Search results for: fasting blood glucose

Authors: Anahita Izadyar, My Ni Van, Kayleigh Amber Rodriguez, Ilwoo Seok, Elizabeth E. Hood

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Using a recombinant enzyme derived from corn and a simple modification, we fabricated a facile, fast, and cost-beneficial biosensor to measure glucose. The Nafion/ Plant Produced Mn Peroxidase (PPMP)– glucose oxidase (GOx)- Bovine serum albumin (BSA) /Au electrode showed an excellent amperometric response to detect glucose. This biosensor is capable of responding to a wide range of glucose—20.0 µM−15.0 mM and has a lower detection limit (LOD) of 2.90µM. The reproducibility response using six electrodes is also very substantial and indicates the high capability of this biosensor to detect a wide range of 3.10±0.19µM to 13.2±1.8 mM glucose concentration. Selectivity of this electrode was investigated in an optimized experimental solution contains 10% diet green tea with citrus containing ascorbic acid (AA), and citric acid (CA) in a wide concentration of glucose at 0.02 to 14.0mM with an LOD of 3.10µM. Reproducibility was also investigated using 4 electrodes in this sample and shows notable results in the wide concentration range of 3.35±0.45µM to of 13.0 ± 0.81 mM. We also used other voltammetry methods to evaluate this biosensor. We applied linear sweep voltammetry (LSV) and this technique shows a wide range of 0.10−15.0 mM to detect glucose with a lower detection limit of 19.5µM. The performance and strength of this enzyme biosensor were the simplicity, wide linear ranges, sensitivities, selectivity, and low limits of detection. We expect that the modified biosensor has the potential for monitoring various biofluids.

Keywords: plant-produced manganese peroxidase, enzyme-based biosensors, glucose, modified gold electrode, glucose oxidase

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Authors: Mojdeh Chelemal Dezfoul Nejad, Ali Mohammadzadeh Shobeagar, Mehrzad Mesbah

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This study was conducted in order to characterize the different levels of dietary vitamin D on some of biochemical parameters of Cyprinus carpio. For this purpose, 180 pieces of Cyprinus carpio with an average weight of 20-25 grams were divided into four treatments and each treatment was divided into three replications and treatments were fed at three different doses (1000 IU, 3000 IU, 5000 IU) of vitamin D for 60 days. The fish were fed 3% of their wet b.wt. per day for a 60 days period. Blood samples were obtained from six fish of each tank at the end of experiment. Based on the results significant difference was observed on the mean amount of total protein, urea, glucose and cholesterol between treatments (p < 0.05). But, there was no significant difference in the mean amount of triglyceride and albumin with the different diets designed for this experiment (p > 0.05).

Keywords: Cyprinus carpio, vitamin D, biochemical parameters, glucose

Procedia PDF Downloads 324

Authors: Oluwadele Joshua Femi, Akinlabi Ebenezer Yemi, Onaopemipo Adeitan, Kazeem Bello, Anthony Ekeocha, Miraim Tawose

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This study was carried out to determine the physiological and stress index of scavenging chickens in LAFARGE (Ewekoro) and Dangote (Ibese) Cement Factories Area of Ogun State. One hundred adult scavenging chickens comprising of 25 chickens from LAFARGE, Dangote and respective adjourning communities (Imasayi and Wasimi) were used. Experimental birds were caught at night on their perch and kept in cages till the next morning. Data were collected on rectal temperature, pulse rate, and respiratory rate of the birds. Also, 5ml blood was collected through the wing vein of the chickens in each location using a sterilized needle and syringe and transported to laboratory for analysis. Significant (P<0.05) highest pulse rate (215.64 beat/minute) and respiratory rate (19.90 breaths/minute) were recorded among scavenging chickens at LAFARGE (Ewekoro) Area and the least (198.61 beat/minute and 16.93 breaths/minute, respectively) at Imasayi. There was no significant (P>0.05) difference in the rectal temperature of the birds in the study area. Significant (P<0.05) differences were also recorded in the Packed Cell Volume (PCV), Hemoglobin (Hb), White Blood Cell (WBC), Monocyte, and Glucose level of the chickens in study area with the highest (P<0.05) Packed Cell Volume (28.06%) and Haemoglobin (4.01g/dl) recorded in Ibese and the least Packed Cell Volume (22.00%) and Haemoglobin (288g/dl) in Imasayi. Highest (P<0.05) Monocyte (4.28%) and glucose (256.53g/dl) were recorded among scavenging chickens at Dangote (Ibese) while the least Monocyte (0.00%) and Glucose (194.53g/dl) was recorded among chickens at Wasimi. Highest (P<0.05) White Blood Cell (6488.89×103µl) was recorded among chickens at Ewekoro and the lowest value in Ibese (4388.44×103µl). There was no significant (P>0.05) difference in the Heterophyl, Lymphocyte, Basophyl and Heterophyl/Lymphocyte ratio of the chickens in the study Area. The study concluded that chickens reared at LAFARGE (Ewekoro) were stressed and had comprised welfare and health status compared to Dangote (Ibese) cement area and other agrarian communities. Effective environmental mitigation programme should be put in place to enhance the welfare of the scavenging chickens in LAFARGE Cement Factory Area.

Keywords: blood, chicken, poisonous substances, pack cell volume, communities

Procedia PDF Downloads 60

Authors: Kim H. K. Choy, Oliva H. K. Chu, W. Y. Keung, B. Lim, Winnie P. Y. Tang

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Background: The prevalence of workplace obesity is rising worldwide; therefore, the workplace is an ideal venue to implement weight control intervention. This pilot randomized controlled trial aimed to develop, implement, and evaluate a nutritional wellness program for obese health care providers working in a hospital. Methods: This hospital-based nutritional wellness program was an 8-week pilot randomized controlled trial for obese health care providers. The primary outcomes were body weight and body mass index (BMI). The secondary outcomes were serum fasting glucose, fasting cholesterol, triglyceride, high-density (HDL) and low-density (LDL) lipoprotein, body fat percentage, and body mass. Participants were randomly assigned to the intervention (n = 20) or control (n = 22) group. Participants in both groups received individual nutrition counselling and nutrition pamphlets, whereas only participants in the intervention group were given mobile phone text messages. Results: 42 participants completed the study. In comparison with the control group, the intervention group showed approximately 0.98 kg weight reduction after two months. Participants in intervention group also demonstrated clinically significant improvement in BMI, serum cholesterol level, and HDL level. There was no improvement of body fat percentage and body mass for both intervention and control groups. Conclusion: The nutritional wellness program for obese health care providers was feasible in hospital settings. Health care providers demonstrated short-term weight loss, decrease in serum fasting cholesterol level, and HDL level after completing the program.

Keywords: weight management, weight control, health care providers, hospital

Procedia PDF Downloads 224

Authors: Anju Joshi, C. N. Tharamani

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Diabetes is one of the leading cause of death at present and remains an important concern as the prevalence of the disease is increasing at an alarming rate. Therefore, it is crucial to diagnose the accurate levels of glucose for developing an efficient therapeutic for diabetes. Due to the availability of convenient and compact self-testing, continuous monitoring of glucose is feasible nowadays. Enzyme based electrochemical sensing of glucose is quite popular because of its high selectivity but suffers from drawbacks like complicated purification and immobilization procedures, denaturation, high cost, and low sensitivity due to indirect electron transfer. Hence, designing a robust enzyme free platform using transition metal oxides remains crucial for the efficient and sensitive determination of glucose. In the present work, manganese doped nickel oxide nanoparticles (Mn-NiO) has been synthesized onto the surface of multiwalled carbon nanotubes using a simple microwave assisted approach for non-enzymatic electrochemical sensing of glucose. The morphology and structure of the synthesized nanostructures were characterized using scanning electron microscopy (SEM) and X-Ray diffraction (XRD). We demonstrate that the synthesized nanostructures show enormous potential for electrocatalytic oxidation of glucose with high sensitivity and selectivity. Cyclic voltammetry and square wave voltammetry studies suggest superior sensitivity and selectivity of Mn-NiO decorated carbon nanotubes towards the non-enzymatic determination of glucose. A linear response between the peak current and the concentration of glucose has been found to be in the concentration range of 0.01 μM- 10000 μM which suggests the potential efficacy of Mn-NiO decorated carbon nanotubes for sensitive determination of glucose.

Keywords: diabetes, glucose, Mn-NiO decorated carbon nanotubes, non-enzymatic

Procedia PDF Downloads 206

Authors: Mehdi Kazemi-Bonchenari, Esmaeil Manidari, Vahid Keshavarz

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This study was carried out to investigate the effects of increased level of sulfur by supplementing magnesium sulfate with or without biologically organic source in dairy cow close-up diets on dry matter intake (DMI) and some blood metabolites. The 24 multiparous close-up Holstein cows averaging body weight 687.94 kg and days until expected calving date 21.89 d were allocated in three different treatments (8 cows per each) in a completely randomized design. The first treatment (T1) has contained 0.21% sulfur (DM basis), the second treatment (T2) has contained 0.41% sulfur which entirely supplied through magnesium sulfate and the third treatment (T3) has contained 0.41% sulfur which supplied through combination of magnesium sulfate and an organic source of sulfur. All the cows were fed same diet after parturition until 21 d. The DMI for both pre-calving (P < 0.001) and post-calving was affected by treatments (P < 0.004) and T2 showed the lowest DMI among treatments. Among the blood metabolites, glucose, calcium, and copper were decreased in T2 (P < 0.05). However, blood concentrations of BHBA, NEFA, urea, CPK, and AST were increased in T2 (P < 0.05). The results of the present study indicate that although magnesium sulfate has negative effect on dairy cow health and performance, a combination of magnesium sulfate and biological organic source of sulfur in close-up diets could have positive effects on DMI and performance of Holstein dairy cows.

Keywords: organic sulfur, dairy cow, intake, blood metabolites

Procedia PDF Downloads 277

Authors: Abdullah S. Alghamdi, Khaled Alghamdi, Richard O. Jenkins, Parvez I. Haris

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Background: Unhealthy diet is one of the modifiable risk factors for developing type 2 diabetes mellitus (T2DM). Improvement in diet can be beneficial for countering diabetes. For example, HbA1c, an important biomarker for diabetes, can be reduced by 1.1% through only alteration in diet. Ramadan fasting has been reported to provide positive health benefits. However, optimal benefits are not achieved, often due to poor dietary habits and lifestyle. There is a need to better understand the dietary habits of people fasting during Ramadan, so that necessary improvements can be made to develop this form of fasting as a non-pharmacological strategy for management and prevention of T2DM. Aim: This study aimed to assess the dietary patterns of Saudi adult patients with T2DM over three different periods (before, during, and after Ramadan) and relate this to HbA1c levels. Methods: This study recruited 82 Saudi with T2DM, who chose to fast during Ramadan, from the Endocrine and Diabetic Centre of Al Iman General Hospital, Riyadh, Saudi Arabia. Ethical approvals for the study were obtained from De Montfort University and Saudi Ministry of Health. Dietary patterns were assessed by a self-administered questionnaire in each period. This assessment included the diet type and frequency. Blood samples were collected in each period for determination of HbA1c. Results: The number of meals per day for the participants significantly decreased during Ramadan (P < 0.001). The consumption of fruit and vegetables significantly increased during Ramadan (P = 0.017). However, the consumption of sugary drinks significantly increased during and after Ramadan (P = 0.005). Approximately 60% of the patients indicated that they ate sugary foods at least once per week. The consumption of bread and rice was reported to be at least two times per week. The consumption of rice significantly reduced during Ramadan (P = 0.002). The mean HbA1c significantly varied between periods (P = 0.001), with lowest level during Ramadan compared to before and after Ramadan. The increase in the consumption of fruits and vegetables had a medium effect size on the reduction in HbA1c during Ramadan. There was a variance of 7.7% in the mean difference in HbA1c levels between groups (who changed their fruit and vegetable consumption) which can be accounted for by the increase in the consumption of fruits and vegetables. Likewise, 9.3% of the variance in the mean HbA1c difference between the groups was accounted for by a decrease in the consumption of rice. Conclusion: The increase in the frequency of fruit and vegetables intake, and especially the reduction in the frequency of rice consumption, during Ramadan produce beneficial effects in reducing HbA1c level. Therefore, further improving the dietary habits of patients with T2DM, such as reducing their sugary drinks intake, may help them to obtain greater benefits from Ramadan fasting in the management of their diabetes. It is recommended that dietary guidance is provided to the public to maximise health benefits through Ramadan fasting.

Keywords: Diabetes, Diet, Fasting, HbA1c, Ramadan

Procedia PDF Downloads 138

Authors: Wantanee Polviset, N. Prakobsaeng, N. Wetchakama, C. Yuangklang

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The aim of this study was to evaluate the effect of soybean oil, palm oil and sunflower oil supplementations in total mixed ration on voluntary feed intake, dry matter (DM) digestibility and blood metabolize in crossbred Thai native x American Brahman Cattle. Three Thai native x American Brahman cattle, one-year-old with liveweight of 116±22.59 kg, were randomly assigned according to a 3 x 3 latin square design. Each period of feeding lasted for 21 days to receive three dietary treatments were soybean oil, palm oil and sunflower oil supplementation at 5%. During the experimental periods, all cattle were fed a diet with total mixed ration containing roughage to concentrate ratio of 40:60 and rice straw was used as a roughage source. Based on the present study, the results revealed that voluntary feed intake (kgDM/head/day) and %BW DM intake were not affected (P>0.05), whereas percentage of dry matter digestibility was greater with the soybean oil supplementation (P<0.01). It was also found that blood glucose, blood urea nitrogen, cholesterol, triglyceride, high density lipoprotein and low density lipoprotein in plasma were similar among treatments. Based on this study, supplementing 5% soybean oil in total mixed ration (TMR) diets was suitable in beef cattle without any effect dry matter digestibility and blood metabolites.

Keywords: plant oils, feed intake, blood metabolize, crossbred Thai native x Brahman cattle

Procedia PDF Downloads 285

Authors: A. Bhattacharya, Sanjay Gupta, S. H. Bodakhe

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HbA1c is form of the haemoglobin that is used to measure the average plasma glucose concentration over prolonged periods of time. It is formed in a non-enzymatic glycation pathway by hemoglobin's exposure to plasma glucose. In diabetes mellitus, higher amounts of glycated hemoglobin, indicating poorer control of blood glucose levels, have been associated with cardiovascular disease, nephropathy, and retinopathy. Guraku’s basic components are nicotine and jaggery, jaggery is made up of sugarcane so can have a diabetogenic potential which is exacerbated in presence of nicotine. This work had done with the aim to find correlation between Diabetic cataract, HbA1c and Guraku. Subjects were enrolled according to the inclusion and exclusion criteria. In this study total 75 subjects were included. In the study it was found that people consuming Guraku had a high level of HbA1c thus are more prone to the development of diabetic cataract. Male subjects are the more than female subjects. Most of the subjects belong to the lower socioeconomical class and not very educated. It could be concluded that this type of study could be useful in indentifying number of subjects suffering from diabetic cataract whose condition get worse by use of nicotine product like Guraku and preventive measure to be taken in prevention of this type of diabetic complication.

Keywords: diabetic cataract, HbA1c, Guraku, diabetogenic potential

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Authors: Maha Deif, Alaa Eldin Hassan, Eman Shaat, Nesrine Elazhary, Eman Magdy

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Background: Erythropoietin (EPO) is a hematopoietic factor with multiple protective effects. The aim of the present study was to investigate the potential effect of EPO administration on renal functions and hypoxia inducible factor 1-alpha (HIF-1a) in diabetic rat kidneys. Methodology: The current study was carried out on 40 male albino rats divided into four groups (n= 10 in each). Group I served as normal control, group II was the diabetic control, group III rats received EPO on the same day of diagnosis of diabetes mellitus (DM), while group IV received the first dose of EPO 2 weeks after the diagnosis of DM. Results: The results showed that EPO supplementation leads to a significant decrease in serum urea, urinary protein and creatinine clearance as well as a significant increase in renal HIF-1a in group III and IV rats compared to the diabetic control group (group II). However, fasting blood glucose was significantly decreased in group III as compared to the diabetic control group in the third week, but no significant difference was reported in the fourth week among groups II, III and IV. Conclusion: EPO administration leads to the improvement of renal functions and increased levels of HIF-1a in diabetic rats.

Keywords: erythropoietin, diabetic nephropathy, hypoxia-inducible factor1-alpha, renal functions

Procedia PDF Downloads 261

Authors: Apichai Sawisit, Sirima Suvarnakuta Jantama, Sunthorn Kanchanatawee, Lonnie O. Ingram, Kaemwich Jantama

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Escherichia coli KJ122 was engineered to produce succinate from glucose using the wild type GalP for glucose uptake instead of the native phosphotransferase system (ptsI mutation). This strain ferments 10% (w/v) xylose poorly. Mutants were selected by serial transfers in AM1 mineral salts medium with 10% (w/v) xylose. Evolved mutants exhibited a similar improvement, co-fermentation of an equal mixture of xylose and glucose. One of these, AS1600a, produced 84.26±1.37 g/L succinate, equivalent to that produced by the parent (KJ122) strain from 10% glucose (85.46±1.78 g/L). AS1600a was sequenced and found to contain a mutation in galactose permease (GalP, G236D). Expressing the galP* mutation gene in KJ122ΔgalP resembled the xylose utilization phenotype of the mutant AS1600a. The strain AS1600a and KJ122ΔgalP (pLOI5746; galP*) also co-fermented a mixture of glucose, xylose, arabinose, and galactose in sugarcane bagasse hydrolysate for succinate production.

Keywords: xylose, furfural, succinate, sugarcane bagasse, E. coli

Procedia PDF Downloads 360

Authors: Bahlous Afef, Bouzid Kahena, Bardkis Ahlem, Mrad Mehdi, Kalai Eya, Sonia Bahri, Abdelmoula Jaouida

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Diabetic nephropathy is the first cause of chronic renal failure and hemodialysis use in several countries including Tunisia. The role of hypertension (HT) as major risk factor for nephropathy is undeniable. The aim of our study was to determine the relationship between blood pressure and nephropathy in a population of diabetic type 2 recently discovered. Materials and methods: We conducted a prospective study focused on 60 patients with type 2 diabetes recently discovered (<5 years). Each patient have benefited from: -a full clinical examination with measurement of blood pressure - exploring a blood-glucose control and renal function -urinary exploration with the determination of proteinuria microalbuminumie of 24 hours with a immunoturbidimetric method using Architect (ABBOTT CI 8200). Results and discussion: Hypertension was present in 46.7% of cases. Twenty patients, 35% of the study population showed nephropathy. Four of these patients (6.66% of cases) had proteinuria, while 16 (26.6% of patients) had microalbuminuria (> 30mg/24 hours). Systolic blood pressure was significantly (p < 0.05) associated with the presence of nephropathy (139 +19.44) vs. for the group with normal renal function (128.65 +15.12 mmHg). Conclusion: The etiology of diabetic nephropathy is multifactorial. However, systolic blood pressure and glycemic control remains the major risk factors. Better glycemic control and treatment of hypertension allowed preventing and slowing the progression of diabetic nephropathy.

Keywords: hypertension, nephropathy, hemodialysis, diabetes

Procedia PDF Downloads 292

Authors: Nasiruddin Khan, Nasser M. Al-Daghri, Dara A. Al-Disi, Asim Al-Fadda, Mohamed Al-Seif, Gyanendra Tripathi, A. L. Harte, Philip G. Mcternan

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For the last few decades, the prevalence of type 2 diabetes mellitus (T2DM) in the Kingdom of Saudi Arabia (KSA) is increasing alarmingly high and is unprecedented at 31.6 %. Preventive measures should be taken to curb down the increasing incidence. In this prospective, 3-month study, we aimed to determine whether dietary modification program would confer favorable affects among overweight and obese adult Saudi women with or without T2DM. A total of 92 Saudi women [18 healthy controls, 24 overweight subjects and 50 overweight or obese patients with early onset T2DM were included in this prospective study. Baseline anthropometrics and fasting blood samples were taken at baseline and after 3 months. Fasting blood sugar and lipid profile were measured routinely. A 500 Kcal deficit energy diet less than their daily recommended dietary allowances were prescribed to all participants. After 3 months of follow-up visit, significant improvements were observed in both the overweight and DMT2 group as compared to baseline with decreased mean BMI [Overweight Group 28.54±1.49 versus 27.95±2.25, p<0.05; DMT2 group 35.24±7.67 versus 35.04±8.07, p<0.05] and hip circumference [Overweight group 109.67±5.01 versus 108.07±4.07, p<0.05; DMT2 group 112.3±13.43 versus 109.21±12.71, p<0.01]. Moreover, in the overweight group, baseline HDL-cholesterol was significantly associated with protein intake and inversely associated with carbohydrate intake in controls. In the DMT2 group, carbohydrate intake at baseline was significantly associated with BMI. A 3-month 500kcal/day deficit dietary modification alone is probably effective among adult overweight or obese Saudi females without or with T2DM. Longer prospective studies are to determine whether the dietary intervention alone can reduce progression of T2DM among high-risk adult Arabs.

Keywords: diet, lipid, obesity, T2DM

Procedia PDF Downloads 448

Authors: Saman Ahani, Aboozar Dehghan, Roham Vali, Hamid Salehian, Amin Ebrahimi

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Pulsed-wave (PW) doppler ultrasonography is a non-invasive, relatively accurate imaging technique that can measure blood speed. The imaging could be obtained via the common carotid artery, as one of the main vessels supplying the blood of vital organs. In horses, factors such as susceptibility to depression of the cardiovascular system and their large muscular mass have rendered them vulnerable to changes in blood speed. One of the most important factors causing blood velocity changes is the administration of anesthetic drugs, including Ketamine and Acepromazine. Thus, in this study, the Pulsed-wave doppler technique was performed to assess the highest blood velocity in the common carotid artery following administration of Ketamine and Acepromazine. Six male and six female healthy Kurdish horses weighing 351 ± 46 kg (mean ± SD) and aged 9.2 ± 1.7 years (mean ± SD) were housed under animal welfare guidelines. After fasting for six hours, the normal blood flow velocity in the common carotid artery was measured using a Pulsed-wave doppler ultrasonography machine (BK Medical, Denmark), and a high-frequency linear transducer (12 MHz) without applying any sedative drugs as a control group. The same procedure was repeated after each individual received the following medications: 1.1, 2.2 mg/kg Ketamine (Pfizer, USA), and 0.5, 1 mg/kg Acepromizine (RACEHORSE MEDS, Ukraine), with an interval of 21 days between the administration of each dose and/or drug. The ultrasonographic study was done five (T5) and fifteen (T15) minutes after injecting each dose intravenously. Lastly, the statistical analysis was performed using SPSS software version 22 for Windows and a P value less than 0.05 was considered to be statistically significant. Five minutes after administration of Ketamine (1.1, 2.2 mg/kg) in both male and female horses, the blood velocity decreased to 38.44, 34.53 cm/s in males, and 39.06, 34.10 cm/s in females in comparison to the control group (39.59 and 40.39 cm/s in males and females respectively) while administration of 0.5 mg/kg Acepromazine led to a significant rise (73.15 and 55.80 cm/s in males and females respectively) (p<0.05). It means that the most drastic change in blood velocity, regardless of gender, refers to the latter dose/drug. In both medications and both genders, the increase in doses led to a decrease in blood velocity compared to the lower dose of the same drug. In all experiments in this study, the blood velocity approached its normal value at T15. In another study comparing the blood velocity changes affected by Ketamine and Acepromazine through femoral arteries, the most drastic changes were attributed to Ketamine; however, in this experiment, the maximum blood velocity was observed following administration of Acepromazine via the common carotid artery. Therefore, further experiments using the same medications are suggested using Pulsed-wave doppler measuring the blood velocity changes in both femoral and common carotid arteries simultaneously.

Keywords: Acepromazine, common carotid artery, horse, ketamine, pulsed-wave doppler ultrasonography

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Authors: Ramin Amirmardfar

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Throughout Necessary oxygen should be supplied for all cells of a mammal at any moment through blood to make it possible remain alive all cells the mammal’s body. In case a mammal’s bulk is large, there is a farther distance between cells in different tissues and mammals’ heart. Therefore red blood cells in bulky mammal’s body should be capable of conveying oxygen to farther distances. To make it practical, oxygen should be glued red blood cells tenaciously. In other words, cohesion strength of oxygen to red blood cell of bulky mammal’s blood should be much more than the same of small mammal’s blood. In mammal’s bodies, the controlling factor of amount of cohesion of oxygen to red blood cell, are organic phosphates (like DPG). The less DPG in red blood cells of a mammal, the more cohesion of oxygen to red blood cell (at the same rate). As much as oxygen is glued more tenacious to red blood cells, oxygen could been carried to farther distance and as much as oxygen could be conveyed to farther points of heart, bulk of mammal could be larger at the same rate.

Keywords: mammals size, animals size, organic phosphates, DPG, red blood cell, metabolism

Procedia PDF Downloads 327

Authors: Pramod Somvanshi, Manu Tomar, K. V. Venkatesh

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Insulin and glucagon are responsible for homeostasis of key plasma metabolites like glucose, amino acids and fatty acids in the blood plasma. These hormones act antagonistically to each other during the secretion and signaling stages. In the present work, we analyze the effect of macronutrients on the response from integrated insulin and glucagon signaling pathways. The insulin and glucagon pathways are connected by DAG (a calcium signaling component which is part of the glucagon signaling module) which activates PKC and inhibits IRS (insulin signaling component) constituting a crosstalk. AKT (insulin signaling component) inhibits cAMP (glucagon signaling component) through PDE3 forming the other crosstalk between the two signaling pathways. Physiological level of anabolism and catabolism is captured through a metric quantified by the activity levels of AKT and PKA in their phosphorylated states, which represent the insulin and glucagon signaling endpoints, respectively. Under resting and starving conditions, the phosphorylation metric represents homeostasis indicating a balance between the anabolic and catabolic activities in the tissues. The steady state analysis of the integrated network demonstrates the presence of a bistable response in the phosphorylation metric with respect to input plasma glucose levels. This indicates that two steady state conditions (one in the homeostatic zone and other in the anabolic zone) are possible for a given glucose concentration depending on the ON or OFF path. When glucose levels rise above normal, during post-meal conditions, the bistability is observed in the anabolic space denoting the dominance of the glycogenesis in liver. For glucose concentrations lower than the physiological levels, while exercising, metabolic response lies in the catabolic space denoting the prevalence of glycogenolysis in liver. The non-linear positive feedback of AKT on IRS in insulin signaling module of the network is the main cause of the bistable response. The span of bistability in the phosphorylation metric increases as plasma fatty acid and amino acid levels rise and eventually the response turns monostable and catabolic representing diabetic conditions. In the case of high fat or protein diet, fatty acids and amino acids have an inhibitory effect on the insulin signaling pathway by increasing the serine phosphorylation of IRS protein via the activation of PKC and S6K, respectively. Similar analysis was also performed with respect to input amino acid and fatty acid levels. This emergent property of bistability in the integrated network helps us understand why it becomes extremely difficult to treat obesity and diabetes when blood glucose level rises beyond a certain value.

Keywords: bistability, diabetes, feedback and crosstalk, obesity

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Authors: Muhammad Fiaz Qamar, Ahmad Faraz, Muhammad Arfan Zaman, Kazim Ali, Waleed Akram

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Babesiosis is reflected as the most important disease of cattle that are transmitted by arthropods. In Pakistan, its prevalence is up to 29% in the cattle and buffalo population in different regions. Cattle show a long lasting and durable immunity by giving an infection of B.bovis, B. bigemina, or Babesiadivergens. this is used in cattle to immunize them in a few countries as anti-babesiosis vaccine. Development of frozen vaccine allows for complete testing after production of each batch, However, once thawed, its reduced its shelf life, frozen vaccines are more difficult to transport as well as expensive to produce as compared to chilled vaccine. The contamination of blood derived vaccine has the potential risk that makes pre-production and post-production quality control necessary. For the trail master seed production of whole blood frozen bivalent Babesia(Babesiabovis and Babesiabigemina), 100 blood samples of Babesial positive suspected cattle was taken and processed for separation microscopic detection and rectification by PCR. Vaccine passages were done to reduce the parasitaemiasis in live calves. After 8 passages, parasitemia of Babesia reduced from 80% to 15%. Infected donor calf’s blood was taken by jugular cannulation by using preservative free lithium heparin as an anticoagulant (5 International Units IU heparin/ml blood). In lab, parasite containing blood was mixed in equal volumes with 3 M glycerol in PBS supplemented with 5 mM glucose (final concentration of glycerol 1.5 M) at 37°C. The mixture was then equilibrized at 37°C for 30 minutes and were dispensed in required containers (e.g., 5 ml cryovials).

Keywords: distribution, babesia, primer sequences, PCV

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Authors: Sudkate Chaiyo, Weena Siangproh, Orawon Chailapakul, Kurt Kalcher

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In the present work, a simple and sensitive non-enzymatic electrochemical detection of glucose in disposable paper-based sensor was developed at ionic liquid/graphene/cobalt phthalocyanine composite (IL/G/CoPc) modified electrode. The morphology of the fabricated composite was characterized and confirmed by scanning electron microscopy and UV-Vis spectroscopy. The UV-Vis spectroscopy results confirmed that the G/CoPc composite formed via the strong π–π interaction between CoPc and G. Amperometric i-t technique was used for the determination of glucose. The response of glucose was linear over the concentration ranging from 10 µM to 1.5 mM. The response time of the sensor was found as 30 s with a limit of detection of 0.64 µM (S/N=3). The fabricated sensor also exhibited its good selectivity in the presence of common interfering species. In addition, the fabricated sensor exhibited its special advantages such as low working potential, good sensitivity along with good repeatability and reproducibility for the determination of glucose.

Keywords: glucose, paper-based sensor, ionic liquid/graphene/cobalt phthalocyanine composite, electrochemical detection

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Authors: Ndou Rembuluwani Moddy, Daniel Ter Goon, Takalani Grace Tshitangano, Lindelani Fhumudzani Mushaphi

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Obesity is a global public health problem. The study aimed to determine the risk factors associated with and the consequences of obesity among residents of Tshikota, Makhado Municipality, Limpopo Province, South Africa. A cross-sectional study involving 318 randomly selected adults aged 18-45 years residing at Tshikota, Makhado Local Municipality, South Africa. Sociodemographic information includes age, gender, educational level, occupation, behavioral lifestyle, environmental, psychological, and family history. Anthropometric, blood pressure, and blood glucose measurements followed standard procedure. The prevalence of obesity and overweight was 35.5% and 28.6%, respectively. About 75.2% of obese do not engage in physical activity. Most participants (63.5%) take meals three times a day, and 19.2% do not skip breakfast. Most participants do not have access to fruits and vegetables. Participants who were pre-hypertensive were 92(28.9%) and 32(10.1%) were in Stage 1 hypertension. Of the participants with Class 1 obesity, 40.9% were pre-hypertensive, and 15.2% were in Stage 1 hypertension. In Class 2 obesity, 37.8% were pre-hypertensive, and 26.7% were in Stage 1 hypertension. There was a significant difference between BMI and blood pressure among participants (p=0.00). About 6.1% of the participants in Class 1 obesity were at high risk, and 3.0% were at very high risk of glucose levels. Regarding cholesterol levels, 65 (20.4%) were at borderline, and 17(5.3%) were at high risk. There was no significant difference in BMI and cholesterol levels among participants (p= 0.20). The prevalence of obesity and overweight was high among residents of this setting. Age, marital and educational status, and employment were significantly associated with obesity. An obesity awareness campaign is crucial, and the availability of supermarkets and full-service grocery stores would provide accessibility to healthy food such as fruits and vegetables.

Keywords: obesity, overweight, risk factors, adults.

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Authors: Muhammad Umair Arshad, Saima Ishtiaq, Ali Imran

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Different polysaccharides contribute towards the management of glycemic and satiety and consequently manage the metabolic syndrome. In the present study, we compared the postprandial glycemic and satiety responses of different dietary polysaccharides when added to milk (2% Milk Fat). The objective of this study was to evaluate different polysaccharides against postprandial glucose, appetite responses, and food intake at subsequent meals. In a repeated measures crossover design, 30 females (18–30 years) consumed 250 ml milk with 2% M.F. (control), or milk with carrageenan (2.5 g), guar gum (2.5 g) and alginate (2.5 g), followed by an ad libitum pizza meal after 120 min. Alginate and guar gum addition resulted in lower caloric intake at subsequent pizza meal. The post-treatment (0–120 min) glucose and average appetite were suppressed by alginate and guar gum (p < 0.0001), with a more pronounced effect of guar gum. However, alginate resulted in lower blood glucose (p < 0.0001) compared with control and carrageenan during post-treatment. Alginate and guar gum, added milk, and other beverages would be beneficial in the short-term regulation of postprandial glycemia and satiety.

Keywords: satiety, glycemic control, milk polysaccharides, food intake

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Authors: Bitewulign Mekonnen

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Context: This scientific paper focuses on the use of near-infrared (NIR) spectroscopy to determine glucose concentration in aqueous solutions accurately and rapidly. The study compares six different machine learning methods for predicting glucose concentration and also explores the development of a deep learning model for classifying NIR spectra. The objective is to optimize the detection model and improve the accuracy of glucose prediction. This research is important because it provides a comprehensive analysis of various machine-learning techniques for estimating aqueous glucose concentrations. Research Aim: The aim of this study is to compare and evaluate different machine-learning methods for predicting glucose concentration from NIR spectra. Additionally, the study aims to develop and assess a deep-learning model for classifying NIR spectra. Methodology: The research methodology involves the use of machine learning and deep learning techniques. Six machine learning regression models, including support vector machine regression, partial least squares regression, extra tree regression, random forest regression, extreme gradient boosting, and principal component analysis-neural network, are employed to predict glucose concentration. The NIR spectra data is randomly divided into train and test sets, and the process is repeated ten times to increase generalization ability. In addition, a convolutional neural network is developed for classifying NIR spectra. Findings: The study reveals that the SVMR, ETR, and PCA-NN models exhibit excellent performance in predicting glucose concentration, with correlation coefficients (R) > 0.99 and determination coefficients (R²)> 0.985. The deep learning model achieves high macro-averaging scores for precision, recall, and F1-measure. These findings demonstrate the effectiveness of machine learning and deep learning methods in optimizing the detection model and improving glucose prediction accuracy. Theoretical Importance: This research contributes to the field by providing a comprehensive analysis of various machine-learning techniques for estimating glucose concentrations from NIR spectra. It also explores the use of deep learning for the classification of indistinguishable NIR spectra. The findings highlight the potential of machine learning and deep learning in enhancing the prediction accuracy of glucose-relevant features. Data Collection and Analysis Procedures: The NIR spectra and corresponding references for glucose concentration are measured in increments of 20 mg/dl. The data is randomly divided into train and test sets, and the models are evaluated using regression analysis and classification metrics. The performance of each model is assessed based on correlation coefficients, determination coefficients, precision, recall, and F1-measure. Question Addressed: The study addresses the question of whether machine learning and deep learning methods can optimize the detection model and improve the accuracy of glucose prediction from NIR spectra. Conclusion: The research demonstrates that machine learning and deep learning methods can effectively predict glucose concentration from NIR spectra. The SVMR, ETR, and PCA-NN models exhibit superior performance, while the deep learning model achieves high classification scores. These findings suggest that machine learning and deep learning techniques can be used to improve the prediction accuracy of glucose-relevant features. Further research is needed to explore their clinical utility in analyzing complex matrices, such as blood glucose levels.

Keywords: machine learning, signal processing, near-infrared spectroscopy, support vector machine, neural network

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Authors: Jeonghyun Chang, Taegeun Lee, Heungsup Sung, Yoon-Seon Lee, Youn-Jung Kim, Mi-Na Kim

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Background: Safety device has been applied to improve safety and performance of blood culture practice. BD vacutainer® Safety-Lokᵀᴹ blood collection sets with pre-attached holder (Safety-Lok) (BD, USA) was evaluated in the emergency room (ER) of a tertiary care hospital. Methods: From April to June 2017, interns and nurses in ER were surveyed for blood culture practices with a questionnaire before and after 2 or 3 weeks of experience of Safety-Lok. All of them participated in exercise workshop for 1 hour combined with video education prior to the initial survey. The blood volume, positive and contamination rates of Safety-Lok-drawn (SD) blood cultures were compared to those of overall blood cultures. Results: Eighteen interns and 30 nurses were enrolled. As a result of the initial survey, interns had higher rates of needlestick incidence (27.8%), carriage of the blood-filled syringe with needle (88.9%) and lower rates of vacutainer use (38.9%) than nurses (13.3%, 53.3%, and 60.0%). Interns preferred to use safety devices (88.9%) rather than nurses (40.0%). The number of overall blood cultures and SD blood cultures was 9,053 and 555, respectively. While the overall blood volume of aerobic bottles was 2.6±2.1 mL, those of SD blood cultures were 5.0±3.0 mL in aerobic bottles and 6.0±3.0 mL in anaerobic bottles. Positive and contamination rates were 6.5% and 0.72% with SD blood cultures and 6.2% and 0.3% with overall blood cultures. Conclusions: The introduction of the safety device would encourage healthcare workers to collect adequate blood volume as well as lead to safer practices in the ER.

Keywords: blood culture, needlestick, safety device, volume

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Authors: Chetna Tyagi. G. B. V. S. Lakshmi, Ambuj Tripathi, D. K. Avasthi

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Graphene oxide provides a good host matrix for preparing nanocomposites due to the different functional groups attached to its edges and planes. Being biocompatible, it is used in therapeutic applications. As enzyme-based biosensor requires complicated enzyme purification procedure, high fabrication cost and special storage conditions, we need enzyme-less biosensors for use even in a harsh environment like high temperature, varying pH, etc. In this work, we have prepared both enzyme-based and enzyme-less graphene oxide-based biosensors for glucose detection using glucose-oxidase as enzyme and gold nanoparticles, respectively. These samples were characterized using X-ray diffraction, UV-visible spectroscopy, scanning electron microscopy, and transmission electron microscopy to confirm the successful synthesis of the working electrodes. Electrochemical measurements were performed for both the working electrodes using a 3-electrode electrochemical cell. Cyclic voltammetry curves showed the homogeneous transfer of electron on the electrodes in the scan range between -0.2V to 0.6V. The sensing measurements were performed using differential pulse voltammetry for the glucose concentration varying from 0.01 mM to 20 mM, and sensing was improved towards glucose in the presence of gold nanoparticles. Gold nanoparticles in graphene oxide nanocomposite played an important role in sensing glucose in the absence of enzyme, glucose oxidase, as evident from these measurements. The selectivity was tested by measuring the current response of the working electrode towards glucose in the presence of the other common interfering agents like cholesterol, ascorbic acid, citric acid, and urea. The enzyme-less working electrode also showed storage stability for up to 15 weeks, making it a suitable glucose biosensor.

Keywords: electrochemical, enzyme-less, glucose, gold nanoparticles, graphene oxide, nanocomposite

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Authors: Eunseop Yeom

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Many cardiovascular diseases, such as thrombosis and atherosclerosis, can change biochemical molecules in plasma and red blood cell. These alterations lead to excessive increase of blood viscosity contributing to peripheral vascular diseases. In this study, a simple microfluidic-based method is used to measure blood viscosity. Microfluidic device is composed of two parallel side channels and a bridge channel. To estimate blood viscosity, blood samples and reference fluid are separately delivered into each inlet of two parallel side channels using pumps. An interfacial line between blood samples and reference fluid occurs by blocking the outlet of one side-channel. Since width for this interfacial line is determined by pressure ratio between blood and reference flows, blood viscosity can be estimated by measuring width for this interfacial line. This microfluidic-based method can be used for evaluating variations in the viscosity of animal models with cardiovascular diseases under flow conditions.

Keywords: blood viscosity, microfluidic chip, pressure, shear rate

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Authors: Fahad Raj Khan, Suleman Khan

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There is a lack of consensus about the predictive value of raised blood glucose levels in terms of major adverse cardiac events (MACEs) in non-diabetic patients admitted for acute decompensated heart failure. The purpose of this research was to examine the long-term prognosis of acute decompensated heart failure (ADHF) in non-diabetic persons who had increased blood glucose levels, i.e., stress hyperglycemia, at the time of their ADHF hospitalization. The research involved 650 non-diabetic patients. Based on their admission stress hyperglycemia, they were divided into two groups.ie with and without (SHGL). The two groups' one-year outcomes for major adverse cardiac events (MACEs) were compared, and key predictors of MACEs were discovered. For statistical analysis, the two-tailed Mann-Whitney U test, Fisher's exact test, and binary logistic regression analysis were utilized. SHGL was found in 353 (54.3%) individuals. It was more frequent in men than in women. About 27% of patients with SHGL had previously been admitted for ADHF. Almost 62% were hypertensive, whereas 14 % had CKD. MACEs were significantly predicted by SHGL, HTN, prior hospitalization for ADHF, CKD, and cardiogenic shock upon admission. SHGL at the time of ADHF admission, independent of DM status, may be a predictive indication of MACEs.

Keywords: stress hyperglycemia, acute heart failure, major adverse cardiac events, MACEs

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Authors: Geetha Jayapathy, Lakshmi Muthukrishnan, Manoj Kumar Reddy Pulim , Radhika Raman

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Introduction: Ketones are the end products of fatty acid metabolism and a source of energy for vital organs such as the brain, heart and skeletal muscles. Ketones are produced in excess when glucose is not available as a source of energy or it cannot be utilized as in diabetic ketoacidosis. Children admitted in the emergency department often have starvation ketosis which is not clinically manifested. Decision on admission of children to the emergency room with subtle signs can be difficult at times. Point of care blood ketone testing can be done at the bedside even in a primary level care setting to supplement and guide us in our management decisions. Hence this study was done to explore the utility of this simple bedside parameter as a supplement in assessing pediatric patients presenting to the emergency department. Objectives: To estimate blood ketones of children admitted in the emergency department. To analyze the significance of blood ketones in various disease conditions. Methods: Blood ketones using point of care testing instrument (ABOTTprecision Xceed Pro meters) was done in patients getting admitted in emergency room and in out-patients (through sample collection centre). Study population: Children aged 1 month to 18 years were included in the study. 250 cases (In-patients) and 250 controls (out-patients) were collected. Study design: Prospective observational study. Data on details of illness and physiological status were documented. Blood ketones were compared between the two groups and all in patients were categorized into various system groups and analysed. Results: Mean blood ketones were high in in-patients ranging from 0 to 7.2, with a mean of 1.28 compared to out-patients ranging from 0 to 1.9 with a mean of 0.35. This difference was statistically significant with a p value < 0.001. In-patients with shock (mean of 4.15) and diarrheal dehydration (mean of 1.85) had a significantly higher blood ketone values compared to patients with other system involvement. Conclusion: Blood ketones were significantly high (above the normal range) in pediatric patients who are sick requiring admission. Patients with various forms of shock had very high blood ketone values as found in diabetic ketoacidosis. Ketone values in diarrheal dehydration were moderately high correlating to the degree of dehydration.

Keywords: admission, blood ketones, paediatric emergencies, point of care testing

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Authors: Isaac Gbadura Adanlawo, Olusola Olalekan Elekofehinti

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This work investigated the antihyperglycemic, antioxidant and antihyperlipidemic effects of saponins from the fruit of Solanum anguivi, a plant generally used in folk medicine to treat diabetes and hypertension and to compare its effect with metformin in streptozotocin (STZ)-induced diabetic rats. Diabetes was induced in albino rats by administration of STZ (65 mg/kg) intraperitoneally. Saponin (40 and 100 mg/kg) was administered by oral gavage once daily for 21 days. Metformin (200 mg/kg b.w.) was administered as the positive control. The effect of saponin on blood glucose, serum lipids and enzymatic antioxidants defense systems, like superoxide dismutase (SOD), catalase (CAT), as well as MDA levels in serum, liver and pancreas were studied. Saponins from S. anguivi fruits reduced the blood glucose, total cholesterol (TC), triglycerides (TG) and low-density lipoprotein (LDL) levels in STZ-diabetic rats. They also significantly abolished the increase in MDA level in serum, liver and pancreas of diabetic rats. The activities of SOD and CAT in serum, liver and pancreas were significantly increased as well as concentration of HDL in the serum. Metformin had the same effect as saponin but saponins seems to be more potent in reducing serum TC, TG, LDL, and MDA, and increasing SOD and CAT. Conclusions: These results suggest that saponins from S. anguivi fruits have anti-diabetic and antihypercholesterolemic, antihypertriglyceridemic antiperoxidative activities mediated through their antioxidant properties. Also, saponins appeared to have more hypolipidemic, antiperoxidative and antioxidant activity than metformin.

Keywords: saponin, diabetes, metformin, streptozotocin, Solanum anguivi

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Authors: Tania Mannan, Rahelee Zinnat, Fatema Jebunnesa, Israt Ara Hossain

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Background: Exposure to arsenic has known toxic effects but the effect on pregnancy outcomes is not as widely documented especially in women with diabetes. Growing evidence has suggested a potential role of arsenic exposure in the development of gestational diabetes mellitus (GDM). Therefore, we aimed to investigate the association of urinary arsenic (UAs) with birth outcomes in GDM subjects. Methods: Under an observational cross-sectional design a total of 263 GDM subjects (age in years, M±SD, 21±3.7) residing in an arsenic affected area of Bangladesh, were subjected to a 2 sample OGTT at the third trimester of gestation. Among them, 73 GDM and 190 non-GDM subjects enrolled in this study. Clinical and anthropometric measurements were done by standard techniques. Degree of chronic arsenic exposure was assessed by the level of UAs level. According to World Health Organization (WHO) criteria, GDM was diagnosed and neonatal outcomes using APGAR (Activity Pulse Grimace Appearance Respirations) Score, birth weight and size were assessed by a specialist obstetrician. Serum glucose was measured by the Glucose Oxidase method and UAs level was determined by ultraviolet/visible spectrophotometry. Result: Out of the 263 pregnant women, 28% developed GDM. Urinary Arsenic was significantly higher in the GDM as compared to the non-GDM group [UAs, µg/l, M±SD (range), 204.2±67.0 (67.0-377.0) vs 77.3±38.1 (22.0-99.0), p < 0.001]. Activity Pulse Grimace Appearance Respirations Score of the neonates from GDM mothers was significantly lower compared to the neonates from non-GDM mothers [APGAR Score, M±SD, 4.7±0.8 vs. 6.4±0.7, p<0.001]. Pearson’s correlation analysis in GDM subjects revealed that UA levels were found to have a significant positive correlation with both fasting and postprandial serum glucose levels (p < 0.001) and (p < 0.001) respectively. Again, a significant inverse correlation of UAs with birth weight and size was observed (p < 0.001). The APGAR Score of the neonates were found to have a significant negative correlation (p < 0.001) with UAs level. Conclusion: The effect of chronic arsenic exposure is associated with glucose intolerance during pregnancy and it also adversely affects birth outcomes. The study suggests further research on the impact of total arsenic exposure on pregnancy outcomes.

Keywords: APGAR score, arsenic exposure, birth outcome, gestational diabetes mellitus,

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Authors: Salima Taleb, Kafila Boulaba, Ahlem Yousfi, Nada Taleb, Difallah Basma

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Introduction: Obesity is recognized as a cardiovascular risk factor. It is associated with cardio-metabolic diseases. Its prevalence is increasing significantly in both rich and poor countries. However, there are obese people who have no metabolic disturbance. So we think obesity is not always a risk factor for an abnormal metabolic profile that increases the risk of cardiometabolic problems. However, there is no definition that allows us to identify the individual group Metabolically Healthy but Obese (MHO). Objective: The objective of this study is to evaluate the relationship between MHO and some factors associated with it. Methods: A longitudinal study is a prospective cohort study of 600 participants aged ≥18 years. Metabolic status was assessed by the following parameters: blood pressure, fasting glucose, total cholesterol, HDL cholesterol, LDL cholesterol, and triglycerides. Body Mass Index (BMI) was calculated as weight (in kg) divided by height (m2), BMI = Weight/(Height)². According to the BMI value, our population was divided into four groups: underweight subjects with BMI <18.5 kg/m2, normal weight subjects with BMI = 18.5–24.9 kg/m², overweight subjects with BMI=25–29.9 kg/m², and obese subjects who have (BMI ≥ 30 kg/m²). A value of P < 0.05 was considered significant. Statistical processing was done using the SPSS 25 software. Results: During this study, 194 (32.33%) were identified as MHO among 416 (37%) obese individuals. The prevalence of the metabolically unhealthy phenotype among normal-weight individuals was (13.83%) vs. (37%) in obese individuals. Compared with metabolically healthy normal-weight individuals (10.93%), the prevalence of diabetes was (30.60%) in MHO, (20.59%) in metabolically unhealthy normal weight, and (52.29%) for metabolically unhealthy obese (p = 0.032). Blood pressure was significantly higher in MHO individuals than in metabolically healthy normal-weight individuals and in metabolically unhealthy obese than in metabolically unhealthy normal weight (P < 0.0001). Familial coronary artery disease does not appear to have an effect on the metabolic status of obese and normal-weight patients (P = 0.544). However, waist circumference appears to have an effect on the metabolic status of individuals (P < 0.0001). Conclusion: This study showed a high prevalence of metabolic profile disruption in normal-weight subjects and a high rate of overweight and/or obese people who are metabolically healthy. To understand the physiological mechanism related to these metabolic statuses, a thorough study is needed.

Keywords: metabolically health, obesity, factors associated, cardiovascular diseases

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Authors: Boris Nemzer, Tania Reyes-Izquierdo, Zbigniew Pietrzkowski

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Glucosinolate is a family of glucosides that can be found in a family of brassica vegetables. Upon the damage of the plant, glucosinolate breakdown by an internal enzyme myrosinase (thioglucosidase; EC 3.2.3.1) into isothiocyanates, such as sulforaphane. Sulforaphane is formed by glucoraphanin cleaving the sugar off by myrosinase and rearranged. Sulforaphane nitrile is formed in the same reaction as sulforaphane with the active of epithiospecifier protein (ESP). Most common food processing procedure would break the plant and mix the glucoraphanin and myrosinase together, and the formed sulforaphane would be further degraded. The purpose of this study is to understand the glucoraphanin/sulforaphane and the myrosinase activity of broccoli seeds germinated at a different time and technological processing conditions that keep the activity of the enzyme to form sulforaphane. Broccoli seeds were germinated in the house. Myrosinase activities were tested as the glucose content using glucose assay kit and measured UV-Vis spectrophotometer. Glucosinolates were measured by HPLC/DAD. Sulforaphane was measured using HPLC-DAD and GC/MS. The 6 hr germinated sprouts have a myrosinase activity 32.2 mg glucose/g, which is comparable with 12 and 24 hour germinated seeds and higher than dry seeds. The glucoraphanin content in 6 hour germinated sprouts is 13935 µg/g which is comparable to 24 hour germinated seeds and lower than the dry seeds. GC/MS results show that the amount of sulforaphane is higher than the amount of sulforaphane nitrile in seeds, 6 hour and 24 hour germinated seeds. The ratio of sulforaphane and sulforaphane nitrile is high in 6 hour germinated seeds, which indicates the inactivated ESP in the reaction. After evaluating the results, the short time germinated seeds can be used as the source of glucoraphanin and myrosinase supply to form potential higher sulforaphane content. Broccoli contains glucosinolates, glucoraphanin (4-methylsulfinylbutyl glucosinolate), which is an important metabolite with health-promoting effects. In the pilot clinical study, we observed the effects of a glucosinolates/glucoraphanin-rich extract from short time germinated broccoli seeds on blood adenosine triphosphate (ATP), reactive oxygen species (ROS) and lactate levels. A single dose of 50 mg of broccoli sprouts extract increased blood levels of ATP up to 61% (p=0.0092) during the first 2 hours after the ingestion. Interestingly, this effect was not associated with an increase in blood ROS or lactate. When compared to the placebo group, levels of lactate were reduced by 10% (p=0.006). These results indicate that broccoli germinated seed extract may positively affect the generation of ATP in humans. Due to the preliminary nature of this work and promising results, larger clinical trials are justified.

Keywords: broccoli glucosinolates, glucoraphanin, germinated seeds, myrosinase, adenosine triphosphate

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