Strategies for Multiplexed Electrochemical Sensor Development

Detection of multiple biomarkers for disease diagnosis or treatment monitoring has received a lot of attention due to their potential impact on clinical decision making. Electrochemical biosensors have become one of the preferred detection approaches, due to the simplicity of the accompanying instrumentation. This chapter will explore how electrochemical sensors can be utilized for detection of multiple analytes by integration of sensors into microfluidic microsystems. Some key fabrication technologies for such devices will be presented utilizing polymer microfabrication, paper-based approaches, and the use of printed circuit boards. Next, the use of electrode arrays will be presented along with some commercial platforms, outlining plausible paths towards a successful electrochemical multiplexed sensor. Novel approaches based on microbeads and various labels will then be introduced along with various strategies and technologies utilized to achieve ultrasensitive multiplexed detection

Noise Stress-Induced Changes in mRNA Levels of Corticotropin-Releasing Hormone Family Molecules and Glucocorticoid Receptors in the Rat Brain

Noise is a widespread stress resource that may lead to detrimental effects on the health. However, the molecular basis of the stress response caused by noise remains elusive. We have studied the effects of acute and chronic noise stress on stress-related molecules in the hypothalamus and hippocampus and also corticosterone responses. Sprague Dawley rats were randomized into control, acute and chronic noise stress groups. While the chronic noise stress group animals were exposed to 100 dB white noise for 4 h/a day during 30 days, the acute noise stress group of animals was exposed to the same level of stress once for 4 h. The expression profiles of corticotropin-releasing hormone (CRH), CRH1, CRH2 receptors and glucocorticoid receptor (GR) mRNAs were analysed by RT-PCR. Chronic noise stress up-regulated CRH mRNA levels in the hypothalamus. Both acute and chronic noise increased CRH-R1 mRNA in the hypothalamus but decreased it in the hippocampus. GR mRNA levels were decreased by chronic noise stress in the hippocampus. The present results suggest that while corticosterone responses have habituated to continuous noise stress, the involvement of CRH family molecules and glucocorticoid receptors in the noise stress responses are different and structure specific

The effects of Saccharomyces cerevisiae extract on the weight of some organs, liver, and pancreatic digestive enzyme activity in breeder hens fed diets contaminated with aflatoxins

The effects of the Saccharomyces cerevisiae extract on some organ, liver, and pancreatic digestive enzymes in breeder hens fed on aflatoxin (AF)-contaminated feed were investigated. Forty-eight 58-wk-old Ross 308 breeder hens were used. The hens were fed diets containing 0 or 100 mu g of AF/kg and 0 or 1 g of S. cerevisiae/kg in a 2 x 2 factorial arrangement of treatments. Although serum alkaline phosphatase levels were significantly higher, serum alkaline aminotransferase (P = 0.068) and gamma-glutamyltransferase (P = 0.067) levels tended to increase (P < 0.05) in hens fed the AF-contaminated diet than those of hens fed the uncontaminated diet. Both AF and S. cerevisiae extract increased (P < 0.001) pancreatic amylase activity, but the effect was not additive, resulting in an AF x S. cerevisiae extract interaction (P < 0.001). a-Amylase activity in duodenum was lower (P < 0.001) in hens fed the AF-contaminated diet. Duodenum a-amylase activity was higher (P = 0.024), but jejunum a-amylase activity was lower in S. cerevisiae extract-supplemented hens than that of nonsupplemented hens. There was a significant interaction between AF and S. cerevisiae extract on pancreatic and duodenal lipase activity. Pancreatic lipase activity decreased in hens fed the AF-contaminated diet. However, S. cerevisiae supplementation extract minimized this effect of AF on pancreatic lipase activity. Duodenal lipase activity was decreased in hens fed the AF-contaminated diet without S. cerevisiae extract supplementation. However, there were not any significant differences between hens fed the AF-contaminated diet and hens fed the uncontaminated diet after S. cerevisiae extract supplementation. Pancreatic trypsin activity was higher (P = 0.044) in hens fed the AF-contaminated diet than that of hens fed the uncontaminated diet. There was a significant interaction between AF and S. cerevisiae extract on pancreatic chymotrypsin activity. It was increased in hens fed the AF-contaminated diet without S. cerevisiae extract supplementation. However, S. cerevisiae extract supplementation counteracted this negative effect of AF on pancreatic chymotrypsin activity. The treatments did not result in any change in duodenal chymotrypsin activity, but S. cerevisiae supplementation decreased (P < 0.05) jejunal chymotrypsin activity. In conclusion, our results showed that addition of 1 g/kg of S. cerevisiae extract reduces the toxic effects of AF on pancreatic lipase and chymotrypsin activity. Therefore, it may be useful to supplement feedstuff with S. cerevisiae extract to reduce the effects of AF in laying breeder hens

The effects of Enterococcus faecium NCIMB10415 on the development of pancreas and small intestine and on activity of pancreatic digestive enzymes in broiler chickens

The effects of Enterococcus faecium NCIMB 10415 on the development of pancreas and small intestine, and on activity of some pancreatic digestive enzymes were investigated in broiler chickens. In total, 168 Ross 308 female broiler chickens were used. The chickens were assigned to two groups, control and experimental. Enterococcus faecium NCIMB10415 was added by 1.23x10(9) CFU/kg to the diet of the experimental group animals for a period of 42 days. Feed intake and body weights of animals were recorded weekly and feed efficiency was calculated. On days 0, 7,14, 21, 28, 35 and 42 twelve chickens from each group were sacrificed and the development of pancreas and small intestine was investigated. Activities of a-amylase, lipase, trypsin and chymotrypsin in pancreas and contents of duodenum, jejunum and ileum were also determined. Body weight, feed intake and feed efficiency were not significantly different between treatments. Enterococcus faecium NCIMB10415 had no effect on weight and allometric growth of pancreas, duodenum, jejunum and ileum. Activities of a-amylase, lipase, trypsin and chymotrypsin in the intestinal content were lower in the probiotic group (P<0.05) than in the control group. The results of the study suggest that the supplementation of Enterococcus faecium NCIMB10415 to the diet of broilers at a level of 1.23x109 CFU/kg does not affect fattening performance and does not have a negative effect on the development of the gas-trointestinal tract. However, it might affect enzyme biosynthesis and secretion from pancreas indirectly

The effects of Enterococcus faecium Cernelle 68 (SF 68) on output properties and some haematological parameters in broilers

To investigate the effects of Enterococcus faecium Cernelle 68 on output properties and some haematological and biochemical properties in broiler chicks. 130,male Ross-308 broiler chick's were, used. The animals were divided into two groups as control and experimental 35 mg/kg Enterococcus faecium Cernelle 68 was supplemented to diet of the experimental group. Body weight gain. food consumption and feed efficiency ratio were determined on day 14, 28, 42 and 49. Also on the same days, red blood cell (RBC), white blood, cell (WBC), thrombocyte counts, packed cell,volume (PCV), haemoglobin (Hb) amount and serum creatine,kinase (CK), lactate dehydrogenase (LDH), aspartate aminotransferase (AST), alanine aminotransferase (ALT), total protein, albumin, triglyceride, cholesterol, and glucose levels were determined from the blood samples of randomly selected 15 animals from both control and experimental groups. After blood samples were taken, carcass weight, small intestine weight and ileum pH of the animals were determined during autopsy. It has been determined that, food consumption was less in probiotic group than in control, and feed efficiency ratio of the probiotic group was higher than that of the control group, but these differences were not statistically significant. The carcass weight of the probiotic group was higher on day 49 (P < 0,05). It has been determined that, the probiotic had no effect on small intestine weight and ileum pH.,No statistically significant change was observed in RBC, WBC, thrombocyte, PCV, and Hb values. AST and ALT levels of the probiotic group decreased statistically on day 49 (P < 0,05). The cholesterol level of the probiotic group was statistically lower than that of the control group on day 14 (P < 0,05). These results show that Enterococcus faecium Cernelle 68 will bring economic advantage to the breeders by improving feed efficiency ratio and carcass weight, and it is safe for the host animal and it is well tolerated by the organism

Effects of Saccharomyces cerevisiae extract on haematological parameters, immune function and the antioxidant defence system in breeder hens fed aflatoxin contaminated diets

1. The study was conducted to investigate the efficacy of Saccharomyces cerevisiae extract (SC) on haematological parameters, immune function, and the antioxidant defence system in breeder hens fed a diet contaminated with low level aflatoxin (AF)

Freezing and storage of leukodepleted erythrocyte suspensions

Studies on the frozen storage of human blood products have benefited veterinary transfusion medicine in recent years, but the long-term cryopreservation of canine red blood cells (RBCs) has not yet been thoroughly investigated. Further, no studies are available with respect to the frozen storage of leukocyte-depleted canine red blood cells (LD-RBCs). The objective of the current study was to investigate time-dependent effects of long-term frozen storage on leukocyte-depleted canine RBCs. Twelve healthy adult dogs meeting the criteria for blood transfusion were used in the study. Whole blood samples (450 +/- 45 ml) collected from each dog were centrifuged for 5 min at 22 degrees C and 4200 x g in a cryogenic microcentrifuge and concentrated RBC (pRBC) suspensions were obtained. Leukocyte depletion was achieved by filtration (2.6 log(10)). Then, the filtrated samples were prewashed three times in 0.9% NaCl solution and were allocated into three subgroups to be evaluated at three different time points (Day 0, Month 4 and Month 6). The samples for cryopreservation were subjected to glycerolisation and then stored at -80 degrees C for 4- and 6-month periods. At the end of this period pRBC units were thawed by manual agitation in a water bath maintained at 36-38 degrees C, centrifuged and then washed in a consecutive series of 12%, 1.6% and 0.9% of NaCl + 0.2 dextrose solutions. 2,3-Diphosphoglycerate (2,3-DPG), adenosine triphosphate (ATP), supernatant haemoglobin (SupHb), sodium (Na+) and potassium (K+) levels, residual glycerol concentrations and haemograms of thawed and deglycerolised pRBC samples were evaluated together with those of Day 0. Sterility tests were performed on all samples for bacterial contamination. No statistically significant differences were noted except for Hct and SupHb levels. No bacterial contamination was noted in any of the samples on the basis of sterility tests. It was found that the described glycerolisation procedure could be a method of choice in the cryopreservation of leukocyte-depleted pRBCs (LD-pRBCs) since no negative effect was observed on the quality of the products and long-term frozen storage did not cause RBC destruction

Erythropoietin Protects the Kidney by Regulating the Effect of TNF-α in L-NAME-Induced Hypertensive Rats.

Background/Aims: Hypertension is the leading cause of death worldwide. Chronic high blood pressure induces inflammation. Tumor necrosis factor (TNF)-α plays a major role in inflammation and also depresses the synthesis of erythropoietin, which exerts protective effects on tissue; however, the mechanism is still unclear. We investigated the protective effect of erythropoietin against tissue damage caused by hypertension in the kidney and whether this effect was suppressed by TNF-α. Methods: First, we detected the optimum chronic dose for darbepoetin-α (Depo), which is a long-acting erythropoietin analog for rats. We separated 60 female adult rats into 6 groups: control, Nω-nitro-L-arginine methyl ester hydrochloride (L-NAME), L-NAME+Depo, L-NAME+Remicade (an anti-TNF-α antibody), L-NAME+Depo+Remicade, Depo, and control. After 1 month of treatment, we measured cardiovascular parameters, took blood samples, sacrificed the rats, and removed kidneys for analyses. Results: The apoptotic index and the plasma and kidney mRNA levels of TNF-α increased in the L-NAME group and decreased in all other treatment groups. Macrophage accumulation increased in the L-NAME and L-NAME+Remicade groups, while it decreased in the Depo group. The mRNA abundance of TNF receptor 1 (TNFR1) decreased slightly in the Depo group and TNFR2 increased significantly in the same group. Conclusion: Erythropoietin protects kidney tissue against hypertension by preventing the apoptotic effects of TNF-α by blocking macrophage accumulation, decreasing TNF-α levels, and switching the TNF-α receptors from the apoptotic receptor TNFR1 to the proliferative receptor TNFR2