Search results for: blood
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 367

Search results for: blood

217 Anti-Diabetic Effect of Bryophyllum pinnatum Leaves

Authors: E. F. Aransiola, M.O. Daramola, E. O. Iwalewa, A. M. Seluwa, O. O. Olufowobi

Abstract:

Diabetes is a chronic metabolic disorder that affects the quality of life in terms of physical health, social and psychological well-being. In spite of the enormous progress in the treatment of diabetes using existing commercial drugs, such as, insulin and oral hypoglycemic agents, the quest and search for new drugs is imperative due to several limitations of the commercial drugs. In addition, the existing diabetic drugs are expensive and unaffordable by the rural populace in the developing countries. The present study demonstrates the anti-diabetic property of aqueous extract of Bryophyllum pinnatum (BP) leaves using diabetic rats (albino rats) as models. At the same time, the anti-diabetic effect of the aqueous extract was compared to that of a sample containing a mixture of the extract and a commercial diabetic medicine, glibenclamide. A specified dosage of aqueous extract of Bryophyllum pinnatum (BP) leaves was administered on the experimental diabetic rats, and their BGL was measured and recorded. The results showed a significant drop in the BGL of the diabetic rats to a value close to normal blood glucose level within 120 minutes when only aqueous extract from BP leaves was used. When a sample containing a mixture of the aqueous extract and glibenclamide was administered, a further drop in BGL was observed. Therefore, the results reveal that aqueous extract of Bryophyllum pinnatum leaves have significant anti-diabetic properties, and that the performance of the existing drugs (glibenclamide) could be enhanced with the use of the aqueous extract.

Keywords: Anti-diabetics, Bryophyllum pinnatum, Blood glucose level, albino rats.

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216 The Effect of X-Ray on Plasma and Erythrocyte Concentration of Zn and Cu in Radiology Staff of Tehran Oil Hospital

Authors: L. Nekoozad, M. Salehi Barough, B. Salmasian

Abstract:

Introduction: Some parameters should be considered to investigate the chronic effects of radiation absorption in radiation workers. Trace elements are parameters which small changes in them can cause significant effects on live systems. The role of trace element concentration in human health is significant. These elements play an important role in the developing and functioning of the immune system, cellular respiration, and oxidation processes. Considering the importance and necessity of this issue and few studies, measurements of concentration changes of these elements due to the absorbed dose are important. Purpose: This study aimed to determine the biological effects of occupational dose absorption on plasma and erythrocyte concentration of Zn and Cu in the radiology staff of Tehran Oil Hospital. Material and methods: In this analytical-comparative study, 72 people have entered. 36 people (18 males and 18 females) were selected as radiology staff in the diagnostic and therapeutic departments of Tehran Oil Hospital. And 36 people (18 males and 18 females) were selected as general section staff in the same hospital as a control group. Radiology and control groups’ age and sex were matched. 10 ml of venous blood was taken from all people.  An atomic absorption spectrometer was used to obtain zinc and copper plasma concentrations. Levine test was used to compare these results validity. Results: The mean concentrations of copper and zinc were measured as 0.951 and 0.754 mg/L in the plasma phase and 3.2  and 0.401 mg/L in the RBC phase for the radiology group.  Copper and zinc average concentrations, respectively 0.976 and 0.813 mg/L in the plasma phase and 2.906 and 0.476 mg/L in the RBC phase, were measured for the control group. These elements Concentrations in the plasma phase were significantly different from that of the control group, but the concentrations in the red blood cell phase did not show a significant difference compared to the control group. In comparison, a separate comparison between men and women in the experimental and control groups showed a significant difference in the values of the elements mentioned. With a significant increase in samples, a better justification than the available statistical results can be extracted. Conclusions: Within this study results, chronic occupational probabilistic absorption destructive effects (even within the permitted range) on blood trace element concentration have been confirmed.

Keywords: Chronic absorption, atomic absorption spectrometry, radiology staff, trace element concentration.

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215 Shannon-Weaver Biodiversity of Neutrophils in Fractal Networks of Immunofluorescence for Medical Diagnostics

Authors: N.E.Galich

Abstract:

We develop new nonlinear methods of immunofluorescence analysis for a sensitive technology of respiratory burst reaction of DNA fluorescence due to oxidative activity in the peripheral blood neutrophils. Histograms in flow cytometry experiments represent a fluorescence flashes frequency as functions of fluorescence intensity. We used the Shannon-Weaver index for definition of neutrophils- biodiversity and Hurst index for definition of fractal-s correlations in immunofluorescence for different donors, as the basic quantitative criteria for medical diagnostics of health status. We analyze frequencies of flashes, information, Shannon entropies and their fractals in immunofluorescence networks due to reduction of histogram range. We found the number of simplest universal correlations for biodiversity, information and Hurst index in diagnostics and classification of pathologies for wide spectra of diseases. In addition is determined the clear criterion of a common immunity and human health status in a form of yes/no answers type. These answers based on peculiarities of information in immunofluorescence networks and biodiversity of neutrophils. Experimental data analysis has shown the existence of homeostasis for information entropy in oxidative activity of DNA in neutrophil nuclei for all donors.

Keywords: blood and cells fluorescence in diagnostics ofdiseases, cytometric histograms, entropy and information in fractalnetworks of oxidative activity of DNA, long-range chromosomalcorrelations in living cells.

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214 Understanding the Nature of Blood Pressure as Metabolic Syndrome Component in Children

Authors: Mustafa M. Donma, Orkide Donma

Abstract:

Pediatric overweight and obesity need attention because they may cause morbid obesity, which may develop metabolic syndrome (MetS). Criteria used for the definition of adult MetS cannot be applied for pediatric MetS. Dynamic physiological changes that occur during childhood and adolescence require the evaluation of each parameter based upon age intervals. The aim of this study is to investigate the distribution of blood pressure (BP) values within diverse pediatric age intervals and the possible use and clinical utility of a recently introduced Diagnostic Obesity Notation Model Assessment Tension (DONMA tense) Index derived from systolic BP (SBP) and diastolic BP (DBP) [SBP+DBP/200]. Such a formula may enable a more integrative picture for the assessment of pediatric obesity and MetS due to the use of both SBP and DBP. 554 children, whose ages were between 6-16 years participated in the study; the study population was divided into two groups based upon their ages. The first group comprises 280 cases aged 6-10 years (72-120 months), while those aged 10-16 years (121-192 months) constituted the second group. The values of SBP, DBP and the formula (SBP+DBP/200) covering both were evaluated. Each group was divided into seven subgroups with varying degrees of obesity and MetS criteria. Two clinical definitions of MetS have been described. These groups were MetS3 (children with three major components), and MetS2 (children with two major components). The other groups were morbid obese (MO), obese (OB), overweight (OW), normal (N) and underweight (UW). The children were included into the groups according to the age- and sex-based body mass index (BMI) percentile values tabulated by WHO. Data were evaluated by SPSS version 16 with p < 0.05 as the statistical significance degree. Tension index was evaluated in the groups above and below 10 years of age. This index differed significantly between N and MetS as well as OW and MetS groups (p = 0.001) above 120 months. However, below 120 months, significant differences existed between MetS3 and MetS2 (p = 0.003) as well as MetS3 and MO (p = 0.001). In comparison with the SBP and DBP values, tension index values have enabled more clear-cut separation between the groups. It has been detected that the tension index was capable of discriminating MetS3 from MetS2 in the group, which was composed of children aged 6-10 years. This was not possible in the older group of children. This index was more informative for the first group. This study also confirmed that 130 mm Hg and 85 mm Hg cut-off points for SBP and DBP, respectively, are too high for serving as MetS criteria in children because the mean value for tension index was calculated as 1.00 among MetS children. This finding has shown that much lower cut-off points must be set for SBP and DBP for the diagnosis of pediatric MetS, especially for children under-10 years of age. This index may be recommended to discriminate MO, MetS2 and MetS3 among the 6-10 years of age group, whose MetS diagnosis is problematic.

Keywords: Blood pressure, children, index, metabolic syndrome, obesity.

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213 Nutrient Modelling to Fabricate Dairy Milk Constituents: Let Milk Serve More Than a Food Item

Authors: M.Aasif Shahzad, N.Mukhtar, M.Sarwar

Abstract:

Dietary macro and micro nutrients in their respective proportion and fractions present a practical potential tool to fabricate milk constituents since cells of lactating mammary glands obtain about 80 % of milk synthesis nutrients from blood, reflecting the existence of an isotonic equilibrium between blood and milk. Diverting milk biosynthetic activities through manipulation of nutrients towards producing milk not only keeping in view its significance as natural food but also as food item which prevents or dilutes the adverse effects of some diseases (like cardiovascular problem by saturated milk fat intake) has been area of interest in the last decade. Nutritional modification / supplementation has been reported to enhance conjugated linoleic acid, fatty acid type and concentration, essential fatty acid concentration, vitamin B12& C, Se, Cu, I and Fe which are involved to counter the health threats to human well being. Synchronizing dietary nutrients aimed to modify rumen dynamics towards synthesis of nutrients or their precursors to make their drive towards formulated milk constituents presents a practical option. Formulating dietary constituents to design milk constituents will let the farmers, consumers and investors know about the real potential and profit margins associated with this enterprise. This article briefly recapitulates the ways and means to modify milk constituents keeping an eye on human health and well being issues, which allows milk to serve more than a food item.

Keywords: Nutritional modification, fabricating milk composition, human health.

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212 Enhanced Differentiation of Stromal Cells and Embryonic Stem Cells with Vitamin D3

Authors: Mayada Alqaisi, Nasser Al-Shanti, Quiyu Wang, William S. Gilmore

Abstract:

In-vitro mouse co-culture of E14 embryonic stem cells (ESCs) and OP9 stromal cells can recapitulate the earliest stages of haematopoietic development, not accessible in human embryos, supporting both haemogenic precursors and their primitive haematopoietic progeny. 1α, 25-Dihydroxy-vitamin D3 (VD3) has been demonstrated to be a powerful differentiation inducer for a wide variety of neoplastic cells, and could enhance early differentiation of ESCs into blood cells in E14/OP9 co-culture. This study aims to ascertain whether VD3 is key in promoting differentiation and suppressing proliferation, by separately investigating the effects of VD3 on the proliferation phase of the E14 cell line and on stromal OP9 cells.The results showed that VD3 inhibited the proliferation of the cells in a dose-dependent manner, quantitatively by decreased cell number, and qualitatively by alkaline-phosphatase staining that revealed significant differences between VD3-treated and untreated cells, characterised by decreased enzyme expression (colourless cells). Propidium-iodide cell-cycle analyses showed no significant percentage change in VD3-treated E14 and OP9 cells within their G and S-phases, compared to the untreated controls, despite the increased percentage of G-phase compared to the S-phase in a dosedependent manner. These results with E14 and OP9 cells indicate that adequate VD3 concentration enhances cellular differentiation and inhibits proliferation. The results also suggest that if E14 and OP9 cells were co-cultured andVD3-treated, there would be furtherenhanced differentiation of ESCs into blood cells.

Keywords: Differentiation, embryonic stem cells, OP9 stromal cells, , 25-dihydroxy-vitamin D3

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211 Test Method Development for Evaluation of Process and Design Effect on Reinforced Tube

Authors: Cathal Merz, Gareth O’Donnell

Abstract:

Coil reinforced thin-walled (CRTW) tubes are used in medicine to treat problems affecting blood vessels within the body through minimally invasive procedures. The CRTW tube considered in this research makes up part of such a device and is inserted into the patient via their femoral or brachial arteries and manually navigated to the site in need of treatment. This procedure replaces the requirement to perform open surgery but is limited by reduction of blood vessel lumen diameter and increase in tortuosity of blood vessels deep in the brain. In order to maximize the capability of these procedures, CRTW tube devices are being manufactured with decreasing wall thicknesses in order to deliver treatment deeper into the body and to allow passage of other devices through its inner diameter. This introduces significant stresses to the device materials which have resulted in an observed increase in the breaking of the proximal segment of the device into two separate pieces after it has failed by buckling. As there is currently no international standard for measuring the mechanical properties of these CRTW tube devices, it is difficult to accurately analyze this problem. The aim of the current work is to address this discrepancy in the biomedical device industry by developing a measurement system that can be used to quantify the effect of process and design changes on CRTW tube performance, aiding in the development of better performing, next generation devices. Using materials testing frames, micro-computed tomography (micro-CT) imaging, experiment planning, analysis of variance (ANOVA), T-tests and regression analysis, test methods have been developed for assessing the impact of process and design changes on the device. The major findings of this study have been an insight into the suitability of buckle and three-point bend tests for the measurement of the effect of varying processing factors on the device’s performance, and guidelines for interpreting the output data from the test methods. The findings of this study are of significant interest with respect to verifying and validating key process and design changes associated with the device structure and material condition. Test method integrity evaluation is explored throughout.

Keywords: Buckling, coil reinforced thin-walled tubes, fracture, test method.

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210 Coalescence of Insulin and Triglyceride/High Density Lipoprotein Cholesterol Ratio for the Derivation of a Laboratory Index to Predict Metabolic Syndrome in Morbid Obese Children

Authors: Orkide Donma, Mustafa M. Donma

Abstract:

Morbid obesity is a health threatening condition particularly in children. Generally, it leads to the development of metabolic syndrome (MetS) characterized by central obesity, elevated fasting blood glucose (FBG), triglyceride (TRG), blood pressure values and suppressed high density lipoprotein cholesterol (HDL-C) levels. However, some ambiguities exist during the diagnosis of MetS in children below 10 years of age. Therefore, clinicians are in the need of some surrogate markers for the laboratory assessment of pediatric MetS. In this study, the aim is to develop an index, which will be more helpful during the evaluation of further risks detected in morbid obese (MO) children. A total of 235 children with normal body mass index (N-BMI), with varying degrees of obesity; overweight (OW), obese (OB), MO as well as MetS participated in this study. The study was approved by the Institutional Ethical Committee. Informed consent forms were obtained from the parents of the children. Obesity states of the children were classified using BMI percentiles adjusted for age and sex. For the purpose, tabulated data prepared by WHO were used. MetS criteria were defined. Systolic and diastolic blood pressure values were measured. Parameters related to glucose and lipid metabolisms were determined. FBG, insulin (INS), HDL-C, TRG concentrations were determined. Diagnostic Obesity Notation Model Assessment Laboratory (DONMALAB) Index [ln TRG/HDL-C*INS] was introduced. Commonly used insulin resistance (IR) indices such as Homeostatic Model Assessment for IR (HOMA-IR) as well as ratios such as TRG/HDL-C, TRG/HDL-C*INS, HDL-C/TRG*INS, TRG/HDL-C*INS/FBG, log, and ln versions of these ratios were calculated. Results were interpreted using statistical package program (SPSS Version 16.0) for Windows. The data were evaluated using appropriate statistical tests. The degree for statistical significance was defined as 0.05. 35 N, 20 OW, 47 OB, 97 MO children and 36 with MetS were investigated. Mean ± SD values of TRG/HDL-C were 1.27 ± 0.69, 1.86 ± 1.08, 2.15 ± 1.22, 2.48 ± 2.35 and 4.61 ± 3.92 for N, OW, OB, MO and MetS children, respectively. Corresponding values for the DONMALAB index were 2.17 ± 1.07, 3.01 ± 0.94, 3.41 ± 0.93, 3.43 ± 1.08 and 4.32 ± 1.00. TRG/HDL-C ratio significantly differed between N and MetS groups. On the other hand, DONMALAB index exhibited statistically significant differences between N and all the other groups except the OW group. This index was capable of discriminating MO children from those with MetS. Statistically significant elevations were detected in MO children with MetS (p < 0.05). Multiple parameters are commonly used during the assessment of MetS. Upon evaluation of the values obtained for N, OW, OB, MO groups and for MO children with MetS, the [ln TRG/HDL-C*INS] value was unique in discriminating children with MetS.

Keywords: Children, index, laboratory, metabolic syndrome, obesity.

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209 Dry Needling Treatment in 38 Cases of Chronic Sleep Disturbance

Authors: P. Gao, Z. Q. Li, Y. G. Jin

Abstract:

In the past 10 years, computers and cellphones have become one of the most important factors in our lives, and one which has a tremendously negative impact on our muscles. Muscle tension may be one of the causes of sleep disturbance. Tension in the shoulders and neck can affect blood circulation to the muscles. This research uses a dry needling treatment to reduce muscle tension in order to determine if the strain in the head and shoulders can influence sleep duration. All 38 patients taking part in the testing suffered from tinnitus and have been experiencing disturbed sleep for at least one to five years. Even after undergoing drug therapy treatments and traditional acupuncture therapies, their sleep disturbances have not shown any improvement. After five to 10 dry needling treatments, 24 of the patients reported an improvement in their sleep duration. Five patients considered themselves to be completely recovered, while 12 patients experienced no improvement. This study investigated these pathogenic and therapeutic problems. The standard treatment for sleep disturbances is drug-based therapy; the results of most standard treatments are unfortunately negative. The result of this clinical research has demonstrated that: The possible cause of sleep disturbance for a lot of patients is the result of tensions in the neck and shoulder muscles. Blood circulation to those muscles is also influenced by the duration of sleep. Hypertonic neck and shoulder muscles are considered to impact sleeping patterns and lead to disturbed sleep. Poor posture, often adopted while speaking on the phone, is one of the main causes of hypertonic neck and shoulder muscle problems. The dry needling treatment specifically focuses on the release of muscle tension.

Keywords: Dry needling, sleep disturbance, sleep duration, muscle tension, trigger points.

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208 Growth Performance and Economy of Production of Pullets Fed on Different Energy Based Sources

Authors: O. A. Anjola, M. A. Adejobi, A. Ogunbameru, F. P. Agbaye, R. O. Odunukan

Abstract:

This experiment was conducted for 8 weeks to evaluate the growth performance and economics of pullets fed on different dietary energy sources. A total of 300 Harco black was used for this experiment. The birds were completely randomized and divided into four diet treatment groups. Each treatment group had three replicates of twenty-five birds per replicate. Four diets containing maize, spaghetti, noodles, and biscuit was formulated to represent diet 1, 2, 3 and 4 respectively. Diet 1 containing maize is the control, while diet 2, 3, and 4 contains spaghetti, noodles, and biscuit waste meal at 100% replacement for maize on weight for weight basis. Performance indices on Feed intake, body weight, weight gain, feed conversion ratio (FCR) and economy of production were measured. Blood samples were also collected for heamatology and serum biochemistry assessment. The result of the experiment indicated that different dietary energy source fed to birds significantly (P < 0.05) affect feed intake, body weight, weight gain, and feed conversion ratio (FCR). The best cost of feed per kilogram of body weight gain was obtained in Spaghetti based diet (₦559.30). However, the best performance were obtained from diet 1(maize), it can be concluded that spaghetti as a replacement for maize in diet of pullet is most economical and profitable for production without any deleterious effects attached. Blood parameters of birds were not significantly (p > 0.05) influenced by the use of the dietary energy sources used in this experiment.

Keywords: Growth performance, spaghetti, noodles, biscuit, profit, hematology and serum biochemistry.

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207 On the Design of Shape Memory Alloy Locking Mechanism: A Novel Solution for Laparoscopic Ligation Process

Authors: Reza Yousefian, Michael A. Kia, Mehrdad Hosseini Zadeh

Abstract:

The blood ducts must be occluded to avoid loss of blood from vessels in laparoscopic surgeries. This paper presents a locking mechanism to be used in a ligation laparoscopic procedure (LigLAP I), as an alternative solution for a stapling procedure. Currently, stapling devices are being used to occlude vessels. Using these devices may result in some problems, including injury of bile duct, taking up a great deal of space behind the vessel, and bile leak. In this new procedure, a two-layer suture occludes a vessel. A locking mechanism is also required to hold the suture. Since there is a limited space at the device tip, a Shape Memory Alloy (SMA) actuator is used in this mechanism. Suitability for cleanroom applications, small size, and silent performance are among the advantages of SMA actuators in biomedical applications. An experimental study is conducted to examine the function of the locking mechanism. To set up the experiment, a prototype of a locking mechanism is built using nitinol, which is a nickel-titanium shape memory alloy. The locking mechanism successfully locks a polymer suture for all runs of the experiment. In addition, the effects of various surface materials on the applied pulling forces are studied. Various materials are mounted at the mechanism tip to compare the maximum pulling forces applied to the suture for each material. The results show that the various surface materials on the device tip provide large differences in the applied pulling forces.

Keywords: Laparoscopic surgery, ligation process, locking mechanism, Shape Memory Alloy (SMA) actuator.

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206 An Index for the Differential Diagnosis of Morbid Obese Children with and without Metabolic Syndrome

Authors: Mustafa M. Donma, Orkide Donma

Abstract:

Metabolic syndrome (MetS) is a severe health problem caused by morbid obesity, the severest form of obesity. The components of MetS are rather stable in adults. However, the diagnosis of MetS in morbid obese (MO) children still constitutes a matter of discussion. The aim of this study was to develop a formula, which facilitated the diagnosis of MetS in MO children and was capable of discriminating MO children with and without MetS findings. The study population comprised MO children. Age and sex-dependent body mass index (BMI) percentiles of the children were above 99. Increased blood pressure, elevated fasting blood glucose (FBG), elevated triglycerides (TRG) and/or decreased high density lipoprotein cholesterol (HDL-C) in addition to central obesity were listed as MetS components for each child. Two groups were constituted. In the first group, there were 42 MO children without MetS components. Second group was composed of 44 MO children with at least two MetS components. Anthropometric measurements including weight, height, waist and hip circumferences were performed during physical examination. BMI and homeostatic model assessment of insulin resistance (HOMA-IR) values were calculated. Informed consent forms were obtained from the parents of the children. Institutional Non-Interventional Clinical Studies Ethics Committee approved the study design. Routine biochemical analyses including FBG, insulin (INS), TRG, HDL-C were performed. The performance and the clinical utility of Diagnostic Obesity Notation Model Assessment Metabolic Syndrome Index (DONMA MetS index) [(INS/FBG)/(HDL-C/TRG)*100] was tested. Appropriate statistical tests were applied to the study data. p value smaller than 0.05 was defined as significant. MetS index values were 41.6 ± 5.1 in MO group and 104.4 ± 12.8 in MetS group. Corresponding values for HDL-C values were 54.5 ± 13.2 mg/dl and 44.2 ± 11.5 mg/dl. There was a statistically significant difference between the groups (p < 0.001). Upon evaluation of the correlations between MetS index and HDL-C values, a much stronger negative correlation was found in MetS group (r = -0.515; p = 0.001) in comparison with the correlation detected in MO group (r = -0.371; p = 0.016). From these findings, it was concluded that the statistical significance degree of the difference between MO and MetS groups was highly acceptable for this recently introduced MetS index. This was due to the involvement of all of the biochemically defined MetS components into the index. This is particularly important because each of these four parameters used in the formula is a cardiac risk factor. Aside from discriminating MO children with and without MetS findings, MetS index introduced in this study is important from the cardiovascular risk point of view in MetS group of children.

Keywords: Fasting blood glucose, high density lipoprotein cholesterol, insulin, metabolic syndrome, morbid obesity, triglycerides.

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205 Unsupervised Segmentation Technique for Acute Leukemia Cells Using Clustering Algorithms

Authors: N. H. Harun, A. S. Abdul Nasir, M. Y. Mashor, R. Hassan

Abstract:

Leukaemia is a blood cancer disease that contributes to the increment of mortality rate in Malaysia each year. There are two main categories for leukaemia, which are acute and chronic leukaemia. The production and development of acute leukaemia cells occurs rapidly and uncontrollable. Therefore, if the identification of acute leukaemia cells could be done fast and effectively, proper treatment and medicine could be delivered. Due to the requirement of prompt and accurate diagnosis of leukaemia, the current study has proposed unsupervised pixel segmentation based on clustering algorithm in order to obtain a fully segmented abnormal white blood cell (blast) in acute leukaemia image. In order to obtain the segmented blast, the current study proposed three clustering algorithms which are k-means, fuzzy c-means and moving k-means algorithms have been applied on the saturation component image. Then, median filter and seeded region growing area extraction algorithms have been applied, to smooth the region of segmented blast and to remove the large unwanted regions from the image, respectively. Comparisons among the three clustering algorithms are made in order to measure the performance of each clustering algorithm on segmenting the blast area. Based on the good sensitivity value that has been obtained, the results indicate that moving kmeans clustering algorithm has successfully produced the fully segmented blast region in acute leukaemia image. Hence, indicating that the resultant images could be helpful to haematologists for further analysis of acute leukaemia.

Keywords: Acute Leukaemia Images, Clustering Algorithms, Image Segmentation, Moving k-Means.

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204 Clinical Signs of Neonatal Calves in Experimental Colisepticemia

Authors: Samad Lotfollahzadeh

Abstract:

Escherichia coli (E. coli) is the most isolated bacteria from blood circulation of septicemic calves. Given the prevalence of septicemia in animals and its economic importance in veterinary practice, better understanding of changes in clinical signs following disease, may contribute to early detection of disorder. The present study has been carried out to detect changes of clinical signs in induced sepsis in calves with E. coli. Colisepticemia has been induced in 10 twenty-day old healthy Holstein- Frisian calves with intravenous injection of 1.5 X 109 colony forming units (cfu) of O111:H8 strain of E. coli. Clinical signs including rectal temperature, heart rate, respiratory rate, shock, appetite, sucking reflex, feces consistency, general behavior, dehydration and standing ability were recorded in experimental calves during 24 hours after induction of colisepticemia. Blood culture was also carried out from calves four times during experiment. ANOVA with repeated measure is used to see changes of calves’ clinical signs to experimental colisepticemia, and values of P≤ 0.05 was considered statistically significant. Mean values of rectal temperature and heart rate as well as median values of respiratory rate, appetite, suckling reflex, standing ability and feces consistency of experimental calves increased significantly during study (P<0.05). In the present study median value of shock score was not significantly increased in experimental calves (P> 0.05). The results of present study showed that total score of clinical signs in calves with experimental colisepticemia increased significantly, although score of some clinical signs such as shock did not change significantly.

Keywords: Calves, Clinical signs scoring, E. coli O111:H8, Experimental colisepticemia,

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203 The Importance of Erythrocyte Parameters in Obese Children

Authors: Orkide Donma, M. Metin Donma, Burcin Nalbantoglu, Birol Topcu, Feti Tulubas, Murat Aydin, Tuba Gokkus, Ahmet Gurel

Abstract:

Increasing prevalence of childhood obesity has increased the interest in early and late indicators of gaining weight. Cell blood counts may be indicators of pro-inflammatory states. The aim was to evaluate associations of hematological parameters, including hematocrit (HTC), hemoglobin, blood cell counts and their indices with the degree of obesity in pediatric population. A total of 249; -139 morbidly obese (MO), 82 healthy normal weight (NW) and 28 overweight (OW) children were included into the scope of the study. WHO BMI-for age percentiles were used to form age- and sexmatched groups. Informed consent forms and the Ethics Committee approval were obtained. Anthropometric measurements were performed. Hematological parameters were determined. Statistical analyses were performed using SPSS. The degree for statistical significance was p≤0.05. Significant differences (p=0.000) between waist-to-hip ratios and head-to- neck ratios (hnrs) of MO and NW children were detected. A significant difference between hnrs of OW and MO children (p=0.000) was observed. Red cell distribution width (RDW) was higher in OW children than NW group (p=0.030). Such finding couldn’t be detected between MO and NW groups. Increased RDW was prominent in OW children. The decrease in mean corpuscular hemoglobin concentration (MCHC) values in MO children was sharper than the values in OW children (p=0.006 vs p=0.042) compared to those in NW group. Statistically higher HTC levels were observed between MO-NW (p=0.014), but none between OW-NW. Though the cause-effect relationship between obesity and erythrocyte indices still needs further investigation, alterations in RDW, HTC, MCHC during obesity may be of significance in the early life.

Keywords: Anthropometry, children, erythrocytes, obesity.

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202 Cubic Splines and Fourier Series Approach to Study Temperature Variation in Dermal Layers of Elliptical Shaped Human Limbs

Authors: Mamta Agrawal, Neeru Adlakha, K.R. Pardasani

Abstract:

An attempt has been made to develop a seminumerical model to study temperature variations in dermal layers of human limbs. The model has been developed for two dimensional steady state case. The human limb has been assumed to have elliptical cross section. The dermal region has been divided into three natural layers namely epidermis, dermis and subdermal tissues. The model incorporates the effect of important physiological parameters like blood mass flow rate, metabolic heat generation, and thermal conductivity of the tissues. The outer surface of the limb is exposed to the environment and it is assumed that heat loss takes place at the outer surface by conduction, convection, radiation, and evaporation. The temperature of inner core of the limb also varies at the lower atmospheric temperature. Appropriate boundary conditions have been framed based on the physical conditions of the problem. Cubic splines approach has been employed along radial direction and Fourier series along angular direction to obtain the solution. The numerical results have been computed for different values of eccentricity resembling with the elliptic cross section of the human limbs. The numerical results have been used to obtain the temperature profile and to study the relationships among the various physiological parameters.

Keywords: Blood Mass Flow Rate, Metabolic Heat Generation, Fourier Series, Cubic splines and Thermal Conductivity.

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201 Impact of Standardized Therapeutic Hypothermia Protocol on Neurological Performance after Resuscitation from Cardiac Arrest

Authors: Tahsien Mohamed Okasha, Warda Youssef Mohamed Morsy, Hanan Elsayed Zaghla

Abstract:

We hypothesized that post cardiac arrest patients with Glasgow Coma Scale (GCS) score of less than 8 and who will be exposed to therapeutic hypothermia protocol will exhibit improvement in their neurological performance. 17 subjects were enrolled in this study all over one year. The study was carried out using Quasi-experimental research design. Four tools were used for data collection of this study: Demographic and medical data sheet, Post cardiac arrest health assessment sheet, Bedside Shivering Assessment Scale (BSAS), and Glasgow Pittsburgh cerebral performance category scale (CPC). The mean age was X̅ ± SD = 53 ± 8.122 years, 47.1% were arrested because of cardiac etiology. 35.3% subjects were initially arrested in form of ventricular tachycardia (VT), 23.5% initially arrested in form of ventricular fibrillation (VF), and 29.4% in form of A-Systole. Favorable neurological outcome was seen among 70.6%. There was significant statistical difference in WBC, Platelets, blood gases value, random blood sugar. Also, initial arrest rhythm, etiology of cardiac arrest, and shivering status were significantly correlated with cerebral performance categories score. Therapeutic hypothermia has positive effects on neurological performance among post cardiac arrest patients with GCS score of less than 8. Replication of the study on larger probability sample, with randomized control trial design is recommended with further study for suggesting nursing protocol for patients undergoing therapeutic hypothermia is recommended.

Keywords: Therapeutic hypothermia, neurological performance, after resuscitation from cardiac arrest, initial arrest rhythm.

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200 Pre-Clinical Studying of Antitumor Ramon Preparation: Acute Toxicity

Authors: Raissa A. Muzychkina, Irina M. Korulkina, Dmitriy Yu. Korulkin

Abstract:

In article the data of acute toxicity for pre-clinical researches of Ramon preparation is described. Ramon effects to clinical characteristics of blood, cardio-vascular system, hepatotoxic and diuretic effects were studied.

Keywords: Cancer, toxicity, antitumor activity, pre-clinical testing, anthraquinones, phytopreparation, Ramon.

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199 Relationship between Trauma and Acute Scrotum: Test Torsion and Epididymal Appendix Torsion

Authors: Saimir Heta, Kastriot Haxhirexha, Virtut Velmishi, Nevila Alliu, Ilma Robo

Abstract:

Background: Testicular rotation can occur at any age. The possibility to save the testicle is the fastest possible surgical intervention which is indicated by the presence of acute pain even at rest. The time element is more important to diagnose and proceed further with surgical intervention. Testicular damage is a consequence which mainly depends on the moment of onset of symptoms, at the time when the symptoms are diagnosed, the earliest action to be performed is surgical intervention. Sometimes medical tests are needed to confirm a diagnosis, or to help identify another cause for symptoms; for example, the urine test, that is used to check for infection, associated with the scrotal ultrasound test. Control of blood flow to the longitudinal supply vessels of the testicles is indicated. The sign that indicates testicular rotation is a reduction in blood flow. This is the element which is distinguished from ultrasound examination. Surgery may be needed to determine if the patient’s symptoms are caused by the rotation of the testis or any other condition. Discussion: As a surgical intervention of the emergency, the torsion of the test depends very much on the duration of the torsion, as the success in the life of the testicle depends on the fastest surgical intervention. From the previous clinic, it is noted that in any case presented to the pediatric patient diagnosed with testicular rotation, there is always a link with personal history that the patient refers to the presence of a previous episode of testicular trauma. Literature supports this fact very logically. Conclusions: Salvation without testicular atrophy depends closely on establishing the diagnosis of testicular rotation as soon as possible. Following the logic above, it can be said that the diagnosis for rotation should be performed as soon as possible, to avoid consequences that will not be favorable for the patient.

Keywords: Acute scrotum, testicular torsion, newborns, infants, clinical presentation.

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198 Electronic System Design for Respiratory Signal Processing

Authors: C. Matiz C., N. Olarte L., A. Rubiano F.

Abstract:

This paper presents the design related to the electronic system design of the respiratory signal, including phases for processing, followed by the transmission and reception of this signal and finally display. The processing of this signal is added to the ECG and temperature sign, put up last year. Under this scheme is proposed that in future also be conditioned blood pressure signal under the same final printed circuit and worked.

Keywords: Conditioning, Respiratory Signal, Storage, Teleconsultation.

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197 Clinical Parameters Response to Low-Level Laser versus Monochromatic Near-Infrared Photo Energy in Diabetic Patients with Peripheral Neuropathy

Authors: Abeer A. Abdelhamed

Abstract:

Background: Diabetic sensorimotor polyneuropathy (DSP) is one of the most common microvascular complications of type 2 diabetes. Loss of sensation is thought to contribute to a lack of static and dynamic stability and increased risk of falling. Purpose: The purpose of this study was to compare the effects of low-level laser (LLL) and monochromatic near-infrared photo energy (MIRE) on pain, cutaneous sensation, static stability, and index of lower limb blood flow in diabetic patients with peripheral neuropathy. Methods: Forty diabetic patients with peripheral neuropathy were recruited for participation in this study. They were divided into two groups: The MIRE group, which contained 20 patients, and the LLL group, which contained 20 patients. All patients who participated in the study had been subjected to various physical assessment procedures, including pain, cutaneous sensation, Doppler flow meter, and static stability assessments. The baseline measurements were followed by treatment sessions that were conducted twice a week for six successive weeks. Results: The statistical analysis of the data revealed significant improvement of pain in both groups, with significant improvement in cutaneous sensation and static balance in the MIRE group compared to the LLL group; on the other hand, the results showed no significant differences in lower limb blood flow between the groups. Conclusion: LLL and MIRE can improve painful symptoms in patients with diabetic neuropathy. On the other hand, MIRE is also useful in improving cutaneous sensation and static stability in patients with diabetic neuropathy.

Keywords: Diabetic neuropathy, Doppler flow meter, –Lowlevel laser, Monochromatic near-infrared photo energy.

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196 Pharmaceutical Applications and Clinical Efficiency of Anti-Inflammatory Ramon Preparation

Authors: Dmitriy Yu. Korulkin, Raissa A. Muzychkina

Abstract:

The Ramon preparation is received from a plant; it is destined for external treatment of inflammations in post-surgery period. The Ramon is a biogenic immune stimulator accelerating metabolism, contributing to improvement of blood indexes, having general tonic, anti-inflammatory and bactericidal effect.

Keywords: Anti-inflammatory, anthraquinones, bactericidal activity, Ramon.

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195 The Cooperation among Insulin, Cortisol and Thyroid Hormones in Morbid Obese Children and Metabolic Syndrome

Authors: Orkide Donma, Mustafa M. Donma

Abstract:

Obesity, a disease associated with a low-grade inflammation, is a risk factor for the development of metabolic syndrome (MetS). So far, MetS risk factors such as parameters related to glucose and lipid metabolisms as well as blood pressure were considered for the evaluation of this disease. There are still some ambiguities related to the characteristic features of MetS observed particularly in pediatric population. Hormonal imbalance is also important, and quite a lot information exists about the behaviour of some hormones in adults. However, the hormonal profiles in pediatric metabolism have not been cleared yet. The aim of this study is to investigate the profiles of cortisol, insulin, and thyroid hormones in children with MetS. The study population was composed of morbid obese (MO) children without (Group 1) and with (Group 2) MetS components. WHO BMI-for age and sex percentiles were used for the classification of obesity. The values above 99 percentile were defined as morbid obesity. Components of MetS (central obesity, glucose intolerance, high blood pressure, high triacylglycerol levels, low levels of high density lipoprotein cholesterol) were determined. Anthropometric measurements were performed. Ratios as well as obesity indices were calculated. Insulin, cortisol, thyroid stimulating hormone (TSH), free T3 and free T4 analyses were performed by electrochemiluminescence immunoassay. Data were evaluated by statistical package for social sciences program. p<0.05 was accepted as the degree for statistical significance. The mean ages±SD values of Group 1 and Group 2 were 9.9±3.1 years and 10.8±3.2 years, respectively. Body mass index (BMI) values were calculated as 27.4±5.9 kg/m2 and 30.6±8.1 kg/m2, successively. There were no statistically significant differences between the ages and BMI values of the groups. Insulin levels were statistically significantly increased in MetS in comparison with the levels measured in MO children. There was not any difference between MO children and those with MetS in terms of cortisol, T3, T4 and TSH. However, T4 levels were positively correlated with cortisol and negatively correlated with insulin. None of these correlations were observed in MO children. Cortisol levels in both MO as well as MetS group were significantly correlated. Cortisol, insulin, and thyroid hormones are essential for life. Cortisol, called the control system for hormones, orchestrates the performance of other key hormones. It seems to establish a connection between hormone imbalance and inflammation. During an inflammatory state, more cortisol is produced to fight inflammation. High cortisol levels prevent the conversion of the inactive form of the thyroid hormone T4 into active form T3. Insulin is reduced due to low thyroid hormone. T3, which is essential for blood sugar control- requires cortisol levels within the normal range. Positive association of T4 with cortisol and negative association of it with insulin are the indicators of such a delicate balance among these hormones also in children with MetS.

Keywords: Children, cortisol, insulin, metabolic syndrome, thyroid hormones.

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194 Haematology and Serum Biochemical Profile of Laying Chickens Reared on Deep Litter System with or without Access to Grass or Legume Pasture under Humid Tropical Climate

Authors: E. Oke, A. O. Ladokun, J. O. Daramola, O. M. Onagbesan

Abstract:

There has been a growing interest on the effects of access to pasture on poultry health status. However, there is a paucity of data on the relative benefits of grass and legume pastures. An experiment was conducted to determine the effects of rearing systems {deep litter system (DL), deep litter with access to legumes (LP) or grass (GP) pastures} haematology and serum chemistry of ISA Brown layers. The study involved the use of two hundred and forty 12 weeks old pullets. The birds were reared until 60 weeks of age. Eighty birds were assigned to each treatment; each treatment had four replicates of 20 birds each. Blood samples (2.5 ml) were collected from the wing vein of two birds per replicate and serum chemistry and haematological parameters were determined. The results showed that there were no significant differences between treatments in all the parameters considered at 18 weeks of age. At 24 weeks old, the percentage of heterophyl (HET) in DL and LP were similar but higher than that of GP. The ratio of H:L was higher (P<0.05) in DL than those of LP and GP while LP and GP were comparable. At week 38 of age, the percentage of PCV in the birds in LP and GP were similar but the birds in DL had significantly lower level than that of GP. In the early production phase, serum total protein of the birds in LP was similar to that of GP but higher (P<0.05) than that of DL. At the peak production phase (week 38), the total protein in GP and DL were similar but significantly lower than that of LP. The albumin level in LP was greater (P<0.05) than GP but similar to that of DL. In the late production phase, the total protein in LP was significantly higher than that of DL but similar to that of GP. It was concluded that rearing chickens in either grass or legume pasture did not have deleterious effects on the health of laying chickens but improved some parameters including blood protein and HET/lymphocyte.

Keywords: Rearing systems, Stylosanthes, Cynodon serum chemistry, haematology, hen.

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193 Haemodynamics Study in Subject Specific Carotid Bifurcation Using FSI

Authors: S. M. Abdul Khader, Anurag Ayachit, Raghuvir Pai, K. A. Ahmed, V. R. K. Rao, S. Ganesh Kamath

Abstract:

The numerical simulation has made tremendous advances in investigating the blood flow phenomenon through elastic arteries. Such study can be useful in demonstrating the disease progression and hemodynamics of cardiovascular diseases such as atherosclerosis. In the present study, patient specific case diagnosed with partially stenosed complete right ICA and normal left carotid bifurcation without any atherosclerotic plaque formation is considered. 3D patient specific carotid bifurcation model is generated based on CT scan data using MIMICS-4.0 and numerical analysis is performed using FSI solver in ANSYS-14.5. The blood flow is assumed to be incompressible, homogenous and Newtonian, while the artery wall is assumed to be linearly elastic. The two-way sequentially coupled transient FSI analysis is performed using FSI solver for three pulse cycles. The hemodynamic parameters such as flow pattern, Wall Shear Stress, pressure contours and arterial wall deformation are studied at the bifurcation and critical zones such as stenosis. The variation in flow behavior is studied throughout the pulse cycle. Also, the simulation results reveal that there is a considerable increase in the flow behavior in stenosed carotid in contrast to the normal carotid bifurcation system. The investigation also demonstrates the disturbed flow pattern especially at the bifurcation and stenosed zone elevating the hemodynamics, particularly during peak systole and later part of the pulse cycle. The results obtained agree well with the clinical observation and demonstrates the potential of patient specific numerical studies in prognosis of disease progression and plaque rupture.

Keywords: Fluid-Structure Interaction, arterial stenosis, Wall Shear Stress, Carotid Artery Bifurcation.

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192 Relationship between Iron-Related Parameters and Soluble Tumor Necrosis Factor-Like Weak Inducer of Apoptosis in Obese Children

Authors: Mustafa M. Donma, Orkide Donma, Savas Guzel

Abstract:

Iron is physiologically essential. However, it also participates in the catalysis of free radical formation reactions. Its deficiency is associated with amplified health risks. This trace element establishes some links with another physiological process related to cell death, apoptosis. Both iron deficiency and iron overload are closely associated with apoptosis. Soluble tumor necrosis factor-like weak inducer of apoptosis (sTWEAK) has the ability to trigger apoptosis and plays a dual role in the physiological versus pathological inflammatory responses of tissues. The aim of this study was to investigate the status of these parameters as well as the associations among them in children with obesity, a low-grade inflammatory state. The study was performed on groups of children with normal body mass index (N-BMI) and obesity. 43 children were included in each group. Based upon age- and sex-adjusted BMI percentile tables prepared by the World Health Organization, children whose values varied between 85 and 15 were included in N-BMI group. Children, whose BMI percentile values were between 99 and 95, comprised obese (OB) group. Institutional ethical committee approval and informed consent forms were taken prior to the study. Anthropometric measurements (weight, height, waist circumference, hip circumference, head circumference, neck circumference) and blood pressure values (systolic blood pressure and diastolic blood pressure) were recorded. Routine biochemical analyses, including serum iron, total iron binding capacity (TIBC), transferrin saturation percent (Tf Sat %) and ferritin, were performed. sTWEAK levels were determined by enzyme-linked immunosorbent assay. study data were evaluated using appropriate statistical tests performed by the statistical program SPSS. Serum iron levels were 91 ± 34 mcrg/dl and 75 ± 31 mcrg/dl in N-BMI and OB children, respectively. The corresponding values for TIBC, Tf Sat %, ferritin were 265 mcrg/dl vs. 299 mcrg/dl, 37.2 ± 19.1% vs. 26.7 ± 14.6%, and 41 ± 25 ng/ml vs 44 ± 26 ng/ml. In N-BMI and OB groups, sTWEAK concentrations were measured as 351 ng/L and 325 ng/L, respectively (p > 0.05). Correlation analysis revealed significant associations between sTWEAK levels and iron related parameters (p < 0.05) except ferritin. In conclusion, iron contributes to apoptosis. Children with iron deficiency have decreased apoptosis rate in comparison with that of healthy children. sTWEAK is an inducer of apoptosis. OB children had lower levels of both iron and sTWEAK. Low levels of sTWEAK are associated with several types of cancers and poor survival. Although iron deficiency state was not observed in this study, the correlations detected between decreased sTWEAK and decreased iron as well as Tf Sat % values were valuable findings, which point out decreased apoptosis. This may induce a proinflammatory state, potentially leading to malignancies in the future lives of OB children.

Keywords: Apoptosis, children, iron-related parameters, obesity, soluble tumor necrosis factor-like weak inducer of apoptosis.

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191 Effect of Dietary Supplementation of Different Levels of Black Seed (Nigella Sativa L.) on Growth Performance, Immunological, Hematological and Carcass Parameters of Broiler Chicks

Authors: R. S. Shewita, A. E. Taha

Abstract:

This experiment was conducted to investigate the effect of dietary supplementation of different levels of black seed (Nigella sativa L.) on the performance and immune response of broiler chicks. A total 240 day-old broiler chicks were used and randomly allotted equally into six experimental groups designated as 1, 2, 3, 4, 5 and 6 having black seed at the rate of 0, 2, 4, 6, 8 and 10 g /kg diet respectively. The study was lasted for 42 days. Average body weight, weight gain, relative growth rate, feed conversion, antibody titer against Newcastle disease, phagocytic activity and phagocytic index, some blood parameters(GOT, GPT, Glucose, Cholesterol, Triglyceride, Total protein, Albumen, WBCs, RBCs, Hb and PCV), dressing percentage, weight of different body organs, abdominal fat weight, were determined. It was found that, N. Sativa significantly improved final body weight, total body gain and feed conversion ratio of groups 2 and 3 when compared with the control group. Higher levels of N. Sativa did not improve growth performance of the chicks. Non significant differences were observed for antibody titer against Newcastle virus, WBCs count, serum GOT, glucose level, dressing %, relative liver, spleen, heart and head percentages. Lymphoid organs (Bursa and Thymus) improved significantly with increasing N. Sativa level in all supplemented groups. Serum cholesterol, triglyceride and visible fat % significantly decreased with Nigella sativa supplementation while serum GPT level significantly increased with nigella sativa supplementation.

Keywords: Nigella Sativa, broiler, growth, carcass traits, serum, blood

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190 Neutrophil-to-Lymphocyte Ratio: A Predictor of Cardiometabolic Complications in Morbid Obese Girls

Authors: Mustafa M. Donma, Orkide Donma

Abstract:

Obesity is a low-grade inflammatory state. Childhood obesity is a multisystem disease, which is associated with a number of complications as well as potentially negative consequences. Gender is an important universal risk factor for many diseases. Hematological indices differ significantly by gender. This should be considered during the evaluation of obese children. The aim of this study is to detect hematologic indices that differ by gender in morbid obese (MO) children. A total of 134 MO children took part in this study. The parents filled an informed consent form and the approval from the Ethics Committee of Namik Kemal University was obtained. Subjects were divided into two groups based on their genders (64 females aged 10.2±3.1 years and 70 males aged 9.8±2.2 years; p ≥ 0.05). Waist-to-hip as well as head-to-neck ratios and body mass index (BMI) values were calculated. The children, whose WHO BMI-for age and sex percentile values were > 99 percentile, were defined as MO. Hematological parameters [haemoglobin, hematocrit, erythrocyte count, mean corpuscular volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration, red blood cell distribution width, leukocyte count, neutrophil %, lymphocyte %, monocyte %, eosinophil %, basophil %, platelet count, platelet distribution width, mean platelet volume] were determined by the automatic hematology analyzer. SPSS was used for statistical analyses. P ≤ 0.05 was the degree for statistical significance. The groups included children having mean±SD value of BMI as 26.9±3.4 kg/m2 for males and 27.7±4.4 kg/m2 for females (p ≥ 0.05). There was no significant difference between ages of females and males (p ≥ 0.05). Males had significantly increased waist-to-hip ratios (0.95±0.08 vs 0.91±0.08; p=0.005) and mean corpuscular hemoglobin concentration values (33.6±0.92 vs 33.1±0.83; p=0.001) compared to those of females. Significantly elevated neutrophil (4.69±1.59 vs 4.02±1.42; p=0.011) and neutrophil-to-lymphocyte ratios (1.70±0.71 vs 1.39±0.48; p=0.004) were detected in females. There was no statistically significant difference between groups in terms of C-reactive protein values (p ≥ 0.05). Adipose tissue plays important roles during the development of obesity and associated diseases such as metabolic syndrom and cardiovascular diseases (CVDs). These diseases may cause changes in complete blood cell count parameters. These alterations are even more important during childhood. Significant gender effects on the changes of neutrophils, one of the white blood cell subsets, were observed. The findings of the study demonstrate the importance of considering gender in clinical studies. The males and females may have distinct leukocyte-trafficking profiles in inflammation. Female children had more circulating neutrophils, which may be the indicator of an increased risk of CVDs, than male children within this age range during the late stage of obesity. In recent years, females represent about half of deaths from CVDs; therefore, our findings may be the indicator of the increasing tendency of this risk in females starting from childhood.

Keywords: Children, gender, morbid obesity, neutrophil-to-lymphocyte ratio.

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189 A Novel Multiplex Real-Time PCR Assay Using TaqMan MGB Probes for Rapid Detection of Trisomy 21

Authors: Mehrdad Hashemi, Mitra Behrooz Aghdam, Reza Mahdian, Ahmad Reza Kamyab

Abstract:

Cytogenetic analysis still remains the gold standard method for prenatal diagnosis of trisomy 21 (Down syndrome, DS). Nevertheless, the conventional cytogenetic analysis needs live cultured cells and is too time-consuming for clinical application. In contrast, molecular methods such as FISH, QF-PCR, MLPA and quantitative Real-time PCR are rapid assays with results available in 24h. In the present study, we have successfully used a novel MGB TaqMan probe-based real time PCR assay for rapid diagnosis of trisomy 21 status in Down syndrome samples. We have also compared the results of this molecular method with corresponding results obtained by the cytogenetic analysis. Blood samples obtained from DS patients (n=25) and normal controls (n=20) were tested by quantitative Real-time PCR in parallel to standard G-banding analysis. Genomic DNA was extracted from peripheral blood lymphocytes. A high precision TaqMan probe quantitative Real-time PCR assay was developed to determine the gene dosage of DSCAM (target gene on 21q22.2) relative to PMP22 (reference gene on 17p11.2). The DSCAM/PMP22 ratio was calculated according to the formula; ratio=2 -ΔΔCT. The quantitative Real-time PCR was able to distinguish between trisomy 21 samples and normal controls with the gene ratios of 1.49±0.13 and 1.03±0.04 respectively (p value <0.001). These results represent the presence of 3 copies of target gene in DS samples Vs 2 copies in normal controls. The results of quantitative Real-time PCR were in complete agreement with results of cytogenetic analysis. This study confirms previous reports regarding successful implementation of quantitative Real-time PCR for detection of trisomy 21. However, the assay has been improved by using MGB probes and more accurate data analysis. This assay, in particular, when performed in combination with another molecular assay such as QF-PCR or MLPA, can be used as a reliable technique for rapid prenatal diagnosis of trisomy 21.

Keywords: Trisomy 21, Real-time PCR, MGB-TaqMan Probes, Gene Dosage.

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188 Biomolecules Based Microarray for Screening Human Endothelial Cells Behavior

Authors: Adel Dalilottojari, Bahman Delalat, Frances J. Harding, Michaelia P. Cockshell, Claudine S. Bonder, Nicolas H. Voelcker

Abstract:

Endothelial Progenitor Cell (EPC) based therapies continue to be of interest to treat ischemic events based on their proven role to promote blood vessel formation and thus tissue re-vascularisation. Current strategies for the production of clinical-grade EPCs requires the in vitro isolation of EPCs from peripheral blood followed by cell expansion to provide sufficient quantities EPCs for cell therapy. This study aims to examine the use of different biomolecules to significantly improve the current strategy of EPC capture and expansion on collagen type I (Col I). In this study, four different biomolecules were immobilised on a surface and then investigated for their capacity to support EPC capture and proliferation. First, a cell microarray platform was fabricated by coating a glass surface with epoxy functional allyl glycidyl ether plasma polymer (AGEpp) to mediate biomolecule binding. The four candidate biomolecules tested were Col I, collagen type II (Col II), collagen type IV (Col IV) and vascular endothelial growth factor A (VEGF-A), which were arrayed on the epoxy-functionalised surface using a non-contact printer. The surrounding area between the printed biomolecules was passivated with polyethylene glycol-bisamine (A-PEG) to prevent non-specific cell attachment. EPCs were seeded onto the microarray platform and cell numbers quantified after 1 h (to determine capture) and 72 h (to determine proliferation). All of the extracellular matrix (ECM) biomolecules printed demonstrated an ability to capture EPCs within 1 h of cell seeding with Col II exhibiting the highest level of attachment when compared to the other biomolecules. Interestingly, Col IV exhibited the highest increase in EPC expansion after 72 h when compared to Col I, Col II and VEGF-A. These results provide information for significant improvement in the capture and expansion of human EPC for further application.

Keywords: Cardiovascular disease, cell microarray platform, cell therapy, endothelial progenitor cells, high throughput screening.

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