Search results for: biochemical changes

Authors: Fariha Salman, Helmet Steinberg, Hiba Al-Zubeidi

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OBJECTIVE: Polycystic ovary syndrome (PCOS) is the most common cause of infertility, with a prevalence of 5-10 % in women of reproductive age. There is evidence for differences between adult and pediatric endocrinologists and other specialists in their approach to the diagnosis and management of PCOS. METHODS: A survey consisting of 37 questions was distributed among adult and pediatric endocrinologists, aiming to understand the current practice for the diagnosis and management of PCOS. A total of 100 responses were available for final analysis; 36 % from adult endocrinologists (AE) and 64 % from pediatric endocrinologists(PE). RESULTS: The majority (64%) of respondents to the survey were endocrinologists from a multispecialty group. For both adults and adolescents with PCOS, the most commonly reported presenting symptoms were menstrual irregularities, obesity and hirsutism. The most common features used for diagnostic criteria were clinical or biochemical hyperandrogenism and ovulatory dysfunction. Most AE and PE screened for PCOS with total testosterone (83%) and free Testosterone (71%), screening for prolactin excess in 70 % and congenital adrenal hyperplasia (83 %), 66 % of AE will obtain pelvic US for evaluation vs 45 % of PE. Only 20 % of all respondents will obtain a midluteal progesterone for documentation of anovulation. In terms of treatment of hyperandrogenism and menstrual irregularities in adolescents, the most common form used is oral contraceptive pills, followed by metformin, then spironolactone. A similar approach was used in adults however the use of spironolactone was higher, 53 % vs 21 % in adolescents. The most common modality used for infertility was lifestyle interventions followed by metformin and clomiphene citrate. Screening of OSA and depression was not done by most of the endocrinologists (never + sometimes), 72 % and 76 %, respectively. Though screening for diabetes/metabolic syndrome and insulin resistance was done by most of the endocrinologists (always+often), 95 and 68 %, respectively. DISCUSSION: There are multiple diagnostic criteria used for PCOS diagnosis, however, given the wide variation in presentation and approach to diagnosis in adults and adolescents, there has not been a consensus on which is the gold standard criteria. CONCLUSION: Our survey showed the most common trends in diagnosing and treating PCOS among adult and pediatric endocrinologists. Further studies and trials need to be conducted to compare different treatment modalities used for hyperandrogenism, menstrual irregularities and infertility, as PCOS, if not treated earlier, can lead to long-term complications.

Keywords: PCOS, adolescents, diagnosis, treatment

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Authors: Baishali Deb, Hari Om Pandey, Shrilla Elangbam, Mukesh Singh, Ayon Tarafdar, A. K. S. Tomar, A. K. Pandey, Triveni Dutt

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The goal of the present investigation was to determine the impact of social stress on behavioural characteristics, physiological responses, and haemato-biochemical indicators under various social environments in Tharparkar cattle. Serum cortisol and faecal cortisol metabolites analysis were used to determine the stress level of Tharparkar cattle. Social isolation and social mixing were the two different social circumstances used to evaluate the animals. In both the experiments i.e., social isolation and social mixing, the lying period of animals decreased significantly (p<0.05) while standing period significantly (p<0.05) increased. Frequency and duration of activities like idling, walking, exploration, oral manipulation, and elimination increased significantly (p<0.05) in Tharparkar cattle after being subjected to social isolation and social mixing. Time spent in grooming (self-grooming and allo-grooming) in respect to social isolation significantly increased during isolation and post-reunion, whereas there was a significant (p<0.05) decline in the grooming behaviour especially allo-grooming during mixing of the animals. Feeding and rumination time also decreased significantly (p<0.05) in animals during both the experiments. Physiological parameters such as respiration rate, heart rate and pulse rate increased during the treatment periods. There was no significant difference in the haematological parameters for both the experiments. There was significant (p<0.05) increase in serum cortisol and faecal cortisol metabolites (FCM) concentration in animals subjected to social stress. Therefore, it can be concluded that social stress strongly impacts the behaviour and physiological parameters of the animals, causing stress and nervousness, proving that social stress is a valid psychological stress in animals. The higher concentration of FCM in Tharparkar cattle subjected to social stress, further supported by higher serum cortisol and behaviour manifestations, suggest that FCM could be used to assess stress response as a non-invasive method.

Keywords: social stress, fecal cortisol metabolites, non-invasive, animal welfare, behaviour

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Authors: Hwang Ahreum, Kang Taejoon, Kim Bongsoo

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Nanosensors for high sensitive detection of diseases have been widely studied to improve the quality of life. Here, we suggest robust nano-plasmonic diagnostic sensor using cysteine tagged protein G (Cys3-protein G) and ultraflat, ultraclean and single-crystalline Au nanoplates. Protein G formed on an ultraflat Au surface provides ideal background for dense and uniform immobilization of antibodies. The Au is highly stable in diverse biochemical environment and can immobilize antibodies easily through Au-S bonding, having been widely used for various biosensing applications. Especially, atomically smooth single-crystalline Au nanomaterials synthesized using chemical vapor transport (CVT) method are very suitable to fabricate reproducible sensitive sensors. As the C-reactive protein (CRP) is a nonspecific biomarker of inflammation and infection, it can be used as a predictive or prognostic marker for various cardiovascular diseases. Cys3-protein G immobilized uniformly on the Au nanoplate enable CRP antibody (anti-CRP) to be ordered in a correct orientation, making their binding capacity be maximized for CRP detection. Immobilization condition for the Cys3-protein G and anti-CRP on the Au nanoplate is optimized visually by AFM analysis. Au nanoparticle - Au nanoplate (NPs-on-Au nanoplate) assembly fabricated from sandwich immunoassay for CRP can reduce zero-signal extremely caused by nonspecific bindings, providing a distinct surface-enhanced Raman scattering (SERS) enhancement still in 10-18 M of CRP concentration. Moreover, the NP-on-Au nanoplate sensor shows an excellent selectivity against non-target proteins with high concentration. In addition, comparing with control experiments employing a Au film fabricated by e-beam assisted deposition and linker molecule, we validate clearly contribution of the Au nanoplate for the attomolar sensitive detection of CRP. We expect that the devised platform employing the complex of single-crystalline Au nanoplates and Cys3-protein G can be applied for detection of many other cancer biomarkers.

Keywords: Au nanoplate, biomarker, diagnostic sensor, protein G, SERS

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Authors: Rozeena Baig, R. Rehana Rehman, Rifat Ahmed

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Background: Osteoporosis, characterized by low bone mineral density, poses a global health concern. Diagnosis increases the likelihood of developing osteoporosis, a multifactorial disorder marked by low bone mass, elevating the risk of fractures in the lumbar spine, femoral neck, hip, vertebras, and distal forearm, particularly in postmenopausal women due to bone loss influenced by various pathophysiological factors. Objectives: The aim is to investigate the association of serum cytokine, bone turnover marker, bone mineral density and TLR4 gene polymorphism in pre and post-menopausal women and to find if any of these can be the potential predictor of osteoporosis in postmenopausal women. Material and methods: The study participants consisted of Group A (n=91) healthy pre-menopausal women and Group B (n=102) healthy postmenopausal women having ≥ 5 years’ history of menopause. ELISA was performed for cytokine (TNFα) and bone turnover markers (carboxytelopeptides), respectively. Bone Mineral Density (BMD)was measured through a dual X-ray absorptiometry (DEXA) scan. Toll-like Receptors 4 (TLR4) gene polymorphisms (A896G; Asp299Gly) and (C1196T; Thr399Ile) were investigated by PCR and Sanger sequencing. Results: Statistical analysis reveals a positive correlation of age and BMI with T scores in the premenopausal group, whereas in post-menopausal group found a significant negative correlation between age and T-score at hip (r = - 0.352**), spine (r = - .306**), and femoral neck (r = - 0.344**) and a significant negative correlation of BMI with TNF-α (- 0.316**). No association and significant differences were observed for TLR4 genotype and allele frequencies among studied groups However, both SNPs exhibited significant association with each other. Conclusions: This study concludes that BMI, BMD and TNF-α are the potential predictors of osteoporosis in post-menopausal women. However, CTX and TLR4 gene polymorphism did not appear as potential predictors of bone loss in this study and apparently cannot help in predicting bone loss in post-menopausal women.

Keywords: osteoporosis, post-menopausal, pre-menopausal woemn, genetics mutaiont, TLR4 genepolymorphsum

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Authors: Kehinde Joseph Awosan, Balarabe Adami Isah, Edzu Usman Yunusa, Sarafadeen Adeniyi Arisegi, Izuchukwu Obasi, Oluchi Solomon-Anucha

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Background: Metabolic syndrome (MetS) is prevalent in patients with diabetes mellitus and a significant risk for major cardiovascular events. Identifying its burden and peculiarities is crucial to preventing complications among those at risk. Aim: This study was conducted to determine the prevalence and predictors of metabolic syndrome among diabetes clinic attendees in Sokoto, Nigeria. Materials and Methods: A cross-sectional study was conducted among 365 patients with type 2 diabetes attending the diabetes clinic of Specialist Hospital, Sokoto, Nigeria. A structured questionnaire was used to obtain data on the respondents’ socio-demographic variables, treatment history, and lifestyle. Blood pressure and anthropometric measurements (including weight, height, and waist circumference) were done for the patients. Likewise, biochemical assessment (including fasting plasma glucose, high-density lipoprotein cholesterol (HDL-c), and triglyceride (TG) was done. Metabolic syndrome was defined according to the National Cholesterol Education Program Adult Treatment Panel III (NCEP ATP III). Data were analyzed using the IBM Statistical Package for Social Sciences (SPSS) version 25. Results: The ages of the patients ranged from 30 to 78 (mean = 50.9 ±11.7) years. The overall prevalence of MetS was 57.3%, with a higher prevalence in females (68.1%) than males (43.0%). The most common components of MetS observed were hypertension (69.2%), and elevated fasting plasma glucose (65.7%); while the predictors of MetS were age > 50 years (OR 6.960, 95% CI: 3.836-12.628, p < 0.001), female sex (OR 2.300, 95% CI: 1.355-3.903, p = 0.002), physical activity (OR 0.214, 95% CI: 0.126-0.363, p < 0.001), and overweight/obesity (OR 3.356, 95% CI: 1.838-6.127, p < 0.001). Conclusion: Metabolic syndrome is prevalent among patients with type 2 diabetes in Sokoto, Nigeria, and the predictors were age > 50 years, female sex, physical activity, and overweight/obesity. Diabetes care providers should screen their patients for MetS to prevent adverse cardiovascular events.

Keywords: prevalence, predictors, metabolic syndrome, diabetes

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Authors: Kathryn L. Duncan, Charles A. Kuntz, Alessandra C. Santamaria, James O. Simcock

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Access to megavoltage radiation therapy for small animals is limited in many locations around the world. This can preclude the use of palliative radiation therapy for the treatment of appendicular osteosarcoma in dogs. The objective of this study was to retrospectively assess the adverse effects and survival times of dogs with appendicular osteosarcoma that were treated with hypofractionated orthovoltage radiation therapy and adjunctive carboplatin chemotherapy administered via a single subcutaneous infusion. Medical records were reviewed retrospectively to identify client-owned dogs with spontaneously occurring appendicular osteosarcoma that was treated with palliative orthovoltage radiation therapy and a single subcutaneous infusion of carboplatin. Data recorded included signalment, tumour location, results of diagnostic imaging, haematologic and serum biochemical analyses, adverse effects of radiation therapy and chemotherapy, and survival times. Kaplan-Meier survival analysis was performed, and log-rank analysis was used to determine the impact of specific patient variables on survival time. Twenty-three dogs were identified that met the inclusion criteria. Median survival time for dogs was 182 days. Eleven dogs had adverse haematologic effects, 3 had adverse gastrointestinal effects, 6 had adverse effects at the radiation site and 7 developed infections at the carboplatin infusion site. No statistically significant differences were identified in survival times based on sex, tumour location, development of infection, or pretreatment serum alkaline phosphatase. Median survival time and incidence of adverse effects were comparable to those previously reported in dogs undergoing palliative radiation therapy with megavoltage or cobalt radiation sources and conventional intravenous carboplatin chemotherapy. The use of orthovoltage palliative radiation therapy may be a reasonable alternative to megavoltage radiation in locations where access is limited.

Keywords: radiotherapy, veterinary oncology, chemotherapy, osteosarcoma

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Authors: Denise Schrama, Claudia Raposo, Pedro Rodrigues

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Background: Food allergies are conducted by a hypersensitive response of the immune system. These allergies are a global concern for the public health. Consumption of fish is increasing worldwide as it is a healthy meat with high nutritional value. Unfortunately, fish can cause adverse immune-mediate reactions, affecting part of the population with higher incidence in children. β-parvalbumin, a small, highly conserved stable, calcium or magnesium binding muscle protein is the main fish allergen. In fish-allergic patients, cross-reactivity between different fish species exist due to recognition of highly identical protein regions. Enolases, aldolases, or fish gelatin are other identified fish allergens in some fish species. With no available cure for fish allergies, clinical management is only based on an avoidance diet aiming at the total exclusion of offending food. Methods: Mediterranean fish (S. aurata and D. labrax) were fed specifically designed diets, enriched in components that target the expression or inactivation of parvalbumin (creatine and EDTA, respectively). After 90 days fish were sampled and biological tissues were excised. Proteomics was used to access fish allergens characterization and expression in muscle while IgE assays to confirm the lower allergenic potential are conducted in patients with history of fish allergies. Fish welfare and quality of flesh were established with biochemical, texture and sensorial analysis. Results: Fish welfare shows no major impact between diets. In case of creatine supplementation in D. labrax proteomic analysis show a slight decrease in parvalbumin expression. No accumulation of this compound was found in muscle. For EDTA supplementation in S. aurata IgE assay show a slight decrease in allergenicity when using sera of fish allergic patients. Conclusion: Supplementation with these two compounds seems to change slightly the allergenicity of the two mean Mediterranean species.

Keywords: fish allergies, fish nutrition, proteomics, aquaculture

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Authors: Malgorzata Rudnicka, Waldemar Karcz

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Introduction: Naphthoquinones are widely occurring organic compounds produced by bacteria, fungi, and plants. They can act as the functional components of biochemical systems (plastoquinone) as well as biologically active substances, which have a negative impact on environmental processes. Naphthoquinones seem to act through two mechanisms: a covalent modification of biological molecules at their nucleophilic sites or by generation of reactive oxygen species (ROS) connected with redox cycling. Investigating the effect of naphthoquinones (1,4-naphthoquinone, lawsone and naphthazarin) on the elongation growth, membrane potential and the level of oxidative stress of maize cells seems to be important due to the possibility of using these substances as herbicides. Methods: All experiments were performed on etiolated maize coleoptile segments. Simultaneous measurements of elongation growth and pH of the incubation medium were carried out using an angular position transducer, allowing a precise record of the growth kinetics. To compare the oxidative stress level induced by all tested naphthoquinones, the changes in malondialdehyde content, as well as superoxide dismutase and catalase activities were measured. In order to measure the membrane potential of parenchymal cells the standard electrophysiology technique was used. Results: Naphthoquinones such as: 1,4-naphthoquinone, lawsone and naphthazarin were studied. It was found that all of the naphthoquinones diminished the growth of the maize coleoptile cells depending on the type of naphthoquinones and their concentration. Interestingly, naphthazarin at the intermediate concentration was less toxic compared to the others. In addition, the effect of naphthoquinones on the oxidative stress was dependent on their concentration as well. Superoxide dismutase and catalase activities were changed in the presence of higher concentrations of naphthoquinones. Similar interrelations were observed for membrane potential changes. Conclusion: It can be concluded that naphthoquinones tested differ in their toxic effect on the growth of maize coleoptile cells. Furthermore, naphthoquinones can be distinguish considering the oxidative stress level and membrane potential changes. The results presented here give new insight into the possible opportunities of practical usage of naphthoquinones for herbicides improvement.

Keywords: growth rate, membrane potential, naphthoquinones, oxidative stress

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Authors: Mansouri Ouarda, Abdennour Cherif, Saidi Malika

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Since the industrial revolution, many anthropogenic activities have caused environmental, considerable and overall changes. The lead represents a very dangerous disruptive for the functioning of the body. In this context the current study aims at evaluating a natural therapy by the use of the plant grass in wheat (Triticum durum) against the toxicity of lead in rat wistar male. The rats were divided into three groups: the control group, the group treated with 600 mg /kg food of lead only (Pb) is the group treated with the combination of 600 mg/kg of food and 9g/rat /day of the plant grass in wheat (Pb-bl). The duration of the treatment is 6 weeks. The results of the biometrics of the organs (thyroid, kidney, testis and epididymis) show no significant difference between the three groups. The dosage of a few parameters and hormonal biochemical shows a decrease in the concentration of the hormone T3 and TSH levels among the group pb alone compared to the control and Pb-Bl. These results have been confirmed by the study of histological slices. A morphological changes represented by a shrinking volume of vesicles with the group treated with Pb alone. A return to the normal state of the structure of the follicles was observed. The concentration in serum testosterone, urea and creatinine was significantly increased among the group treated by Pb only in relation to the control and Pb-Bl. whereas the rate of glucose did not show any significant difference. The histology study of the kidney, testis and epididymal weights show no modification at the group Pb-bl comparing to the control. The parenchyma of the kidney shows a dilation of tubes distal and proximal causing a tubular nephropathy for the batch processed by Pb only. The testicles have marked a destruction or absence of germ cells and the light of some seminiferous are almost empty. Conclusion: The supplementation of the plant Triticum durum has caused a considerable improvement which ensures the return of parameters investigated in the normal state.

Keywords: creatinine, glucose, histological sections, T3, TSH, testosterone

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Authors: Mohd Asharf Ganie, Aafia Rashid, Mohd Afzal Zargar, Showkat Ali Zargar, Syed Mudasar, Tabasum Parvaiz, Zafar Amin Shah

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Accumulating evidence suggests that Vitamin D deficiency (VDD) might at least contribute to the metabolic co-morbidities in PCOS. Hence, we aimed to study the effect of vitamin D supplementation in co-prescription with insulin sensitizers like metformin and pioglitazone on clinical, hormonal and metabolic parameters in women with PCOS. In this open label randomized, controlled trial a total of 120 women with PCOS diagnosis (AE-PCOS 2009 Criteria) were assigned to four treatment groups (n= 30 in each): group I (metformin 1 gm/day in combination with cholecalciferol 4000 IU/day), group II (pioglitazone 30 mg/day in combination with cholecalciferol 4000 IU/day), group III (metformin 1 gm /day) and group IV (pioglitazone 30 mg/day). Vitamin D supplementation was given as 60,000 units every two weeks for 24 weeks. All the subjects were routinely evaluated for clinical, biochemical, hormonal and insulin sensitivity parameters in addition to various safety parameters especially serum calcium levels at baseline and after 24 weeks of the treatment. Our results indicate that 95.5% of PCOS women were vitamin D deficient at baseline. Serum 25 (OH) D levels increased significantly (p < 0.001) in groups I and II without any adverse effects after 24 weeks of oral administration of 4000 IU cholecalciferol daily. However, serum 25 (OH) D levels remained unchanged in group III and IV. By six months, number of menstrual cycles per year increased whereas Ferriman-Gallwey score, serum total testosterone and HOMA-IR decreased significantly (P < 0.001) in the treatment groups supplemented with cholecalciferol as compared to those treated either drug alone. No significant beneficial changes were observed on weight, BMI, blood pressure, glucose tolerance and serum lipids in any of the groups supplemented with cholecalciferol. We conclude that daily dose of 4000 IU cholecalciferol might be a useful adjunct in complex treatment of PCOS with fewer adverse events. Furthermore, pioglitazone and cholecalciferol combination seems to be marginally better although there was no statistical significance.

Keywords: PCOS, vitamin D supplementation, insulin resistance, spironolactone, metformin, pioglitazone

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Authors: Diana C. El Assal, Fatima Monteiro, Caroline May, Peter Barbuti, Silvia Bolognin, Averina Nicolae, Hulda Haraldsdottir, Lemmer R. P. El Assal, Swagatika Sahoo, Longfei Mao, Jens Schwamborn, Rejko Kruger, Ines Thiele, Kathrin Marcus, Ronan M. T. Fleming

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Degeneration of substantia nigra pars compacta dopaminergic neurons is one of the hallmarks of Parkinson's disease. These neurons have a highly complex axonal arborisation and a high energy demand, so any reduction in ATP synthesis could lead to an imbalance between supply and demand, thereby impeding normal neuronal bioenergetic requirements. Synaptic mitochondria exhibit increased vulnerability to dysfunction in Parkinson's disease. After biogenesis in and transport from the cell body, synaptic mitochondria become highly dependent upon oxidative phosphorylation. We applied a systems biochemistry approach to identify the metabolic pathways used by neuronal mitochondria for energy generation. The mitochondrial component of an existing manual reconstruction of human metabolism was extended with manual curation of the biochemical literature and specialised using omics data from Parkinson's disease patients and controls, to generate reconstructions of synaptic and somal mitochondrial metabolism. These reconstructions were converted into stoichiometrically- and fluxconsistent constraint-based computational models. These models predict that Parkinson's disease is accompanied by an increase in the rate of glycolysis and a decrease in the rate of oxidative phosphorylation within synaptic mitochondria. This is consistent with independent experimental reports of a compensatory switching of bioenergetic pathways in the putamen of post-mortem Parkinson's disease patients. Ongoing work, in the context of the SysMedPD project is aimed at computational prediction of mitochondrial drug targets to slow the progression of neurodegeneration in the subset of Parkinson's disease patients with overt mitochondrial dysfunction.

Keywords: bioenergetics, mitochondria, Parkinson's disease, systems biochemistry

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Authors: Pierre Mintom, William Djeukeu Asongni, Michelle Moni, William Dakam, Christine Fernande Nyangono Biyegue.

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Background: In order to prevent congestive heart failure, control of hypertension and hypercholesterolemia is necessary because those risk factors frequently occur in combination. Objective: The aim of the study is to determine the prevalence and risk factors of hypertension, hypercholesterolemia and double burden HTA-Hypercholesterolemia in patients with congestive heart failure. Methodology: A database of 98 patients suffering from congestive heart failure was used. The latter were recruited from August 15, 2017, to March 5, 2018, in the Cardiology department of Deido District Hospital of Douala. This database provides information on sociodemographic parameters, biochemical examinations, characteristics of heart failure and food consumption. ESC/ESH and NCEP-ATPIII definitions were used to define Hypercholesterolemia (total cholesterol ≥200mg/dl), Hypertension (SBP≥140mmHg and/or DBP≥90mmHg). Double burden hypertension-hypercholesterolemia was defined as follows: total cholesterol (CT)≥200mg/dl, SBP≥140mmHg and DBP≥90mmHg. Results: The prevalence of hypertension (HTA), hypercholesterolemia (hyperchol) and double burden HTA-Hyperchol were 61.2%, 66.3% and 45.9%, respectively. No sociodemographic factor was associated with hypertension, hypercholesterolemia and double burden, but Male gender was significantly associated (p<0.05) with hypercholesterolemia. HypoHDLemia significantly increased hypercholesterolemia and the double burden by 19.664 times (p=0.001) and 14.968 times (p=0.021), respectively. Regarding dietary habits, the consumption of rice, peanuts and derivatives and cottonseed oil respectively significantly (p<0.05) exposed to the occurrence of hypertension. The consumption of tomatoes, green bananas, corn and derivatives, peanuts and derivatives and cottonseed oil significantly exposed (p<0.05) to the occurrence of hypercholesterolemia. The consumption of palm oil and cottonseed oil exposed the occurrence of the double burden of hypertension-hypercholesterolemia. Consumption of eggs protects against hypercholesterolemia, and consumption of peanuts and tomatoes protects against the double burden. Conclusion: hypercholesterolemia associated with hypertension appears as a complicating factor of congestive heart failure. Key risk factors are mainly diet-based, suggesting the importance of nutritional education for patients. New management protocols emphasizing diet should be considered.

Keywords: risk factors, hypertension, hypercholesterolemia, congestive heart failure

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Authors: Nanh Lovanh, John Loughrin, Kimberly Cook, Phil Silva, Byung-Taek Oh

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In the era of sustainability, utilization of livestock wastes as soil amendment to provide micronutrients for crops is very economical and sustainable. It is well understood that livestock wastes are comparable, if not better, nutrient sources for crops as chemical fertilizers. However, the large concentrated volumes of animal manure produced from livestock operations and the limited amount of available nearby agricultural land areas necessitated the need for volume reduction of these animal wastes. Composting of these animal manures is a viable option for biomass and pathogenic reduction in the environment. Nevertheless, composting also increases the potential loss of available nutrients for crop production as well as unwanted emission of anthropogenic air pollutants due to the loss of ammonia and other compounds via volatilization. In this study, we examine the emission of ammonia and nitrous oxide from swine manure windrows to evaluate the benefit of biomass reduction in conjunction with the potential loss of available nutrients. The feedstock for the windrows was obtained from swine farm in Kentucky where swine manure was mixed with wood shaving as absorbent material. Static flux chambers along with photoacoustic gas analyzer were used to monitor ammonia and nitrous oxide concentrations during the composting process. The results show that ammonia and nitrous oxide fluxes were quite high during the initial composting process and after the turning of each compost pile. Over the period of roughly three months of composting, the biochemical oxygen demand (BOD) decreased by about 90%. Although composting of animal waste is quite beneficial for biomass reduction, composting may not be economically feasible from an agronomical point of view due to time, nutrient loss (N loss), and potential environmental pollution (ammonia and greenhouse gas emissions). Therefore, additional studies are needed to assess and validate the economics and environmental impact of animal (swine) manure composting (e.g., crop yield or impact on climate change).

Keywords: windrow, swine manure, ammonia, nitrous oxide, fluxes, management

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Authors: S. El Ballal, H. El Sabbagh, M. Abd El Gaber, A. Eisa, A. Al Gamal

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Cadmium is one of the most harmful heavy metals able to induce severe injury. In this study, sixty four male Sprague Dawley rats weighing (70-80 gm) were used. Rats were divided into 4 groups each group of 16 rats. Group A: served as control and received commercial ration and distilled water Group B: cadmium chloride was administered orally in water at dose of 300 ppm cadmium (560 mg/L as CdCl2). Group C: Animals received cadmium in drinking water in addition to administration of N-acetylcysteine (NAC) orally at a dose of 150 mg/kg body weight, equivalent to 1500 ppm in food. Group D: Animals received cadmium in drinking water in addition to administration of alpha lipoic acid (ALA) orally at a dose of 150 mg/kg body weight, equivalent to 1500 ppm in food. The experiment was continued for 2 months. Collection of blood and tissue samples was performed at 2, 4, 6, 8 weeks. Blood sample were collected for serum biochemical analysis including malondialdehyde (MDA), total antioxidants, aspartate aminotransferase (AST), alanine aminotransferase (ALT), total protein, albumin, urea and uric acid. Tissue specimens were collected for histopathological examination including liver, kidney, brain and testis. Histopathological examination revealed that cadmium choloride induces pathological alterations which increased in severity with time. The use of NAC and ALA can ameliorate toxic effect of CdCl2. The results showed significant decrease MDA and significant increase total antioxidants in group C and D compared to group B, Liver enzymes include AST and ALT showed significant decrease. Regarding to results of total protein and albumin, they revealed significant increase. Urea and uric acid showed significant decrease. From our study we conclude that NAC and ALA have protective effect against cadmium toxicity.

Keywords: ALA, cadmium, histopathology, NAC

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Authors: Parvin Samadi Pakchin, Reza Saber, Hossein Ghanbari, Yadollah Omidi

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Ovarian cancer is one of the leading cause of mortality among the gynecological malignancies, and it remains the one of the most prevalent cancer in females worldwide. Tumor markers are biochemical molecules in blood or tissues which can indicates cancers occurrence in the human body. So, the sensitive and specific detection of cancer markers typically recruited for diagnosing and evaluating cancers. Recently extensive research efforts are underway to achieve a simple, inexpensive and accurate device for detection of cancer biomarkers. Compared with conventional immunoassay techniques, electrochemical immunosensors are of great interest, because they are specific, simple, inexpensive, easy to handling and miniaturization. Moreover, in the past decade nanotechnology has played a crucial role in the development of biosensors. In this study, a signal-off electrochemical immunosensor for the detection of CA125 antigen has been developed using chitosan-gold nanoparticles (CS-AuNP) and multi-wall carbon nanotubes (MWCNT) composites. Toluidine blue (TB) is used as redox probe which is immobilized on the electrode surface. CS-AuNP is synthesized by a simple one step method that HAuCl4 is reduced by NH2 groups of chitosan. The CS-AuNP-MWCNT modified electrode has shown excellent electrochemical performance compared with bare Au electrode. MWCNTs and AuNPs increased electrochemical conductivity and accelerate electrons transfer between solution and electrode surface while excessive amine groups on chitosan lead to the effective loading of the biological material (CA125 antibody) and TB on the electrode surface. The electrochemical, immobilization and sensing properties CS-AuNP-MWCNT-TB modified electrodes are characterized by cyclic voltammetry, electrochemical impedance spectroscopy, differential pulse voltammetry and square wave voltammetry with Fe(CN)63−/4−as an electrochemical redox indicator.

Keywords: signal-off electrochemical biosensor, CA125, ovarian cancer, chitosan-gold nanoparticles

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Authors: Rumana Keyani, Haleema Sadia, Asia Nosheen, Rabia Naz, Humaira Yasmin, Sidra Zahoor

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Tomato is a significant crop of the world and is one of the staple foods of Pakistan. A vast number of plant pathogens from simple viruses to complex parasites cause diseases in tomatoes but fungal infection in our country is quite high. Sometimes the symptoms are too harsh destroying the crop altogether. Countries like our own with continuously increasing massive population and limited resources cannot afford such an economic loss. There is an array of morphological, genetic, biochemical and molecular processes involved in plant resistance mechanisms to biotic stress. The study of different metabolic pathways like Jasmonic acid (JA) pathways and most importantly signaling molecules like ROS/RNS and their redoxin enzymes i.e. TRX and NRX is crucial to disease management, contributing to healthy plant growth. So, improving tolerance in crop plants against biotic stresses is a dire need of our country and world as whole. In the current study, fungal pathogenic strains Alternaria solani and Rhizoctonia solani were used to inoculate tomatoes to check the defense responses of tomato plant against these pathogens at molecular as well as phenotypic level with jasmonic acid and spermidine pretreatment. All the growth parameters (root and shoot length, dry and weight root, shoot weight measured 7 days post-inoculation, exhibited that infection drastically declined the growth of the plant whereas jasmonic acid and spermidine assisted the plants to cope up with the infection. Thus, JA and Spermidine treatments maintained comparatively better growth factors. Antioxidant assays and expression analysis through real time quantitative PCR following time course experiment at 24, 48 and 72 hours intervals also exhibited that activation of JA defense genes and a polyamine Spermidine helps in mediating tomato responses against fungal infection when used alone but the two treatments combined mask the effect of each other.

Keywords: fungal infection, jasmonic acid defence, tomato, spermidine

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Authors: Lubna Azmi, Ila Shukla, Shyam Sundar Gupta, Padam Kant, C. V. Rao

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To investigate the chemoprotective potential of NBRIQU16 chemotype isolated from Argyreia speciosa (Family: Convolvulaceae) on N-methyl-N-nitro-N-nitrosoguanidine (MNNG) and NaCl-induced gastric carcinomas in Wistar rats. Forty-six male 6-week-old Wistar rats were divided into two groups. Thirty rats in group A were fed with a diet supplemented with 8 % NaCl for 20 weeks and simultaneously given N-methyl-N’-nitro-N-nitrosoguanidine (MNNG) in drinking water at a concentration of 100 ug/ml for the first 17 weeks. After administration of the carcinogen, 200 and 400 mg/kg of NBRIQU16 were administered orally once a day throughout the study. From week 18, these rats were given normal water. From week 21, these rats were fed with a normal diet for 15 weeks. Group B containing 16 rats was fed standard diet for thirty-five days. It served as control. Ten rats from group A were sacrificed after 20 weeks. Scarification of remaining animals was conducted after 35 weeks. Entire stomach and some part of the duodenum were incised parallel to the greater curvature, and the samples were collected. After opening the stomach location and size of tumors were recorded. The number of tumors with their locations and sizes were recorded. Expression of survivin was examined by recording the Immunohistochemistry of the specimens. The treatment with NBRIQU16 significantly reduced the nodule incidence and nodule multiplicity in the rats after MNNG administration. Surviving expression in glandular stomachs of normal rats, of rats in middle induction period, in adenocarcinomas and NBRIQU16 treated tissues adjacent to tumor were 0, 42.0 %, 79.3%, and 36.4 %, respectively. Expression of survivin was significantly different as compared to the normal rats. Histological observations of stomach tissues too correlated with the biochemical observations.These finding powerfully supports that NBRIQU16 chemopreventive effect by suppressing the tumor burden and restoring the activities of gastric cancer marker enzymes on MNNG and NaCl-induced gastric carcinomas in Wistar rats.

Keywords: Argyreia speciosa, gastric carcinoma, immunochemistry, NBRIQU16

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Authors: Noura El-Ahmady El-Naggar

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Among the antitumour drugs, bacterial enzyme L-asparaginase has been employed as the most effective chemotherapeutic agent in pediatric oncotherapy especially for acute lymphoblastic leukemia. Glutaminase free L-asparaginase producing actinomycetes were isolated from soil samples collected from Egypt. Among them, a potential culture, strain NEAE-119, was selected and identified on the basis of morphological, cultural, physiological and biochemical properties, together with 16S rDNA sequence as Streptomyces olivaceus NEAE-119 and sequencing product(1509 bp) was deposited in the GenBank database under accession number KJ200342. The optimization of different process parameters for L-asparaginase production by Streptomyces olivaceus NEAE-119 using Plackett–Burman experimental design and response surface methodology was carried out. Fifteen nutritional variables (temperature, pH, incubation time, inoculum size, inoculum age, agitation speed, dextrose, starch, L-asparagine, KNO3, yeast extract, K2HPO4, MgSO4.7H2O, NaCl and FeSO4. 7H2O) were screened using Plackett–Burman experimental design. The most positive significant independent variables affecting enzyme production (temperature, inoculum age and agitation speed) were further optimized by the central composite face-centered design -response surface methodology. As a result, a medium of the following formula is the optimum for producing an extracellular L-asparaginase in the culture filtrate of Streptomyces olivaceus NEAE-119: Dextrose 3g, starch 20g, L-asparagine 10g, KNO3 1g, K2HPO4 1g, MgSO4.7H2O 0.1g, NaCl 0.1g, pH 7, temperature 37°C, agitation speed 200 rpm/min, inoculum size 4%, v/v, inoculum age 72 h and fermentation period 5 days.

Keywords: Streptomyces olivaceus NEAE-119, glutaminase free L-asparaginase, production, Plackett-Burman design, central composite face-centered design, 16S rRNA, scanning electron microscope

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Authors: Kalbiye Konanc, Ergin Ozturk

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To examine the effects of an ethanol liquid extract obtained from raw bee propolis (PE) on fattening performance and physiology such as vaccine-antibody relationship, microbial profile, immune status and some blood parameters of broiler chickens were used a total of 600 broiler (Ross 308) chicks, obtained from eggs of 288, 38-weeks-old broiler breeding. There were 6 groups: CC (Parent-Control and Offspring-Control, CP (Parent-Control and Offspring-propolis extract, Cip (Parent-Control and Offspring-in-ovo propolis extract), Cis (Parent-Control and Chickens-in-ovo saline), PeC (Parent-propolis extract and Offspring-Control), PeP (Parent-Propolis extract and Offspring-Propolis extract). Each group was consisted of 10 replications with 10 broiler offspring, and the experiment was lasted for 6 weeks with ethanol-extracted propolis concentration is 400 ppm/kg diet. While the highest feed consumptions at 0-21 days and 0-42 days were found in PeC, the best feed conversion ratio at 0-42 days was found in CP group. The live weight gains were found not to be different among the groups. The highest alanine aminotransferase activities were found in CC and CP and aspartate aminotransferase activities in PeP and PeC groups. The highest triglyceride and total antioxidant levels were found highest in CC and the highest total oxidant level in Cip group. IgA level in hatched eggs and IgM value after slaughtering were highest in Cip group. The best immune response was obtained for 21st day Newcastle Disease vaccine in CC and Cis groups and for 28th day Infectious Bursal Disease vaccine in CP group. The highest total aerobic microorganism and the lowest total fungi count were found in PeP group. In conclusion, it was determined that in-ovo propolis ethanol extract (Cip) increased the maternal antibody levels, that had not consistent effects on blood biochemical parameters except for triglyceride, that led to decrease in E. coli counts and that it can provide strong immune response against Infectious Bursal Disease.

Keywords: bee propolis, in-ovo feeding, immune parameters, poultry, maternal antibody, microorganisms

Procedia PDF Downloads 291

Authors: Shital Bangar, Chetana Mandavia

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A field experiment was carried out in Factorial Randomized Block Design (FRBD) with three replications at Instructional Farm Krushigadh, Junagadh Agricultural University, Junagadh, India to assess the biochemical parameters of wheat in order to assess the thermotolerance. Nine different wheat varieties GW 433, GW 431, HI 1571, GW 432, RAJ 3765, HD 2864, HI 1563, HD 3091 and PBW 670 sown in timely and late sown conditions (i.e., 22 Nov and 6 Dec 2012) were analysed. All the varieties differed significantly with respect to grain yield morpho-physiological parameters and growth indices for time of sowing, varieties and varieties x time of sowing interactions. The observations on morpho-physiological parameters viz., germination percentage, canopy temperature depression and growth indices viz., leaf area index (LAI), leaf area ratio (LAR) were recorded. Almost all the morpho-physiological parameters, growth indices and grain yield studied were affected adversely by late sowing, registering reduction in their magnitude. Germination percentage was reduced under late sown condition but variety PBW 670 was the best. Varieties GW 432 performed better with respect to canopy temperature depression while sown late. Under late sown condition, variety GW 431 recorded higher LAI while HI 1563 had maximum LAR. Considering yield performance, HD 2864 was best under timely sown condition, while GW 433 was best under late sown condition. Varieties HI 1571, GW 433 and GW 431 could be labelled as thermo-tolerant because there was least reduction in grain yield under late sown condition (1.75 %, 7.90 % and13.8 % respectively). Considering correlation coefficient, grain yield showed very strong significant positive association with germination percentage. Leaf area ratio was strongly and significantly correlated with grain yield but in negative direction. Canopy temperature depression and leaf area index also had positive correlation with grain yield but were non-significant.

Keywords: growth indices, morpho-physiological parametrs, thermo-tolerance, wheat

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Authors: Didem Ildırar, Serap Fındık

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Baker’s yeast industry uses molasses as a raw material. Molasses wastewater contains high molecular weight polymers called melanoidins. Melanoidins are obtained after the reactions between the amino acids and carbonyl groups in molasses. The molasses wastewater has high biochemical and chemical oxygen demand and dark brown color. If it is discharged to receiving bodies without any treatment, it prevents light penetration and dissolved oxygen level of the surface water decreases. Melanoidin compounds are toxic effect to the microorganism in water and there is a resistance to microbial degradation. Before discharging molasses wastewater, adequate treatment is necessary. In addition to changing environmental regulations, properties of treated wastewater must be improved. Advanced oxidation processes can be used to improve existing properties of wastewater. Sonochemical oxidation is one of the alternative methods. Sonochemical oxidation employs the use of ultrasound resulting in cavitation phenomena. In this study, decolorization and chemical oxygen demand removal (COD) of baker’s yeast effluent was investigated by using ultrasound. Baker’s yeast effluent was supplied from a factory which is located in the north of Turkey. An ultrasonic homogenizator was used for this study. Its operating frequency is 20kHz. SnO2/TiO2 catalyst has been used as sonocatalyst. The effects of the composite preparation method, mixing time while composite prepared, the molar ratio of SnO2/TiO2, the calcination temperature, and time, the catalyst amount were investigated on the treatment of baker’s yeast effluent. . According to the results, the prepared composite SnO2/TiO2 by using ultrasonic probe gave a better result than prepared composite by using an ultrasonic bath. Prepared composite by using an ultrasonic probe with a 4:1 molar ratio treated at 800°C for 60min gave a better result. By using this composite, optimum catalyst amount was 0.2g/l. At these conditions 26.6% decolorization was obtained. There was no COD removal at the studied conditions.

Keywords: baker’s yeast effluent, COD, decolorization, sonocatalyst, ultrasonic irradiation

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Authors: Sunidhi Syal, Rasangi Tennakoon, Patrick O'Donoghue

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Polyglutamine (polyQ) diseases are a group of diseases related to neurodegeneration caused by repeats of the amino acid glutamine (Q) in the DNA, which translates into an elongated polyQ tract in the protein. The pathological explanation is that the polyQ tract forms cytotoxic aggregates in the neurons, leading to their degeneration. There are no cures or preventative efforts established for these diseases as of today, although the symptoms of these diseases can be relieved. This study specifically focuses on Huntington's disease, which is a type of polyQ disease in which aggregation is caused by the extended cytosine, adenine, guanine (CUG) codon repeats in the huntingtin (HTT) gene, which encodes for the huntingtin protein. Using this principle, we attempted to create six models, which included mutating wildtype tRNA alanine variant tRNA-AGC-8-1 to have glutamine anticodons CUG and UUG so serine is incorporated at glutamine sites in poly Q tracts. In the process, we were successful in obtaining tAla-8-1 CUG mutant clones in the HTTexon1 plasmids with a polyQ tract of 23Q (non-pathogenic model) and 74Q (disease model). These plasmids were transfected into mouse neuroblastoma cells to characterize protein synthesis and aggregation in normal and mistranslating cells and to investigate the effects of glutamines replaced with alanines on the disease phenotype. Notably, we observed no noteworthy differences in mean fluorescence between the CUG mutants for 23Q or 74Q; however, the Triton X-100 assay revealed a significant reduction in insoluble 74Q aggregates. We were unable to create a tAla-8-1 UUG mutant clone, and determining the difference in the effects of the two glutamine anticodons may enrich our understanding of the disease phenotype. In conclusion, by generating structural disruption with the amino acid alanine, it may be possible to find ways to minimize the toxicity of Huntington's disease caused by these polyQ aggregates. Further research is needed to advance knowledge in this field by identifying the cellular and biochemical impact of specific tRNA variants found naturally in human genomes.

Keywords: Huntington's disease, polyQ, tRNA, anticodon, clone, overlap PCR

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Authors: Jordan Chamarande, Lisiane Cunat, Corentine Alauzet, Catherine Cailliez-Grimal

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Gut microbiota (GM) is now considered a new organ mainly due to the microorganism’s specific biochemical interaction with its host. Although mechanisms underlying host-microbiota interactions are not fully described, it is now well-defined that cell surface molecules and structures of the GM play a key role in such relation. The study of surface structures of GM members is also fundamental for their role in the establishment of species in the versatile and competitive environment of the digestive tract and as a potential virulence factor. Among these structures are capsular polysaccharides (CPS), fimbriae, pili and lipopolysaccharides (LPS), all well-described for their central role in microorganism colonization and communication with host epithelium. The health-promoting Parabacteroides distasonis, which is part of the core microbiome, has recently received a lot of attention, showing beneficial properties for its host and as a new potential biotherapeutic product. However, to the best of the authors’ knowledge, the cell surface molecules and structures of P. distasonis that allow its maintain within the GM are not identified. Moreover, although P. distasonis is strongly recognized as intestinal commensal species with benefits for its host, it has also been recognized as an opportunistic pathogen. In this study, we reported gene clusters potentially involved in the synthesis of the capsule, fimbriae-like and pili-like cell surface structures in 26 P. distasonis genomes and applied the new RfbA-Typing classification in order to better understand and characterize the beneficial/pathogenic behaviour related to P. distasonis strains. In context, 2 different types of fimbriae, 3 of pilus and up to 14 capsular polysaccharide loci, have been identified over the 26 genomes studied. Moreover, the addition of data to the rfbA-Type classification modified the outcome by rearranging rfbA genes and adding a fifth group to the classification. In conclusion, the strain variability in terms of external proteinaceous structure could explain the inter-strain differences previously observed in P. distasonis adhesion capacities and its potential pathogenicity.

Keywords: gut microbiota, Parabacteroides distasonis, capsular polysaccharide, fimbriae, pilus, O-antigen, pathogenicity, probiotic, comparative genomics

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Authors: Sher Mohammed, Purushottam Lal, Pawan K. Kasera

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The Thar Desert of Rajasthan covers a wide geographical area spreading between 23.3° to 30.12°, North latitude and 69.3◦ to 76◦ Eastern latitudes; having a unique spectrum of arid zone vegetation. This desert is spreading over 12 districts having a rich source of economically important/threatened plant diversity interacting and growing with adverse climatic conditions of the area. Due to variable geological, physiographic, climatic, edaphic and biotic factors, the arid zone medicinal flora exhibit a wide collection of angiosperm families. The herbal diversity of this arid region is medicinally important in household remedies among tribal communities as well as in traditional systems. The on-going increasing disturbances in natural ecosystems are due to climatic and biological, including anthropogenic factors. The unique flora and subsequently dependent faunal diversity of the desert ecosystem is losing its biotic potential. A large number of plants have no future unless immediate steps are taken to arrest the causes, leading to their biological improvement. At present the potential loss in ecological amplitude of various genera and species is making several plant species as red listed plants of arid zone vegetation such as Commmiphora wightii, Tribulus rajasthanensis, Calligonum polygonoides, Ephedra foliata, Leptadenia reticulata, Tecomella undulata, Blepharis sindica, Peganum harmala, Sarcostoma vinimale, etc. Mostly arid zone species are under serious pressure against prevailing ecosystem factors to continuation their life cycles. Genetic, molecular, cytological, biochemical, metabolic, reproductive, germination etc. are the several points where the floral diversity of the arid zone area is facing severe ecological influences. So, there is an urgent need to conserve them. There are several opportunities in the field to carry out remarkable work at particular levels to protect the native plants in their natural habitat instead of only their in vitro multiplication.

Keywords: ecology, evaluation, xerophytes, economically, threatened plants, conservation

Procedia PDF Downloads 267

Authors: Salma A. El-Marasy, Rehab F. Abdel-Rahman, Reham M. Abd-Elsalam

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The burden of stroke is intensely increasing worldwide. Brain injury following transient or permanent focal cerebral ischemia develops ischemic stroke as a consequence of a complex series of pathophysiological events. The aim of this study is to evaluate the possible neuroprotective effect of a dipeptidyl peptidase-4 inhibitor, vildagliptin, independent on its insulinotropic properties in non-diabetic rats subjected to cerebral ischemia. Anaesthetized Wistar rats were subjected to either left middle cerebral artery occlusion (MCAO) or sham operation followed by reperfusion after 30 min of MCAO. The other three groups were orally administered vildagliptin at 3 dose levels (2.5, 5, 10 mg/kg) for 3 successive weeks before subjected to left focal cerebral ischemia/reperfusion and till the end of the study. Neurological deficit scores and motor activity were assessed 24h following reperfusion. 48h following reperfusion, rats were euthanized and their left brain hemispheres were harvested and used in the biochemical, histopathological, and immunohistochemical investigations. Vildagliptin pretreatment improved neurological score deficit, locomotor activity and motor coordination in MCAO rats. Moreover, vildagliptin reduced malondialdehyde (MDA), elevated reduced glutathione (GSH), phosphotylinosital 3 kinase (PI3K), phosphorylated of protein kinase B (p-AKT), and mechanistic target of rapamycin (mTOR) brain contents in addition to reducing protein expression of caspase-3. Also, vildagliptin showed a dose-dependent attenuation in neuronal cell loss and histopathological alterations in MCAO rats. This study proves that vildagliptin exerted the neuroprotective effect in a dose-dependent manner as shown in amelioration of neuronal cell loss and histopathological damage in MCAO rats, which may be mediated by attenuating neuronal and motor deficits, it’s anti-oxidant property, activation of PI3K/AKT/mTOR pathway and its anti-apoptotic effect.

Keywords: caspase-3, cerebral ischemia, dipeptidyl peptidase-4 inhibitor, oxidative stress, PI3K/AKT/mTOR pathway, rats, vildagliptin

Procedia PDF Downloads 156

Authors: Lavrentiadou S. N., Angelou V., Chatzimisios K., Papazoglou L.

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The aim of this study was the isolation and culture of keratinocytes and fibroblasts from feline skin to ultimately create an artificial engineered skin (including dermis and epidermis) useful for the effective treatment of large cutaneous deficits in cats. Epidermal keratinocytes and dermal fibroblasts were freshly isolated from skin biopsies using an 8 mm biopsy punch obtained from 8 healthy cats that had undergone ovariohysterectomy. The owner’s consent was obtained. All cats had a complete blood count and a serum biochemical analysis and were screened for feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) preoperatively. The samples were cut into small pieces and incubated with collagenase (2 mg/ml) for 5-6 hours. Following digestion, cutaneous cells were filtered through a 100 μm cell strainer, washed with DMEM, and grown in DMEM supplemented with 10% FBS. The undigested epidermis was washed with DMEM and incubated with 0.05% Trypsin/0.02% EDTA (TE) solution. Keratinocytes recovered in the TE solution were filtered through a 100 μm and a 40 μm cell strainer and, following washing, were grown on a collagen type I matrix in DMEM: F12 (3:1) medium supplemented with 10% FΒS, 1 μm hydrocortisone, 1 μm isoproterenol and 0.1 μm insulin. Both fibroblasts and keratinocytes were grown in a humidified atmosphere with 5% CO2 at 37oC. The medium was changed twice a week and cells were cultured up to passage 4. Cells were grown to 70-85% confluency, at which point they were trypsinized and subcultured in a 1:4 dilution. The majority of the cells in each passage were transferred to a freezing medium and stored at -80oC. Fibroblasts were frozen in DMEM supplemented with 30% FBS and 10% DMSO, whereas keratinocytes were frozen in a complete keratinocyte growth medium supplemented with 10% DMSO. Both cell types were thawed and successfully grown as described above. Therefore, we can create a bank of fibroblasts and keratinocytes, from which we can recover cells for further culture and use for the generation of skin equivalent in vitro. In conclusion, cutaneous cell isolation and cell culture and expansion were successfully developed. To the authors’ best knowledge, this is the first study reporting isolation and culture of keratinocytes and fibroblasts from feline skin. However, these are preliminary results and thus, the development of autologous-engineered feline skin is still in process.

Keywords: cat, fibroblasts, keratinocytes, skin equivalent, wound

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Authors: Chakrapani Pullagummi, Arun Jyothi Bheemagani, B. Chandra Sekhar Singh, Prem Kumar, A. Roja Rani

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Diabetes mellitus describes a metabolic disorder of multiple aetiology characterized by chronic hyperglycaemia with disturbances of carbohydrate, fat and protein metabolism resulting from defects in insulin secretion and its action. The objective of present study was alloxan induced diabetes in S.D (Sprague Dawley) rats, treated with leaf extract of Andrographis paniculata and bark extract of Erythrina indica. Plant extract treated rats were analyzed biochemically and molecularly. on normal and diabetic rats. The changes in MDA (lipid peroxidation) and glucose (by GOD method) levels in blood of both normal and diabetic rat were analyzed. Diabetes induced rats were treated with methanolic extracts of Andrographis paniculata leaf and Erythrina indica bark which are of medicinal importance. Later after inducing diabetes the rats were treated with medicinal plant extracts, Andrographis paniculata leaf and Erythrina indica bark which are well known for their anti diabetic and antioxidative property in order to control the glucose and MDA levels. The blood plasma of diabetic and normal rats was analyzed for the levels of MDA (lipid peroxidation) and glucose levels. Results of this study suggested that the Andrographis paniculata leaf and Erythrina indica can be used as a potential natural antidiabetic agent for treating and postponing the appearance of complications that arise due to Diabetes. Molecular study deals with the analysis of binding mechanism of 2 selected natural compounds from Andrographis and Erythrina extracts against the novel target for type T2D namely PPAR-γ compared with Rosiglitazone (standard compound). The results revealed that most of the selected herbal lead compounds were effective targets against the receptors. These compounds showed favorable interactions with the amino acid residues thereby substantiating their proven efficacy as anti-diabetic compounds.

Keywords: andrographis paniculata, erythrina indica, alloxan, lipid peroxidation, blood glucose level, PPAR-γ

Procedia PDF Downloads 477

Authors: Bouchiha Hanene, Rouabhi Rachid, Bouchama Khaled, Djebar Berrebbah Houraya, Djebar Mohamed Reda

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Recently, some natural products such as essentials oils (EOs) have been used in the fields as alternative to synthetic compounds, to minimize the negative impacts to the environment. This fact has led to questions about the possible impact of EOs on ecosystems. Currently in toxicology, the use of alternative models can help to understand the mechanisms of toxic action, at different levels of organization of ecosystems. Algae, protozoa and bacteria form the base of the food chain and protozoan cells are used as bioindicators often of pollution in environment. Unicellular organisms offer the possibility of direct study of independent cells with specific characteristics of individual cells and whole organisms at the same time. This unicellular facilitates the study of physiological processes, and effects of pollutants at the cellular level, which makes it widely used to assess the toxic effects of various xenobiotics. This study aimed to verify the effects of EOs of one famous plant used tremendously in our folk medicine, namely Artemisia herba alba in causing acute toxicity (24 hours) and chronic (15 days) toxicity for model cellular (Paramecium sp). To this end, cellular’s of paramecium were exposed to various concentrations (Three doses were chosen) of EOs extracted from plant (Artemisia herba alba). In the first experiment, the cellular s cultures were exposed for 48 hours to different concentrations to determine the median lethal concentration (DL50). We followed the evolution of physiological parameters (growth), biochemical (total proteins, respiratory metabolism), as well as the variations of a bio marker the GSH. Our results highlighted a light inhibition of the growth of the protozoa as well as a disturbance of the contents of total proteins and a reduction in the reduced rate of glutathione. The polarographic study revealed a stimulation of the consumption of O2 and this at the treated cells.

Keywords: essential oils, protozoa, bio indicators, toxicity, Growth, bio marker, proteins, polarographic

Procedia PDF Downloads 346

Authors: Simon Nyarko

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This study examined the microbial flora contamination of the Ghanaian paper currency notes and antibiotic resistance in Ejura Municipal, Ashanti Region, Ghana. This is a descriptive cross-sectional study designed to assess the profile of microflora contamination of the Ghanaian paper currency notes and antibiotic-resistant in the Ejura Municipality. The research was conducted in Ejura, a town in the Ejura Sekyeredumase Municipal of the Ashanti region of Ghana. 70 paper currency notes which were freshly collected from the bank, consisting of 15 pieces of GH ¢1, GH ¢2, and GH ¢5, 10 pieces of GH ¢10 and GH ¢20, and 5 pieces of GH ¢50, were randomly sampled from people by exchanging their money in usage with those freshly secured from the bank. The surfaces of each GH¢ note were gently swabbed and sent to the lab immediately in sterile Zip Bags and sealed, and tenfold serial dilution was inoculated on plate count agar (PCA), MacConkey agar (MCA), mannitol salt agar (MSA), and deoxycholate citrate agar (DCA). For bacterial identification, the study used appropriate laboratory and biochemical tests. The data was analyzed using SPSS-IBM version 20.0. It was found that 95.2 % of the 70 GH¢ notes tested positive for one or more bacterial isolates. On each GH¢ note, mean counts on PCA ranged from 3.0 cfu/ml ×105 to 4.8 cfu/ml ×105. Of 124 bacteria isolated. 36 (29.03 %), 32 (25.81%), 16 (12.90 %), 20 (16.13%), 13 (10.48 %), and 7 (5.66 %) were from GH¢1, GH¢2, GH¢10, GH¢5, GH¢20, and GH¢50, respectively. Bacterial isolates were Escherichia coli (25.81%), Staphylococcus aureus (18.55%), coagulase-negative Staphylococcus (15.32%), Klebsiella species (12.10%), Salmonella species (9.68%), Shigella species (8.06%), Pseudomonas aeruginosa (7.26%), and Proteus species (3.23%). Meat shops, commercial drivers, canteens, grocery stores, and vegetable shops contributed 25.81 %, 20.16 %, 19.35 %, 17.74 %, and 16.94 % of GH¢ notes, respectively. There was 100% resistance of the isolates to Erythromycin (ERY), and Cotrimoxazole (COT). Amikacin (AMK) was the most effective among the antibiotics as 75% of the isolates were susceptible to it. This study has demonstrated that the Ghanaian paper currency notes are heavily contaminated with potentially pathogenic bacteria that are highly resistant to the most widely used antibiotics and are a threat to public health.

Keywords: microflora, antibiotic resistance, staphylococcus aureus, culture media, multi-drug resistance

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Authors: Akram Sadat Jafari Roodbandi, Farzaneh Akbari, Vafa Feyzi, Zahra Zare, Zohreh Foroozanfar

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Introduction: Circadian rhythm or sleep-awake cycle in 24 hours is one of the important factors affecting the physiological and psychological characteristics in humans that contribute to biochemical, physiological and behavioral processes and helps people to set up brain and body for sleep or active awakening during certain hours. The purpose of this study was to investigate the relationship between the characteristics of circadian rhythms on the sleep quality and sleepiness according to their demographic characteristics such as age. Methods: This cross-sectional descriptive-analytic study was carried out among the general population of Kerman, aged 15-84 years. After dividing the age groups into 10-year demographic characteristics questionnaire, the type of circadian questionnaire, Pittsburgh sleep quality questionnaire and Euporth sleepiness questionnaire were completed in equal numbers between men and women of that age group. Using cluster sampling with effect design equal 2, 1300 questionnaires were distributed during the various hours of 24 hours in public places in Kerman city. Data analysis was done using SPSS software and univariate tests and linear regressions at a significance level of 0.05. Results: In this study, 1147 subjects were included in the study, 584 (50.9%) were male and the rest were women. The mean age was 39.50 ± 15.38. 133 (11.60%) subjects from the study participants had sleepiness and 308 (26.90%) subjects had undesirable sleep quality. Using linear regression test, sleep quality was the significant correlation with sex, hours needed for sleep at 24 hours, chronic illness, sleepiness, and circadian rhythm amplitude. Sleepiness was the meaningful relationship with marital status, sleep-wake schedule of other family members and the stability of circadian rhythm. Both women and men, with age, decrease the quality of sleep and increase the rate of sleepiness. Conclusion: Age, sex, and type of circadian people, the need for sleep at 24 hours, marital status, sleep-wake schedule of other family members are significant factors related to the sleep quality and sleepiness and their adaptation to night shift work.

Keywords: circadian type, sleep quality, sleepiness, age, shift work

Procedia PDF Downloads 157