Antibodies to bluetongue and epizootic hemorrhagic disease viruses from white-tailed deer blood samples dried on paper strips.

The feasibility of using dried blood samples for serologic testing of white-tailed deer (Odocoileus virginianus) for antibodies to bluetongue virus (BTV) and epizootic hemorrhagic disease virus (EHDV) was tested with matched samples of serum and eluted dried whole blood. Results from matched serum virus neutralization (SN) tests indicated that a 1-ml elution from a 1- x 2-cm section of filter paper strip containing dried blood approximated a 1:10 serum dilution. Neutralizing antibody titers detected from 34 matched titrations of serum and dried blood samples were equivalent in 25 (74%) titrations and were within a single dilution in the remaining nine (26%) titrations. Eluted blood samples from SN-positive deer, however, did not produce detectable precipitin lines on agar gel immunodiffusion tests for antibodies to either BTV or EHDV. In a trial using serum and dried blood samples from 108 hunter-killed deer from five locations in Georgia (USA), antibody prevalence and serotype distribution results were similar. Use of dried blood samples for serologic testing for antibodies to BTV and EHDV provides a reliable alternative to serum but should be considered only when serum collection is not feasible.     

Selenium in Hungary

Selenium (Se) status of Hungarian rock-soil systems has been poorly investigated. The goal of this study was to get general information on the main features of Se geochemistry, and to have a brief look into the human Se status in Hungary, developing an appropriate method for a forthcoming systematic regional investigation. A multistage program was applied: 1. stream sediment sampling; 2. investigation of rock-soil profiles typical in Hungary; and 3. extraction of mobile Se fractions from soils. Human serum samples from apparently healthy blood donors residing in both urban and rural areas were collected from three districts in Hungary. It was concluded that Hungarian rock-soil systems, especially the acid igneous rocks and the widely distributed young sediments (loess and sand formations) with most of the agricultural activity, are low in Se, and the mean serum Se level of the blood samples were also low.     

Hematological and serum chemistry profiles of free-ranging southern two-toed sloths in French Guiana.

Free-ranging southern two-toed sloths (Choloepus didactylus) were translocated during the flooding of a forest at a hydroelectric dam site in French Guiana. Over an 11 mo period blood samples were collected from 90 sloths (38 males, 52 females) in order to determine hematological and serum chemistry reference values. Mean values and range of values were calculated for 13 hematological and 21 serum chemistry parameters. Variations associated with sex, age and reproductive status were identified. Males had a significantly lower red blood cell count than females. Immature animals had more monocytes while adults had more neutrophils and higher mean corpuscular hemoglobin concentration. Aspartate aminotransferase and triglyceride values were higher in young than in adult sloths but uric acid was lower. Lactating females showed lower red blood cells count and iron levels than non-lactating females. These profiles will help to provide reliable baseline data for medical evaluation of sloths.     

A comparison between the concentration of prostaglandin F in human plasma and serum

The concentration of prostaglandin F (PGF) has been measured in serum and plasma samples prepared under different conditions from the antecubital vein blood of 4 non-pregnant and 7 pregnant women. Prostaglandin F concentrations were less than 41 pg/ml in 19 samples of serum or plasma prepared by centrifugation within 30 minutes of collection. When the blood was allowed to clot at room temperature for 24 hours, highly variable, but usually markedly increased concentrations of PGF (<30 - 3020 pg/ml) were found in the serum. Plasma obtained from blood which stood at 23°C for 24 hours contained undetectable amounts of PGF in 4 out of 6 samples and less than 75 pg/ml in the 2 remaining samples. Plasma and serum obtained from blood which stood at 4°C for 24 hours contained less than 45 pg PGF/ml. These results show that (i) incubation of blood at room temperature may markedly elevate concentrations of PGF in serum, (ii) plasma samples rather than serum should be used for measurements of PGF concentrations.     

Serum biochemical values in fallow deer (<Emphasis Type="Italic">Dama dama</Emphasis> L.) from different habitats in Croatia

Serum biochemical values were determined in blood samples collected from 48 shot fallow deer from the Brijuni islands and 45 sedated fallow deer (Dama dama L.) from hunting grounds in the continental part of Croatia. The parameters were compared with regard to age, sex and habitat. Statistically significant differences were found for serum total protein concentration, alkaline phosphatase, aspartate aminotransferase and alanine aminotransferase activity between young and adult island deer, as well as for total protein, triacylglyceride, cholesterol concentration and alkaline phosphatase activity between young and adult continental deer. In young animals, island males had higher albumin concentrations, while continental males had higher alkaline phosphatase values than females. In adult animals, island males had a higher blood urea nitrogen concentration, while continental males had higher albumin and cholesterol concentrations, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and creatine kinase activities. In this group, males exhibited lower bilirubin and triacylglyceride concentrations than females. Our results indicate that besides age, sex and sampling method, nutritional and environmental factors should be considered when evaluating serum biochemical parameters of fallow deer     

Young plasma reverses age‐dependent alterations in hepatic function through the restoration of autophagy

Recent studies showing the therapeutic effect of young blood on aging‐associated deterioration of organs point to young blood as the solution for clinical problems related to old age. Given that defective autophagy has been implicated in aging and aging‐associated organ injuries, this study was designed to determine the effect of young blood on aging‐induced alterations in hepatic function and underlying mechanisms, with a focus on autophagy. Aged rats (22 months) were treated with pooled plasma (1 ml, intravenously) collected from young (3 months) or aged rats three times per week for 4 weeks, and 3‐methyladenine or wortmannin was used to inhibit young blood‐induced autophagy. Aging was associated with elevated levels of alanine transaminase and aspartate aminotransferase, lipofuscin accumulation, steatosis, fibrosis, and defective liver regeneration after partial hepatectomy, which were significantly attenuated by young plasma injections. Young plasma could also restore aging‐impaired autophagy activity. Inhibition of the young plasma‐restored autophagic activity abrogated the beneficial effect of young plasma against hepatic injury with aging. In vitro, young serum could protect old hepatocytes from senescence, and the antisenescence effect of young serum was abrogated by 3‐methyladenine, wortmannin, or small interfering RNA to autophagy‐related protein 7. Collectively, our data indicate that young plasma could ameliorate age‐dependent alterations in hepatic function partially via the restoration of autophagy.     

Hypothalamus-hypophysis-thyroid axis function in healthy aging

There is an increased frequency of dysthyroidism in elderly people. We investigated whether there are differences among healthy young middle-aged and elderly people in the 24 hour secretory profiles of TRH, TSH and free thyroxine. The study was carried out on fifteen healthy young, middle-aged subjects (range 36-55 years, mean age±s.e. 44.1±1.7) and fifteen healthy elderly subjects (range 67-79 years, mean age±s.e. 68.5±1.2). TRH, TSH and free thyroxine serum levels were measured in blood samples collected every four hours for 24 hours. The area under the curve (AUC), the mean of 06:00h-10:00h-14:00h and the mean of 18:00h-22:00h-02:00h hormone serum levels and the presence of circadian rhythmicity were evaluated. A normal circadian rhythmicity was recognizable for TRH and TSH in young, middle-aged subjects and for TSH in elderly subjects. Elderly subjects presented lower TSH levels, whereas there was no statistically significant difference in TRH and free thyroxine serum levels between young, middle-aged and elderly subjects. Aging is associated with an altered TSH secretion.     

一种动态检测小鼠微量血中T细胞受体重排删除环含量的方法

目的:通过检测比较外周血(颈动脉大量采血和微量隐静脉采血)、胸腺组织和脾脏组织等不同成分或组织中T细胞受体重排删除环(T cell receptor rearrangement excision circles,TREC)的含量,并建立一种通过微量外周血PCR预扩增的方法,评估胸腺输出功能。方法:采用C57BL/6小鼠作为实验对象,分成幼年组(5周龄,n=10)和成年组(15周龄,n=10)。经过隐静脉采血及颈动脉采血后处死小鼠,取小鼠胸腺器官和脾脏器官,提取基因组DNA(g DNA),隐静脉微量血g DNA通过PCR预扩增,提纯后再和颈动脉血、胸腺组织和脾脏组织g DNA一起进行实时定量PCR,分析各组织成分TREC的表达水平及差别。结果:幼年组胸腺组织及外周血中TREC的含量比成年组高,且二者趋势基本一致;而脾脏组织结果与其相反;幼年组小鼠胸腺组织中TREC的含量比脾脏组织中高,成年组小鼠结果与之相反;微量外周血经过预扩增后再进行实时定量PCR的结果与直接定量PCR结果一致,且更高效。结论:研究提供了一种新型高效的动态检测T细胞受体重排删除环和检测胸腺输出功能的方法。     

Determination of mercury in human serum and packed blood cells by neutron activation analysis

A method is described for the determination of mercury in human blood serum and packed blood cells employing neutron activation analysis. Great attention was devoted to the collection and manipulation of the samples. The accuracy and precision of the method were tested by analyzing biological reference materials and by comparing the concentrations measured in a number of serum samples to those obtained by another, independent technique (cold vapor atomic absorption spectrometry) in the same samples. The article reports the levels measured in blood serum and packed blood cells samples from 15 adult volunteers, as well as the figures determined in a “second-generation” biological reference material (freeze-dried human serum), prepared and conditioned at the University of Ghent.     

SOD2 gene polymorphism and response of oxidative stress parameters in young wrestlers to a three-month training

The aim of the study was to analyse the effect of Val 16Ala polymorphism in SOD2 gene on oxidative stress parameters and lipid profile of the blood during a three-month wrestling training. The study included 53 Polish young wrestlers. Blood samples were collected at the beginning of the programme and following three months of the training. The list of analysed parameters included erythrocyte and serum activities of superoxide dismutase (SOD), whole blood glutathione peroxidase (GPx) activity, total glutathione (tGSH) level, concentration of lipid hydroperoxides (LHs), total antioxidant capacity (TAC) and creatine kinase (CK) activity in the serum, as well as lipid profile parameters: triglycerides (TG), total cholesterol (TC), high-density (HDL-C), and low-density lipoprotein cholesterol (LDL-C). Three-month training resulted in a decrease in CK activity, an increase in serum SOD activity, as well as in unfavourable changes in serum lipid profile: an increase in TC, LDL-C, and TG, and a decrease in HDL-C. Aside from CK activity, all these changes seemed to be associated with presence of Val allele. Prior to the training programme, subjects with Ala/Ala genotype presented with lower levels of LHs, lower whole blood GPx activity, and lower serum concentrations of TC than the individuals with Ala/Val genotype. Both prior to and after three-month training, higher levels of tGSH were observed in Val/Val genotype as compared to Ala/Val genotype carriers. Moreover, multiple regression analysis demonstrated that SOD2 genotype was a significant predictor of pre-training whole blood GPx activity and erythrocyte SOD activity (Val/Val?>?Ala/Val?>?Ala/Ala). Altogether, these findings suggest that Val 16Ala polymorphism in SOD2 gene contributes to individual variability in oxidative stress status and lipid profile of the blood in young wrestlers, and may modulate biochemical response to training.     

Postprandial lipemia in young men and women of contrasting training status.

This study compared the postprandial triacylglycerol (TAG) response to a high-fat meal in trained and untrained normolipidemic young adults after 2 days' abstinence from exercise. Fifty-three subjects (11 endurance-trained men, 9 endurance-trained women, 10 sprint/strength-trained men, 11 untrained men, 11 untrained women) consumed a meal (1.2 g fat, 1.1 g carbohydrate, 66 kJ per kg body mass) after a 12-h fast. Venous blood samples were obtained in the fasted state and at intervals until 6 h. Postprandial responses were the areas under the plasma or serum concentration-vs.-time curves. Neither fasting TAG concentrations nor the postprandial TAG response differed between trained and untrained subjects. The insulinemic response was 29% lower in endurance-trained men than in untrained men [mean difference -37.4 (95% confidence interval -62.9 to -22.9) microIU/ml x h, P = 0.01]. Responses of plasma glucose, serum insulin, and plasma nonesterified fatty acids were all lower for endurance-trained men than for untrained men. These findings suggest that, in young adults, no effect of training on postprandial lipemia can be detected after 60 h without exercise. The effect on postprandial insulinemia may persist for longer.     

Serum lipids and glucose as associated with hemoglobin levels and copper and zinc intake in young adults

The association of copper and zinc intake with serum total cholesterol, HDL-cholesterol, triglycerides, and glucose concentrations were studied in young adults (N=59). Three-day diet records, hair, fasting blood, and overnight urine samples were collected from each subject. Higher hemoglobin concentrations, diet zinc intake, and serum copper:zinc ratios were associated with lower HDL-cholesterol concentrations. Higher hemoglobin and hair copper concentrations were associated with higher cholesterol concentrations. Higher serum copper concentrations were associated with higher triglyceride concentrations and with lower glucose concentrations. Greater hematocrits were associated with lower triglyceride concentrations. Hemoglobin levels had the strongest relationship with cholesterol and HDL-cholesterol. These results appeared inconsistent with the hypothesis on copper:zinc imbalance and reports from animal studies.     

Age-related features of protein synthesis rhythm in hepatocytes. Effects of extracellular medium

Cell interactions have been studied in cultures pf hepatocytes from young and old rats. The rhythm of protein synthesis is an index of cell interaction and synchronization in culture, while the amplitude of oscillations characterized cell cooperation in an aggregate rhythm. The mean rhythm amplitude in the culture of hepatocytes from old rats is twice lower than that from young rats. Gangliosides (mixture, bovine brain gangliosides) and alpha1-adrenomimetic phenylephrine enhanced synchronization of cultures of the cells from old rats and increased the amplitude of oscillations to the level of young animals. Addition of rat blood serum (10%) to the medium revealed the rhythm of protein synthesis in the culture, asynchronous in the control, i.e., led to their synchronization. In media with young and old rat blood sera, oscillations were intense, with high amplitudes, and low, respectively. Addition of bovine brain gangliosides to a medium with old rat blood serum increased the amplitudes of oscillations to a level of the rhythm stimulated by the young rat serum. Thus, the cells of old animals can fully perceive synchronizing factors and, in the case of their increased concentration, the rhythm of protein synthesis in old animals did not differ from that in young rats. Current data on biochemical mechanisms underlying intercellular cooperation in the formation of population rhythm of protein synthesis have been discussed.     

Ochratoxin A in porcine blood and in consumed feed samples

The aim of the study was to estimate occurrence of ochratoxin A (OA) in feeds and the metabolite residues in porcine blood serum in Poland. Samples were collected in the period from February to May, 1999, in the southern Wielkopolska region. Altogether 40 and 45 samples of feed and porcine blood serum, respectively, were analyzed for OA. Percentage of samples contaminated with OA, both in case of feeds and blood, collected in the winter season was considerably higher than that for the spring season. The percentages for feeds were as follows: 47.6 and 26.3 %, while for porcine serum: 66.7 and 50.0 %, respectively winter and spring. In 25 % of cases ochratoxin A was present in both types of investigated material (feed, blood), whereas in 27.5 % of samples this metabolite was detected in blood only, or in 7.5 % only in the feed. The presence of OA was found neither in the feed nor in the serum in 40 % of all cases. In subgroups (feed, blood) the concentration in the whole collective of positive samples were in the range 0.3–13.5 ng/g and 0.3–69.5 ng/ml, respectively, while median values were 2.3 ng/g and 6.0 ng/ml. Only one feed and three porcine serum samples, were found to be contaminated at concentration levels higher than 10 ng/g or 10 ng/ml.     

Sequencing a specific kinetoplast DNA fragment of Leishmania donovani for polymerase chain reaction amplification in diagnosis of leishmaniasis in bone marrow and blood samples

A set of oligonucleotide primers I and II was developed by analyzing the specificity of a cloned kinetoplast DNA (kDNA) fragment of Leishmania donovani and sequencing the fragment. Polymerase chain reaction (PCR) was conducted with the primers to amplify a minicircle kDNA fragment (297 bp) to detect L. donovani in the bone marrow (22 samples), whole blood (16 samples), and serum (17 samples) of 22 patients with visceral leishmaniasis. All of 22 patients were diagnosed by microscopic identification. Control samples of bone marrow, whole blood, and serum were obtained from patients with leukemia and from healthy volunteers. In addition, 12 dogs were infected with L. donovani promastigotes for the PCR test. The total number of patients positive by PCR testing was 95.5% (21/22), with 91.0% (20/22) from the bone marrow, 68.8% (11/16) from the blood, and 29.4% (5/17) from the sera. Similar results were obtained in infected dogs. No amplification products were seen in control samples from humans or dogs. Our results suggest that PCR may be useful in detecting kDNA in the bone marrow and blood of patients with visceral leishmaniasis.     

The immunization of adults against respiratory syncytial virus infection

The results of the comparative study of the immunological effectiveness of experimental samples of respiratory syncytial (RS) viral vaccine, prepared from a live attenuated strain and introduced in a single administration to young adults by the intranasal, intradermal and combined intranasal-intradermal) routes, are presented. The effectiveness of intranasal immunization was inversely related to the level of previously existing humoral (serum, secretory) antibodies. Intradermal immunization enhanced the frequency of the formation of serum antibodies in persons having had such antibodies before the introduction of RS vaccine. The most active formation of serum and secretory antibodies was ensured by the combined (intranasal-intradermal) method of the administration of live RS vaccine which proved to be particularly effective in persons having had antibodies in the blood and secretions prior to immunization.     

Determination of vitamin A in dried human blood spots by high-performance capillary electrophoresis with laser-excited fluorescence detection

We have developed a high-performance capillary electrophoresis (HPCE) method to analyze the retinol (vitamin A) concentration as retinol-retinol binding protein (holo-RBP) from microvolumes of serum (5–10 μl) or one to two drops (∼20 μl) of blood collected and air-dried on blood collection filter paper. A 0.64-cm diameter disk was cut from the dried whole blood specimens and the samples were dissolved in a pretreatment buffer and filtered. Filtrate was injected onto the HPCE column for analysis. The separation was carried out in a 60 cm × 50 μm I.D. fused-silica capillary and the running voltage was 20 kV. A HeCd laser with a wavelength of 325 nm was used for excitation, and the fluorescence of the holo-RBP complex was monitored at 465 nm by a photodiode. A virtual linear relationship was obtained for the retinol concentrations between HPCE and HPLC for 28 serum samples, 19 dried venous blood samples and 9 capillary dried blood spot samples, indicating that valid measures of serum retinol can be obtained from one to two drops of capillary blood collected on filter paper. The absolute detection limit for retinol by HPCE is below 3 μg/l. The method is very useful for vitamin A level screening, especially for children and premature new-born babies.     

浙江省献血员HGV和TTV感染情况的调查

应用逆转录套式聚合酶链反应(RT-Nested PCR)和半套式聚合酶链反应(Semi-nested PCR)分别检测来自浙江省3个地区165例献血员血清标本中的庚型肝炎病毒(HGV)RNA和输血传播性病毒(TTV)DNA.14.6%(24/165)和12.7%(21/165)的血清标本分别检出HGV RNA和TTV DNA,其中3.6%的血清标本(6/165)可同时检出HGV RNA和TTV DNA.实验结果表明,浙江省献血员中HGV和TTV的感染率较高.     

Fecal dehydroepiandrosterone (DHEA) immunoreactivity as a noninvasive index of circulating DHEA activity in young male laboratory rats

Evidence suggests that dehydroepiandrosterone (DHEA) plays a key role in stress and coping responses. Fecal sampling permits assessment of hormone-behavior interactions reliably and effectively, but no previous study has compared circadian- or stress-dependent alterations between serum DHEA and its fecal metabolites. In the current study, young (28 d of age) male rats were assigned to either an experimental (n = 6) or control (n = 6) group. Rats in the experimental group were exposed to a forced swim test to assess their behavioral and physiologic response to an environmental stressor; blood samples were drawn before the test (baseline), immediately after the test, and at 2 later time points. Only fecal samples were collected from control animals. Fecal DHEA and corticosterone metabolites were monitored in all animals for 24 h. DHEA metabolites in control rats exhibited significant diurnal variation, showing a similar temporal pattern as that of corticosterone metabolites. In addition, fecal and serum DHEA levels were highly correlated. Significant peaks in both DHEA and corticosterone metabolite levels were detected. These data suggest that measures of fecal DHEA can provide a complementary, noninvasive method of assessing adrenal gland function in rats.     

Demonstration of stimulatory effects of platelet-derived growth factor on cultivated rat arterial smooth muscle cells : Differences between cells from young and adult animals

The effects of platelet-derived growth factor (PDGF) on DNA synthesis and proliferation in cultures of arterial smooth muscle cells obtained from young and adult rats, respectively, were measured. Addition of 10-20 ng/ml of PDGF to medium MCDB 104 induced DNA synthesis in quiescent cultures of cells from young animals to a similar extent as 10-20% whole blood serum (WBS). PDGF further stimulated proliferation of the cells in medium MCDB 104, although less markedly than 10% WBS. Antibodies against PDGF partially inhibited the growth response after stimulation with serum. This shows that PDGF is a major growth factor in serum for these cells and that PDGF can promote entrance into and passage through S phase and mitosis independent o plasma factors. Cells from adult animals were also found to respond to PDGF, although a higher concentration (25 ng/ml) was required to obtain a maximum effect. These cells, however, responded better than cells from young animals to stimulation with serum. Further, antibodies against PDGF did not inhibit the growth-stimulatory effect of serum to any appreciable extent. Thus, serum contains growth factors other than PDGF that stimulate preferentiaLly the proliferation of smooth muscle cells from adult animals.