Increased plasma zonulin in patients with sepsis

Introduction:

Zonulin is a eukaryotic protein structurally similar to Vibrio cholerae’s zonula occludens toxin. It plays an important role in the opening of small intestine tight junctions. The loss of gut wall integrity during sepsis might be pivotal and has been described in various experimental as well as human studies. Increased levels of zonulin could be demonstrated in diseases associated with increased intestinal inflammation, such as celiac disease and type 1 diabetes. We therefore investigated the role of plasma levels of zonulin in patients with sepsis as a non-invasive marker of gut wall integrity.

Materials and methods:

Plasma level of zonulin was measured in 25 patients with sepsis, severe sepsis or septic shock according to ACCP/SCCM criteria at the first day of diagnosed sepsis. 18 non-septic post-surgical ICU-patients and 20 healthy volunteers served as control. Plasma levels were determined by using commercially available ELISA kit. Data are given as median and interquartile range (IQR).

Results:

Significantly higher plasma concentration of zonulin were found in the sepsis group: 6.61 ng/mL (IQR 3.51–9.46), as compared to the to the post-surgical control group: 3.40 ng/mL (IQR 2.14–5.70) (P = 0.025), as well as to the healthy group: 3.55 ng/mL (IQR 3.14–4.14) (P = 0.008).

Conclusion:

We were able demonstrate elevated levels of plasma zonulin, a potential marker of intestinal permeability in septic patients. Increased zonulin may serve as an additional mechanism for the observed increased intestinal permeability during sepsis and SIRS.     

Increased plasma vaspin concentration in patients with sepsis: an exploratory examination

Introduction

Vaspin (visceral adipose tissue-derived serpin) was first described as an insulin-sensitizing adipose tissue hormone. Recently its anti-inflammatory function has been demonstrated. Since no appropriate data is available yet, we sought to investigate the plasma concentrations of vaspin in sepsis.

Materials and methods

57 patients in intensive care, fulfilling the ACCP/SCCM criteria for sepsis, were prospectively included in our exploratory study. The control group consisted of 48 critically ill patients, receiving intensive care after trauma or major surgery. Patients were matched by age, sex, weight and existence of diabetes before statistical analysis. Blood samples were collected on the day of diagnosis. Vaspin plasma concentrations were measured using a commercially available enzyme-linked immunosorbent assay.

Results

Vaspin concentrations were significantly higher in septic patients compared to the control group (0.3 (0.1-0.4) ng/mL vs. 0.1 (0.0-0.3) ng/mL, respectively; P < 0.001). Vaspin concentration showed weak positive correlation with concentration of C-reactive protein (CRP) (r = 0.31, P = 0.002) as well as with SAPS II (r = 0.34, P = 0.002) and maximum of SOFA (r = 0.39, P < 0.001) scoring systems, as tested for the overall study population.

Conclusion

In the sepsis group, vaspin plasma concentration was about three-fold as high as in the median surgical control group. We demonstrated a weak positive correlation between vaspin and CRP concentration, as well as with two scoring systems commonly used in intensive care settings. Although there seems to be some connection between vaspin and inflammation, its role in human sepsis needs to be evaluated further.Key words: adipocytokine, inflammation, vaspin, CRP, intensive care     

Platelet factor 3 availability time,plasma phenol and phenolic acids in patients with uremia

Platelet factor 3 availability time (PF3 AT), Prothrombin time (PT), Plasma phenol, phenolic acids, blood urea and serum creatinine were estimated in 31 uremic patients. Significant increase (P<0.01) in PF3 AT and plasma phenolic acid was seen in 100 per cent of cases, while the increase in plasma phenol was seen in 90.3% of cases. The increase in PF3 AT was not uniformly proportional to the increase in plasma phenol or phenolic acids in all cases. Increase in PF3 AT was significant in cases of uremia with bleeding diathesis compared to the cases without bleeding diathesis. Increase in PF3 AT after addition of phenol and urea together to normal platelet rich plasma (PRP) in vitro was greater than the increase in PF3 AT after the addition of phenol or urea alone. Significant decrease (P<0.01) in PF3 AT, plasma phenol, plasma phenolic acids, blood urea and serum creatinine was seen in uremic patients after haemodialysis indicating that the retained toxic metabolites which increase PF3 AT are dializable substances.     

X-linked adreno leukodistrophy: Profiles of very long chain fatty acids in plasma and fibroblasts in eigth Serbian patients

X-linked adrenoleukodistrophy is a severe neurodegenerative disorder with impaired very long chain fatty acid metabolism. The disease associated ABCD1 gene encodes a peroxisomal membrane protein which belongs to the superfamily of ATP-binding cassette transporters. We investigated eight male X-ALD patients diagnosed among 142 suspected patients referred for investigation. Plasma levels of very long chain fatty acids were measured at our laboratory using capillary gas chromatography. Eight cases of childhood X-ALD were diagnosed. This is the first published series of Serbian patients with X-ALD. In addition, diagnosis identifies carriers, which could be benefit for genetic counselling and prenatal diagnosis.     

Relationship between serum interleukin-1beta levels and acute phase response proteins in patients with familial Mediterranean fever

Introduction:

The aim of this study was to investigate whether serum levels of interleukin-1β (IL–1β) has any possible correlation on inflammatory parameters such as C-reactive protein (CRP), erythrocyte sedimentation rate (ESR) and fibrinogen concentration in patients with familial Mediterranean fever (FMF) patients during attack-free period.

Materials and methods:

The serum levels of IL-1β, as an indicator of cytokines status, and the acute phase response proteins, CRP, ESR and fibrinogen levels were evaluated in 35 attack-free patients with FMF and 25 healthy volunteers.

Results:

Serum IL-1β levels were significantly higher in patients with FMF than control subjects (P = 0.018). There was no statistically significant difference in the serum levels of ESR, CRP and fibrinogen between two groups (P = 0.181, P = 0.816, P = 0.686, respectively). There was a significant correlation between IL-1β and CRP (r = 0.513, P = 0.002) values of FMF group.

Conclusions:

In conclusion, our results confirm the presence of increased IL-1β levels in FMF patients during attack-free period. Serum IL-1β values seems to correlate with CRP levels. The elevation of IL-1β levels may be important in monitoring subclinical inflammation of attack free period in FMF patients.     

Investigation of ABCA1 C69T polymorphism in patients with type 2 diabetes mellitus

Introduction:

Non insulin dependent diabetes mellitus is the most common type of diabetes. Genetic factors, lipid profiles, hypertension are potential risk factors for diabetes mellitus. Adenosine binding cassette transporter proteins 1 (ABCA1) plays a role in cholesterol metabolism, especially high density lipoprotein (HDL-cholesterol). There are multiple mechanisms by which HDL-cholesterol can be atheroprotective, it is clear that the relative activity of ABCA1 plays a major role. We aimed to investigate association of ABCA1 C69T gene polymorphism with lipid levels in Turkish type 2 diabetic patients.

Materials and methods:

After isolation of DNA by ethanol precipitation we determined ABCA1 gene polymorphism by using polimerase chain reaction - restriction fragment lenght polymorphism (PCR-RFLP) method in 107 type 2 diabetic patients and 50 healthy controls.

Results:

We have observed that the frequency of TT genotype is significantly higher in healthy controls compared to patients (14% vs. 3%; P = 0.008). Also frequency of T allele was higher in controls than in patients (34% vs. 21%; P = 0.020; OR (95% CI) = 0.52 (0.30–0.88)). There was no association of lipid levels and ABCA1 C69T polymorphism subgroups.

Conclusion:

We have found significantly higher frequency of both T allele and genotype in control group when compared to patients that made us think that T allele may be a protective factor against diabetes mellitus. But, we could not find a relationship between genotypes and lipid concentrations in our two groups. Larger studies will help us to understand the relationship between ABCA1 C69T genotype and lipid parameters in diabetes mellitus.     

Seminal plasma levels of 15-F2α-isoprostane, malondialdehyde and total homocysteine in normozoospermic and asthenozoospermic males

It has been proposed that oxidative stress plays an important role in male infertility. The aims of this study were to compare seminal plasma levels of 15-F2t-isoprostane (8-iso-PGF2α), malondialdehyde (MDA), and total (sum of free and bound) homocysteine (tHcy) from normozoospermic vs. asthenozoospermic men, and to examine the relationships between tHcy and lipid peroxidation products. The study was a case-control study with a simple random sampling. The case group was consisted of 15 asthenozoospermic males. This group was compared with 15 normozoospermic men. Seminal plasma levels of 15-F2α-isoprostane and tHcy were measured using commercially available enzyme immunoassay (EIA) kits. MDA levels were determined by the thiobarbituric acid (TBA) assay. The Mann-Whitney U test was used to compare two groups. Coefficients of correlation were calculated using Spearman’s correlation analysis. All hypothesis tests were two-tailed with statistical significance assessed at the p value <0.05 level. MDA levels were higher in asthenozoospermic subjects than in control subjects (0.72±0.06 μM vs. 0.40±0.06 μM; p<0.05). No differences were seen in 15-F2α-isoprostane levels in asthenozoospermic subjects and controls (65.00±3.20 pg/ml vs. 58.17±4.12 pg/ml; p>0.05). Interestingly, tHcy levels were to be slightly higher in asthenozoospermic subjects than in controls (6.18±1.17 μM vs. 4.8±0.52 μM). Sperm motility was inversely correlated with seminal plasma 15-F2α-isoprostane and MDA levels, respectively (p<0.05). In summary, seminal plasma levels of 15-F2α-isoprostane and tHcy showed no significant difference between normozoospermic and asthenozoospermic men. Sperm motility was not correlated with seminal plasma levels of tHcy. No relationship was found between tHcy and lipid peroxidation.     

Increased serum concentrations of Soluble CD40 Ligand as a prognostic marker in patients with Acute Coronary Syndrome

CD40-CD40L interaction plays a significant role in the pathogenesis of atherosclerosis and coronary artery disease. The clinical predictive value of Soluble CD40 Ligand (sCD40L) was evaluated in patients with Acute Coronary Syndrome (ACS) and Non-Cardiac Chest Pain (NCCP). The levels of serum soluble CD 40 ligand were measured by ELISA in 485 patients admitted to emergency care unit, of which 89 patients were diagnosed as NCCP. The levels of sCD40L were significantly increased in patients with ACS when compared to controls and NCCP. Receiver Operator Characteristic (ROC) Curve analysis showed sCD40L to be a good discriminator between patients with ischemic heart disease and patients without ischemic heart disease. The area under the curve was found to be 0.940 with 95% CI (0.915 to 0.960) (P<0.0001). The cut off value from the ROC curve was 2.99 ng/ml, above which sCD40L was considered to be positive. Combined assessment of sCD40L, Troponin I and CK-MB enhanced the risk prediction and early classification of patients. sCD40L seems to be a promising biomarker for identification and risk stratification for patients with acute coronary syndrome.     

Effect of hemodialysis on circulating cystatin c levels in patients with end stage renal disease

Plasma cystatin C is an emerging parameter to assess kidney function. Its utility in assessing the adequacy of hemodialysis in patients with end-stage-renal disease has however not been established with certainty. This study was therefore carried out to assess the usefulness of serum cystatin C estimation in patients undergoing low flux membrane hemodialysis. Serum creatinine and cystatin C were estimated in 20 patients before and after undergoing hemodialysis. The mean serum creatinine decreased from a pre-dialysis value of 7.72 mg/dL to a post-dialysis value of 2.90 mg/dL. On the contrary, the mean serum cystatin C levels were found to increase from a pre-dialysis value of 5.97 mg/L to a post-dialysis value of 8.25 mg/L. Therefore, serum cystatin C cannot be used to monitor dialysis adequacy. It however, serves as a surrogate marker of the inadequacy of low flux membrane bicarbonate hemodialysis in clearing low molecular weight proteins from the circulation.     

Effect of corticosteroids on elevated intracellular sodium, plasma lipid peroxide levels and reduced Na+, K+-ATPase activity in patients of bronchial asthma

In view of several reports that there is a lack of balance in free radicals in case of bronchial asthma (1) the effect of free radicals on cell membrane was studied by estimating the membrane bound protein Na⁺, K⁺-ATPase activity and the intracellular sodium level in patients of bronchial asthma before and after a short course (one week) of oral corticosteroid (prednisolone 0.75–1mg/kg body weight) therapy. Results showed that there is a definite statistically significant rise in free radical level and intracellular sodium level and a significant lowering of Na⁺, K⁺-ATPase activity in case of untreated bronchial asthma. After short course of therapy with oral corticosteroids, the free radical level and intracellular sodium level decreased significantly, together with a significant rise of the Na⁺, K⁺-ATPase activity. Also, a significant negative correlation (r=−0.74) between the lipid peroxide level and the Na⁺,-K⁺-ATPase activity was found in these cases.     

Lipid peroxidation in erythrocytes of patients with type 2 diabetes mellitus

Lipid peroxidation by measurement of Thiobarbituric Acid Reactive Substances (TBARS) and antioxidant status by determining the activities of the enzymes, Catalase (Cat), Glutathione Peroxidase (GPx) and Superoxide Dismutase (SOD) and the level of Reduced Glutathione (GSH) in the erythrocytes of patients with type 2 Diabetes Mellitus were investigated. It was observed that the level of TBARS in the erythrocytes was increased by 50% showing a significant generation of free radicals in the erythrocytes of these patients. The activities of both Cat and SOD were enhanced while that of GPx was not altered. The level of GSH was also not changed.     

Comparative analysis of RBC membrane fatty acids, proteins and glycophorin in patients with heterozygous beta thalassemia and iron deficiency anemia

Membrane lipid and protein composition was compared in erythrocytes from iron deficiency anemia (IDA) and heterozygous beta thalassemia patients. The study was planned to correlate the influence of iron deficiency with the intrinsic defect of the heterozygous condition on the membrane structural integrity as well as to investigate whether there are differences in membrane changes between the two conditions. Results indicate high levels of saturated fatty acids and low unsaturated fatty acids in both disorders although arachidonic acid and the unsaturation index were lower in heterozygous thalassemia than IDA. Nevertheless, neither of the conditions provoked any alterations in membrane protein or glycophorin suggesting alterations in the lipid moiety only. Present findings indicate that irrespective to the etiology, both, iron deficiency and the heterozygous condition show a common pattern of lipid derangement, which may in turn result in increased membrane rigidity and decreased cellular deformability.     

Immunization with <Emphasis Type="Italic">Wuchereria bancrofti</Emphasis> Glutathione-S-transferase Elicits a Mixed Th1/Th2 Type of Protective Immune Response Against Filarial Infection in Mastomys

Lymphatic filariasis is a mosquito borne parasitic infection and can severely affect the normal working ability of an individual. Currently there is no vaccine available to prevent this infection and the development of a potential vaccine could effectively support the on-going mass drug administration program by World Health Organization (WHO). Filarial parasites have complex mechanisms to modulate the host immune responses against them. The glutathione-S-transferases (GST) are the important enzymes effectively involved to counteract the oxidative free radicals produced by the host. In the present study, we have shown that the mastomys which are fully permissible rodents for Brugia malayi when immunized with Wuchereria bancrofti recombinant GST (rWbGST) could induce 65.5 % in situ cytotoxicity against B. malayi infective (L₃) larvae. There was a balanced Th1/Th2 immune response in the vaccinated animals, characterized by higher levels of WbGST-specific IgG1 and IgG2a antibodies and pronounced IFN-γ, IL-10 and IL-4 cytokines production by the spleen cells.