Pro-Gastrin-Releasing Peptide as a Marker of Small Cell Lung Cancer.

The serum levels of pro-gastrin-releasing peptide (proGRP), neuron-specific enolase (NSE), and chromogranin A (CgA) were studied in 69 patients with small cell lung cancer and 50 apparently healthy donors. A significant increase of all studied biochemical markers was revealed in small cell lung cancer patients, while the highest diagnostic efficiency was demonstrated by proGRP compared to NSE and CgA. ProGRP is a promising biochemical marker of small cell lung cancer, especially sensitive in patients with distant metastases (in the brain, liver, and bones).

Assessment of iron metabolism disorders and adequate treatment of anemic syndrome in patients with breast cancer on the background of adjuvant chemotherapy.

Early and adequate correction of the anemic syndrome (AS) of cancer patients can prevent deterioration in the quality of life and be considered as a reserve for increasing the effectiveness of treatment for breast cancer (BC). The aim of the study was to assess the status of iron using modern methods of ferrokinetics in breast cancer patients on the background of adjuvant chemotherapy for early diagnosis and adequate treatment of AS. The object of the study included 21 breast cancer patients with a relatively favorable prognosis, with luminal types A and B (Her 2 / neu positive or negative), three times negative type. The examination was carried out in the postoperative period, against the background of adjuvant chemotherapy. The main metabolites of ferrokinetics were studied: hepcidin 25 (GP25); ferritin (FR); soluble transferrin receptors (rRTP); transferin (TRF); iron (Fe); erythropoietin (EPO); CRP and IL-6 indicators. AC correction was performed (ferinject, epotin-alpha, B12). 10 (47.6%) patients with breast cancer had AS. Most of them were diagnosed with IDA with microcytic, hypochromic characteristics of erythrocytes, low concentration of FR, Fe, GP25, IL-6, CRP, and high levels of TRP and rRTP. Functional iron deficiency (FDF) was established in some patients. In contrast to patients with IDA, they had a high concentration of FR, CRP and significant production of GP25, IL-6. The EPO level was not optimal for the majority of patients with AS. In isolated cases, during treatment with recombinant erythropoietins, a deficiency of vitamin B12 (cyanocobalamin) was revealed. The rational use of iron preparations, vitamins, and recombinant forms of EPO made it possible to restore Fe metabolism, stabilize the hemoglobin level, and also improve the condition of most breast cancer patients. The obtained data on IL-6, GP25, CRP indicate a certain relationship between them in the development of anemia with VDF in breast cancer patients and the need for further study of the characteristics of iron metabolism in cancer patients.

The role of interleukin-6 and hepcidin 25 in the pathogenesis of anemic syndrome associated with malignant neoplasms with breast cancer patients before neoadjuvant chemotherapy.

A study of interleukin-6 (IL-6), hepcidin-25 (GP-25) was conducted in 22 patients with breast cancer before neoadjuvant chemotherapy and in 27 healthy women in the control group. Significant expression of the GP-25 protein was revealed in breast cancer patients, compared to control. The rates were high both in patients with anemic sindrome (AS) and without it (p <0.01). Latent iron deficiency, AS, IDA and functional iron deficiency (FJ) were more often detected in patients with stage III disease. A significant difference in the parameters of GP-25 and IL-6 was noted, the indicators were higher in patients with stage III (p <0.01). No close correlation was found between IL-6, GP-25 and other acute-phase proteins (FR, CRP) at the initial stages of AS formation. On the contrary, a positive correlation was observed in patients with IDA and FJ between IL-6 and all acute-phase proteins (GP-25, FR, CRP). However, a small number of observations do not allow an unambiguous conclusion about the role of IL-6 and GP-25 expression in the development of AS in cancer patients with breast cancer and requires further study.

Glial Fibrillary Acidic Protein in the Diagnosis and Prognosis of Malignant Glial Tumors.

Serum levels of glial fibrillar acidic protein (GFAP) were analyzed in 317 patients with primary and metastatic tumors of the brain, 78 patients with neurological diseases, and 66 normal subjects. A significant increase in the basal level of GFAP was typical of patients with glioblastomas in comparison with other groups (patients with astrocytomas, cerebral metastases, benign tumors, non-tumor diseases, and healthy subjects). An association of GFAP levels with unfavorable prognosis of overall survival in patients with glioblastoma was revealed. The data attest to high specificity and sensitivity of GFAP as a biochemical marker of glioblastoma.

[Development of strategic approaches to modern diagnosis of anemic syndrome in patients with breast cancer.]

A study of the clinical analysis of blood and major metabolites of ferrokinetics in 107 breast cancer patients before treatment was conducted. In 31 (28.9%) patients revealed anemic syndrome (AS). A feature of the AS is pronounced microcytosis, erythrocyte hypochromia and low hemoglobin content in reticulocytes. Most often (n = 22; 71%) there was iron deficiency (IDA), which was characterized by a low concentration of iron (F), ferritin (FR), hepcidin 25 (GP25), interleukin-6 (IL-6) and high - soluble transferrin receptors (rTFR), transferrin (TRF). In 9 (29%) patients with AS, on the basis of a high concentration of FR, GP25, IL-6, the anemia of chronic disease (AHZ) with functional iron deficiency (FDI) was established. In 23 (74.2%) patients with AS, the was a deficiency of erythropoietin (EPO), the lowest rates were found in the group of patients with a common tumor process and FDI, with less in patients with IDA.

Immunochemical Diagnosis of Multiple Myeloma.

The diagnostic potentialities of complex immunochemical analysis of the serum and daily urine were evaluated in 118 patients with multiple myeloma. In 95 patients, we observed secretion of monoclonal intact immunoglobulins with heavy chains G (N=69), A (N=19), and M (N=4) and biclonal secretion of paraproteins G and A (N=3). Bence-Jones protein was detected in the sera and daily urine of 16 patients and Bence-Jones proteinuria alone was detected in 3 patients. The diagnostic sensitivity of serum immunoelectrophoresis in multiple myeloma is 94.1%. Analysis of paraproteinuria is particularly important in Bence-Jones myeloma, when paraprotein excretion may be not associated with paraproteinemia. Complex study by immunoelectrophoretic and immunoturbidimetric methods in multiple myeloma increases the diagnostic sensitivity to 99.2%.

Free Immunoglobulin Light Chains as Criteria of Extracorporeal Hemocorrection in Patients with Monoclonal Gammopathies.

Elimination of free immunoglobulin light chains with the use of EMic2 selective filters was carried out in 12 patients with monoclonal gammopathies and high production of free immunoglobulin light chains. We showed that extracorporeal detoxification for direct removal of excessive free immunoglobulin light chains from the circulation is advisable for these patients, irrespective of the presence and severity of renal insufficiency. Rapid reduction or elimination of free light chains of immunoglobulins in the course of selective extracorporeal elimination helps to prevent the development of irreversible renal failure and to perform adequate antitumor therapy.

[TYPING OF LEPTOSPIRA SPP. STRAINS BASED ON 16S rRNA].

AIM: Comparative typing of Leptospira spp. strain collection based on analysis of 16S RNA fragment. MATERIALS AND METHODS: 2 pairs of primers were used for PCR, that jointly flank 1423b.p. sized fragment. Sequences of Leptospira spp. strain 16S rRNA, presented in the international database, were used for phylogenetic analysis. RESULTS: A high similarity, including interspecies, of the 16S fragment in Leptospira spp. strains was shown independently of the source, serovar and serogroup. Heterogeneity of the primary matrix, spontaneous mutations of hotspots and erroneous nucleotide couplings, characteristic for 16S sequence of pathogenic Leptospira spp. strains, are discussed. Molecular-genetic characteristic of certain reference Leptospira spp. strains by 16S sequence is obtained. CONCLUSION: Results of the studies give evidence on expedience of introduction into clinical practice of identification of Leptospira spp. by 16S sequence directly from the clinical material, that would allow to significantly reduce identification time, dismiss complex type-specific sera and other labor-intensive methods.

Chromogranin As a Biochemical Marker of Neuroendocrine Tumors.

Comparative study of plasma chromogranin A levels was carried out in 227 patients with neuroendocrine tumors of various locations and 66 normal subjects, men and women, by standard ELISA. In patients with tumors of all locations (pancreas, stomach, small and large intestine, and lungs), the glycoprotein levels were significantly (p<0.000001) higher than in controls. The patients demonstrated high variability of chromogranin A levels; the maximum concentrations were detected in patients with neuroendocrine tumors of the stomach and lung. The highest median values of chromogranin A were found in patients with tumors of the small intestine, large intestine, and pancreas. A relationship between secretion of chromogranin A and dissemination and activity of the neuroendocrine tumor was detected, which was the maximum in patients with metastases in the liver and carcinoid syndrome. High diagnostic sensitivity of chromogranin A was shown: 85.8% for the total group of patients; specificity was 98.5%. These data confirmed high efficiency of chromogranin A as the marker of neuroendocrine tumors. Detection of this marker promotes more accurate diagnosis and evaluation of dissemination of the neuroendocrine tumors.

[The glio-fibrillar acid protein in blood serum of patients with brain tumors and neurological diseases of non-tumorous etiology].

The glio-fibrillar acid protein is considered as a biomarker of astroglyal pathology in case of neurological diseases. The glio-fibrillar acidprotein is intracytoplasmatic filamentous protein specific for cellular skeleton of mature astrocytes playing important role in their differentiation. According publications' data, the highest serum concentrations of glio-fibrillar acid protein are detected in case of neurolepsia, meningitis, febrile schizophrenia, encephalitis, Alzheimer's disease, multiple sclerosis, stroke, opened cranial-cerebral injury and other critical conditions related to damage of hematoencephalic barrier. In the recent years, publications appeared considering analysis of glio-fibrillar acid protein in blood of patients with primary and metastatic brain tumors. Purpose of study To compare levels of glio-fibrillar acid protein in blood serum of patients with primary and metastatic brain tumors and also patients with neurological diseases of non-tumorous etiology and healthy individuals. Material and methods. The analysis was applied to levels of glio-fibrillar acid protein in blood serum of neurooncologic patients (n=175) and also patients with pathology of brain of non-tumorous etiology i.e. degenerate and cerebrovascular diseases (n=38). The control group consisted of 52 healthy individuals with no principal differences in gender and age from examined groups of patients. The concentration of glio-fibrillar acid protein was established in blood serum using immune enzyme analysis in reagents of "BioVendor" firm. Results and discussion. The analysis of results of detection of glio-fibrillar acid protein demonstrated that in patients with primary malignant brain tumors the initial levels of marker were reliably higher than compared with groups of healthy individuals, patients with benign neoplasms, patients with cerebral metastases, and also in case of non-tumorous pathology. The levels of glio-fibrillar acid protein achieve their maximum in case of glioblastoma. The obtained results permit assuming existence of relationship between increasing of content of glio-fibrillar acid protein in blood serum and damage of hematoencephalic barrier permeability in case of development of malignant neoplasms of brain. Conclusion. The obtained data testify higher specificity and sensitivity of glio-fibrillar acid protein as a biochemical marker of glioblastoma appropriate to be applied in case of primary examination of patients with brain affection.

[The diagnostic significance of biochemical markers.]

INTRODUCTION: The modern diagnostic of neuroendocrine tumors is based on analysis of compounds produced by tumor cells (peptides, amines, hormones). PURPOSE OF STUDY: To evaluate diagnostic effectiveness of Chromogranin A, serotonin and its metabolite 5-hydroxyindolilacetic acid as biochemical markers of neuroendocrine tumors as biochemical markers of neuroendocrine tumors. MATERIAL AND METHODS: The detection of Chromogranin A and serotonin in blood serum and 5-hydroxyindolilacetic acid in day urine was applied to 330 patients with neuroendocrine tumors of lungs, pancreas, stomach, small and large intestines and also to 115 healthy males and females. The detection was implemented by enzyme-linked immunosorbent assay based on testsystems "Chromogranin A ELISA Kit" (Dako A/S, Denmark), "Serotonin ELISA" and 5-HIAA ELISA (IBL International GMBH, Germany). RESULTS AND DISCUSSION: The levels of Chromogranin A at all localizations of neuroendocrine tumors reliably (p<0.000001) exceeded corresponding control index. In case of serotonin and 5-hydroxyindolilacetic acid reliable differences were established in all groups except neuroendocrine tumors of stomach. The dependence is established between secretion of markers from prevalence and activity of neuroendocrine tumors. The corresponding levels were higher in patients with metastases in liver and under carcinoid syndrome as compared with patients without corresponding clinical manifestations. The evaluation of diagnostic significance of Chromogranin A, serotonin and 5-hydroxyindolilacetic acid was applied considering threshold levels calculated according results of their detection in control group (33 ng/ml, 320 ng/ml and 60 mkmol/day correspondingly). The high diagnostic sensitivity of Chromogranin A was demonstrated that amounted to 80.6% at specificity 98.5% on the whole in group of patients with neuroendocrine tumors. The serotonin and 5-hydroxyindolilacetic acid manifested comparable diagnostic sensitivity only in patients with carcinoid syndrome (72.5 and 60.3%). CONCLUSION: The obtained data substantiate high effectiveness of Chromogranin A as a marker of neuroendocrine tumors. The detection of this marker contributes into enhancement of accuracy of diagnostic and evaluation of prevalence of tumors of neuroendocrine nature. The serotonin and 5-hydroxyindolilacetic acid are markers of carcinoid syndrome.

Chemokine Receptors CXCR3 and CCR6 and Their Ligands in the Liver and Blood of Patients with Chronic Hepatitis C.

We performed a comprehensive analysis of CCR6 and CXCR3 chemokine receptors and their ligands CCL20/MIP-3α, CXCL9/MIG, CXCL10/IP-10, and CXCL11/ITAC in the liver and blood of patients with chronic hepatitis C at different stages of the disease. TaqMan PCR was used to determine mRNA gene expression of chemokines and their receptors in liver specimens, xMAP multiplex analysis was performed to estimate the concentration of chemokines in blood plasma, and fl ow cytofluorometry was used to evaluate CCR6 and CXCR3 expression on peripheral blood lymphocyte populations. In the liver of patients with hepatitis C, mRNA expression of CXCL10, CCR6, and CXCR3 genes increases with fibrosis progression in the liver tissue. In the plasma, concentrations of all studied chemokines increased depending on the stage of liver fibrosis, CCR6 and CXCR3 expression was changed in various lymphocyte populations. Thus, chemokines are involved in the immunopathogenesis and fibrogenesis in chronic viral hepatitis C. The results suggest using these chemokines in the diagnosis and prognosis of the disease.

[PRODUCTION OF CERTAIN PRO- AND ANTI-INFLAMMATORY CYTOKINES DURING STIMULATION BY LIVE AND INACTIVATED LEPTOSPIRA PATHOGENS IN THE MODEL OF HUMAN WHOLE BLOOD].

AIM: Study of the ability of clinical isolates of leptospira to cause production of certain pro- and antiinflammatory cytokines in the model of human whole blood. MATERIALS AND METHODS: Leptospira interrogans strain was taken for the experiment. Cytokine content was determined by a method based on xMAP technology using a standard panel, composed of 9 analytes: TNF-α, MCP-1, IL-8, IL-4, IL-6, IL-10, IL-IRa, IL- 12 (p70), IFN-γ. RESULTS: An optimal concentration of L. interrogans was selected for stimulation of human whole blood--1 x 10(6) leptospirae/ml. For the first time in the model of human whole blood it was determined, that at early stages of incubation IFN-γ, IL-12(p70), IL-4 and IL-1Ra are more actively produced; at later stages (6 hour incubation)--IL-8 and TNF-α. CONCLUSION: A differential pattern of cytokine production stimulation was shown in the model of human whole blood by live and inactivated leptospirae.

Biochemical markers of bone metabolism in breast cancer.

The relationship between biochemical markers of bone resorption (C-terminal telopeptide, deoxypyridinoline, tartrate-resistant acid phosphatase 5b) and osteosynthesis (bone alkaline phosphatase) and clinical manifestations of bone metastases were studied in 239 patients with breast cancer with and without signs of bone involvement. High levels of C-terminal telopeptide and bone alkaline phosphatase were associated with poor prognosis: more severe skeletal complications and lesser survival values. These biochemical markers can be used for prognosis and optimization of therapy for bone metastases in patients with breast cancer.

Components of the RANK/RANKL/OPG system, IL-6, IL-8, IL-16, MMP-2, and calcitonin in the sera of patients with bone tumors.

Serum levels of sRANKL, RANK, OPG, IL-8, IL-6, IL-16, MMP-2, and calcitonin were measured by ELISA in patients with malignant, borderline, and benign bone tumors and in healthy individuals (control). Serum levels of RANK, OPG, IL-8, IL-6, and the OPG/sRANKL ratio were significantly higher, while the level of MMP-2 was significantly lower in patients with bone tumors than in controls. Serum concentration of IL-16 in osteosarcoma patients was significantly lower than in chondrosarcoma patients. No significant differences between bone sarcomas of different differentiation were detected for any of the studied markers. Calcitonin level depended on the tumor location and type.

Biochemical markers of brain tumours.

The paper presents the results of neurospecific proteins S-100 and glial fibrillary acidic protein (GFAP) determination in blood serum samples of 145 neuro-oncology patients and 69 healthy people. The significant elevation of S-100 and GFAP was revealed in glioblastoma (G IV) patients compare to the patients with anaplastic astrocytoma (G III), benign meningioma (GI), celebral metastasis and healthy controls. The concentration of S-100 in blood serum of patients with anaplastic astrocytoma, benign meningioma, and celebral metastasis did not significantly differ among themselves, and in relation to the control group there was a significantly increase only in patients with cerebral metastasis. GFAP was characterized by high frequency of its detection in patients with glioblastoma (83%) compare to other groups of patients and healthy donors, in which it was practically undetectable. These data suggest the possibility of using GFAP as a marker of glioblastoma and S-100- as an additional biochemical criteria of cerebral lesions in oncology patients.

Granulosa cell tumors of the ovary and inhibin B.

We present the results of immunoenzyme detection of inhibin B in blood serum of patients with adult-type granulosa cell tumors of the ovary. Blood concentration of inhibin B at the end of menstrual cycle in patients with tumor relapse was significantly higher than in patients during remission and in virtually healthy women. The increase in inhibin B concentration preceded clinical manifestation of the disease relapse by 2-13 months, which demonstrates high diagnostic sensitivity of this marker and suggests that it can be recommended for the use in diagnostics and monitoring of granulosa cell tumors of the ovaries.

Neurospecific proteins in the serum of patients with brain tumors.

Neurospecific proteins S-100 and GFAP were measured in the serum of 145 patients with neural tumors and 69 healthy individuals. In patients with glyoblastomas, the concentrations of S-100 and GFAP were significantly higher than in patients with anaplastic astrocytomas, benign meningiomas, and brain metastases and in healthy individuals. Serum S-100 concentrations in patients with anaplastic astrocytomas, benign meningiomas, and brain metastases were similar; significant difference from the control was found only for patients with cerebral metastases. A specific feature of GFAP was high incidence of its detection in patients with glioblastomas (83%) compared to other groups of patients with neural tumors and healthy volunteers who demonstrated practically zero level of this protein. These findings attest to the possibility of using S-100 as an additional biochemical criterion of brain involvement in tumor patients and GFAP as a glioblastoma marker.

Serum angiogenic factors in patients with neuroendocrine tumors of abdominal organs.

The initial (before treatment) levels of VEGF, endostatin, and tumor necrosis factor-beta (TNF-beta) were measured in the sera of 20 patients with malignant and benign neuroendocrine tumors of the abdominal organs and 25 healthy controls. The initial levels of VEGF, endostatin, and TNF-beta in the total group of patients with neuroendocrine tumors of abdominal organs did not differ from the control. A significant difference was detected between the mean serum concentrations of endostatin in patients with benign and malignant neuroendocrine tumors: 64.1+/-14.7 and 107.8+/-14.1 ng/ml, respectively (p=0.043). The content of VEGF, endostatin, and TNF-beta did not correlate with patients' gender and age. A direct correlation between endostatin, TNF-beta concentrations and maximum size of the primary tumor was detected in patients with malignant neuroendocrine tumors. Direct correlations between the initial levels of VEGF and endostatin and an inverse correlation between VEGF and TNF-beta concentrations were detected in patients with benign neuroendocrine tumors. Relapse-free survival was significantly lower in patients with serum endostatin concentration >100 ng/ml and TNF-beta<30 pg/ml.

Synergy of gene-mediated immunoprophylaxis and microbeam radiation therapy for advanced intracerebral rat 9L gliosarcomas.

PURPOSE: Microbeam radiation therapy (MRT), a novel experimental radiosurgery that largely spares the developing CNS and other normal tissues, is tolerated well by developing animals and palliates advanced 9LGS tumors. This report, to our knowledge, is the first demonstration that gene-mediated immunotherapy (GMIMPR) enhances the efficacy of MRT for advanced 9LGS tumors. METHODS: Seventy-six male Fischer 344 rats were implanted ic with 10(4)9LGS cells on d0. By d14, the cells had generated approximately approximately 40 mm3 ic 9LGS tumours, experimental models for therapy of moderately aggressive human malignant astrocytomas. Each of the 14 untreated (control) rats died from a large (>100 mg) ic tumor before d29 (median, d21). On d14, the remaining 62 rats were given deliberately suboptimal microbeam radiation therapy (MRT) by a single lateral exposure of the tumor-bearing zone of the head to a 10.1 mm-wide, approximately approximately 11 mm-high array of 20-39 microm-wide, nearly parallel beams of synchrotron wiggler-generated radiation (mainly approximately 50-150 keV X-rays) that delivered 625 Gy peak skin doses at approximately approximately 211 microm ctc intervals in approximately approximately 300 ms either without additional treatments (MRT-only, 25 rats), with post-MRT GMIMPR (MRT+GMIMPR, 23 rats: multiple sc injections of irradiated (clonogenically-disabled) GM-CSF gene-transfected 9LGS cells), or with post-MRT IMPR (MRT+IMPR, 14 rats: multiple sc injections of irradiated (clonogenically-disabled) 9LGS cells. RESULTS: The median post-implantation survivals of rats in the MRT-only, MRT+GMIMPR and MRT+IMPR groups were over twice that of controls; further, approximately approximately 20% of rats in MRT-only and MRT+IMPR groups survived >1 yr with no obvious disabilities. Moreover, over 40% of MRT+GMIMPR rats survived >1 yr with no obvious disabilities, a significant (P<0.04) increase over the MRT-only and MRT+IMPR groups. SIGNIFICANCE: These data suggest that the combination of MRT+GMIMPR might be better than MRT only for unifocal CNS tumors, particularly in infants and young children.