Blood sampling

Background. During haemodialysis (HD), polymorphonuclear cells (PMNs) and platelets are activated and release various granule products, including myeloperoxidase (MPO) and platelet factor 4 (PF4). MPO triggers the generation of reactive oxygen species, leading to irreversible protein, carbohydrate and lipid modification. PF4 probably also contributes to oxidative stress. As previously shown, HD-induced PMN degranulation is almost completely abolished during citrate anticoagulation, most probably due to its calcium chelation ability. Methods. In the present study, apart from HDinduced PMN and platelet degranulation, oxidative stress was analysed during three modes of anticoagulation. Heparin, dalteparin and citrate (HD hep , HD dal and HD cit ) were compared in a randomized, crossover fashion in eight chronic HD patients. Multiple blood samples were taken during the third HD session of each modality, from both the afferent and efferent line. Besides the degranulation markers MPO and PF4, various markers of oxidative stress were measured, including oxidized low-density lipoprotein (ox-LDL), malondialdehyde (MDA) and carboxymethyllysine (CML). Results. During HD hep and HD dal , marked degranulation was observed shortly after the start of HD. In contrast, during HD cit , PF4 and MPO levels remained unaltered, suggesting no release at all. After 1 week of HD cit , ox-LDL levels were markedly reduced [median 26% (3-65%), P ¼ 0.01], if compared with HD hep and HD dal . As regards MDA and CML, no differences were found.

Objective: To measure the frequency of depression and its risk factors in patients under going hemodialysis. Methodology: It is a cross-sectional prospective study conducted at Hemodialysis unit of Shalamar Hospital and Shaikh Zayed Hospital, Lahore from 1 st January 2006 to 30 th April 2006. All patients getting regular hemodialysis for more than three months were included. Beck’s Depression Inventory- II (BDI-II; adapted in Urdu) was administered on all the patients who were able to read or understand it. Blood sample were drawn at the same time for routine hematological, biochemical parameters and viral markers (Anti HCV and HbsAg). Diagnosis was made as per Diagnostic and Statistical Manual of Mental Disorders, fourth edition (DSM IV) for correlation of psychological variables with clinical, hematological and biochemical parameters. Results: Eighty nine patients were enrolled which included fifty two (58.4%) were male and seventy seven (86.5%) were married. Major causes of renal...

Trypanosome infections in their natural hosts are frequently di.cult to detect by microscopy\ and culture methods are unreliable and not suitable for all species of Trypanosoma[ A nested PCR strategy for detecting and identifying Trypanosoma species\ suitable for detecting both known and unknown trypanosomes\ is presented[ Thirty!two blood samples from 12 species of Australian birds and mammals were screened by a nested PCR for the presence of Trypanosoma sp[ ssrRNA[ Three infections were detected\ one in an eastern grey kangaroo "Macropus giganteus#\ one in a common wombat "Vombatus ursinus# and one in a platypus "Ornithorhynchus anatinus#[ The kangaroo and wombat are new host records for Trypanosoma sp[^the platypus parasite was Trypanosoma binneyi[ The three parasites could be distinguished by restriction fragment length polymorphisms of the ampli_ed fragment of the ssrRNA gene[ The kangaroo and wombat parasites were also isolated in a semi!solid blood agar medium[ The culture forms of the kangaroo trypanosome had an expanded~agellar sheath in which structures similar to hemidesmosomes were detected by EM[ The nested PCR was at least as sensitive as culture\ and analysis of the PCR products gave parasite!speci_c _ngerprints[ Therefore this method could be suitable for rapidly screening host animals for the presence of trypanosomes and identifying the infecting strain[ Þ 0888 Australian Society for Parasitology[ Published by Elsevier Science Ltd[ All rights reserved[ Keywords] Macropus giganteus^Ornithorhynchus anatinus^Ortholfersia macleayi^Trypanosoma binneyi^Trypanosoma cruzi^Vombatus ursinus^Guanidine thiocyanate

O objetivo do estudo foi examinar a influência de dois diferentes protocolos de treinamento de força sobre parâmetros antropométricos (peso, IMC, massa gorda), funcionais (teste de 1-RM e teste de repetições máximas) e relacionados ao sistema endócrino (concentração de testosterona e de cortisol) e ao sistema imunológico (concentração de glutamina e de IgG). Participaram do estudo 12 homens treinados (27,4 ± 4,8 anos). Esses indivíduos foram aleatoriamente divididos em dois grupos, que posteriormente foram submetidos a dois protocolos de treinamento distintos: Múltiplas séries (MS) e Tri-set (TS). Amostras de sangue foram coletadas antes e depois de uma sessão de exercício de força, no início e no final do período de oito semanas de treinamento. Não foram observadas alterações nos parâmetros morfo-funcionais (com exceção do teste de repetições máximas para o agachamento). Com relação aos parâmetros endócrinos, foi observado que o TS provocou aumento significativo do cortisol, imedia...

Purpose: To date, the exact cause of abnormal exercise response in chronic fatigue syndrome (CFS) remains to be revealed, but evidence addressing intracellular immune deregulation in CFS is growing. Therefore, the aim of this cross-sectional study was to examine the interactions between several intracellular immune variables and exercise performance in CFS patients. Methods: After venous blood sampling, subjects (16 CFS patients) performed a maximal exercise stress test on a bicycle ergometer with continuous monitoring of cardiorespiratory variables. The following immune variables were assessed: the ratio of 37 kDa Ribonuclease (RNase) L to the 83 kDa native RNase L (using a radiolabeled ligand/receptor assay), RNase L enzymatic activity (enzymatic assay), protein kinase R activity assay (comparison Western blot), elastase activity (enzymatic-colorimetric assay), the percent of monocytes, and nitric oxide determination (for monocytes and lymphocytes; flow cytometry, live cell assay). Results: Forward stepwise multiple regression analysis revealed 1) that elastase activity was the only factor related to the reduction in oxygen uptake at a respiratory exchange ratio (RER) of 1.0 (regression model: R 2 ϭ 0.53, F (1,14) ϭ 15.5, P Ͻ 0.002; elastase activity P Ͻ 0.002); 2) that the protein kinase R activity was the principle factor related to the reduction in workload at RER ϭ 1.0; and 3) that elastase activity was the principle factor related to the reduction in percent of target heart rate achieved. Conclusion: These data provide evidence for an association between intracellular immune deregulation and exercise performance in patients with CFS. To establish a causal relationship, further study of these interactions using a prospective longitudinal design is required.

Aim. To verify the rationale of a pelvic stop-flow technique for the perfusion of high-doses of mitomycin C and anthacyclines in patients with inoperable, recurrent pelvic cancer.Methods. The stop-flow technique was realized by using percutaneous double-balloon arterial–venous catheters that selectively isolate the pelvic vascular section and a perfusion provided by an extracorporeal pump for 20 min. Ten patients (pts) with unresectable pelvic recurrence from colon-rectal cancer were treated with a combination of Mitomycin C (MMC, 20 mg/sqm) plus doxorubicin (DOXO, 75 mg/sqm; 8 pts) or epirubicin (EPI, 75 mg/sqm; 2 pts) infused into the isolated pelvic compartment. Blood samples were collected from the extracorporeal vascular flow and from peripheral plasma, and analysed for drug quantitation.Results. During the procedure, there were no technical or hemodynamic complications, and no deaths occurred during surgery or in the postoperative period. MMC and DOXO peak levels measured in the extracorporeal system which irrotates the tumor area, were on average 21.6 (range: 4.3–44.3, MMC) and 17.2 (range: 1.8–48.4, DOXO) times higher than those observed in the peripheral blood. Similarly; the area under concentration (AUC) versus time curves measured in the pelvic compartment during stop-flow perfusion were 19.9 (range: 3.8–45.0, MMC) and 13.4 (range: 1.2–26.6, DOXO) times higher than the corresponding value in peripheral circulation. The drug percentage eliminated in the ultra filtrate was only 7.7% (MMC) and 0.9% (DOXO), and the plasmatic AUC0–24 were similar to those observed with iv bolus of equivalent drug doses. Minimal systemic and local toxicities were observed. One complete pathological and 2 partial responses were observed; pain remission in 8/10 patients. Median survival was 12 months (8–31).Conclusion. The endo-arterial administration into the local vasculature produces high pelvic-systemic concentration gradients during the stop-flow perfusion with limited local and systemic toxicity. The encouraging clinical results suggest further evaluation.

The mutual influences of wheat selenium (Se) and n-3 polyunsaturated fatty acids (n-3 PUFA) on plasma Se and indicators of increased oxidative stress were investigated in a randomized, double-blind study with 31 women (23.5 +/- 3.4 yr). Groups 1 and 2 ingested 5.4 g n-3 PUFA daily (as ethyl esters), whereas groups 3 and 4 received placebo capsules. Groups 2 and 3 received 3 slices of high-Se bread daily, providing 115 micrograms Se, in addition to the 77 +/- 26 micrograms Se in the diet. Groups 1 and 4 received placebo slices. Blood samples were drawn at baseline and at 3 and 6 wk. Serum Se concentrations increased in both groups given Se-enriched bread, but significantly less in subjects given n-3 PUFA (group 2). There were no changes in the plasma ratio alpha-tocopherol:mg cholesterol or plasma ascorbic acid levels. In group 1, plasma-conjugated dienes and thiobarbituric acid-reactive substances (TBARS) rose by 130% (p < 0.005) and 126% (p < 0.005), respectively. Two-way ANO...

Objective. To investigate whether formocresol, in Buckley's original formulation, is mutagenic in vivo to lymphocyte cultures obtained from the peripheral blood of children aged from 5 to 10 years old. These children were recruited from those attending the dental clinics of Recife City Council and the University of Pernambuco School of Dentistry, Brazil.

There is accumulating evidence that the metabolism of several trace elements is altered in diabetes mellitus and that these nutrients might have specific roles in the pathogenesis and progress of this disease. The aim of present study was to compare the level of essential ...

The aim of this study was to evaluate the negative effects of the Pringle maneuver on pancreatic tissue with respect to the time of performing the maneuver. Moreover, the efficacy of octreotide therapy on pancreatic changes at the time of the Pringle maneuver was assessed. Fifty male Wistar Albino rats were randomized into 5 groups. The groups were formed as follows: Group A: sham operation, Group B: Pringle maneuver for 30 min plus octreotide (PM30-OCT), Group C: Pringle maneuver for 60 min plus octreotide (PM60-OCT) and Group D: Pringle maneuver for 30 min plus 0.9 % saline solution (PM30-SS), Pringle maneuver for 60 min plus 0.9 % saline solution (PM60-SS, Group E). Blood samples for the evaluation of both amylase and lipase levels were taken via the portal vein. Levels of glutathione, glutathione reductase, catalase, myeloperoxidase, nitric oxide, xanthine oxidase, malondialdehyde, tumour necrosis factor alpha (TNF-alpha), and interleukin-1 beta (IL-1 beta) were assessed in the ...

The stability of interleukin 6 (IL-6), its soluble receptor (sIL-6R), IL-10 and CC16 or uteroglobin (an endogenous cytokine inhibitor) in human serum was examined using an accelerated stability testing protocol according to the Arrhenius equation. Further, the effect of time delay between blood sampling and sample processing, clotting temperature and repeated freeze-thaw cycles on serum levels of these proteins were determined. Paired serum samples were stored at 4 C, 20 C, 30 C and 40 C for 1 to 21 days. We found that IL-6 and CC16 concentrations did not change at 4 C, 20 C and 30 C. Interleukin-6 concentrations significantly declined after 11 days at 40 C. The concentrations of sIL-6R and IL-10 did not change at 4 C but significantly decreased at 20 C (after 21 and 14 days respectively), 30 C and 40 C (after 1 day at both temperatures for both cytokines). Arrhenius-plots indicated that sIL-6R and IL-10 are stable for at least several years at 20 C and 70 C, respectively. Since their relative stability, no Arrhenius-plot could be calculated for IL-6 and CC16. The concentrations of the proteins examined were not significantly altered by repeated freeze-thaw cycles, nor by extended clotting times at 4 C or 20 C. We conclude that serum samples for the determination of IL-6, sIL-6R and CC16 can be stored at 20 C for several years, but for IL-10 determinations, storage at 70 C is recommended.

There are several convincing reports showing relationships between the area under the curve of ultra®lterable concentration versus time (AUC) and pharmacodynamics of carboplatin. It is advisable, in treated patients, to check the AUC that is eectively delivered as compared with the prescribed AUC. To this end, limited sampling strategy seems to be an adequate approach since it limits the constraints of repeated blood sampling for both patients and nursing sta. A¯exible limited sampling method for assessing ultra®lterable carboplatin AUC was developed. This method was based on a Bayesian estimation of carboplatin clearance using the NON linear Mixed Eect Model (NONMEN) program and a large pharmacokinetic and covariates database (103 patients). The optimal sampling design was a two-sample schedule (1 and 4 h after the end of infusion). During a prospective evaluation, it allowed an adequate estimation of carboplatin clearance with a non-signi®cant bias (À4.5%) and a good precision (9%). In a second stage, this method was clinically applied to monitor carboplatin AUC in a group of 5 patients with metastatic germ cell tumours treated with intensi®ed high dose carboplatin-based chemotherapy for 4 days. Dosage adjustments were performed according to daily controls of their AUC in order to obtain a total AUC of 20 mg/mlÂmin. By using this strategy all patients eectively received a total AUC very close to this targeted AUC, thus proving the clinical usefulness of this limited sampling method. #

Canine monocytic ehrlichiosis caused by Ehrlichia canis is of veterinary importance worldwide. In Thailand, there has been little information available on E. canis and its phylogeny. The objective of this study was to characterize and establish molecular structure and phylogeny of Thai Ehrlichia and Anaplasma strains. Genus-specific primers for Ehrlichia and Anaplasma were used to amplify the 16S rRNA gene from naturally infected canine blood samples, and these amplicon sequences were compared with other sequences from GenBank. Both homology and secondary structure analysis of 16S rRNA sequences indicated that they were novel E. canis and A. platys strains. Phylogenetic analysis revealed that the Thai E. canis strain was closely related and formed a single cluster with E. canis from different countries. A. platys found in this study showed close relationship with earlier report of A. platys from Thailand. To our knowledge this report represents the first molecular characterization of the nearly complete 16S rRNA gene from E. canis in dogs from Thailand.

Celastrol, a natural substance isolated from plant extracts used in traditional Chinese medicine, has been extensively investigated as a possible drug for treatment of cancer, autoimmune diseases, and protein misfolding disorders. Although studies focusing on celastrol's effects in specific cellular pathways have revealed a considerable number of targets in a diverse array of in vitro models there is an essential need for investigations that can provide a global view of its effects. To assess cellular effects of celastrol and to identify target proteins as biomarkers for monitoring treatment regimes, we performed large-scale quantitative proteomics in cultured human lymphoblastoid cells, a cell type that can be readily prepared from human blood samples. Celastrol substantially modified the proteome composition and 158 of the close to 1800 proteins with robust quantitation showed at least a 1.5 fold change in protein levels. Up-regulated proteins play key roles in cytoprotection with a prominent group involved in quality control and processing of proteins traversing the endoplasmic reticulum. Increased levels of proteins essential for the cellular protection against oxidative stress including heme oxygenase 1, several peroxiredoxins and thioredoxins as well as proteins involved in the control of iron homeostasis were also observed. Specific analysis of the mitochondrial proteome strongly indicated that the mitochondrial association of certain antioxidant defense and apoptosis-regulating proteins increased in cells exposed to celastrol. Analysis of selected mRNA transcripts showed that celastrol activated several different stress response pathways and dose response studies furthermore showed that continuous exposure to sub-micromolar concentrations of celastrol is associated with reduced cellular viability and proliferation. The extensive catalog of regulated proteins presented here identifies numerous cellular effects of celastrol and constitutes a valuable biomarker tool for the development and monitoration of disease treatment strategies.

This pilot study of ethyl-eicosapentaenoate (ethyl-EPA) in the treatment of Alzheimer's disease aimed to estimate the magnitude of any change in measures of cognition during a 12-week treatment period. A simple linear design was used in which each patient had a baseline period of 12 weeks without treatment, followed by 12 weeks' treatment with ethyl-EPA. Blood samples were taken both before and after the treatment period to measure erythrocyte membrane fatty acids. Assessments comprised cognitive measures and visual analogue ratings of overall assessment of functioning.

BACKGROUND: The aim of this study was to provide a model-based analysis of the pharmacokinetics of remifentanil in infants and children undergoing cardiac surgery with cardiopulmonary bypass (CPB). METHODS: We studied nine patients aged 0.5 to 4 years who received a continuous remifentanil infusion via a computer-controlled infusion pump during cardiac surgery with mildly hypothermic CPB were studied. Arterial blood

OBJECTIVE The role of leptin in the acute stress response to extracorporeal circulation has been well documented, however, the relationship between leptin and zinc has not been investigated previously. We aimed to research the circulating leptin, zinc, and copper levels before, during, and after the extracorporeal circulation, and effect of preoperative zinc administration to these. METHODS Twenty patients who were taken to elective coronary artery bypass grafting operations using extracorporeal circulation were taken to this research and divided into two equal groups (n1, n2). In both groups blood samples were taken just before the operation (T0), at the end of operation (T1), and at the first postoperative day (T2). In the second group (n2) oral zinc (50 mg, once a day) was administered to patients for 5 days, preoperatively. The serum leptin, zinc, and copper levels were studied. RESULTS In group n1 circulating leptin levels were significantly increased at T2 when compared to T0 ...

A double-center, open-label, two-way crossover study was conducted in 24 healthy volunteers to assess the bioequivalence of a combined lamivudine/zidovudine tablet related to a reference and test drug products. The volunteers were randomly assigned to receive one lamivudine/zidovudine combination tablet of reference or test product with 7-days washout period between. Blood samples were collected up to 36 h post dose. Pharmacokinetic parameters were estimated. Drug products were bioequivalent if 90% confidence intervals for the ratio of least squares (CI 90%) means are under plasma concentration-time curve (AUC 0-τ) and absorption rate (C max) fell within 80 to 125% for log-transformed parameters. Test and reference products present data of AUC 0-τ , C max referents to lamivudine and data of ASC ∞ referents to zidovudine, in agreement of these limits. The result of C max (CI 90%) to zidovudine was: 116% (90-141%), it has confirm that the zidovudine has high individual variability of absorption.

The main objective of the present study was to examine the dietary intake and physiological status of members of the US National Figure Skating Dance Team. Study participants included seven female and seven male figure skating dance pairs. Three-day food records were collected from each dancer and analyzed using a computerized nutrient database. Height, body weight and skinfold measurements were completed on each dancer. Body fat was estimated using the Jackson et al prediction equations. Fasting blood samples were collected for biochemical analysis. Total energy intake of female dancers (1416 kcal/d, 339 kjoules/day) was considerably lower than the dietary recommendations, while the male dancers appeared to have adequate energy intake (2837 kcal/d, 679 kjoules/day). Carbohydrate intake of both male and female dancers was below the recommendations, 44% and 51%, respectively, whereas total fat and protein intake of both male and female dancers were higher than the dietary recommendations. All dancers met at least 2/3 rd of the dietary recommendations for all micronutrients, except folate and zinc intake of the female dancers, which were 68% and 62% of the recommended values. Although, blood lipid concentrations and other physiological parameters of the overall group were within normal ranges, two of the dancers (14% of the dancers) had elevated blood cholesterol concentrations and low iron status. The results of the present study suggest that these elite figure skating dancers, especially females, may not be consuming adequate amounts of energyyielding nutrients and those that influence bone health. Future studies are needed to examine the effects of nutritional status on performance of these figure skating dancers and to develop strategies to improve the nutritional status of these elite dancers.

Whole body hyperthermia (WBH) has been used as an adjunct to radio-/chemotherapy in patients with various malignant diseases. Although clear evidence is still missing, it has been hypothesized that an activation of the immune system might contribute to the therapeutic effect of WBH. To examine whether a treatment with 60-minute 41.8°C WBH as an adjunct to chemotherapy (WBH-CT) induces an activation of T cells, blood samples were collected at numerous time points before and up to 48 h post-treatment. The aim of this study was to examine the effect of WBH-CT on the expression of a broad range of activation markers on peripheral blood lymphocytes (PBL), on serum cytokines and intracellular cytokine levels in T cells, and the capacity of these cells to proliferate. Immediately after 41.8°C WBH-CT treatment, a drastic increase in peripheral natural killer (NK) cells (P<0.05) and CD56 + cytotoxic T lymphocytes (CTL; P<0.01) in the patients' peripheral blood was observed. At 5 h post-treatment, the percentages of both effector cell types had returned to baseline levels. This transient phenomenon was accompanied by a short period of reduced T cell activity, indicated by diminished serum levels of soluble interleukin-2 receptors (sIL-2R) at 3 h post-WBH-CT (P<0.05) and decreased lymphocytic proliferation at the same point in time. This first phase was followed by a marked but short-lived increase in the patients' serum levels of interleukin-6 (IL-6; P<0.01) during the first 5 h following treatment, with a subsequent decrease to baseline levels at 24 h and significantly increased serum levels of tumor necrosis factor-a (TNF-a) at 0 h (P<0.01), 3 h (P<0.05), 5 h (P<0.05) and 24 h (P<0.01) post-WBH-CT. The third phase of the immunological consequences of WBH-CT consisted of an increase in the percentage of peripheral cytotoxic T lymphocytes (CTL) expressing CD56, reaching a maximum at 48 h post-WBH (P<0.01). Furthermore, the percentage of CD4 + T cells expressing the T cell activation marker CD69 increased nearly twofold over time, reaching its maximum at 48 h (P<0.05). As an additional marker for T cell activation, serum levels of sIL-2R increased markedly (P<0.01), reaching maximum levels at the same point in time. Elevated intracellular concentrations of interferon-gamma (IFN-c) and/or TNF-a in CD8 + T cells were found in 4 out of 5 patients at 24 h post-WBH-CT. Since similar changes were not observed in patients receiving chemotherapy alone, this is the first study to provide evidence for prolonged WBH-CT-induced activation of human T cells.

The study investigated the influence of prolonged physical stress during survival training with food and fluid deprivation on the serum concentrations of erythropoietin (EPO). A group of 29 male subjects [mean age 22.2 (SD 2.8) years, height 1.78 (SD 0.06) m, and body mass (rob) 73.5 (SD 8.6) kgl were studied for 5 days of multifactorial stress including restricted water intake (11 H20"day-1) and food intake (628 kJ' day-1) combined with physical exercise (estimated energy expenditure approximately 24 000 kJ" day-l) and sleep deprivation (20 h within 5 days). Blood samples were taken before (T1), after 72 h (T2) and 120h (T3) of physical stress, and after 48 h, (T4) and 72 h (T5) of recovery. The samples were analysed for EPO, and concentrations of serum iron (Fe), haptoglobin (Hapto), transferrin (Trans), ferritin (Fer), haemoglobin (Hb) and packed cell volume (PCV). The mb had decreased by 6.77kg at T3 (P<0.01) and 0.68 kg at T5. The EPO and Hapto decreased during the survival training (P <0.01) and increased during the recovery period (P < 0.01). The Fe increased during the survival training (P <0.01) and remained above the control concentrations during recovery (P <0.01). The Hapto decreased during the survival training (P < 0.01) and remained below control concentration at T4 and T5 (P <0.01). The Trans decreased continuously over the week (P <0.01). The Fer increased during the survival training (P <0.01) and

The correct tidal volume during cardiopulmonary resuscitation (CPR) is presently debated. While the European Resuscitation Council (ERC) and American Heart Association (AHA) previously recommended a tidal volume of 800 -1200 ml, the ERC has recently reduced this to 400-600 ml. In a prospective, randomised study of 17 non-traumatic out-of-hospital cardiac arrest patients intubated and mechanically ventilated 12 min − 1 with 100% oxygen, we have therefore compared arterial blood gases generated with tidal volumes of 500 and 1000 ml. Mean time from cardiac arrest to arrival of the ambulance was 13 9 8 and 14 98 min in the two groups, respectively. Arterial blood samples were taken percutaneously 5 and 10 -15 min after onset of the mechanical ventilation and analysed instantly. Pa CO 2 was significantly higher for a tidal volume of 500 than 1000 ml at both 5 and 10-15 min, 7.4892.23 versus 3.70 9 0.83 kPa (P =0.002) and 7.45 9 1.19 versus 3.98 9 1.58 kPa (P B 0.001). The pH was lower for 500 than 1000 ml at 10-15 min, 7.0190.10 versus 7.20 9 0.17 (P =0.034), with a strong trend in the same direction at 5 min (P= 0.06). There was adequate oxygenation with no differences in Pa O 2 or BE at any time between the two groups, and no significant differences in any blood gas variables between the 5-and 10 -15-min samples. We conclude that arterial normocapnia is not achieved with either tidal volume during advanced life support with non-rebreathing ventilation at 12 min − 1 . What ventilation volume is required for CO 2 removal and oxygenation during basic life support with mouth-to-mouth ventilation cannot be extrapolated from the present data. In that situation the risk of gastric inflation, regurgitation and aspiration must also be taken into account.

Carbon dioxide (CO,) insufflation with increased intraabdominal pressure (IAP) has adverse hemodynamic, pulmonary, and renal effects. To avoid these problems, an abdominal wall lift method with a retractor was used to provide the surgical view without CO2 insufflation. Twenty-six patients undergoing elective laparoscopic cholecystectomy were randomly allocated to either the gasless, retractor group, or conventional CO, pneurnoperitoneum group (CPP). Hemodynamic data, ventilatory variables, urine output, urine oxygen tension, and blood samples for determining stress hormones were collected throughout the perioperative period. Patients in the retractor group had lower mean arterial pressure, heart rate, and central venous pressure (P < 0.001). They also had higher pulmonary dynamic compliance and needed a lower minute volume of ventilation to achieve normocarbia (P < 0.001). Urine output and oxygen tension in urine were higher (P < 0.05) with the retractor method than with CPP. Increase in plasma renin activity (P < 0.05) and decrease in core temperature (P < 0.001) were smaller with the gasless method than with CPP. The gasless method for laparoscopic cholecystectomy might be beneficial, especially in patients with compromised cardiorespiratory or renal function. Implications:

The effect of dietary docosahexaenoic acid (22:6n-3, DHA) on the metabolism of oleic, linoleic, and linolenic acids was investigated in male subjects (n = 6) confined to a metabolic unit and fed diets containing 6.5 or <0.1 g/d of DHA for 90 d. At the end of the diet period, the subjects were fed a mixture of deuterated triglycerides containing 18:1n-9[d6], 18:2n-6[d2], and 18:3n-3[d4]. Blood samples were drawn at 0, 2, 4, 6, 8, 12, 24, 48, and 72 h. Methyl esters of plasma total lipids, triglycerides, phospholipids, and cholesterol esters were analyzed by gas chromatography-mass spectrometry. Chylomicron triglyceride results show that the deuterated fatty acids were equally well absorbed and diet did not influence absorption. Compared to the low-DHA diet (LO-DHA), clearance of the labeled fatty acids from chylomicron triglycerides was modestly higher for subjects fed the high DHA diet (HI-DHA). DHA supplementation significantly reduced the concentrations of most n-6[d2] and n-3[d4] long-chain fatty acid (LCFA) metabolites in plasma lipids. Accumulation of 20:5n-3[d4] and 22:6n-3[d4] was depressed by 76 and 88%, respectively. Accumulations of 20:3n-6[d2] and 20:4n-6[d2] were both decreased by 72%. No effect of diet was observed on acyltransferase selectivity or on uptake and clearance of 18:1n-9[d6], 18:2n-6[d2], and 18:3n-3[d4]. The results indicate that accumulation of n-3 LCFA metabolites synthesized from 18:3n-3 in typical U.S. diets would be reduced from about 120 to 30 mg /d by supplementation with 6.5 g/d of DHA. Accumulation of n-6 LCFA metabolites synthesized from18:2n-6 in U.S. diets is estimated to be reduced from about 800 to 180 mg/d. This decrease is two to three times the amount of n-6 LCFA in a typical U.S. diet. These results support the hypothesis that health benefits associated with DHA supplementation are the combined result of reduced accretion of n-6 LCFA metabolites and an increase in n-3 LCFA levels in tissue lipids.

Unrelated hematopoietic stem cell transplantation (HSCT) is a recognized therapy for hematological diseases and over 8 million HLA-typed donors are ready to donate. Increased international exchanges and rapid requests through the Bone Marrow Donor Worldwide (BMDW) ask for standardized quality assurance. Since no such standards have been established to date, we tested a pilot program in order to evaluate donor availability and quality of HLA typing of the Swiss Registry.The 18500 donors of the registry have been analyzed by serology for HLA-AB and by molecular typing for HLA-DR. Through three successive annual quality control (QC) exercises, a total of 114 donor requests were sent to 13 blood transfusion centers responsible for donor recruitment asking for a blood sample. Donors were randomly selected according to recruitment periods (to homozygosity for HLA-A and/or -B antigens. An additional 80 frozen blood samples from the repository corresponding to the three periods (n ¼ 26) and to the 2001 period (n ¼ 54) were also included in the HLA study. HLA-AB typings were done by polymerase chain reaction-sequence specific primers (PCR-SSP) and all discrepancies were retyped. The results showed that 79 samples provided by 69.3% of the requested donors were received within 14 days, and 19 samples (16.7%) were received in 414 days. Altogether, an 86% rate of donor availability was observed, independent of the recruitment period. Among the requested donors, 16 (14%) were not available: for medical reasons (two), for personal reasons (eight), for loss (one), and for an unknown reason (five). The HLA-A/B DNA typing results of 166 homozygous and 12 heterozygous blood samples showed that 437/439 (99.5%) of the assigned A/B antigens were correct.

We present data on geographic variation in polychlorinated biphenyl (PCB) congeners in adult female polar bears (Ursus maritimus) from Svalbard eastward to the Chukchi Sea. Blood samples from 90 free-living polar bears were collected in 1987±1995. Six PCB congeners, penta to octa chlorinated , were selected for this study. Dierences between areas were found in PCB levels and congener patterns. Bears from Franz Josef Land (11,194 ng/g lipid weight) and the Kara Sea (9,412 ng/g lw) had similar SPCB levels and were higher than all other populations (Svalbard 5,043 ng/g lw, East Siberian Sea 3,564 ng/g lw, Chukchi Sea 2,465 ng/g lw). Svalbard PCB levels were higher than those from the Chukchi Sea. Our results, combined with earlier ®ndings, indicate that polar bears from Franz Josef Land and the Kara Sea have the highest PCB levels in the Arctic. Decreasing trends were seen eastwards and westwards from this region. Of the congeners investigated in the present study, the lower chlorinated PCBs are increasing and the high chlorinated PCBs are decreasing from Svalbard eastward to the Chukchi Sea. Dierent pollution sources, compound transport patterns and regional prey dierences could explain the variation in PCB congener levels and patterns between regions.

Canine piroplasmosis is an emerging disease worldwide, with multiple species of piroplasm now recognised to infect dogs. A nested polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay was developed for the detection and differentiation of each of the piroplasm species currently known to infect dogs on the basis of the 18S ribosomal RNA gene. The assay can potentially amplify and discriminate between Theileria annae, Theileria equi, Babesia conradae, Babesia gibsoni, Babesia sp. (Coco) and each of the Babesia canis subspecies. Non-canine piroplasm species can also potentially be detected using the described assay, however amplification of Neospora caninum was also observed. The PCR was found to have a high detection limit, capable of detecting a 2.7 Â 10 À7 % parasitaemia or the equivalent of 1.2 molecules of target DNA when using DNA extracted from whole EDTA blood and detected a parasitaemia of 2.7 Â 10 À5 % using blood applied to both Flinders Technology Associates (FTA 1 ) cards and IsoCode TM Stix. The application of blood samples to filter paper may greatly assist in piroplasm identification in regions of the world where local technologies for molecular characterisation are limited. The assay reported here has the potential to be standardised for routine screening of dogs for piroplasmosis. #

In the early 1970s an increased prevalence of gallstones was reported in adults and children with immunoglobulin deficiency. As the advent of ultrasonography has largely changed the diagnostic approach to gallstones, we have reevaluated the prevalence of cholelithiasis in a group including 37 patients with common variable immunodeficiency and seven patients with other forms of primary immunodeficiency. All patients were receiving intravenous gammaglobulin replacement since 1983 or, in more recent cases, soon after the diagnosis was made, and therefore had relatively few infections. All patients underwent a hepatobiliary ultrasonogram and blood sampling. Data were compared, after age and sex standardization, with those obtained by the GREPCO in a free-living population of 1239 men and 1081 women. Only two women with immunoglobulin deficiency had gallstones. One of these was obese and had had one pregnancy. Both were asymptomatic. None of the patients studied had a history of cholecystectomy or evidence of biliary sludge. Thus, the observed prevalence rates of gallstone disease were 8.7% in women and 0% in males, respectively, against expected values of 9.5% and 3.2%. Immunodeficient patients expressed several putative risk factors for gallstones in the low range (body mass index, total and HDL cholesterol, and blood glucose in both sexes, and triglycerides in men). We conclude that gallstone disease is not more frequent in patients with immunodeficiency syndromes undergoing immunoglobulin therapy than in the general population.

Feed efficiency is a trait of enormous importance in any animal production sector including aquaculture. Individuals that are more feed efficient need to use less feed to achieve similar growth rates as compared to less efficient individuals. Considering that feed represents the largest cost of production and one of the main causes for the ecological footprint of a farm, more knowledge on the extent of individual differences in feed efficiency and underlying physiological processes are needed. This study is the first to investigate individual differences in feed efficiency, measured as residual feed intake and its relationship with feeding behaviour and stress response in one of the most important farmed fish, the Nile tilapia. The individual feed intake of twenty-four juvenile of Nile tilapia was followed during 57 days. Fish were fed until apparent satiation with a commercial feed twice per day. Individual feed behavior was registered once per week and consisted on measuring the latency to start feeding (LAT, min), total feeding time (TFT, min) and the number of feeding acts (NFA, min). Blood samples for plasma cortisol was taken at the end of the experiment (control, indicative of undisturbed levels) and after a stress test (netting), 15 days after the previous sampling. Individual growth rates and residual feed intake were determined at the end of the experiment. Results show pronounced individual differences in residual feed intake as well as a significant correlation with both feeding behavior (feeding latency, total feeding time and number of feeding acts) and stress (cortisol) response. Cortisol levels obtained after the stress test and feeding behavior could explain differences in residual feed intake by 4%. These results show that individuals with higher feeding activity and higher cortisol response are less efficient fish. This suggests that individual differences in feeding activity and stress response explain part of the differences in feed efficiency by explaining variance in maintenance energy expenditure.

Hair sheep ewes (St. Croix White and Barbados Blackbelly) were used to evaluate 3 methods of estrus synchronization for use with transcervical artificial insemination (TAI). To synchronize estrus, ewes (n = 18) were treated with PGF2¢ (15 mg, im) 10 d apart, with controlled internal drug release (CIDR) devices containing 300 mg progesterone for 12 d (n = 18), or with intravaginal sponges containing 500 mg progesterone for 12 d (n = 18). On the day of the second PGF2a injection or at CIDR or sponge removal, sterile rams were placed with the ewes. jugular blood samples were collected from the ewes at 6-h intervals until the time of ovulation, and daily for 16 d after estrus (Day 0). Plasma was harvested and stored at-20°C until LH, and progesterone concentrations were determined by RIA. There was no difference (P > 0.10) in time to estrus among the CIDR-, PGF2c¢-or sponge-treated ewes. All of the ewes in the CIDR group and 94.4% of the sponge treated ewes exhibited estrus by 36 h after ram introduction, while only 72.2% of PGF2a-treated ewes showed signs of estrus by this time (P < 0.06). The time from ram introduction to ovulation was not different (P > 0.10) among the CIDR-, PGF2c¢-or sponge-treated ewes. The time to the preovulatory LH surge was similar (P > 0.10) among CIDR, PGF2c¢ and sponge treated ewes. Progesterone levels through Day 16 after the synchronized estrus were not different (P > 0.10) among treatment groups. Hair sheep ewes (n = 23) were synchronized using PGF2c¢ and bred by TAI using frozenthawed semen 48 h after the second injection. The conception rate to TAI was 2/23 (8.7%) and produced 3 ram lambs. In a subsequent trial, 17 ewes were synchronized with CIDR devices and bred by TAI using frozen-thawed semen 48 h after CIDR removal, resulting in a conception rate of 52.9 % (9/17). It is possible to synchronize estrus in hair sheep using either CIDRs, sponges or PGF2c¢. Even though there were no significant differences in the timing of ovulation Or the LH surge among the treatment groups, a higher conception rate was achieved in ewes synchronized with CIDR devices during the second trial. This may reflect an increase in the skill level of the TAI technidan.

Objective The homocysteine level is considered an important cardiocerebral risk factor. Our aim was to investigate the impact of endurance training on the homocysteine cycle in élite and non-élite athletes. Methods The level of physical activity was measured using a SenseWear armband in five healthy marathon runners. Two blood samples, the first under basal conditions and the second after running a half marathon, were obtained in all subjects by venepuncture. Biochemical parameters, including total homocysteine levels, were assessed. Results Armband data indicated that each athlete ran the half marathon keeping a race pace close to their anaerobic threshold speed. Although the run times were very different between élite and non-élite athletes, a statistically significant increase in plasma homocysteine levels (p=0.026) was observed in all runners. Conclusion Endurance training for 75-100 min held at a race pace close to the anaerobic threshold speed induces an increase in serum plasma homocysteine level.

is an X-linked primary immunodeficiency that is usually associated with thrombocytopenia and eczema. The very variable phenotype of WAS results from defects in the WAS protein (WASP), the function of which is not well understood. In many cases causative mutations have now been identified in the WAS gene. Attempts have been made to correlate the nature of the mutations with the severity of the disease. In this study we investigated mutations in 13 patients with WAS and analyzed the expression of WASP in patient blood samples by immunoblot analysis. We found that despite extensive variation in the nature of the mutations in patients with severe WAS symptoms, none express the protein. However, in 1 patient with a mild clinical phenotype WASP expression was detected. Such an analysis could be used as an initial screening procedure for the diagnosis of WAS prior to genotypic analysis.

A reduced nitric oxide availability is a hallmark of endothelial dysfunction occurring early in atherosclerosis. Recently, we have shown that plasma nitrite mirrors acute changes in endothelial nitric oxide synthase activity in various mammals, including humans. Here, we examined the hypothesis that plasma nitrite levels are reduced in humans with endothelial dysfunction and the decrease is correlated with increasing numbers of cardiovascular risk factors (RF). Plasma nitrite concentrations were quantified by flow-injection analysis. The coefficient of variation for repeated measurements of plasma nitrite was <8%, and heart rate and blood pressure at the time of blood sampling had no significant effect on nitrite values measured (n = 10). Baseline levels of plasma nitrite followed a normal distribution in each group studied and decreased progressively with increasing numbers of cardiovascular risk factors (n = 351, p < 0.001): 351 T 13 (0 RF), 261 T 10 (1 RF), 253 T 11 (2 RF), 222 T 18 (3 RF), and 171 T 29 nmol/L (4 RF). Intima media thickness (IMT) and flow-mediated dilation (FMD) were determined via ultrasound. Plasma nitrite and FMD levels were lower, whereas IMT was greater in individuals with endothelial dysfunction (n = 12) compared to healthy volunteers (n = 12). Nitrite correlated significantly with FMD (r = 0.56, p < 0.001) and inversely with IMT (r = À0.49, p < 0.01). Plasma nitrite levels are reliably measurable in humans, indicate endothelial dysfunction, and correlate with cardiovascular risk factors. Future studies are necessary to identify the prognostic relevance of plasma nitrite determination in patients suffering from cardiovascular disease. D 2005 Elsevier Inc. All rights reserved.

Received: 27 February 2009 / Accepted: 31July 2009 / Published (online): 01 November 2009 ... Association between neuromuscular tests and kumite performance on the Brazilian Karate National Team ... Hamilton Roschel ✉1, Mauro Batista 1, Rodrigo Monteiro 1, Romulo C. ...

Objective: To determine the prevalence and factors associated with Diabetic Retinopathy (DR) of type 1 diabetes mellitus in Thailand. Material and Method: A cross-sectional, multicenter, hospital-based study was carried out from April to December 2003. Diabetic patients in diabetic clinics of 11 tertiary centers were registered. Retinopathy was evaluated by the ophthalmologists. Results: Seven thousand one hundred and nineteen diabetic patients received retinal examination. The number of patients with type 1 diabetes was 347. The prevalence of DR in type 1 diabetes was 21.6% (75). This consisted of Non-Proliferative DR (NPDR) 10.9% (38) and Proliferative DR (PDR) 10.7%. Patients with DR were significantly older, predominantly female, longer duration of diabetes, had higher BMI, systolic Blood Pressure (BP), diastolic BP, serum creatinine, and TriGlycerides (TG) levels than those without DR. Both groups of patients were not different in term of plasma glucose and glycosylated hemoglobin levels. Although the patients with DR had a higher percentage of overt proteinuria than those without DR, there was no difference in percentage of patients with positive microalbuminuria in both groups. This may be explained by limitation of data (only 16% had results of microalbuminuria and 19% had results of proteinuria). After adjusted for duration of diabetes, serum creatinine and smoking status, factors (adjusted odds ratio [95% confidence interval]) associated with DR were duration of diabetes 5-9.9 years (4.0 [1.49-10.91]), 10-14.9 years (6.86 [2.45-19.20]), 15-19.9 years (21.13 [7.22-61.78]), ≥ 20 years (22.15 [7.32-66.99]) when compared with duration of diabetes less than 5 years, serum creatinine > 2 mg/dl (6.0 [2.09-17.22]) when compared with creatinine less than 2 mg/dl. From the presented model, age, gender, systolic BP > 140 mmHg, diastolic BP > 90 mmHg, serum TG and smoking status were not factors associated with DR. Conclusion: Diabetic retinopathy affects about one fifth of type 1 diabetic patients in our study. The authors found the factors associated with DR in type 1 DM were duration of diabetes and serum creatinine. Regular screening for DR and more aggressive management of metabolic factors should be done to reduce the prevalence of DR.

The inXuence of extremely low temperatures on the human body and physiological reactions are not fully recognized. It has been postulated that cryostimulation could modify immunological reactions, leukocytes mobilization and levels of cytokines. The aim of this research was to estimate the inXuence of a ten sessions 3-min-long exposures to cryogenic temperature (¡130°C) on the white blood cell (WBC) count, level of IL6 and the total oxidative and antioxidative status in 15 young, clinically healthy men. Blood samples were obtained in the morning before cryostimulation, again 30 min after treatment and the next day in the morning, both during the Wrst and tenth session. The WBC count, level of IL6 and total lipid peroxides as the total oxidative status and the total antioxidative status (TAS), were measured. After completing a total of ten whole-body therapy sessions a signiWcant increase in WBC count, especially lymphocytes and monocytes was noted. There was an increase in level of IL6 after Wrst and the last cryostimulation the most pronounced after tenth session. On the contrary the TAS level decreased signiWcant after the treatment. It was concluded that repeated expositions to extremely low temperatures use in cryostimulation have mobilization eVect on immunological system.

The objectives of this study were to define the pharmacodynamics of etoposide and to develop potentially useful models (1) to estimate the plasma clearance using a limited number of samples and (2) to describe the relationship between clearance and the dose-limiting toxicity. A total of 17 patients with extensive-stage small-cell lung cancer were treated with 150 mg/m2 etoposide daily for 3 consecutive days and with 100 rag/m2 cisplatin on day 3 only. Both drugs were given intravenously over 1 h. Treatment was repeated every 21 days for up to six courses. All patients were newly diagnosed (no previous chemotherapy or irradiation) and had a performance status of 0-2. Six patients achieved a complete response as confirmed by repeat bronchoscopy and five patients showed a partial response, for an overall objective response rate of 65% (95% confidence interval, 38%-87%). The median survival was 8 months (range, 1-24+ months). The dose-limiting toxicity was neutropenia. Etoposide pharmacokinetics were measured during the first course and determinations were repeated during courses 3 or 4 and 6. Complete blood counts were obtained weekly. Correlations for etoposide clearance and hematologic toxicities were evaluated for 17 initial courses and for an overall number of 33 courses. Pharmacodynamic correlations were significant for graded hematologic toxicities, as well as nadirs of leukocytes, neutrophils, and platelets for the initial courses and for all courses. To reduce the requirement for numerous blood samples, a limited sampling model was developed to estimate the area under the concentration versus time curve (AUC) with the following equation: AUC = 15.45+3.86 x C2+7.10 • C4, where C2 and C4 represent the etoposide concentrations at 2 and 4 h, respectively. The total plasma clearance was calculated as the dose divided by the AUC; correlations with toxicity were better for clearance expressed in milliliters per minute than for that expressed in milliliters per minute per square meter of body surface area. The absolute neutrophil count at the nadir (ANCn) can be estimated by the following pharmacodynamic model, which is based on 33 courses: ANCn =~3.399+0.024 XEcl,