The influence of the low-energy defibrillating impulse on the endothelin-1 concentration in the left heart ventricle and blood serum of the healthy rabbits.

The influence of low-energy defibrillation on changes in the ET-1 levels in the myocardium and on disruptions in coronary blood flow and microcirculation being their consequence still remains unclear. Myocardial microcirculatory dysfunction is considered as a significant cause underlying myocardial dysfunction in post-cardiac arrest syndrome. This study is aimed at evaluating time-dependent changes in ET-1 levels in serum and the heart of a healthy rabbit following the application of a low-energy two-phase shock impulse. The research was conducted in 35 healthy rabbits at the age of 36 - 42 weeks and with body mass from 3200 to 4150 grams. The animals were divided in a randomized way into four groups depending on the dose of the electrical energy planned for the application during the experiment. The life parameters of the animals were monitored with the application of BeneView T5 patient monitor. The concentration of endothelin-1 in the groups was measured before, 15 and 360 minutes after the application of the low-energy double-phase straight-lined electrical impulse. A transthoracic low-energy defibrillation shock impulse causes a long-term increase in the endothelin-1 levels in the heart muscle and blood serum in a healthy rabbit. The increase in ET-1 levels results from the effect of electrical energy, independently of consequences of the ischemia/reperfusion injury. The increase in the endothelin-1 levels may lead to capillary blood flow abnormalities in the heart, contributing to the development of its dysfunction in the course of postresuscitation disease.

Vitamins B12 and D deficiencies and macro- and microelement disturbances among diabetic elderly patients.

Vitamin and mineral disturbances may interfere with glucose metabolism. Elderly persons with diabetes type 2 (T2DM) are more prone to mineral disturbances and vitamin deficiencies. The aim of this study was to analyze concentrations of vitamins B12 and D and macro- and microelements among diabetic elderly patients. The study enrolled 347 patients with T2DM of whom 247 were elderly (median 76 years of age) (SenDM group) and 100 younger T2DM (median 59 years of age) (Y-DM group), and 320 patients aged 65 years and above without T2DM (mean 77 years of age) - Sen-nonDM - as a control group. Patient clinical and biochemical characteristics were recorded (drugs taken and glucose concentration, glycated hemoglobin level, complete blood count, concentration of Na, K, Ca, Fe and serum vitamins D and B12 levels). All elderly patients had insufficient/deficient vitamin D concentration. Vitamin B12 levels were below the reference limit for 15.6% of the SenDM group. No significant differences in Na, K, were observed among the investigated groups. 30.7% of the SenDM were Fe-deficient. In the SenDM group, vitamin B12-deficient patients did not develop macrocytic anaemia while Fe-deficient patients with T2DM tended to develop microcytic anaemia. The prevalence of vitamin deficiencies in elderly patients with T2DM is clinically relevant. Elderly patients with T2DM are clinically predisposed to Fe deficiencies. We suggest to monitor vitamin B12 and Fe concentration toward developing a full clinical picture as it may accelerate the treatment options and improve elderly patients' outcome.

Impact of low-energy shock impulse on nitric oxide serum levels in healthy rabbits.

Nitric oxide (NO) has emerged as a potent effector molecule for a post-resuscitation disease as a result of the low-energy defibrillation. In this article, the authors anlysed the changes of endogenous nitric oxide levels in serum of 35 healthy rabbits after the transthoracic application of low-energy two-phase shock impulse. The animals were randomised into four groups according to the electrical energy dose to be applied during the experiment. Life parameters of the animals were monitored using the BeneViev T5 patient monitor. The nitric oxide concentration in the groups was measured before, 15 and 360 minutes after applying the low-energy two-phase linear electrical impulse. From 15 to 360 minutes the decrease in nitric oxide concentration was observed. The effect was dependent on single impulse and accumulated energy volumes. The highest decrease of NO concentration was observed in 15 minutes following the application of two electrical impulses of accumulated energy of 8 J/kg/bm. After 360 minutes following the application, a significant increase in nitric oxide levels was recorded. The changes in the nitric oxide levels in the blood serum 15 minutes after the defibrillation may initiate the vasomotor disorders as a result of the low-energy influence of the shock impulse.

Body composition in healthy older persons: role of the ratio of extracellular/total body water.

The aim of this study was to identify the best prognostic parameters for quickly assessing fluid volume status in the context of nutritional status and water balance in older persons and to facilitate decision-making of the general practitioner (GP). This pilot study was conducted with 142 volunteers aged 60 years or older who were Polish students of the University of the Third Age. Inclusion and exclusion criteria for the study were defined. Assessment tools included: the Mini Nutritional Assessment questionnaire (MNA®) and the anthropometric measurements. Weight and body composition analysis were determined by Bioelectrical Impedance Analysis (BIA) using the Tanita MC-780 multi frequency segmental Body Composition Analyzer. According to the MNA scale, 89.2% of the sample was wellnourished and 10.8% were at risk of malnutrition. A total of 47.1% participants had normal body mass index, 20.6% were overweight, and 32.3% were obese. The BIA showed that females had more fat mass (FM) compared to males (35.84% vs 23.90%), while men had more free fat mass (FFM) and total body water (TBW; 61.16% vs 45.22% and 53.31% vs 45.22%respectively). There were no statistically significant differences in FM, FFM, and TBW by age. The ratio of Extracellular to Total Body Water (ECW/TBW) was higher in women than in men (46.76% vs 43.66%). Of all measures, only ECW/TBW increased significantly with age and sex, especially after 65 years. We propose that ECW/TBW may be used as the first, simple, and fast indicator of water volume status in the context of nutritional status and water balance in older subjects. Systematic control of the ECW/TBW by GP or nurse may increase senior independence, resulting in longer self-maintenance at home and reduced hospital admissions.

Epigenetic silencing of endothelin-3 in colorectal cancer.

Endothelins are expressed in a variety of human tissue and are involved in the processes as proliferation, migration and differentiation. The signal transduction pathway is a result of the endothelin-1-3 (ET1-3) binding to their receptors (ETAR, ETBR). ET-3 is a new candidate tumour suppressor gene, which is often downregulated or silenced in human cancer.The aim of the study was to examine DNA methylation of ET-3 genes in colorectal cancer (CRC) tissue samples in relation to the clinical stage (CS) of cancer. The paper is a continuation of our previously published results, which showed a four-fold transcriptional silencing of the ET-3 gene in the samples of colorectal cancer in comparison to normal tissues.A total of 66 paired CRC and normal (surgical margin) tissue samples were used in the study. The tumour tissues were collected from CRC patients in CS I-IV according the 7th edition of UICC TNM Classification of Malignant Tumours (CS I, n = 8; CS II, n = 20; CS III, n = 27; CS IV, n = 11). Assessment of epigenetic silencing of the ET-3 encoding gene was performed in three steps. The silencing of the ET-3 encoding gene was a result from methylation of the promoter sequence using methylation-specific PCR (MS-PCR). Analyses were performed using primers complementary for a CpG island in the first exon of the gene encoding ET-3. An epigenetic silence through methylation of 7.5% (5/66) in comparison to control was observed, including 10% of CS II (2/20), 7% of CS III (2/27) and 9% of CS IV (1/11). The controls and the samples of tumour in CS I showed no epigenetic silencing via methylation. In conclusion, epigenetic silencing of ET-3 in CRC could play a role in the progression than in the induction process. EDN3 would be a future target for epigenetic therapy in colorectal cancer, but further clinical studies are needed.

Rapid quantification of rice root-associated bacteria by flow cytometry.

UNLABELLED: To understand the mechanism of plant-bacterium interaction, it is critical to enumerate epiphytic bacteria colonizing the roots of the host. We developed a new approach, based on flow cytometry, for enumerating these bacteria and used it with rice plants, 7 and 20 days after colonization with Herbaspirillum rubrisubalbicans and Azospirillum brasilense. The results were compared with those obtained with the traditional plate count method. Both methods gave similar numbers of H. rubrisubalbicans associated with rice roots (c. 10(9) CFU g(-1) ). However, flow cytometry gave a number of viable cells of rice-associated A. brasilense that was approx. 10-fold greater than that obtained with the plate count method. These results suggest that the plate count method can underestimate epiphytic populations. Flow cytometry has the additional advantage that it is more precise and much faster than the plate count method. SIGNIFICANCE AND IMPACT OF THE STUDY: Determination of precise number of root-associated bacteria is critical for plant-bacteria interaction studies. We developed a flow cytometry approach for counting bacteria and compared it with the plate count method. Our flow cytometry assay solves two major limitations of the plate count method, namely that requires long incubation times of up to 48 h and only determines culturable cells. This flow cytometry assay provides an efficient, precise and fast tool for enumerating epiphytic cells.

The profile of melatonin receptors gene expression and genes associated with their activity in colorectal cancer: a preliminary report.

The antiproliferative and immunomodulatory effects of melatonin (MLT) have been demonstrated in a variety of neoplasms including colorectal cancer (CRC). In humans and other mammals, MLT acts on target tissues through membrane and retinoid nuclear receptors. The aim of this study was to evaluate transcription activity of melatonin receptors and genes associated with regulation of their activity in colorectal adenocarcinoma tissues in relation to clinical stage of cancer. A total of 24 pairs of surgically removed tumoral and healthy (marginal) tissue samples from colorectal cancer patients at clinical stages I-II and III-IV were collected. As an additional control, twenty normal samples were tak¬en from people whose large intestine tissues were reported as non-tumoral after colonoscopy. Expression of mRNA genes was studied by microarray HG-U133A analysis. The analysis of gene expression profile was performed using commercially available oligonucleotide microarrays of HG-U133A. High increase of MT1 mRNA expression levels in all cancerous samples vs non-cancerous tissues was observed. The MT2 mRNA expression levels increased slightly in marginal and malignant samples. Among the genes participating in the cascade of signal transfer in cells activated by MLT via melatonin receptors, we found encoding genes (GNA11, OXTR, TPH1) only for differentiating stage III - IV of CRC. Monitoring the expression levels of genes that are related to melatonin receptors may offer a strategy to anticipate tumour development and estimate the molecular changes that occur during carcinogenesis. The mechanism behind this association needs further elucidation.

Expression of ADAM28 and IGFBP-3 genes in patients with colorectal cancer - a preliminary report.

Adamalisynes (ADAMs) play an important role in inter-membrane interactions, cell adhesion and fusion processes and protein shedding from the cell surface. Many reports indicate that members of the ADAMs family are overexpressed in human cancer. The aim of the present study was to evaluate ADAM28 and Insulin Like Growth Factor Binding Protein-3 (IGFBP-3)) gene expression in colorectal carcinoma tissues with regard to the overweight or obese status of the patients using an oligonucleotide microarray technique. Fresh tissue specimens were obtained from colorectal cancer patients during surgical treatment. Eighteen specimens from tumour and 18 normal tissue specimens from colorectal cancer patients at clinical stages III and IV were analysed. The examined patients were divided into two groups; those with BMI greater than or equal to 25 and those with normal BMI. The control group consisted of 18 specimens of non-neoplastic colon tissues, which were divided between overweight/obese and normal body weight patients. The gene transcriptional activity from the specimens was analysed using an oligonucleotide microarray technique. Microarrays and rinsing and marking solutions were prepared according to the procedure in the Gene Expression Analysis Technical Manual. The following conclusions were made: i) change of ADAM28 and IGFBP-3 genes expression are present in the normal tissue in overweight/obese patients with colorectal cancer only; ii) the observed molecular variability of ADAM28 and IGFBP-3 expression may be an initial process of cancer proliferation; iii) the histopathologically normal surgical margin in this group of patients was not equal to the molecular margin.

Analysis of expression profile of gene encoding proteins of signal cascades activated by insulin-like growth factors in colorectal cancer.

The aim of the study is to analyse gene typing with the use of the microarray technique (HG-U133A, Affymetrix), differentiating colorectal cancer tissues from tissues assessed histopathologically as healthy ones among a panel of 93 mRNA of gene encoding proteins involved in the activation of cellular signal transduction pathways by insulin-like growth factors. The study was conducted on a group of 8 colorectal cancer patients. Frozen tumor and healthy specimens from the patients were used in molecular tests. Transcript IGF2 differentiated cancer from healthy tissue. Among the genes participating in the cascade of signal transfer in cells activated by IGF, GRB10, PIK3R3, PIK3R1, and IRS1 were qualified as differentiating transcripts. IRS1 indicated over-expression in tumour. Transcript SMAD2 showed a significant changed in tumour samples (increased expression).

Statistical analysis of differential gene expression in colorectal cancer using CLEAR-test.

CLEAR test provides a novel method of analysis by combining inference for differential expression and variability. Frozen tumor specimens from 14 (3 coded Stage I, 5 Stage II, 2 Stage III and 4 Stage IV) colon cancer patients were obtained. Archived primary tumor samples were collected at the time of surgery and normal colon mucosae (controls specimens) were also collected. The studied transcriptomes were clustered using hierarchical agglomeration with Ward's method and Tchebychev distance. The separable groups of transcriptomes were classified as high clinical stage of adenocarcinoma (HCS; stages II-IV), low clinical stage of adenocarcinoma (LCS; stages I and 3 controls), and two normal colon mucosae (controls N1 and N2). The results of the CLEAR-test algorithm in normal colon specimens and adenocarcinoma specimens with low and high clinical stage showed 50 most and 50 least significant genes. The list of differential genes (p<0.01) in normal colon specimens and adenocarcinoma specimens with low and high clinical stage presented 58 genes.

Ethanol-induced changes in profile of fatty acids in isolated rat hepatocytes and the influence of magnesium ions.

The influence of magnesium and ethanol on the fatty acid content in isolated rat hepatocytes was examined in this study. The isolated liver cells were obtained according to the Selgen method and then subjected to ethanol alone or both ethanol and magnesium activity. MgCl2 was used in two concentrations: 2 and 4 mM. The profile of fatty acids in the hepatocytes was evaluated after 5 hours of incubation. Our results revealed that magnesium ions presented together with ethanol in hepatocyte medium changed the hepatocyte fatty acid profile. The total amounts depended on the concentration of magnesium ions.

Genetic disregulation of gene coding tumor necrosis factor alpha receptors (TNFalpha Rs) in colorectal cancer cells.

The expression of TNF ligand by malignant cells might be a mechanism for tumour immune escape. Genetic disregulation of gene coding TNF receptors was observed in neoplastic disease by an increased number of receptors on tumour cells and ligand-receptor activity. It might cause tumour proliferation and metastatic potential. Structure of TNF receptors influences TNF activity in vivo and structure of TNF R2 gene may suggest post-transcription modification based on alternative splicing. The aim of the study was to analyse the expression of gene coding TNF receptors R2 and R2/R7 (without exon 7) by estimation of mRNA expression of colorectal cancer cells in comparison with surrounding tissue free from neoplastic infiltration and searched for differently spliced TNFalphaR2/R7 isoforms. The study included fifty four patients with histopathologically confirmed adenocarcinoma (Stage III according to the AJC TNM Classification). Tissue samples removed from the tumour region were obtained from colorectal cancer patients undergoing surgical treatment. The samples were divided into two parts. The first one--was routinely examined histopathologically, the second one--was used for RNA extraction and the number of TNF and its receptors mRNA copies were subsequently quantified. The TNF and TNFRII genes expression were estimated based on the number of mRNA copies on 1 microg total RNA. The presence of TNFR2 and TNFR2/R7 isoforms in tumour, normal and metastatic cells was observed. The highest number of mRNA TNF copies and over expressed TNF genes were investigated and significantly noticed in metastatic cells (lymph nodes). The decreased number of TNFR2/R7 mRNA copies in metastatic lymph nodes secondarily influenced the decreased TNF soluble receptors' concentration. In conclusion, the genetic disregulation observed in neoplastic disease usually concerns dysfunction of cytokines receptor genes.

On the holistic approach in cancer biology: tumor necrosis factor, colon cancer cells, chaos theory and complexity.

TNFalpha plays a role in the pathogenesis of septic shock, inflammatory diseases, autoimmune diseases, graft rejection reaction, acute, and chronic respiratory inefficiency among others. Its activity depends on the type of target cells and different regulating factors, but the effect of biological activity is conditioned by specific receptors such as p55 (type I, TNF R55) and p75 (type II, TNF R75). The aim of the study was to answer the following questions: 1) Is it possible to apply elements of non-linear dynamics to assess the level of expression of TNF, TNFRI, TNFRII genes in tumor cells, pathologically unchanged tissue and metastatically changed lymph nodes? 2) Is theoretically anticipated variability of cytokine and its receptors in colorectal carcinoma cells and the immediate vicinity justified in the developed mathematical model? The research material--specimens taken from tumor, unchanged tissue and metastatic lymph nodes--were histopathologically and molecularly analysed. Results of the molecular research were used to develop a mathematical model using the basic studies on the theory of chaos and biological system modelling.

Circadian fluctuations of melatonin, tumor necrosis factor-alpha and its soluble receptors in the circulation of patients with advanced gastrointestinal cancer.

The objective of the study was to investigate the dynamic changes of melatonin (MLT), tumor necrosis factor alpha (TNFalpha), soluble TNFalpha receptors ( type I and type II) in serum of advanced cancer patients during 24 hours. The examined group consisted of 42 patients suffering from advanced gastrointestinal neoplasms (colorectal, gastric and pancreatic cancer). Blood samples were collected 6 times a day (8 a.m., 2 p.m., 6 p.m., 10 p.m., 2 a.m., and again 8 a.m.) as well as in healthy controls. Serum levels of TNFalpha and both its receptors were measured using ELISA type and the radioimmunoassay method was used to assess MLT levels. The circadian rhythm of MLT was altered because MLT reached its peak level at 8.50 a.m. with 5 hours delay in respect to average peak time in healthy humans. The presence of circadian rhythm of TNFalpha was proved (acrophase-1.40 a.m.), and no diurnal rhythm of soluble TNF receptors was observed. The concentration of soluble type I (p-55) receptor was distinctly lower than soluble type II (p-75). The peak of soluble type I receptor value appeared at 10.00 p.m. while the type II receptor reached its minimum level at the same time. Although there was no statistical correlation between the receptor concentrations, the shapes of both curves remained inversely proportional. The present results may suggest the presence of complex self-regulation mechanisms between the neuroendocrine system and the cytokine network in advanced gastrointestinal cancer patients.

Dynamics of diurnal changes in serum concentration of TNF-alpha soluble receptors in gastrointestinal cancer patients.

The objective of the study was to investigate the dynamic changes in soluble TNFalpha p-55 and p-75 receptors in serum of advanced cancer patients during 24 hours. The group examined consisted of 42 patients suffering from advanced gastrointestinal neoplasms (colorectal, gastric and pancreatic cancer). Serum levels of the cytokine and both receptors in cancer patients were measured using ELISA type kits 6 times a day (8.00 a.m., 2.00 p.m., 6.00 p.m., 10.00 p.m., 2.00 a.m. and again 8.00 a.m.) as well as in healthy controls. The levels of TNFalpha and its soluble receptors were substantially increased in the examined group and displayed statistically significant circadian fluctuations. The presence of circadian rhythm of the cytokine was proved (acrophase - 00.36 a.m.), however no diurnal rhythm of soluble TNF receptors was observed. The concentration of p-55 receptor was distinctly lower then p-75. The peak p-55 value appeared at 10.00 p.m. while the p-75 reached its minimum level at the same time. Although there was no statistical correlation between the receptor concentrations the shapes of both curves remained inversely proportional. The present results may suggest the presence of complex self-regulation mechanisms in advanced gastrointestinal cancer patients.

[Soluble tumor necrosis factor receptors--their properties and role in diagnosis and therapy of acute and chronic diseases]. / Rozpuszczalne receptory czynnika martwicy nowotworów--ich wlasciwosci i znaczenie w diagnostyce i terapii schorzen o przebiegu ostrym i przewleklym.

The review presents the properties and significance of soluble forms of tumor necrosis factor-alpha receptors in physiology and pathology of human organism. Special attention was turned to the possibility of practical application of those forms of the receptors in the therapy of diseases caused by excessive activity of TNF-alpha. The significance of the soluble receptors as prognostic factors of disease progression and outcome was stressed.

Diurnal rhythmicity of endogenous TNF alpha concentration in plasma of advanced gastrointestinal cancer patients.

The objective of this study was to test the hypothesis of a diurnal fluctuation of circulating levels of endogenous tumor necrosis factor alpha (TNF alpha) in patients with advanced gastrointestinal cancer. Serum levels of TNF alpha were measured at 8 am., 2 p.m., 6 p.m., 10 p.m., and 2 a.m, in 40 patients with advanced gastro-intestinal cancer. In the control group TNF alpha plasma levels were low and did not exhibit regular patterns. On the contrary serum TNF alpha levels were substantially increased in patients with advanced cancer. Peak value appeared at 2 a.m. In contrast, the levels were low in the afternoon (2 p.m.). The presence of circadian rhythm was found with an acrophase at 0 h 36 min.

Fluctuation of endogenous TNF alpha concentration in plasma in advanced cancer patients.

OBJECTIVES: To test the hypothesis of a fluctuation of circulating levels of endogenous Tumor Necrosis Factor alpha (TNF alpha) in patients with advanced gastrointestinal cancer. METHODS: Serum levels of TNF alpha were measured at 8 a.m., 3 p.m., midnight and 3 a.m. in 21 patients with advanced cancer. RESULTS: Serum TNF alpha levels were substantially increased in patients with advanced cancer as compared to healthy subjects. Peak value appeared at 3 a.m., and gradually declined to very low levels in the afternoon (3 p.m.). Using cosinor least square methods we could not find the circadian rhythm of TNF alpha.