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PubMed1987

[Influence of high environmental temperature on various parameters of blood coagulation in healthy subjects and in thrombosis risk patients].

The effects of environmental hyperthermia (exposure to a hot, dry microclimate) on the human body were investigated with particular reference to certain clotting parameters in healthy subjects and patients at risk of thrombosis. The study covered 70 volunteers, 10 of them clinically healthy (6 males and 4 females) aged 37.7 +/- 9.7 and 60 patients at risk of thrombosis aged 18-60 and divided according to pathology as follows: 26 with ischaemic cardiopathy, 22 with metabolic disorders (12 diabetics, 8 with dyslipidaemia, 2 with hyperuricaemia) and 12 with obliterating arteriopathies of the lower extremities (Fontaine stage 2 and 3). The following standardised protocol was adopted: 2 hours exposure in a controlled climate chamber (40 degrees C, 40-50% humidity, standard air speed 4 m/min, barometric pressure 760 mmHg) for a total of 8 exposures (2 per week for 1 month). This approach was adopted in order to assess not only the effect of each single exposure but also the role of any adaptation to heat. Three blood samples were taken from each subject for each session: the first in basal conditions in a comfortable environment, the second at the end of the 2 hour exposure; the third 30 minutes after the end of the session. Simultaneously samples of arterial blood were taken for pH assays and a spleen echography was performed in basal conditions and at the end of the session for each subject. Each blood sample was tested for several parameters essentially attributable to blood concentration for a broader view of the biological effects of exposure to heart (Ht, blood protein, Nat, K+). The clotting factors under specific study were also assessed (platelet count and volume, beta-thromboglobulin, PF4, von Willebrand Factor VIII, thromboxane B2, fibronectin). Body weight, blood pressure and oral temperature were also measured in all subjects before and after each session. In all subjects both healthy and at risk of thrombosis oral temperature increased (1 +/- 0.4 degrees); on average blood pressure was already higher in basal conditions in the patient group; body weight fell by 900 +/- 120 G in both groups. Ht and blood protein increased significantly in both groups while electrolyte changes were insignificant and blood pH showed a tendency towards acidosis. Clotting parameters revealed a tendency towards thrombophilia in all subjects: platelet count and volume were already higher in the patient group in basal conditions and increased after exposure to hyperthermia. Beta-thromboglobulin, FP4, Factor VIII, thromboxane B2 and fibronectin all increased.(ABSTRACT TRUNCATED AT 400 WORDS)

Adolescent↗
PubMed1985

Subtyping lymphocytes in peripheral blood by immunoperoxidase labeling and light scatter/absorption flow cytometry.

Lymphocyte subpopulations in a whole-blood sample can be detected by adapting mouse monoclonal antibodies (MAbs) and peroxidase (EC 1.11.1.7) labeling to a flow cytometer equipped with a tungsten-halogen light source and scatter/absorption optics (Technicon H6000). In the optimized cytochemical conditions each cell population generates a distinct, well-separated cluster, for accurate "thresholding" of the surface-antigen negative and positive lymphocyte populations in the presence of other leukocytes. After reaction with MAb, the erythrocytes are lysed, and the lymphocytes and other leukocytes are fixed. Biotinylated anti-mouse IgG, used as a bridge, amplifies the response from the avidin-peroxidase label. Granulocytes and monocytes, which have high endogenous peroxidase activity, and the labeled lymphocytes are stained in a specific amount of hydrogen peroxide plus 4-chloro-1-naphthol in 4-(2-hydroxyethyl)-1-piperazine-ethanesulfonic acid buffer. Accuracy and precision are equivalent to those of flow cytometers that measure immunofluorescence (e.g., Ortho Spectrum III), as demonstrated with OKT3, OKT4, OKT8, OKT11, and Leu 12 MAbs.

Adult↗
PubMedDate not supplied

[The isolation and study of the characteristics of a cytopathic strain of the hepatitis A virus].

A fast-growing cytopathic isolate of human hepatitis A virus (strain MB-7) was derived from fecal samples of infected patients and adapted to growth in FRhK-4 cell culture. A positive serum standard against HAV and electron microscopy were used to demonstrate that MB-7 belonged to human hepatitis A virus. The strain MB-7 induced plaque formation in FRhK-4 under agar overlay after 10-12 days of incubation. The PCR products of gene VP1 were cloned in E. coli and its primary structure was determined. MB-7 was shown to have more homology with HAV strains isolated in the USA and China.

Antigens, Viral↗
PubMed1976

[Suitability of the Weidemann test for galactose in mass screening (author's transl)].

With slight practical modification, especially in the preparation of blood samples on filter paper in place of liquid blood samples, it was possible to adapt the determination of galactose dehydrogenase (Weidemann, G. (1971), this Journal 9,527) to mass screening. The technique is described, and the first results are briefly discussed and compared with those obtained over the years in our screening program with proven the Guthrie test (1): The dehydrogenase test is much quicker and shows a higher substrate specificity than the Guthrie test (duration 2 hours compared with 44 hours; false positive results of 16.87 mmol/l galactose or higher 1:42231 compared with 1:3519). It is, however, less sensitive (16.87 mmol/l compared with 1.69 mmol/l galactose), so that a combination of both tests would seem appropriate for the mass screening of new borns.

Alcohol Oxidoreductases↗
PubMed1998

The use of fluorescence methods to monitor unfolding transitions in proteins.

The advantages and some limitations of the use of fluorescence methods for the quantitative determination of the thermodynamics of protein unfolding transitions (i.e., induced by temperature or chemical denaturant) are discussed. Advantages include the sensitivity, multi-dimensional nature of the data, wide amenable concentration range, high signal-to-noise, rapidity of measurement, and adaptability to a variety of sample compartments. Aside from the need for a probe, some problems associated with the method involve the handling of baselines for the pre- and post-transition regions and the difficulty (shared by most other methods) of discerning whether the transition is two-state or multi-state.

Protein Folding↗
PubMed1976

Direct enzymatic determination of urea in plasma and urine with a centrifugal analyzer.

A direct enzymatic micromethod (sample volume, 3mul) has been adapted to the centrifugal analyzer (ENI-GEMSAEC) for measurement of urea in plasma and urine. The method is based on urease (urea amidohydrolase, EC3.5.1.5)/glutamate dehydrogenase [l-glutamate:NAD(P)+oxidoreductase (deaminating), EC1.41.3] coupled reactions, and uses a two-point fixed-time (t(1)=20s,t(2)=50s)kinetic scheme for monitoring the rate of comsumption of NADH at 340 nm. Sensitivity and precision of the method are excellent,and results compare well with those from a commonly used continuous-flow method.

Autoanalysis↗
PubMed1998

The effects of starch and protein degradation rates, hay sources, and feeding frequency on rumen microbial fermentation in a continuous culture system.

Six continuous culture fermenters were used in three experiments to study the effects of dietary starch and protein degradability combination, hay sources, and feeding frequency on fermentation by rumen microorganisms. Experiments 1 and 2 used a 3 x 2 factorial design in which six diets were formulated to contain low (LS), medium (MS), or high starch degradability (HS) in combination with low (LP) or high protein degradability (HP). The dietary combinations were (1) LS + HP, (2) LS + LP, (3) MS + HP, (4) MS + LP, (5) HS + HP, and (6) HS + LP. In experiment 1, pangola was used as the hay source, and in experiment 2, alfalfa hay was used. In experiment 3, two starch degradabilities (LS, MS) and two hay sources (alfalfa, A; pangola, P) were combined with two feeding frequencies (2 X/d, 12 X/d). The dietary combinations were (1) LS + A + 12 X/d, (2) MS + A + 12 X/d, (3) LS + A + 2 X/d, (4) MS + A + 2 X/d, (5) LS + P + 12 X/d, and (6) MS + P + 12 X/d. A CRD design was used for experiment 3. Two rumen-cannulated Holstein cows fed alfalfa hay ad lib were used as donors of rumen fluid for all experiments. Each period was 6 d in length, with 5 d for adaptation and 1 d for sampling. In experiments 1 and 2, the effects of starch degradability on the composition of rumen microorganisms were significant. The MS or HS with HP had the highest total bacterial and protozoal density (P < 0.05). As for VFA, pH and nitrogen products, the effects of starch and protein degradability were not significant. There was no interaction between starch and protein degradability on most of microbial composition (ammonia-N, microbial nitrogen). In experiment 3, 12 X/d feeding frequency (MS + A + 12 X/d vs. MS + A + 12 X/d) resulted in higher pH, which tended to increase bacterial and protozoal density and starch and protein digestibilities. Different hay sources altered the starch and protein synchronization effect on the ammonia-N concentration (mg/dl) and non-ammonia N content (% DM) in the continuous culture system.

Animal Feed↗
PubMed2004

Amino acid analysis in mammalian cell culture media containing serum and high glucose concentrations by anion exchange chromatography and integrated pulsed amperometric detection.

The direct separation detection of amino acids by anion exchange chromatography with integrated pulsed amperometric detection was optimized for the analysis of typical mammalian cell culture broth samples. Existing gradient elution conditions were adapted, considering the additions of peptone (2 g/L) and 10 vol% fetal calf serum to the medium as well as changing concentrations of glucose from 5.5 g/L up to complete consumption. Samples had to be analyzed in two dilutions with water (1:33.3 and 1:200) due to the strongly varying amino acid concentrations in the samples as a result of the medium composition and cell metabolism. The method was validated in a linear working range for the most common amino acids (2.5-7.5 and 1.25-3.75 microM for cystine/cysteine with 15 microl injection volume). The relative standard deviation of the method for all amino acids was less than 5%, with detection limits of less than 0.6 microM and quantitation limits of less than 1.6 microM. As an example, data for the amino acid composition of different media used for the production of inactivated influenza vaccines in cell culture are shown.

Amino Acids↗
PubMedDate not supplied

Attachment from infancy to early adulthood in a high-risk sample: continuity, discontinuity, and their correlates.

This study explores the stability of attachment security and representations from infancy to early adulthood in a sample chosen originally for poverty and high risk for poor developmental outcomes. Participants for this study were 57 young adults who are part of an ongoing prospective study of development and adaptation in a high-risk sample. Attachment was assessed during infancy by using the Ainsworth Strange Situation (Ainsworth & Wittig) and at age 19 by using the Berkeley Adult Attachment Interview (George, Kaplan, & Main). Possible correlates of continuity and discontinuity in attachment were drawn from assessments of the participants and their mothers over the course of the study. Results provided no evidence for significant continuity between infant and adult attachment in this sample, with many participants transitioning to insecurity. The evidence, however, indicated that there might be lawful discontinuity. Analyses of correlates of continuity and discontinuity in attachment classification from infancy to adulthood indicated that the continuous and discontinuous groups were differentiated on the basis of child maltreatment, maternal depression, and family functioning in early adolescence. These results provide evidence that although attachment has been found to be stable over time in other samples, attachment representations are vulnerable to difficult and chaotic life experiences.

Adolescent↗
PubMed1984

Antibodies to dsDNA in connective tissue diseases measured by ultramicro enzyme-linked immunosorbent assay (ELISA) adapted to the chamber analytical technique (CAT).

Human sera (171 samples of patients with connective tissue diseases (CTD) were screened for anti-dsDNA antibodies by ultramicro ELISA adapted to the chamber analytical technique (CAT). The assay offers sensitivity superior to the Crithidia luciliae immunofluorescence technique (CLIFT). Positive results are not restricted to systemic lupus erythematosus (SLE). Nine sera of other CTD patients exhibited antibodies to dsDNA as well. This might be a symptom of a relatively slight immunodisturbance with presence of low-avidity antibodies.

Antibodies, Antinuclear↗
PubMed2025

Harnessing Landscape Genomics to Evaluate Genomic Vulnerability and Future Climate Resilience in an East Asia Perennial.

In this era of rapid climate change, understanding the adaptive potential of organisms is imperative for buffering biodiversity loss. Genomic forecasting provides invaluable insights into population vulnerability and adaptive potential under diverse climatic conditions, thereby facilitating management interventions and bolstering shaping species-specific germplasm conservation strategies. We primarily employed landscape genomics approaches, leveraging single-nucleotide polymorphisms obtained through whole-genome resequencing of 201 individuals across 43 Rheum palmatum complex populations, to pinpoint adaptive variation and its significance in the context of future climates, delineate seed zones, and establish guidelines for ex situ germplasm conservation. The species complex exhibited strong signatures of local adaptation and differential genomic vulnerabilities across its distribution range, with eastern lineage populations facing significant maladaptation risks under future climate scenarios. Using diverse datasets of putatively adaptive loci and climate change scenarios, we delineated three distinct seed zones within the species' range, estimated varying sample sizes per zone to capture most adaptive diversity, and predicted shifts in seed zone centroids ranging from 48.3 to 359.3&#x2009;km from historical distributions to mitigate climate change impacts. Collectively, our findings underscore the importance of integrating genomic and environmental data to forecast the adaptive trajectory of an East Asian perennial under anticipated climate changes, guide seed zone delineation for germplasm conservation and enhance population resilience. These results provide a blueprint for designing targeted conservation strategies and restoration plans in other imperilled species.

Climate Change↗
PubMed1984

In vitro hepatic gluconeogenesis during experimental ketosis produced in steers by 1,3-butanediol and phlorizin.

Adaptations of in vitro incorporation of gluconeogenic substrates into glucose and adaptations of metabolite concentrations of liver to subcutaneous phlorizin and dietary 1,3-butanediol were examined for liver samples from dairy steers. Later, the same adaptations were examined after 6 days of feed restriction. Feeding 1,3-butanediol significantly decreased conversion of carbon-14 of lactate and propionate to glucose and to carbon dioxide. There were no changes of concentrations of hepatic glycogen or triglyceride, and increases were only minor for beta-hydroxybutyrate concentration. Both phlorizin, with or without 1,3-butanediol, and feed restriction significantly increased rates of carbon incorporation into glucose from aspartate, lactate, and propionate but did not change rates of oxidation to carbon dioxide. Phlorizin had no effect on hepatic glycogen or triglyceride concentrations, but feed restriction decreased liver glycogen and increased triglyceride concentrations. Changes associated with either phlorizin treatment or feed restriction are consistent with a decreased ratio of insulin to glucagon of blood plasma. When combined, phlorizin and 1,3-butanediol seem to have some utility for developing a ketosis model.

Acidosis↗
PubMed1984

Rapid serotyping of human rotavirus strains by solid-phase immune electron microscopy.

Nine cell culture-adapted, as well as 30 clinical, human rotavirus (HRV) strains from fecal extracts of children with primary HRV infection were typed by rapid solid-phase immune electron microscopy with protein A and absorbed DS-1 (HRV serotype 2), Wa (serotype 1), and VA70 (assumed serotype 3) rabbit immune sera. As a reference typing test for cell culture-adapted strains, the neutralization assay was used, whereas for noncultivatable strains typing was done for comparison, indirectly, based upon the differential neutralization reactivity of convalescent-phase serum samples from patients with primary HRV infection versus the three reference HRV serotypes. Typing results by solid-phase immune electron microscopy for all strains examined were in complete agreement with those obtained by the neutralization assay, both on cell culture-adapted strains with the three reference rabbit antisera and on three reference HRV strains with human convalescent-phase serum samples. Since adaptation to growth in cell cultures of clinical HRV strains from stool specimens is a time-consuming procedure and is often unsuccessful, solid-phase immune electron microscopy is preferred over the neutralization assay, giving results in about 16 h and also allowing typing of HRV strains from stool specimens low in virus particles. In addition, HRV strains reacting differently from the three reference serotypes may be easily selected by solid-phase immune electron microscopy for further characterization, as was the case for one strain in this study.

Enzyme-Linked Immunosorbent Assay↗
PubMed1987

A procedure for within-trial repeated measurement of gastric emptying in the rat.

A double sampling procedure, developed originally for investigation of liquid phase gastric emptying in humans, is adapted for use in the rat. The double sampling technique permits repeated measurement of gastric volume, allowing a determination of a time course of emptying in a single experimental session. Further, the method allows determination of the amount of gastric secretion, volume emptied into the intestines, and amount of initial gastric load remaining in the stomach. Experiments are presented which: demonstrate the utility of the technique; validate its accuracy in determining gastric volume; indicate the stability of measurements obtained with this procedure; and indicate a procedure for quantitative evaluation of data obtained with this technique. The limitations of the double sampling method are also discussed.

Animals↗
PubMed1999

Effect of a post exercise fat-supplemented diet on muscle glycogen repletion.

Low muscle glycogen at the beginning of exercise may adversely affect performance, increase protein degradation and contribute to the onset of fatigue. As horses are sometimes required to compete on consecutive days both in racing and endurance types of competition, optimal muscle glycogen repletion may improve performance on the day following a race day. The purpose of this experiment was to study the effects of fat supplementation on repletion of muscle glycogen. Twelve Finnhorses performed an exercise test on a treadmill, and 2 and 4 h later they received hay and concentrate (Trial A). Two weeks later these horses performed the same exercise test and were fed the same diet supplemented with either 1000 g of carbohydrate or 400 g of vegetable oil (Trial B). A third trial (Trial C) was 3 weeks later, identical to Trial B, except that the fat group had already been adapted to dietary fat for 3 weeks. Blood samples were analysed for lactate, glucose, glycerol, triglycerides, NEFA, cholesterol, beta-OH-butyrate, insulin and glucagon and muscle samples were analysed for glycogen and triglycerides. The results indicate that in horses not adapted to fat feeding, fat supplementation slows the rate of muscle glycogen repletion, and that after an adaptation period, fat supplementation does not alter the rate of muscle glycogen repletion compared to the rate with a normal diet.

Animals↗
PubMed2001

Metabolic study of the adaptation of the yeast Candida guilliermondii to sugarcane bagasse hydrolysate.

Batch xylitol production from concentrated sugarcane bagasse hydrolysate by Candida guilliermondii was performed by progressively adapting the cells to the medium. Samples were analyzed to monitor sugar and acetic acid consumption, xylitol, arabitol, ethanol, and carbon dioxide production, as well as cell growth. Both xylitol yield and volumetric productivity remarkably increased with the number of adaptations, demonstrating that the more adapted the cells, the better the capacity of the yeast to reduce xylose to xylitol in hemicellulose hydrolysates. Substrate and product concentrations were used in carbon material balances to study in which way the different carbon sources were utilized by this yeast under microaerobic conditions, as well as to shed light on the effect of the progressive adaptation to the medium on its fermentative activity. Such a theoretical means allowed estimation for the first time of the relative contribution of each medium component to the formation of the main products of this fermentation system.

Adaptation, Physiological↗
PubMed2004

Effects of natural plant extracts on ruminal protein degradation and fermentation profiles in continuous culture.

Eight dual-flow continuous culture fermenters were used in four consecutive periods of 10 d to study the effects of six natural plant extracts on ruminal protein degradation and fermentation profiles. Fermenters were fed a diet with a 52:48 forage:concentrate ratio (DM basis). Treatments were no extract (CTR), 15 mg/kg DM of a mixture of equal proportions of all extracts (MIX), and 7.5 mg/kg DM of extracts of garlic (GAR), cinnamon (CIN), yucca (YUC), anise (ANI), oregano (ORE), or pepper (PEP). During the adaptation period (d 1 through 8), samples for ammonia N and VFA concentrations were taken 2 h after feeding. On d 9 and 10, samples for VFA (2 h after feeding), and peptide, AA, and ammonia N concentrations (0, 2, 4, 6, and 8 h after feeding) were also taken. Differences were declared at P < 0.05. During the adaptation period, total VFA and ammonia N concentrations were not affected by treatments. The acetate proportion was higher from d 2 to 6 in CIN, GAR, ANI, and ORE, and the propionate proportion was lower from d 2 to 4 in CIN and GAR, and from d 2 to 5 in ANI and ORE, compared with CTR. However, the proportion of individual VFA (mol/100 mol) was similar in all treatments after d 6, except for valerate in d 9 and 10, which was lower in PEP (2.8 +/- 0.27) compared with CTR (3.5 +/- 0.27). The average peptide N concentration was 31% higher in MIX, and 26% higher in CIN and YUC compared with CTR (6.5 +/- 1.07 mg/100 mL). The average AA N concentration was 17 and 15% higher in GAR and ANI, respectively, compared with CTR (7.2 +/- 0.77 mg/100 mL). The average ammonia N concentration was 31% higher in ANI and 25.5% lower in GAR compared with CTR (5.5 +/- 0.51 mg/100 mL). The accumulation of AA and ammonia N in ANI suggested that peptidolysis and deamination were stimulated. The accumulation of AA N and the decrease in ammonia N in GAR suggests that deamination was inhibited. The accumulation of peptide N and the numerical decrease in AA N in CIN suggest that peptidolysis was inhibited. Results indicate that plant extracts modified ruminal fermentation, but microbes were adapted to some extracts after 6 d of fermentation. Therefore, data from short-term in vitro fermentation studies may lead to erroneous conclusions, and should be interpreted with caution. Careful selection of these additives may allow the manipulation of protein degradation in the rumen.

Acetates↗
PubMed2001

Inorganic nutrient utilisation by "adapted" Pseudomonas putida strain used in the bioremediation of agricultural soil polluted with crude petroleum.

Garden soil samples polluted with crude petroleum were bioremediated by inorganic nutrient monitoring with appropriate adjustment and inoculation with crude oil-adapted strain of Pseudomonasputida (PP) isolated from oil-impacted soils. Soil samples without PP inoculation served as the control samples to compare the abilities of the native soil microflora with the adapted PP strain in biodegrading crude oil pollutant. In the experimental samples, oil concentration and all the inorganic nutrient sources tested decreased more rapidly with a proportional increase in the population densities of both PP and the native soil microflora than were observed in the control samples. This trend was particularly strong for PO4(3-) and NO3- which eventually became limiting both in all the experimental samples and in some control samples. Inoculation of crude oil-impacted agricultural soils by oil -adapted PP strain with nutrient monitoring and adjustment can be effective as bioremediation methods of agricultural land upon pollution with petroleum or petroleum products.

Adaptation, Physiological↗