Seasonal rhythms of "acute phase proteins" in humans

"Acute phase proteins" comprise a group of proteins whose concentrations increase or decrease by at least 25% after a damaging stimulus (burn, trauma, tissue damage, etc.) or during inflammation. We investigated the seasonal variation in the concentrations of several acute phase proteins--alpha1-antichymotrypsin (ACT), alpha1-acid glycoprotein (AGP), transferrin (Tf), alpha2-macroglobulin (alpha2-M), ceruloplasmin (Cp), antitrypsin (AT), and haptoglobin (Hp). Blood samples were collected from 15 healthy volunteers, who were subjected to the seasonal changes in illumination, were drawn at 08:00 h every 3 months (August, November, January/February, March/April, June/July). With the exception of Hp, all acute phase proteins showed an annual rhythm (ANOVA; p < 0.01). Lowest concentrations occurred in the winter months (November through February), with the exception of Tf, which was oppositely phased.     

Serum amyloid-A (SAA) and haptoglobin (Hp) plasma concentrations in newborn calves

The concentrations of 2 major bovine acute-phase proteins, haptoglobin (Hp) and serum amyloid-A (SAA), were measured in plasma obtained shortly after birth from 22 healthy calves. In a separate group of diseased calves (n = 8), Hp and SAA concentrations were measured to determine whether newborn calves (up to 4 d old) are able to produce SAA and Hp. In blood samples taken directly after birth, the Hp plasma concentrations were all below the limit of detection. The mean SAA concentration was independent of weight (r = 0.063), degree of acidosis (r = -0.125), sex (p > 0.05), and were not different in calves born after different types of obstetrical help (p > 0.05). In the group of diseased calves, an increased Hp concentration was measured in only 2 of 8 animals, whereas the mean SAA concentration was significantly higher (p < 0.05) than in the healthy newborn calves. These data suggest that prenatal stress due to parturition does not form a stimulus for the production of acute-phase proteins in the fetal calf. The low Hp plasma concentrations might indicate that either it takes a few days to establish a detectable concentration of this protein, or that Hp production is not fully developed in newborn calves.     

Elevated plasma concentrations of haptoglobin in European brown bears during hibernation

Haptoglobin (Hp), a hemoglobin-binding protein, is known as an acute phase protein and increases during the acute phase of inflammation in most mammals. We reported previously in brown bears that the mean Hp concentrations were higher in blood samples obtained in winter than those in spring. To examine a possible relation of the seasonal variations of Hp to hibernation, in the present study, we measured the plasma concentrations of Hp as well as some other acute phase proteins (alpha(2)-macroglobulin, alpha(1)-antitrypsin, C-reactive protein) in 6 European brown bears (Ursus arctos), from which blood samples were obtained at 5-6 different months of year including February, the time of hibernation. The Hp concentrations showed clear seasonal variations, being highest in February. The alpha(2)-macroglobulin concentrations also showed a similar but much smaller rise in February, but those of alpha(1)-antitrypsin and C-reactive protein did not show any seasonal variations. Our results suggest that the seasonal variation of plasma Hp concentration in brown bears is associated with a hibernation-specific mechanism more than that of acute phase response.     

Lipid peroxidation rate and the antioxidant protection system during a three-day dry immersion experiment

The content of lipid peroxidation products (diene conjugates, malondialdehyde, Schiff bases) and antioxidant defense system indices (the main lipid antioxidant tocopherol and the level of general antioxidant activity) were measured in the blood serum of five male volunteers aged 25?C40 years in a three-day dry immersion experiment. During the immersion test, no deviations of indices from the background values were found. An increase in the tocopherol concentration within 2 h after the beginning of the experiment was the only exception. A significant increase in the concentration of lipid peroxidation products, particularly, diene conjugates, was observed 2 h after immersion completion during the reconditioning period. However, the tocopherol content was significantly lower than the background values. It is concluded that the subjects?? adaptation to simulated microgravity conditions displays no pronounced stress component, whereas bringing back to normal vital functions after exposure to immersion induces a pronounced stress reaction illustrated by a significant increase in the lipid peroxidation product levels against a background of a decrease in the functional activity of the antioxidant defense system.     

Circadian rhythm of acute phase proteins under the influence of bright/dim light during the daytime

We investigated the influence of two different light intensities, dim (100 lx) and bright (5000 lx), during the daytime on the circadian rhythms of selected acute phase proteins of C-reactive protein (CRP), alpha1-acid glycoprotein (AGP), alpha1-antichymotrypsin (ACT), transfferin (TF), alpha2-macroglobulin (alpha2-m), haptoglobin (HP), and ceruloplasmin (CP). Serum samples were collected from 7 healthy volunteers at 4 h intervals during two separate single 24 h spans during which they were exposed to the respective light intensity conditions. A circadian rhythm was detected only in ACT concentration in the bright light condition. The concentration of ACT, a positive acute phase protein (APP), increased (significantly significant differences in the ACT concentration were detected at 14:00 and 22:00 h) and AGP showed a tendency to be higher under the daytime bright compared to dim light conditions. There were no significant differences between the time point means under daytime dim and bright light conditions for alpha2-M, AGP, Tf, Cp, or Hp. The findings suggest that some, but not all, APP may be influenced by the environmental light intensity.     

Molecular associations and surface-active properties of short- and long-N-acyl chain ceramides

The behaviour of N-hexadecanoylsphingosine (Cer16), N-hexanoylsphingosine (Cer6) and N-acetylsphingosine (Cer2) in aqueous media and in lipid-water systems, monolayers and bilayers has been comparatively examined using Langmuir balance and fluorescence techniques. Cer16 behaves as an insoluble non-swelling amphiphile, not partitioning into the air-water interface, thus not modifying the surface pressure of the aqueous solutions into which it is included. By contrast both Cer6 and Cer2 behave as soluble amphiphiles, up to approx. 100 microM. At low concentrations, they become oriented at the air-water interface, increasing surface pressure in a dose-dependent way up to ca. 5 microM bulk concentration. At higher concentrations, the excess ceramide forms micelles, critical micellar concentrations of both Cer6 and Cer2 being in the 5-6 microM range. When the air-water interface is occupied by a phospholipid, 6Cer2 and Cer6 become inserted in the phospholipid monolayer, causing a further increase in surface pressure. This increase is dose dependent, and reaches a plateau at ca. 2 microM ceramide bulk concentration. Both Cer2 and Cer6 become inserted in phospholipid monolayers with initial surface pressures of up to 43 and 46 mN m(-1), respectively, which ensures their capacity to become inserted into cell membranes whose monolayers are estimated to support a surface pressure of about 30 mN m(-1). Both Cer2 and Cer6, but not Cer16, had detergent-like properties, such as giving rise to phospholipid-ceramide mixed micelles, when added to phospholipid monolayers or bilayers. The short-chain ceramides form large aggregates and precipitate at concentrations above approx. 100 microM. These results are relevant in cell physiology studies in which short- and long-chain ceramides are sometimes used as equivalent molecules, in spite of their different biophysical behaviour.     

Effect of gangliosides on the cytotoxic activity of natural killers from Syrian hamsters

The ability of various gangliosides to inhibit the cytotoxic activity of natural killers (NK-cells) from Syrian hamsters towards human lymphoma MOLT-4 cells was studied. The inhibitory effect was found to depend on the structure and concentration of the gangliosides. At concentrations corresponding to those in the blood of tumour-bearing hosts, SiaLacCer and Sia2LacCer inhibited the NK-activity. A significant inhibition was also found for (NeuAc)2GgOse3Cer, whereas NeuGcGgOse3Cer and NeuAcGgOse4Cer were practically inactive. Previously it was shown that Sia2LacCer which is either absent or very low in normal blood, is produced by a number of tumours and that tumour cells "shed" considerable amounts of gangliosides. On this ground, it was proposed that elevated concentrations of SiaLacCer and Sia2LacCer in the blood of tumour-bearing animals may inhibit the NK-activity and thus contribute to the "escape" of tumour cells from host immune surveillance.     

Early fluid and protein shifts in men during water immersion

High precision blood and plasma densitometry was used to measure transvascular fluid shifts during water immersion to the neck. Six men (28-49 years) undertook 30 min of standing immersion in water at 35.0 +/- 0.2 degrees C; immersion was preceded by 30 min control standing in air at 28 +/- 1 degrees C. Blood was sampled from an antecubital catheter for determination of blood density (BD), plasma density (PD), haematocrit (Ht), total plasma protein concentration (PPC), and plasma albumin concentration (PAC). Compared to control, significant decreases (p less than 0.01) in all these measures were observed after 20 min immersion. At 30 min, plasma volume had increased by 11.0 +/- 2.8%; the average density of the fluid shifted from extravascular fluid into the vascular compartment was 1006.3 g.l-1; albumin moved with the fluid and its albumin concentration was about one-third of the plasma protein concentration during early immersion. These calculations are based on the assumption that the F-cell ratio remained unchanged. No changes in erythrocyte water content during immersion were found. Thus, immersion-induced haemodilution is probably accompanied by protein (mainly albumin) augmentation which accompanies the intravascular fluid shift.     

Proteomics-derived cerebrospinal fluid markers of autopsy-confirmed Alzheimer’s disease

The diagnostic performance of several candidate cerebrospinal fluid (CSF) protein biomarkers in neuropathologically confirmed Alzheimer’s disease (AD), non-demented (ND) elderly controls and non-AD dementias (NADD) was assessed. Candidate markers were selected on the basis of initial two-dimensional gel electrophoresis studies or by literature review. Markers selected by the former method included apolipoprotein A-1 (ApoA1), haemopexin (HPX), transthyretin (TTR) and pigment epithelium-derived factor (PEDF), while markers identified from the literature included Aβ1-40, Aβ1-42, total tau, phosphorylated tau, α-1 acid glycoprotein (A1GP), haptoglobin, zinc α-2 glycoprotein (Z2GP) and apolipoprotein E (ApoE). Ventricular CSF concentrations of the markers were measured by enzyme-linked immunosorbent assay (ELISA). The concentrations of Aβ1-42, ApoA1, A1GP, ApoE, HPX and Z2GP differed significantly among AD, ND and NADD subjects. Logistic regression analysis for the diagnostic discrimination of AD from ND found that Aβ1-42, ApoA1 and HPX each had significant and independent associations with diagnosis. The CSF concentrations of these three markers distinguished AD from ND subjects with 84% sensitivity and 72% specificity, with 78% of subjects correctly classified. By comparison, using Aβ1-42 alone gave 79% sensitivity and 61% specificity, with 68% of subjects correctly classified. For the diagnostic discrimination of AD from NADD, only the concentration of Aβ1-42 was significantly related to diagnosis, with a sensitivity of 58%, specificity of 86% and 86% correctly classified. The results indicate that for the discrimination of AD from ND control subjects, measurement of a set of markers including Aβ1-42, ApoA1 and HPX improved diagnostic performance over that obtained by measurement of Aβ1-42 alone. For the discrimination of AD from NADD subjects, measurement of Aβ1-42 alone was superior.     

Ceramide metabolism is affected by obesity and diabetes in human adipose tissue

Ceramide is involved in development of insulin resistance. However, there are no data on ceramide metabolism in human adipose tissue. The aim of our study was to examine sphingolipid metabolism in fat tissue from obese nondiabetic (n = 11), obese diabetic (n = 11), and lean nondiabetic (n = 8) subjects. The content of ceramide (Cer), dihydroceramide (dhCer), sphingosine (SPH), sphinganine (SPA), sphingosine‐1‐phosphate (S1P; pmol/mg of protein), the expression (mRNA) and activity of key enzymes responsible for Cer metabolism: serine palmitoyltransferase (SPT), neutral and acidic sphingomyelinase (nSMase and aSMase, respectively), and neutral and acidic ceramidase (nCDase and aCDase, respectively) were examined in human adipose tissue. The contents of SPA and Cer were significantly lower whereas the content of dhCer was higher in both obese groups than the respective values in the lean subjects. The expression of examined enzymes was elevated in both obese groups. The SPT and CDases activity increased whereas aSMase activity deceased in both obese groups. We have found correlation between adipose tissue Cer content and plasma adiponectin concentration (r = 0.69, P < 0.001) and negative correlation between total Cer content and HOMA‐IR index (homeostasis model of insulin resistance) (r = ?0.67, P < 0.001). We have found that both obesity and diabetes affected pathways of sphingolipid metabolism in the adipose tissue. J. Cell. Physiol. 227: 550–557, 2012. © 2011 Wiley Periodicals, Inc.     

Quantitative proteome analysis of the 20S proteasome of apoptotic Jurkat T cells

Regulated proteolysis plays important roles in cell biology and pathological conditions. A crosstalk exists between apoptosis and the ubiquitin?Cproteasome system, two pathways responsible for regulated proteolysis executed by different proteases. To investigate whether the apoptotic process also affects the 20S proteasome, we performed three independent SILAC-based quantitative proteome approaches: 1-DE/MALDI-MS, small 2-DE/MALDI-MS and large 2-DE/nano-LC?CESI?CMS. Taking the results of all experiments together, no quantitative changes were observed for the ??- and ??-subunits of the 20S proteasome except for subunit ??7. This protein was identified in two protein spots with a down-regulation of the more acidic protein species (??7a) and up-regulation of the more basic protein species (??7b) during apoptosis. The difference in these two ??7 protein species could be attributed to oxidation of cysteine-41 to cysteine sulfonic acid and phosphorylation at serine-250 near the C terminus in ??7a, whereas these modifications were missing in ??7b. These results pointed to the biological significance of posttranslational modifications of proteasome subunit ??7 after induction of apoptosis.     

Iron status in runners of various running specialties.

The blood iron status of 44 male runners of various running specialties (18 sprinters, 13 middle- and 13 long-distance runners) is evaluated by measuring serum ferritin (SF), serum iron (Si), hemoglobin concentration (Hb), hematocrit (Ht), red blood cells content (RBC) and haptoglobin concentration (Hp). The results of these analyses (except Hp) are compared to those obtained in sedentary male subjects (control group) of the same mean age. Mean SF, SI, Hb and Ht measured in athletes are significantly lower than in control group. The remarkably low Hp values obtained in athletes suggests the occurrence of hemolysis. Using unpaired t test, it appears that the blood iron status of these runners does not depend on their running specialty.     

Acute phase proteins, body temperature and urinary melatonin under the influence of bright and dim light intensities during the daytime

Concentrations of five acute phase proteins: C-reactive protein (CRP), alpha 1-antichymotrypsin (ACT), transferin (Tf), alpha 2-macroglobulin (alpha 2-M) and haptoglobin (Hp) as well as glycosylation profiles of alpha 1-antichymotrypsin (ACT) were studied in sera samples with 7 healthy volunteers under the influence of two different light intensities during the daytime dim (100 lx) and bright (3000 lx) light. Concentration of transferin (negative proteins) under the influence of bright light during the daytime decreased significantly. Other proteins have the tendency to increase (positive proteins) under the influence of daytime bright light. The microheterogeneity of ACT did not change under the influence of different light intensities. Melatonin and rectal temperature were also measured simultaneously. Rectal temperature decreased to be lower during the first half of the night and urinary melatonin secretion rate increased to be higher during the night when the subjects spent time under the bright light during the day. Thus, it is concluded that the diurnal bright light exposure may activate some parameters of acute phase proteins, increase nocturnal melatonin secretion and accelerate a fall of rectal temperature during first half period of night sleep.     

Molecular associations and surface-active properties of short- and long-N-acyl chain ceramides

The behaviour of N-hexadecanoylsphingosine (Cer16), N-hexanoylsphingosine (Cer6) and N-acetylsphingosine (Cer2) in aqueous media and in lipid-water systems, monolayers and bilayers has been comparatively examined using Langmuir balance and fluorescence techniques. Cer16 behaves as an insoluble non-swelling amphiphile, not partitioning into the air-water interface, thus not modifying the surface pressure of the aqueous solutions into which it is included. By contrast both Cer6 and Cer2 behave as soluble amphiphiles, up to approx. 100 μM. At low concentrations, they become oriented at the air-water interface, increasing surface pressure in a dose-dependent way up to ca. 5 μM bulk concentration. At higher concentrations, the excess ceramide forms micelles, critical micellar concentrations of both Cer6 and Cer2 being in the 5-6 μM range. When the air-water interface is occupied by a phospholipid, 6Cer2 and Cer6 become inserted in the phospholipid monolayer, causing a further increase in surface pressure. This increase is dose dependent, and reaches a plateau at ca. 2 μM ceramide bulk concentration. Both Cer2 and Cer6 become inserted in phospholipid monolayers with initial surface pressures of up to 43 and 46 mN m⁻¹, respectively, which ensures their capacity to become inserted into cell membranes whose monolayers are estimated to support a surface pressure of about 30 mN m⁻¹. Both Cer2 and Cer6, but not Cer16, had detergent-like properties, such as giving rise to phospholipid-ceramide mixed micelles, when added to phospholipid monolayers or bilayers. The short-chain ceramides form large aggregates and precipitate at concentrations above approx. 100 μM. These results are relevant in cell physiology studies in which short- and long-chain ceramides are sometimes used as equivalent molecules, in spite of their different biophysical behaviour.     

EEG power spectra in simulation of disruptiveness

For analysis of the action of destructive behavior, the EEG power spectra in groups with different levels of disruptiveness at rest and during the modeling of destructive behavior were studied. The destructive processes in the group of less destructive subjects of the experiment are accompanied by the development of activation processes of specific and nonspecific character (??₁-rhythm power loss and ??₂-rhythm power growth). This allows critical assessment of the incoming stimuli, generation of adequate behavioral responses, and inhibition of aggression. In the group of the subjects of the experiment with a high level of destructiveness activation, the criteria are less distinct. The criteria of excessive activation from diencephalic structures are observed (?? rhythm power growth), which may be connected with the development of earlier and less stable mechanisms of behavior regulation and help to select more primitive behavioral models, including aggression.     

Basophils infiltrate human gastric mucosa at sites of Helicobacter pylori infection, and exhibit chemotaxis in response to H. pylori-derived peptide Hp(2-20)

Basophils, which are normally confined to the circulation, can migrate to sites of allergic inflammation. Using the specific mAb, BB1, we detected basophil infiltration of the gastric mucosa of Helicobacter pylori-infected patients affected by moderate and severe gastritis. Basophils were not found in H. pylori-free individuals or in subjects with mild gastritis. The H. pylori-derived peptide, Hp(2-20), was a potent basophil chemoattractant in vitro, whereas the control peptide, Hp1, was ineffective. Basophils from peripheral blood of healthy volunteers expressed mRNA for the formyl peptide receptors, N-formyl-peptide receptor (FPR), FPR-like (FPRL)1, and FPRL2. Preincubation of basophils with FMLP or Hp(2-20) caused complete desensitization to a subsequent challenge with homologous stimulus. Incubation of basophils with a low concentration of FMLP, which binds with high affinity to FPR, but not to FPRL1 or FPRL2, did not affect the chemotactic response to Hp(2-20). In contrast, a high concentration of FMLP, which binds to FPRL1 and FPRL2, reduced the chemotactic response to Hp(2-20). The FPR antagonist, cyclosporin H, prevented chemotaxis induced by FMLP, but not by Hp(2-20). Hp(2-20) could be responsible, at least in part, for basophil infiltration of the gastric mucosa of H. pylori-infected patients presumably through the interaction with FPRL1 and FPRL2.     

Serum haptoglobin concentrations in ringed seals (Pusa hispida) from Svalbard, Norway

Haptoglobin (Hp) levels were measured in blood serum from 185 apparently healthy ringed seals (Pusa hispida) from Svalbard (age range, 1-30 yr) collected during the spring seasons of 2002 through 2004. The Hp concentration was 0.26 +/- 0.02 g/l (mean +/- SE; range, 0.0-1.5 g/l). Maturity status, body condition index (CI), interactions between CI and maturity status, and sex and maturity status all had significant influences on Hp levels. Mature males had significantly higher Hp concentrations compared with mature females (0.30 +/- 0.03 vs. 0.17 +/- 0.01 g/l, F(1,16) = 14.9, P<0.01). No differences were found between Hp levels in immature male and female seals (0.33 +/- 0.06 g/l). The Hp levels increased significantly in immature seals when the CI decreased. Despite the fact that mature males had a significantly lower CI than mature females, no direct correlation was found between Hp concentration and CI among adults. The elevated Hp values found in mature males might be related to injuries inflicted among males while fighting during the mating season or to high stress levels related to mating competition. The Hp concentrations of ringed seals measured in this study are useful baseline data for development of a diagnostic tool for future monitoring of the general health of this and other ringed seal populations.     

Selected contribution: role of spleen emptying in prolonging apneas in humans.

This study addressed the interaction between short-term adaptation to apneas with face immersion and erythrocyte release from the spleen. Twenty healthy volunteers, including ten splenectomized subjects, participated. After prone rest, they performed five maximal-duration apneas with face immersion in 10 degrees C water, with 2-min intervals. Cardiorespiratory parameters and venous blood samples were collected. In subjects with spleens, hematocrit and hemoglobin concentration increased by 6.4% and 3.3%, respectively, over the serial apneas and returned to baseline 10 min after the series. A delay of the physiological breaking point of apnea, by 30.5% (17 s), was seen only in this group. These parameters did not change in the splenectomized group. Plasma protein concentration, preapneic alveolar PCO2, inspired lung volume, and diving bradycardia remained unchanged throughout the series in both groups. Serial apneas thus triggered the hematological changes that have been previously observed after long apneic diving shifts; they were rapidly reversed and did not occur in splenectomized subjects. This suggests that splenic contraction occurs in humans as a part of the diving response and may prolong repeated apneas.     

Morpho-Physiological Responses of Alamo Switchgrass During Germination and Early Seedling Stage Under Salinity or Water Stress Conditions

Switchgrass (Panicum virgatum L.) is a warm perennial grass with valuable characteristics as a biofuel crop. To avoid competition with food crops, biofuel crops will be likely relegated to less productive soils such as marginal lands. Consequently, the salinity and water scarcity problems that commonly affect marginal lands compromise biofuel crop germination, emergence, and seedling establishment. The aims of this study were to study the germination and seedling growth of switchgrass under salinity and water stress and to describe the morpho-anatomical responses of the roots and leaves in the seedlings to these stress conditions. The effect of salt and water stress was assessed using sodium chloride (NaCl) and polyethylene glycol (PEG) 8000 at the same water potentials of ??0.8, ??1.0, and ??1.2 MPa. Seeds were moist prechilled for 7 days at 5 °C and germinated at 30/15 °C (8 h light/16 h dark). NaCl treatments (??0.8 and ??1.0 MPa) delayed germination rates but did not reduce the final germination percentage, whereas at a lower potential (??1.2 MPa), the final germination percentage was diminished. The effects of PEG (??1.0 and ??1.2 MPa) on the germination rate and final percentage were more detrimental than those induced by isosmotic concentrations of NaCl. PEG and NaCl reduced significantly the vigor index of ??0.8 to ??1.2 MPa. The morpho-anatomical changes such as the reduction in the root cross-sectional area and the thickening of the endodermis walls for both stress conditions and aerenchyma formation in the cortex under salinity could significantly contribute in the survival and tolerance during the early seedling stages.