impairing the coagulation function of TF. Several human melanoma lines expressing high levels of either of the two mutant TF molecules were generated by retroviral-mediated transfection of the corresponding TF cDNA into the nonmetastatic parental melanoma line, and the metastatic potential of each transfected line was tested in the SCID mouse model. Metastases occurred in most mice injected with the melanoma lines expressing the extracellular TF mutant but were not detected in most mice injected with the melanoma lines expressing the cytoplasmic TF mutant. Results with the extracellular TF mutant indicate that the metastatic effect of TF in the SCID mouse model does not involve products of the coagulation cascade. Results with the cytoplasmic TF mutant indicate that the cytoplasmic domain of TF is important for the metastatic effect, suggesting that the TF could transduce a melanoma cell signal that promotes metastasis.

Cortisol fecal em ovinos: curva de excreção e estabilidade

O presente estudo foi dividido em dois experimentos, tendo como objetivo determinar a curva de excreção do cortisol fecal e sua estabilidade nas fezes perante exposição à diferentes períodos de tempo e temperatura entre as colheitas e análises, correlacionando os níveis de cortisol fecal com o pico de cortisol sanguíneo. No experimento 1, seis fêmeas mestiças (Dorper x Santa Inês) tiveram suas fezes totais colhidas durante 24 horas após a aplicação do hormônio adrenocorticotrófico (ACTH), além de colheitas de sangue realizadas antes da aplicação do ACTH, 60, 120 e 300 minutos depois; durante as quais foram atribuídos escores de reatividade para cada animal. Logo após as análises foi iniciado o experimento 2, no qual 9 cordeiros mestiços (Dorper x Santa Inês) foram submetidos a uma situação de estresse térmico durante os horários das 11 às 15 horas da tarde, tendo suas fezes colhidas às 23 horas do mesmo dia. Após a colheita, as fezes foram agrupadas e homogeneizadas em três grupos distintos, de onde retiraram-se alíquotas referentes aos tratamentos propostos: três temperaturas (15°, 25° e 35°) e quatro tempos (1, 3, 6 e 12 horas). Os dados da curva de excreção foram analisados por ANOVA, bem como pela correlação entre os valores de cortisol sanguíneo, fecal e reatividade. Para análise da estabilidade foi utilizada ANOVA multifatorial com dois fatores (temperatura e intervalo de tempo). Para avaliação das variáveis comportamentais foi realizada a transformação de escala dos dados para \"arco-seno raiz de porcentagem\", procedendo-se à análise de variância. O modelo estatístico contemplou os efeitos de dia (1, 2 e 3) com análise individual por animal. Os parâmetros de cortisol sanguíneo, frequência respiratória e temperatura retal foram analisados pelo teste t e correlação de Pearson. Todas as comparações de médias foram realizadas por teste F e teste t (PDIFF). A reatividade durante a colheita não exerceu efeito significativo sobre os valores de cortisol sanguíneo, os quais demonstraram médias maiores 60 minutos após a aplicação do ACTH e, após 300 minutos as ovelhas apresentaram níveis de cortisol considerados normais para ovinos sem estresse. Por outro lado, o pico de cortisol nas fezes foi verificado aproximadamente 10 a 12 horas após o pico de cortisol no sangue, não sendo verificadas diminuições significativas nas concentrações que indicassem o retorno aos níveis basais durante o período de 24 horas (P>0,05). Não foram observadas diferenças significativas entre os tempos e temperaturas aos quais as amostras de fezes foram submetidas (P>0,05), verificando-se uma tendência a manutenção da concentração do cortisol fecal em ovinos durante o período de 12 horas após a colheita.

Analytical study of the legal and operational aspects of the Minnesota law entitled "Chemical test for intoxication" - M.S.A. Sec. 169.123. Final report.

National Highway Traffic Safety Administration, Washington, D.C.

Report on a national study of Preliminary Breath Test (PBT) and Illegal Per Se laws; effectiveness of PBT and IPS laws. Final report.

National Highway Traffic Safety Administration, Washington, D.C.

1976 selective enforcement analysis: including a report on the effect of portable breath test devices on detection/apprehension of drinking drivers - Tampa ASAP analytic study no. 3. Final report.

National Highway Traffic Safety Administration, Office of Driver and Pedestrian Programs, Washington, D.C.

A study of self test devices. Technical report.

National Highway Traffic Safety Administration, Washington, D.C.

Interpreting your test results.

"P.O. #X41943"--Colophon.