Quantitative analysis of human immunoregulatory cytokines by electrochemiluminescence method

Quantitative analysis of human immunoregulatory cytokines in physiological media and cell cultures plays an important role in fundamental and clinical research. Here we describe the quantification of interleukin (IL)-2, IL-4, IL-10 and interferon-gamma (IFN-gamma) in human serum and peripheral blood mononuclear cell (PBMC)-conditioned medium by electrochemiluminescence method (ECL). We demonstrate that this approach allows to detect cytokine concentration from 1 pg/ml. The high sensitivity in combination with accuracy and wide range of determined concentration indicates that ECL meets the standards of quantitative analysis of cytokines. Simplicity and short time of procedure, small assay volume and high reproducibility make ECL method competitive in practical use with conventional quantitative methods of cytokine detection.     

The application of an impulse plasma jet for trauma of the liver

The authors made an analysis of the application of a plasma jet with the help of an apparatus of original construction used for treatment of wounds of the liver in 36 patients. This apparatus has a number of advantages as compared with others. The plasma jet gives a reliable homostasis and minimum heat injuries to the liver tissues.     

Adenocarcinoma of the stomach: clinical and immunologic features

Lower natural killer cells activity (NKCA), varying stage of anemia and in vitro depressed colony-forming ability of committed precursors of hemopoiesis were identified in 50 gastric cancer patients. The patients with different patterns of the vegetative nervous system had tumors with a range of adenocarcinoma cell differentiation. A relationship was established between anemia stage, colony-forming ability of committed precursors of hemopoiesis, on the one hand, and vegetative nervous system pattern, stage of tumor cell differentiation and, to a less extent, NKCA, on the other.     

Analysis of the Dynamics of Interneuronal Functional Connections during Conditioned Reflex Activity

The dynamics of interneuronal functional connections were studied in the prefrontal cortex of dogs performing a task consisting of unforeseen remodeling of movement conditioned reflexes. An original method was used to find and classify the temporal patterns of linked spikes coming from several simultaneously recorded neurons. This procedure showed that in 33 pairs of neurons (87% of the total number of pairs showing interneuronal functional connections), parts of the conditioned reflex program were associated with behaviorally significant changes in the functional relationship between the neurons. In different behavioral situations, linked activation of a given pair of cells was restricted to different stages of the performance of the conditioned reflex task. In 17 pairs of neurons, the periods of switching of interneuronal functional connections, i.e., the intervals in which linked spikes were absent during all stages of task performance, were seen only in a particular behavioral context different from the situation in which these same cells generated linked discharges. The specific characteristics of different methods of analyzing the dynamics of interneuronal functional connections in conditions of a dynamic behavioral context and multifactorial determination of conditioned reflex activity are discussed.     

Towards retrievable vascularized bioartificial pancreas: induction and long-lasting stability of polymeric mesh implant vascularized with the help of acidic and basic fibroblast growth factors and hydrogel coating

We seek to improve existing methodologies for allogenic grafting of pancreatic islets. The lack of success of encapsulated transplanted islets inside the peritoneal cavity is presently attributed to poor vascularization of the implant. A thick, fibrotic capsule often surrounds the graft, limiting survival. We have tested the hypothesis that neovascularization of the graft material can be induced by the addition of proper angiogenic factors embedded within a polymeric coat. Biocompatible and nonresorbable meshes coated with hydrophilic polymers were implanted in rats and harvested after 1-, 6-, and 12-week intervals. The implant response was assessed by histological observations on the degree of vascularity, fibrosis, and inflammation. Macrostructural geometry of meshes was conducive to tissue ingrowth into the interstitial space between the mesh filaments. Hydrogel coating with incorporated acidic or basic FGF in an electrostatic complex with polyelectrolytes and/or with heparin provided a sustained slow release of the angiogenic growth factor. Anti-factor VIII and anti-collagen type IV antibodies and a GSL I-B4 lectin were used to measure the extent of vascularization. Vigorous and persistent vascularization radiated several hundred microns from the implant. The level of vascularization should provide a sufficient diffusion of nutrients and oxygen to implanted islets. Based on our observations, stable vascularization may require a sustained angiogenic signal to allow for the development of a permanent implant structure.     

Clinical significance of the determination of R-proteins in the ejaculate of patients with chronic prostatitis

Products of cell receptor (R-proteins) catabolic lysis were measured in ejaculate of 121 patients with chronic prostatitis and 15 healthy males. R-protein levels were higher in the patients (U = 172.5; p = 0.047). When immunological indices of the ejaculate (IgA, sIgA, IgG, IgM, interleukin-8) were measured, it was found that in patients with chronic prostatitis there is a positive correlation between concentration of interleukin-8 and content of R-proteins (r = 0.43; p = 0.028). Positive correlation also exists between R-protein content and prostate volume (r = 0.24; p = 0.034) while negative one--between R-proteins concentration and size of the hypoechogenic sites in the prostate (r = -0.25; p = 0.039), between R-proteins in the ejaculate and serum level of fibrinogen B (r = -0.25; p = 0.021). It is inferred that elevation of ejaculate R-protein level occurs in alteration of the prostate. The test for R-proteins in ejaculate can be helpful in detection of early stages of chronic prostatitis.     

The use of electrochemoluminescent method for detection of cytokines in various media

Electrochemiluminescent (ECL) method was used for cytokine detection with TAG label (ruthenium(II) tris-bipyridine chelate N-hydroxysuccinimide ester). A quantitative ECL detection of IL-4 and IFN-gamma in physiological fluids and cell culture media is described. The results evidence that ECL is compatible to commercial kits or even more effective. The advantages of ECL are simple procedure, no need in washings, small volume of the assay, wide range of cytokine concentrations, high sensitivity, and good reproducibility. The method can be used in research and clinical laboratories.