Search results for: fiducial marker
Commenced in January 2007
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Edition: International
Paper Count: 532

Search results for: fiducial marker

232 Evaluating the Hepato-Protective Activities of Combination of Aqueous Extract of Roots of Tinospora cordifolia and Rhizomes of Curcuma longa against Paracetamol Induced Hepatic Damage in Rats

Authors: Amberkar Mohanbabu Vittalrao, Avin, Meena Kumari Kamalkishore, Padmanabha Udupa, Vinaykumar Bavimane, Honnegouda

Abstract:

Objective: To evaluate the hepato-protective activity of Tinospora cordiofolia (Tc) against paracetamol induced hepatic damage in rats. Methods: The plant stem (test drug) was procured locally, shade dried, powdered and extracted with water. Silymarin was used as standard hepatoprotective drugs and 2% gum acacia as a control (vehicle) against paracetamol (PCT) induced hepatotoxicity. Results and Discussion: The hepato-protective activity of aqueous stem extract was assessed by paracetamol induced hepatotoxicity preventive model in rats. Alteration in the levels of biochemical markers of hepatic damage like AST, ALT, ALP and lipid peroxides were tested in both paracetamol treated and untreated groups. Paracetamol (3g/kg) had enhanced the AST, ALT, ALP and the lipid peroxides in the serum. Treatment of silymarin and aqueous stem extract of Tc (200 and 400mg/kg) extract showed significant hepatoprotective activity by altering biochemical marker levels to the near normal. Preliminary phytochemical tests were done. Aqueous Tc extract showed presence of phenolic compound and flavonoids. Our findings suggested that Tc extract possessed hepatoprotective activity in a dose dependent manner. Conclusions: Tc was found to possess significant hepatoprotective property when treated with PCT. This was evident by decreasing the liver enzymes significantly when treated with PCT as compared to PCT only treated group (P < 0.05). Hence Tinospora cardiofolia could be a good, promising, preventive agent against PCT induced hepatotoxicity.

Keywords: Tinospora cardiofolia, hepatoprotection, paracetamol, silymarin

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231 Diverse Sensitivity to Ultraviolet Radiation of DNA and RNA Viruses

Authors: Nickolay Nosik, Dmitry Nosik, Marina Bochkova, Nina Kondrashina, Olga Lobach

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The bactericidal effect of UV radiation is known for long time and widely used for inactivation of pathogens but for viruses it is not so uniform. Due to a wide variety of viruses their sensitivity to UV radiation is quite different and not quite predictable. The goal of the study was to determine the inactivation kinetics of UV radiation ( 254 nm) of the viruses of social importance (HIV), as well as test-viruses (poliovirus, adenovirus) used for the evaluation of the viral inactivation efficacy of germicides. Methods: DNA viruses- adenovirus, type 5; Herpes simplex virus (HSV), type 1, and RNA viruses–human immunodeficiency virus (HIV), type 1 and poliovirus, type 1 (Sabin strain) were obtained from State collection of viruses ( The D.I. Ivanovsky Institute of Virology). The source of UV radiation was a 15-watt low-pressure mercury vapor lamp (over 60% 254nm). The samples of 5cm2 were placed direct under the UV lamp flow (h-0.3m). Log reduction value was used as a marker for the rate of virus inactivation. Results: The data obtained indicate that poliovirus (one of the viruses most resistant to chemical germicides) and HSV are rather sensitive to UV radiation ( D90 =250-311 J/m2). Adenovirus is much more resistant to UV radiation (750 J/m2 ). The kinetics of adenovirus inactivation : 0 min- 5.0 lg TCID50, 10 min - 5,0, 15 min -4,0, 30 min – 3.5, 60 min – 1,0, 75 min -0,5 lg TCID50, 90 min –virus not detectable. HIV is most resistant to UV radiation among the studied viruses. It takes more than 4 hrs to inactivate the virus on the surface. D90 = 2000 J/m2 Conclusion: The results of the study show that there is no direct dependence between sensitivity to UV light and the size of the virion or presence\absence of the envelope of the virus. Poliovirus and adenovirus are small viruses (20-30nm poliovirus and 70-90nm adenovirus) and both are non-enveloped viruses but adenovirus 3-fold more resistant to UV radiation than poliovirus. It can be expected that viruses with more complicate structure, like Herpes virus (200nm) or HIV (80-100 nm), would be more sensitive to UV light. However, the very high resistance of HIV to UV radiation needs further investigation. The diverse resistance of the different viruses to UV radiation should be taken into the account when UV light is used to inactivate infectious viruses in hospitals and other public environments.

Keywords: HIV, HSV, inhibition of viruses, UV radiation

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230 Diagnostic and Prognostic Use of Kinetics of Microrna and Cardiac Biomarker in Acute Myocardial Infarction

Authors: V. Kuzhandai Velu, R. Ramesh

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Background and objectives: Acute myocardial infarction (AMI) is the most common cause of mortality and morbidity. Over the last decade, microRNAs (miRs) have emerged as a potential marker for detecting AMI. The current study evaluates the kinetics and importance of miRs in the differential diagnosis of ST-segment elevated MI (STEMI) and non-STEMI (NSTEMI) and its correlation to conventional biomarkers and to predict the immediate outcome of AMI for arrhythmias and left ventricular (LV) dysfunction. Materials and Method: A total of 100 AMI patients were recruited for the study. Routine cardiac biomarker and miRNA levels were measured during diagnosis and serially at admission, 6, 12, 24, and 72hrs. The baseline biochemical parameters were analyzed. The expression of miRs was compared between STEMI and NSTEMI at different time intervals. Diagnostic utility of miR-1, miR-133, miR-208, and miR-499 levels were analyzed by using RT-PCR and with various diagnostics statistical tools like ROC, odds ratio, and likelihood ratio. Results: The miR-1, miR-133, and miR-499 showed peak concentration at 6 hours, whereas miR-208 showed high significant differences at all time intervals. miR-133 demonstrated the maximum area under the curve at different time intervals in the differential diagnosis of STEMI and NSTEMI which was followed by miR-499 and miR-208. Evaluation of miRs for predicting arrhythmia and LV dysfunction using admission sample demonstrated that miR-1 (OR = 8.64; LR = 1.76) and miR-208 (OR = 26.25; LR = 5.96) showed maximum odds ratio and likelihood respectively. Conclusion: Circulating miRNA showed a highly significant difference between STEMI and NSTEMI in AMI patients. The peak was much earlier than the conventional biomarkers. miR-133, miR-208, and miR-499 can be used in the differential diagnosis of STEMI and NSTEMI, whereas miR-1 and miR-208 could be used in the prediction of arrhythmia and LV dysfunction, respectively.

Keywords: myocardial infarction, cardiac biomarkers, microRNA, arrhythmia, left ventricular dysfunction

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229 Changes in Behavior and Learning Ability of Rats Intoxicated with Lead

Authors: A. Goma Amira, U. E. Mahrous

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Measuring the effect of perinatal lead exposure on learning ability of offspring is considered as a sensitive and selective index for providing an early marker for central nervous system damage produced by this toxic metal. A total of 35 Sprague-Dawley adult rats were used to investigate the effect of lead acetate toxicity on behavioral patterns of adult female rats and learning ability of offspring. Rats were allotted into 4 groups, group one received 1g/l lead acetate (n=10), group two received 1.5g/l lead acetate (n=10), group three received 2g/l lead acetate in drinking water (n=10), and control group did not receive lead acetate (n=5) from 8th day of pregnancy till weaning of pups. The obtained results revealed a dose-dependent increase in the feeding time, drinking frequency, licking frequency, scratching frequency, licking litters, nest building, and retrieving frequencies, while standing time increased significantly in rats treated with 1.5g/l lead acetate than other treated groups and control. On the contrary, lying time decreased gradually in a dose-dependent manner. Moreover, movement activities were higher in rats treated with 1g/l lead acetate than other treated groups and control. Furthermore, time spent in closed arms was significantly lower in rats given 2g/l lead acetate than other treated groups, while they spent significantly much time spent in open arms than other treated groups which could be attributed to occurrence of adaptation. Furthermore, number of entries in open arms was-dose dependent. However, the ratio between open/closed arms revealed a significant decrease in rats treated with 2g/l lead acetate than the control group.

Keywords: lead toxicity, rats, learning ability, behavior

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228 Effect of Bacillus thuringiensis israelensis against Culex pipiens (Insect: Culicidae) Effect of Bti on Two Non-Target Species Eylais hamata (Acari: Hydrachnidia) and Physa marmorata (Gastropoda: Physidae) and Dosage of Their GST Biomarker

Authors: Meriem Mansouri, Fatiha Bendali Saoudi, Noureddine Soltani

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Biological control presents a means of control for the protection of the environment. Bacillus thuringiensis israelensis Berliner 1915 is an inseticide of biological origin because it is a bacterium of the Bacillaceae family. This biocide has a biological importance, because of its specific larvicidal action against Culicidae, blood-sucking insects, responsible for several diseases to humans and animals through the world. As well as, its high specificity for these insects. Also, the freshwater mites, this necessarily parasitic group for aquatic species such as the Physidae, also have an effective biological control against the Culicidae, because of their voracious predation to the larvae of these insects. The present work aims to study the effects of the biocide Bacillus thuringiensis var israelinsis, against non-target adults of water mites Eylais hamata Koenike, 1897, as well as its associated host species Physa marmorata Fitzinger, 1833. After 12 days of oral treatment of adults with lethal concentration (LC50:0.08µg/ml), determined from essays on 4th instar larvae of Culex pipiens (hematophagous insects). No adverse effect has been recorded for adult individuals of Eylais hamata, contrary, snail Physa marmorata were sensitive for this dose of Bti. In parallel, after treatment at the Bti by LC50, the enzyme stress bio marker glutathione S-transferase, was measured after 24, 48 and 72 hours. The enzymatic activity of GST has increased after 24 and 48 hours following treatment.

Keywords: biological control, Bacillus thuringiensis var israelinsis, culicidae, hydrachnidia, enzymatic activity

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227 Voice of Customer: Mining Customers' Reviews on On-Line Car Community

Authors: Kim Dongwon, Yu Songjin

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This study identifies the business value of VOC (Voice of Customer) on the business. Precisely, we intend to demonstrate how much negative and positive sentiment of VOC has an influence on car sales market share in the unites states. We extract 7 emotions such as sadness, shame, anger, fear, frustration, delight and satisfaction from the VOC data, 23,204 pieces of opinions, that had been posted on car-related on-line community from 2007 to 2009(a part of data collection from 2007 to 2015), and intend to clarify the correlation between negative and positive sentimental keywords and contribution to market share. In order to develop a lexicon for each category of negative and positive sentiment, we took advantage of Corpus program, Antconc 3.4.1.w and on-line sentimental data, SentiWordNet and identified the part of speech(POS) information of words in the customers' opinion by using a part-of-speech tagging function provided by TextAnalysisOnline. For the purpose of this present study, a total of 45,741 pieces of customers' opinions of 28 car manufacturing companies had been collected including titles and status information. We conducted an experiment to examine whether the inclusion, frequency and intensity of terms with negative and positive emotions in each category affect the adoption of customer opinions for vehicle organizations' market share. In the experiment, we statistically verified that there is correlation between customer ideas containing negative and positive emotions and variation of marker share. Particularly, "Anger," a domain of negative domains, is significantly influential to car sales market share. The domain "Delight" and "Satisfaction" increased in proportion to growth of market share.

Keywords: data mining, opinion mining, sentiment analysis, VOC

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226 Morpho-Syntactic Pattern in Maithili Urdu

Authors: Mohammad Jahangeer Warsi

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This is, perhaps, the first linguistic study of Maithili Urdu, a dialect of Urdu language of Indo-Aryan family, spoken by around four million speakers in Darbhanga, Samastipur, Begusarai, Madhubani, and Muzafarpur districts of Bihar. It has the subject–verb–object (SOV) word order and it lacks script and literature. Needless to say, this work is an attempt to document this dialect so that it should contribute to the field of descriptive linguistics. Besides, it is also spoken by majority of Maithili diaspora community. Maithili Urdu does not have its own script or literature, yet it has maintained an oral history of over many centuries. It has contributed to enriching the Maithili, Hindi and Urdu languages and literature very profoundly. Dialects are the contact languages of particular regions, and they have a deep impact on their cultural heritage. Slowly with time, these dialects begin to take shape of languages. The convergence of a dialect into a language is a symbol and pride of the people who speak it. Although, confined to the five districts of northern Bihar, yet highly popular among the natives, it is the primary mode of communication of the local Muslims. The paper will focus on the structure of expressions about Maithili Urdu that include the structure of words, phrases, clauses, and sentences. There are clear differences in linguistic features of Maithili Urdu vis-à-vis Urdu, Maithili and Hindi. Though being a dialect of Urdu, interestingly, there is only one second person pronoun tu and lack of agentive marker –ne. Although being spoken in the vicinity of Hindi, Urdu and Maithili, it undoubtedly has its own linguistic features, of them, verb conjugation is remarkably unique. Because of the oral tradition of this link language, intonation has become significantly prominent. This paper will discuss the morpho-syntactic pattern of Maithili Urdu and will go through a sample text to authenticate the findings.

Keywords: cultural heritage, morpho-syntactic pattern, Maithili Urdu, verb conjugation

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225 Molecular Mechanism on Inflammation and Antioxidant Role of Pterocarpus Marsupiumin in Experimental Hyperglycaemia

Authors: Leelavinothan Pari , Ayyasamy Rathinam

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Diabetes mellitus (DM) is a major and growing public health problem throughout the world. Pterocarpus marsupium (Roxb.) (Family: Fabaceae) is widely used as a traditional medicine to treat various diseases including diabetes. However, the molecular mechanism of Pterocarpus marsupium has not been investigated so far. Two fractions (2.5% and 5%) of extract from the medicinal plant, Pterocarpus marsupium (PME) were conducted in a dose dependent manner in streptozotocin (45 mg/kg b.w.) induced type 2 diabetic rats. Each fraction of PME was administered to diabetic rats intragastrically at a dose of 50, 100 and 200 mg/kg b.w for 45 days. The effective dose 200 mg/kg b.w of 5% fraction was more pronounced in reducing the levels of blood glucose (95.65 mg/dL) and glycosylated hemoglobin (HbA1c) (0.41 mg/g Hb), and increasing the plasma insulin (16.20 µU/mL) level. Moreover, PME (200 mg/kg b.w) significantly ameliorated lipid peroxidation products (thiobarbituric reactive substances, lipid hydroperoxides) enzymatic (superoxide dismutase, catalase and glutathione peroxidase) and non-enzymatic antioxidants (Vitamin C, Vitamin E and reduced glutathione) levels. The altered activities of the key enzymes of lipid metabolism along with the lipid profile in diabetic rats were significantly reverted to near normal levels by the administration of PME 5% 200 mg/kg b.w fraction. PME (200 mg/kg b.w) has the ability to reduce the inflammatory cytokines, such as TNF-α, IL-6 mRNA, as well as protein expression and apoptotic marker, such as caspase-3 enzyme in diabetic hepatic tissue. The above biochemical findings were also supported by histological studies such as improvement in pancreas and liver. Pterocarpus marsupium could effectively reduce the hyperglycemia, oxidative-stress, inflammation and hyperlipedimea in diabetic rats; hence it could be a useful drug in the management of diabetes without any side effects.

Keywords: diabetes mellitus, streptozotocin, Pterocarpus marsupium, lipid peroxidation, Antioxidants, inflammatory cytokines

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224 Use of RAPD and ISSR Markers in Detection of Genetic Variation among Colletotrichum falcatum Went Isolates from South Gujarat India

Authors: Prittesh Patel, Rushabh Shah, Krishnamurthy Ramar, Vakulbhushan Bhaskar

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The present research work aims at finding genetic differences in the genomes of sugarcane red rot isolates Colletotrichum falcatum Went using Random Amplified Polymorphic DNA (RAPD) and interspersed simple sequence repeat (ISSR) molecular markers. Ten isolates of C. falcatum isolated from different red rot infected sugarcane cultivars stalk were used in present study. The amplified bands were scored across the lanes obtained in 15 RAPD primes and 21 ISSR primes successfully. The data were analysed using NTSYSpc 2.2 software. The results showed 80.6% and 68.07% polymorphism in RPAD and ISSR analysis respectively. Based on the RAPD analysis, ten genotypes were grouped into two major clusters at a cut-off value of 0.75. Geographically distant C. falcatum isolate cfGAN from south Gujarat had a level of similarity with Coimbatore isolate cf8436 presented on separate clade of bootstrapped dendrograms. First and second cluster consisted of five and three isolates respectively, indicating the close relation among them. The 21 ISSR primers produced 119 distinct and scorable loci in that 38 were monomorphic. The number of scorable loci for each primer varied from 2 (ISSR822) to 8 (ISSR807, ISSR823 and ISSR15) with an average of 5.66 loci per primer. Primer ISSR835 amplified the highest number of bands (57), while only 16 bands were obtained by primers ISSR822. Four primers namely ISSR830, ISSR845, ISSR4 and ISSR15 showed the highest value of percentage of polymorphism (100%). The results indicated that both of the marker systems RAPD and ISSR, individually can be effectively used in determination of genetic relationship among C falcatum accessions collected from different parts of south Gujarat.

Keywords: Colletotrichum falcatum, ISSR, RAPD, Red Rot

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223 Dynamics and Advection in a Vortex Parquet on the Plane

Authors: Filimonova Alexanra

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Inviscid incompressible fluid flows are considered. The object of the study is a vortex parquet – a structure consisting of distributed vortex spots of different directions, occupying the entire plane. The main attention is paid to the study of advection processes of passive particles in the corresponding velocity field. The dynamics of the vortex structures is considered in a rectangular region under the assumption that periodic boundary conditions are imposed on the stream function. Numerical algorithms are based on the solution of the initial-boundary value problem for nonstationary Euler equations in terms of vorticity and stream function. For this, the spectral-vortex meshless method is used. It is based on the approximation of the stream function by the Fourier series cut and the approximation of the vorticity field by the least-squares method from its values in marker particles. A vortex configuration, consisting of four vortex patches is investigated. Results of a numerical study of the dynamics and interaction of the structure are presented. The influence of the patch radius and the relative position of positively and negatively directed patches on the processes of interaction and mixing is studied. The obtained results correspond to the following possible scenarios: the initial configuration does not change over time; the initial configuration forms a new structure, which is maintained for longer times; the initial configuration returns to its initial state after a certain period of time. The processes of mass transfer of vorticity by liquid particles on a plane were calculated and analyzed. The results of a numerical analysis of the particles dynamics and trajectories on the entire plane and the field of local Lyapunov exponents are presented.

Keywords: ideal fluid, meshless methods, vortex structures in liquids, vortex parquet.

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222 Cerebrum Maturity Damage Induced by Fluoride in Suckling Mice

Authors: Hanen Bouaziz, Françoise Croute, Najiba Zeghal

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In order to investigate the toxic effects of fluoride on cerebrum maturity of suckling mice, we treated adult female mice of Swiss Albinos strain by 500 ppm NaF in their drinking water from the 15th day of pregnancy until the day 14 after delivery. All mice were sacrificed on day 14 after parturition. During treatment, levels of thiobarbituric acid reactive substances, the marker of lipid peroxidation extend, increased, while the activities of the antioxidant enzymes such as glutathione peroxidase, superoxide dismutase and catalase and the level of glutathione decreased significantly in cerebellum compared with those of the control group. These results suggested that fluoride enhanced oxidative stress, thereby disturbing the antioxidant defense of nursing pups. In addition, acetylcholinesterase activity in cerebellum was inhibited after treatment with fluoride. In cerebellum of mice, migration of neurons from the external granular layer to the internal granular layer occurred postnatally. Key guidance signals to these migrating neurons were provided by laminin, an extracellular matrix protein fixed to the surface of astrocytes. In the present study, we examined the expression and distribution of laminin in cerebellum of 14-day-old mice. Immunoreactive laminin was disappeared by postnatal day 14 in cerebellum parenchyma of control pups and was restricted to vasculature despite the continued presence of granular cells in the external granular layer. In contrast, in cerebellum of NaF treated pups, laminin was deposited in organised punctuate clusters in the molecular layer. These data indicated that the disruption of laminin distribution might play a major role in the profound derangement of neuronal migration observed in cerebellum of NaF treated pups.

Keywords: acetylcholinesterase activity, cerebellum, laminin, oxidative stress, suckling mice

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221 Effect of 8 Weeks of Intervention on Physical Fitness, Hepatokines, and Insulin Resistance in Obese Subjects

Authors: Adela Penesova, Zofia Radikova, Boris Bajer, Andrea Havranova, Miroslav Vlcek

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Background: The aim of our study was to compare the effect of intensified lifestyle intervention on insulin resistance (HOMA-IR), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and Fibroblast growth factor (FGF) 21 after 8 weeks of lifestyle intervention. Methods: A group of 43 obese patients (13M/30F; 43.0±12.4 years; BMI (body mass index) 31.2±6.3 kg/m2 participated in a weight loss interventional program (NCT02325804) following an 8-week hypocaloric diet (-30% energy expenditure) and physical activity 150 minutes/week. Insulin sensitivity was evaluated according to the homeostasis model assessment of insulin resistance (HOMA-IR) and insulin sensitivity indices according to Matsuda and Cederholm were calculated (ISImat and ISIced). Plasma ALT, AST, Fetuin-A, FGF 21, and physical fitness were measured. Results: The average reduction of body weight was 6.8±4.9 kg (0-15 kg; p=0.0006), accompanied with a significant reduction of body fat amount of fat mass (p=0.03), and waist circumference (p=0.02). Insulin sensitivity has been improved (IR HOMA 2.71±3.90 vs 1.24±0.83; p=0.01; ISIMat 6.64±4.38 vs 8.93±5.36 p ≤ 0.001). Total, LDL cholesterol, and triglycerides decreased (p=0.05, p=0.04, p=0.04, respectively). Physical fitness significantly improved after intervention (as measure VO2 max (maximal oxygen uptake) (p ≤ 0.001). ALT decreased significantly (0.44±0.26 vs post 0.33±0.18 ukat/l, p=0.004); however, AST not (pre 0.40±0.15 vs 0.35±0.09 ukat/l, p=0.07). Hepatokine Fetuin-A significantly decreased after intervention (43.1±10.8 vs 32.6±8.6 ng/ml, p < 0.001); however, FGF 21 levels tended to decrease (146±152 vs 132±164 pg/ml, p=0.07). Conclusion: 8-weeks of diet and physical activity intervention program in obese otherwise healthy subjects led to an improvement of insulin resistance parameters and liver marker profiles, as well as increased physical fitness. This study was supported by grants APVV 15-0228; VEGA 2/0161/16.

Keywords: obesity, diet, exercice, insulin sensitivity

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220 Design and Creation of a BCI Videogame for Training and Measure of Sustained Attention in Children with ADHD

Authors: John E. Muñoz, Jose F. Lopez, David S. Lopez

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Attention Deficit Hyperactivity Disorder (ADHD) is a disorder that affects 1 out of 5 Colombian children, converting into a real public health problem in the country. Conventional treatments such as medication and neuropsychological therapy have been proved to be insufficient in order to decrease high incidence levels of ADHD in the principal Colombian cities. This work demonstrates a design and development of a videogame that uses a brain computer interface not only to serve as an input device but also as a tool to monitor neurophysiologic signal. The video game named “The Harvest Challenge” puts a cultural scene of a Colombian coffee grower in its context, where a player can use his/her avatar in three mini games created in order to reinforce four fundamental aspects: i) waiting ability, ii) planning ability, iii) ability to follow instructions and iv) ability to achieve objectives. The details of this collaborative designing process of the multimedia tool according to the exact clinic necessities and the description of interaction proposals are presented through the mental stages of attention and relaxation. The final videogame is presented as a tool for sustained attention training in children with ADHD using as an action mechanism the neuromodulation of Beta and Theta waves through an electrode located in the central part of the front lobe of the brain. The processing of an electroencephalographic signal is produced automatically inside the videogame allowing to generate a report of the theta/beta ratio evolution - a biological marker, which has been demonstrated to be a sufficient measure to discriminate of children with deficit and without.

Keywords: BCI, neuromodulation, ADHD, videogame, neurofeedback, theta/beta ratio

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219 Molecular Insights into the Genetic Integrity of Long-Term Micropropagated Clones Using Start Codon Targeted (SCoT) Markers: A Case Study with Ansellia africana, an Endangered, Medicinal Orchid

Authors: Paromik Bhattacharyya, Vijay Kumar, Johannes Van Staden

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Micropropagation is an important tool for the conservation of threatened and commercially important plant species of which orchids deserve special attention. Ansellia africana is one such medicinally important orchid species having much commercial significance. Thus, development of regeneration protocols for producing clonally stable regenerates using axillary buds is of much importance. However, for large-scale micropropagation to become not only successful but also acceptable by end-users, somaclonal variations occurring in the plantlets need to be eliminated. In the light of the various factors (genotype, ploidy level, in vitro culture age, explant and culture type, etc.) that may account for the somaclonal variations of divergent genetic changes at the cellular and molecular levels, genetic analysis of micropropagated plants using a multidisciplinary approach is of utmost importance. In the present study, the clonal integrity of the long term micropropagated A. africana plants were assessed using advanced molecular marker system i.e. Start Codon Targeted Polymorphism (SCoT). Our studies recorded a clonally stable regeneration protocol for A. africana with a very high degree of clonal fidelity amongst the regenerates. The results obtained from these molecular analyses could help in modifying the regeneration protocols for obtaining clonally stable true to type plantlets for sustainable commercial use.

Keywords: medicinal orchid micropropagation, start codon targeted polymorphism (SCoT), RAP), traditional African pharmacopoeia, genetic fidelity

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218 Anti-Phosphorylcholine T Cell Dependent Antibody

Authors: M. M. Rahman, A. Liu, A. Frostegard, J. Frostegard

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The human immune system plays an essential role in cardiovascular disease (CVD) and atherosclerosis. Our earlier studies showed that major immunocompetent cells including T cells are activated by phosphorylcholine epitope. Further, we have determined for the first time in a clinical cohort that antibodies against phosphorylcholine (anti-PC) are negatively and independently associated with the development of atherosclerosis and thus a low risk of cardiovascular diseases. It is still unknown whether activated T cells play a role in anti-PC production. Here we aim to clarify the role of T cells in anti-PC production. B cell alone, or with CD3 T, CD4 T or with CD8 T cells were cultured in polystyrene plates to examine anti-PC IgM production. In addition to mixed B cell with CD3 T cell culture, B cells with CD3 T cells were also cultured in transwell co-culture plates. Further, B cells alone and mixed B cell with CD3 T cell cultures with or without anti-HLA 2 antibody were cultured for 6 days. Anti-PC IgM was detected by ELISA in independent experiments. More than 8 fold higher levels of anti-PC IgM were detected by ELISA in mixed B cell with CD3 T cell cultures in comparison to B cells alone. After the co-culture of B and CD3 T cells in transwell plates, there were no increased antibody levels indicating that B and T cells need to interact to augment anti-PC IgM production. Furthermore, anti-PC IgM was abolished by anti-HLA 2 blocking antibody in mixed B and CD3 T cells culture. In addition, the lack of increased anti-PC IgM in mixed B with CD8 T cells culture and the increased levels of anti-PC in mixed B with CD4 T cells culture support the role of helper T cell for the anti-PC IgM production. Atherosclerosis is a major cause of cardiovascular diseases, but anti-PC IgM is a protection marker for atherosclerosis development. Understanding the mechanism involved in the anti-PC IgM regulation could play an important role in strategies to raise anti-PC IgM. Studies suggest that anti-PC is T-cell independent antibody, but our study shows the major role of T cell in anti-PC IgM production. Activation of helper T cells by immunization could be a possible mechanism for raising anti-PC levels.

Keywords: anti-PC, atherosclerosis, aardiovascular diseases, phosphorylcholine

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217 Anti-Aging Effects of Retinol and Alpha Hydroxy Acid on Elastin Fibers of Artificially Photo-Aged Human Dermal Fibroblast Cell Lines

Authors: Mohammed Jarrar, Shalini Behl, Nadia Shaheen, Abeer Fatima, Reem Nasab

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Skin aging is a slow multifactorial process influenced by both internal as well as external factors. Ultra-violet radiations (UV), diet, smoking and personal habits are the most common environmental factors that affect skin aging. Fat contents and fibrous proteins as collagen and elastin are core internal structural components. The direct influence of UV on elastin integrity and health is crucial on aging of skin by time. The deposition of abnormal elastic material is a major marker in a photo-aged skin. Searching for compounds that may protect against cutaneous photo-damage is highly valued. Retinoids and Alpha Hydroxy Acids protective and or repairing effects of UV have been endorsed by some researchers. For consolidating a better understanding of anti and protective effects of such anti-aging agents, we evaluated the combinatory effects of various dosages of lactic acid and retinol on the dermal fibroblasts elastin levels exposed to UV. The UV exposed cells showed significant reduction in the elastin levels. A combination of drugs with a higher concentration of lactic acid (30-35 mM) and a lower concentration of retinol (10-15mg/mL) showed to work better in enhancing elastin concentration in UV exposed cells. We assume this enhancement could be the result of increased tropo-elastin gene expression stimulated by retinol and lactic acid probably repaired the UV irradiated damage by enhancing the amount and integrity of the elastin fibers.

Keywords: alpha hydroxy acid, elastin, retinol, ultraviolet radiations

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216 Molecular Epidemiology of Anthrax in Georgia

Authors: N. G. Vepkhvadze, T. Enukidze

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Anthrax is a fatal disease caused by strains of Bacillus anthracis, a spore-forming gram-positive bacillus that causes the disease anthrax in animals and humans. Anthrax is a zoonotic disease that is also well-recognized as a potential agent of bioterrorism. Infection in humans is extremely rare in the developed world and is generally due to contact with infected animals or contaminated animal products. Testing of this zoonotic disease began in 1907 in Georgia and is still being tested routinely to provide accurate information and efficient testing results at the State Laboratory of Agriculture of Georgia. Each clinical sample is analyzed by RT-PCR and bacteriology methods; this study used Real-Time PCR assays for the detection of B. anthracis that rely on plasmid-encoded targets with a chromosomal marker to correctly differentiate pathogenic strains from non-anthracis Bacillus species. During the period of 2015-2022, the State Laboratory of Agriculture (SLA) tested 250 clinical and environmental (soil) samples from several different regions in Georgia. In total, 61 out of the 250 samples were positive during this period. Based on the results, Anthrax cases are mostly present in Eastern Georgia, with a high density of the population of livestock, specifically in the regions of Kakheti and Kvemo Kartli. All laboratory activities are being performed in accordance with International Quality standards, adhering to biosafety and biosecurity rules by qualified and experienced personnel handling pathogenic agents. Laboratory testing plays the largest role in diagnosing animals with anthrax, which helps pertinent institutions to quickly confirm a diagnosis of anthrax and evaluate the epidemiological situation that generates important data for further responses.

Keywords: animal disease, baccilus anthracis, edp, laboratory molecular diagnostics

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215 Urine Neutrophil Gelatinase-Associated Lipocalin as an Early Marker of Acute Kidney Injury in Hematopoietic Stem Cell Transplantation Patients

Authors: Sara Ataei, Maryam Taghizadeh-Ghehi, Amir Sarayani, Asieh Ashouri, Amirhossein Moslehi, Molouk Hadjibabaie, Kheirollah Gholami

Abstract:

Background: Acute kidney injury (AKI) is common in hematopoietic stem cell transplantation (HSCT) patients with an incidence of 21–73%. Prevention and early diagnosis reduces the frequency and severity of this complication. Predictive biomarkers are of major importance to timely diagnosis. Neutrophil gelatinase associated lipocalin (NGAL) is a widely investigated novel biomarker for early diagnosis of AKI. However, no study assessed NGAL for AKI diagnosis in HSCT patients. Methods: We performed further analyses on gathered data from our recent trial to evaluate the performance of urine NGAL (uNGAL) as an indicator of AKI in 72 allogeneic HSCT patients. AKI diagnosis and severity were assessed using Risk–Injury–Failure–Loss–End-stage renal disease and AKI Network criteria. We assessed uNGAL on days -6, -3, +3, +9 and +15. Results: Time-dependent Cox regression analysis revealed a statistically significant relationship between uNGAL and AKI occurrence. (HR=1.04 (1.008-1.07), P=0.01). There was a relation between uNGAL day +9 to baseline ratio and incidence of AKI (unadjusted HR=.1.047(1.012-1.083), P<0.01). The area under the receiver-operating characteristic curve for day +9 to baseline ratio was 0.86 (0.74-0.99, P<0.01) and a cut-off value of 2.62 was 85% sensitive and 83% specific in predicting AKI. Conclusions: Our results indicated that increase in uNGAL augmented the risk of AKI and the changes of day +9 uNGAL concentrations from baseline could be of value for predicting AKI in HSCT patients. Additionally uNGAL changes preceded serum creatinine rises by nearly 2 days.

Keywords: acute kidney injury, hemtopoietic stem cell transplantation, neutrophil gelatinase-associated lipocalin, Receiver-operating characteristic curve

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214 Quantitative Trait Loci Analysis in Multiple Sorghum Mapping Populations Facilitates the Dissection of Genetic Control of Drought Tolerance Related Traits in Sorghum [Sorghum bicolor (Moench)]

Authors: Techale B., Hongxu Dong, Mihrete Getinet, Aregash Gabizew, Andrew H. Paterson, Kassahun Bantte

Abstract:

The genetic architecture of drought tolerance is expected to involve multiple loci that are unlikely to all segregate for alternative alleles in a single bi-parental population. Therefore, the identification of quantitative trait loci (QTL) that are expressed in diverse genetic backgrounds of multiple bi-parental populations provides evidence about both background-specific and common genetic variants. The purpose of this study was to map QTL related to drought tolerance using three connected mapping populations of different genetic backgrounds to gain insight into the genomic landscape of this important trait in elite Ethiopian germplasm. The three bi-parental populations, each with 207 F₂:₃ lines, were evaluated using an alpha lattice design with two replications under two moisture stress environments. Drought tolerance related traits were analyzed separately for each population using composite interval mapping, finding a total of 105 QTLs. All the QTLs identified from individual populations were projected on a combined consensus map, comprising a total of 25 meta QTLs for seven traits. The consensus map allowed us to deduce locations of a larger number of markers than possible in any individual map, providing a reference for genetic studies in different genetic backgrounds. The mQTL identified in this study could be used for marker-assisted breeding programs in sorghum after validation. Only one trait, reduced leaf senescence, showed a striking bias of allele distribution, indicating substantial standing variation among present varieties that might be employed in improving drought tolerance of Ethiopian and other sorghums.

Keywords: Drought tolerance , Mapping populations, Meta QTL, QTL mapping, Sorghum

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213 Translating Silence: An Analysis of Dhofar University Student Translations of Elliptical Structures from English into Arabic

Authors: Ali Algryani

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Ellipsis involves the omission of an item or items that can be recovered from the preceding clause. Ellipsis is used as a cohesion marker; it enhances the cohesiveness of a text/discourse as a clause is interpretable only through making reference to an antecedent clause. The present study attempts to investigate the linguistic phenomenon of ellipsis from a translation perspective. It is mainly concerned with how ellipsis is translated from English into Arabic. The study covers different forms of ellipsis, such as noun phrase ellipsis, verb phrase ellipsis, gapping, pseudo-gapping, stripping, and sluicing. The primary aim of the study, apart from discussing the use and function of ellipsis, is to find out how such ellipsis phenomena are dealt with in English-Arabic translation and determine the implications of the translations of elliptical structures into Arabic. The study is based on the analysis of Dhofar University (DU) students' translations of sentences containing different forms of ellipsis. The initial findings of the study indicate that due to differences in syntactic structures and stylistic preferences between English and Arabic, Arabic tends to use lexical repetition in the translation of some elliptical structures, thus achieving a higher level of explicitness. This implies that Arabic tends to prefer lexical repetition to create cohesion more than English does. Furthermore, the study also reveals that the improper translation of ellipsis leads to interpretations different from those understood from the source text. Such mistranslations can be attributed to student translators’ lack of awareness of the use and function of ellipsis as well as the stylistic preferences of both languages. This has pedagogical implications on the teaching and training of translation students at DU. Students' linguistic competence needs to be enhanced through teaching linguistics-related issues with reference to translation and both languages, .i.e. source and target languages and with special emphasis on their use, function and stylistic preferences.

Keywords: cohesion, ellipsis, explicitness, lexical repetition

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212 Gender Estimation by Means of Quantitative Measurements of Foramen Magnum: An Analysis of CT Head Images

Authors: Thilini Hathurusinghe, Uthpalie Siriwardhana, W. M. Ediri Arachchi, Ranga Thudugala, Indeewari Herath, Gayani Senanayake

Abstract:

The foramen magnum is more prone to protect than other skeletal remains during high impact and severe disruptive injuries. Therefore, it is worthwhile to explore whether these measurements can be used to determine the human gender which is vital in forensic and anthropological studies. The idea was to find out the ability to use quantitative measurements of foramen magnum as an anatomical indicator for human gender estimation and to evaluate the gender-dependent variations of foramen magnum using quantitative measurements. Randomly selected 113 subjects who underwent CT head scans at Sri Jayawardhanapura General Hospital of Sri Lanka within a period of six months, were included in the study. The sample contained 58 males (48.76 ± 14.7 years old) and 55 females (47.04 ±15.9 years old). Maximum length of the foramen magnum (LFM), maximum width of the foramen magnum (WFM), minimum distance between occipital condyles (MnD) and maximum interior distance between occipital condyles (MxID) were measured. Further, AreaT and AreaR were also calculated. The gender was estimated using binomial logistic regression. The mean values of all explanatory variables (LFM, WFM, MnD, MxID, AreaT, and AreaR) were greater among male than female. All explanatory variables except MnD (p=0.669) were statistically significant (p < 0.05). Significant bivariate correlations were demonstrated by AreaT and AreaR with the explanatory variables. The results evidenced that WFM and MxID were the best measurements in predicting gender according to binomial logistic regression. The estimated model was: log (p/1-p) =10.391-0.136×MxID-0.231×WFM, where p is the probability of being a female. The classification accuracy given by the above model was 65.5%. The quantitative measurements of foramen magnum can be used as a reliable anatomical marker for human gender estimation in the Sri Lankan context.

Keywords: foramen magnum, forensic and anthropological studies, gender estimation, logistic regression

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211 The Predictive Value of Micro Rna 451 on the Outcome of Imatinib Treatment in Chronic Myeloid Leukemia Patients

Authors: Nehal Adel Khalil, Amel Foad Ketat, Fairouz Elsayed Mohamed Ali, Nahla Abdelmoneim Hamid, Hazem Farag Manaa

Abstract:

Background: Chronic myeloid leukemia (CML) represents 15% of adult leukemias. Imatinib Mesylate (IM) is the gold standard treatment for new cases of CML. Treatment with IM results in improvement of the majority of cases. However, about 25% of cases may develop resistance. Sensitive and specific early predictors of IM resistance in CML patients have not been established to date. Aim: To investigate the value of miR-451 in CML as an early predictor for IM resistance in Egyptian CML patients. Methods: The study employed Real time Polymerase Reaction (qPCR) technique to investigate the leucocytic expression of miR-451 in fifteen newly diagnosed CML patients (group I), fifteen IM responder CML patients (group II), fifteen IM resistant CML patients (group III) and fifteen healthy subjects of matched age and sex as a control group (group IV). The response to IM was defined as < 10% BCR-ABL transcript level after 3 months of therapy. The following parameters were assessed in subjects of all the studied groups: 1- Complete blood count (CBC). 2- Measurement of plasma level of miRNA 451 using real-time Polymerase Chain Reaction (qPCR). 3- Detection of BCR-ABL gene mutation in CML using qPCR. Results: The present study revealed that miR-451 was significantly down-regulated in leucocytes of newly diagnosed CML patients as compared to healthy subjects. IM responder CML patients showed an up-regulation of miR- 451 compared with IM resistant CML patients. Conclusion: According to the data from the present study, it can be concluded that leucocytic miR- 451 expression is a useful additional follow-up marker for the response to IM and a promising prognostic biomarker for CML.

Keywords: chronic myeloid leukemia, imatinib resistance, microRNA 451, Polymerase Chain Reaction

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210 Mouse Knockouts for Elucidating the Role of Cysteine-Rich Angiogenic Inducer 61 in Tendon Development and Maintenance

Authors: Josephine Hai, Jie Jiang, Karen M. Lyons

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Of the musculoskeletal tissues, tendon is least understood in terms of biological development. The current study examines Cysteine-rich angiogenic inducer 61, or CCN1, a member of the CCN family of secreted matricellular proteins that regulate cell behavior via intercellular signaling. Though CCN1 is notable in limiting fibrosis by inducing senescence in fibroblasts, little is known about its role in normal fibrous tissue, where it may be essential to the development of ECM-rich structures like tendons. We found that CCN1 knockout mice (using limb-specific Prx1-Cre) exhibited clubfoot and waddling gaits, a unique phenotype not described in any other mutant to date. Histological analysis showed that the Achilles and patellar tendons, where we previously found high CCN1 expression in adult reporter mice, were thicker and denser in the Prx1-Cre knockouts than in their wildtype littermates. We then hypothesized that CCN1 is required directly in tendon progenitor cells for normal tendon development and generated tendon-specific CCN1 knockout mice using Scx-Cre. We observed similar Achilles/patellar tendon morphology among the Scx-Cre and Prx1-Cre mutants, indicating that the phenotype is a direct result of CCN1’s loss in tendon. To further study phenotype onset and progression, we will histologically characterize these tendons across different developmental time-points. We will also perform RNA-seq and qPCR to analyze tenocyte gene expression and expect fibrotic marker upregulation in the Scx-Cre mutants if CCN1 is required to maintain a normal tendon phenotype. Thus, our study aims to elucidate the molecular mechanisms underlying tendon formation and maintenance. Understanding tendons at the most basic level invites novel approaches to tendon repair.

Keywords: development, matricellular, musculoskeletal, tendon

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209 Anti-TNF: Possibilities of Rising Anti-Phosphorylcholine Antibodies

Authors: Md. Mizanur Rahman, Anquan Liu, Anna Frostegård, Johan Frostegård

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The role of the human immune system is essential in cardiovascular diseases and atherosclerosis. Activated cells in atherosclerosis produce abundant amounts of cytokines, but the exact mechanisms involved in the effects of these inflammatory cytokines are not clear in atherosclerosis. In a large clinical cohort, we have previously determined that antibodies against phosphorylcholine (anti-PC) are negatively and independently associated with both development of atherosclerosis and also a low risk of cardiovascular disease. Further, we reported that rheumatoid arthritis patients who were non-responders to TNF-inhibitors, where those with low anti-PC levels. Upon anti-TNF treatment, anti-PC levels increased. We, therefore, hypothesised that proinflammatory cytokines such as TNF could play a role in anti-PC regulation. Peripheral blood mononuclear cells (PBMC) were cultured with or without TNF and anti-TNF. The cell supernatants were collected after six days for ELISA measurements. In separate experiments, cells were cultured for 24 hours in both polystyrene plates and ELISPOT plates under a similar condition for ELISA and ELISPOT assays respectively. Total RNA was extracted after 6 hours of cell culture to perform RT-qPCR. Cell viability was confirmed by trypan blue staining and MTT assays. ELISA measurements detected less than 40% of anti-PC in TNF-treated cells, in comparison to control cells, whereas anti-PC production was recovered by anti-TNF treatment. ELISPOT assays showed that TNF suppresses anti-PC production by inhibiting anti-PC producing B-cells. In addition, RT-qPCR and ELISA showed that TNF also has effects also on B-cell activation as BAFF expression was inhibited by TNF treatment. Atherosclerosis is a major cause of cardiovascular diseases, but anti-PC is a protection marker for atherosclerosis development. Our findings show that TNF is a negative regulator of anti-PC production. Immune modulation and rising of anti-PC could be of major significance for the patients.

Keywords: anti-PC, Anti-TNF, atherosclerosis, cardiovascular diseases, phosphorylecholine

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208 Phytochemical Screening and Hepatotoxic Effect of Datura metel Linn. Aqueous Seed Extract in Albino Wistar Rats

Authors: I. M. Fakai, A. Abdulhamid, I. Sani, F. Bello, E. O. Olusesi

Abstract:

The phytochemical screening and hepatotoxic effect of Datura metel aqueous seeds extract in Albino Wistar rats were evaluated. Phytochemicals were screened using standard methods. The enzymes activity and liver function indices were also determined using standard methods of analysis. The phytochemicals screening revealed the presence of alkaloid, tannin, glycoside and flavonoid. The organ-body weight decreased significantly (P<0.05) at all the doses of the extract treated groups compared to the control. The activity of alkaline phosphatase decreased significantly (P<0.05) in the liver and increased significantly in the serum at all the doses of the extract treated groups compared to the control. The activity of serum alanine transaminase increased significantly (P<0.05) while there is no significant difference (P>0.05) in the activity liver alanine transaminase at all the doses of the extract treated groups compared to the control. The result also revealed significant increase (P<0.05) in the aspartate transaminase activity in both liver and serum at all doses of the extract treated groups compared to the control. Serum total protein, albumin, globulin, and total bilirubin concentration decreased significantly (P<0.05), while direct bilirubin concentration increased significantly (P<0.05) at all the doses of the extract treated groups compared to the control. The present study therefore revealed that, the present of some phytochemicals in the plant extract attributed the plant to its hepatotoxic effects as revealed in the alteration of marker enzymes and some liver function indices analyzed.

Keywords: datura metel, transaminases, hepatotoxic effect, phytochemicals, rats

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207 Relationship between Matrilin-3 (MATN-3) Gene Single Nucleotide Six Polymorphism, Transforming Growth Factor Beta 2 and Radiographic Grading in Primary Osteoarthritis

Authors: Heba Esaily, Rawhia Eledl, Daila Aboelela, Rasha Noreldin

Abstract:

Objective: Assess serum level of Transforming growth factor beta 2 (TGF-β2) and Matrilin-3 (MATN3) SNP6 polymorphism in osteoarthritic patients Background: Osteoarthritis (OA) is a musculoskeletal disease characterized by pain and joint stiffness. TGF-β 2 is involved in chondrogenesis and osteogenesis, It has found that MATN3 gene and protein expression was correlated with the extent of tissue damage in OA. Findings suggest that regulation of MATN3 expression is essential for maintenance of the cartilage extracellular matrix microenvironment Subjects and Methods: 72 cases of primary OA (56 with knee OA and 16 with generalized OA were compared with that of 18 healthy controls. Radiographs were scored with the Kellgren-Lawrence scale. Serum TGF-β2 was measured by using (ELISA), levels of marker were correlated to radiographic grading of disease and MATN3 SNP6 polymorphism was determined by (PCR-RFLP). Results: MATN3 SNP6 polymorphism and serum level of TGF-β2 were higher in OA compared with controls. Genotype, NN and N allele frequency were higher in patients with OA compared with controls. NN genotype and N allele frequency were higher in knee osteoarthritis than generalized OA. Significant positive correlation between level of TGFβ2 and radiographic grading in group with knee OA, but no correlation between serum level of TGFβ2 and radiographic grading in generalized OA. Conclusion: MATN3 SNP6 polymorphism and TGF-β2 implicated in the pathogenesis of osteoarthritis. Association of N/N genotype with primary osteoarthritis emphasizes on the need for prospective study include larger sample size to confirm the results of the present study.

Keywords: Matrilin-3, transforming growth factor beta 2, primary osteoarthritis, knee osteoarthritis

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206 The Impact of Non-Surgical and Non-Medical Interventions on the Treatment of Infertile Women with Ovarian Reserve Below One and Early Menopause Symptoms

Authors: Flora Tajiki

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This study investigates the effectiveness of non-surgical and non-medical interventions in treating infertile women with severely diminished ovarian reserve (below one), low Anti-Müllerian Hormone (AMH) levels, and symptoms of early menopause. The intervention included yoga, sunlight exposure, vitamin and mineral supplementation, relaxation techniques, and daily prayers performed both before sleep and upon waking. These methods were applied to women who had shown poor response to high-dose fertility treatments, such as IVF and microinjection cycles, leading to low-quality egg production. The focus was on women with severely reduced ovarian reserve and early menopause symptoms, some of whom continued to experience relatively regular menstrual cycles despite the onset of these symptoms. This treatment was aimed at women for whom conventional fertility methods had been ineffective. The study sample consisted of 120 married women, aged 25 to 45, from the provinces of Tehran, Alborz, and western Iran, with 35 participants completing the intervention. Individual factors such as residence, education, employment status, marriage duration, family infertility history, and previous infertility treatments were examined, with income considered as a contextual variable. The results indicate that AMH may not be a definitive marker of ovarian reserve, as lifestyle modifications, such as those implemented in this study, were associated with increased AMH levels, the return of regular menstrual cycles, and successful pregnancies. No short- or long-term complications were reported during the two-year follow-up, highlighting the potential benefits of non-surgical interventions for women with early menopause symptoms and diminished ovarian reserve.

Keywords: anti-müllerian hormone, infertility, ovarian reserve, early menopause, fertility, women’s health, lifestyle modification, pregnancy

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205 Traumatic Osteoarthritis Induces Mechanical Hyperalgesia through IL-1β/TNF-α-Mediated Upregulation of the Sema4D Gene Expression

Authors: Hsiao-Chien Tsai, Yu-Pin Chen, Ruei-Ming Chen

Abstract:

Introduction: Osteoarthritis (OA) is characterized by joint destruction and causes chronic disability. One of the prominent symptoms is pain. Alleviating the pain is necessary and urgent for the therapy of OA patients. However, currently, understanding the mechanisms that drive OA-induced pain remains challenging, which hampers the optimistic management of pain in OA patients. Semaphorin 4D (Sema4D) participates in axon guidance pathway and bone remodeling, thus, may play a role in the regulation of pain in OA. In this study, we have established a rat model of OA to find out the mechanisms of OA-induced pain and to deliberate the roles of Sema4D. Methods: Behavioral changes and the pro-inflammatory cytokines (IL-1β, TNF-α, and IL-17) associated with pain were measured during the development of OA. Sema4D expression in cartilage and synovial membrane at 1, 4, and 12 weeks after inducing OA was analyzed. To assess if Sema4D is related to the neurogenesis in OA as an axon repellant, we analyzed the expression of PGP9.5 as well. Results: Synovitis and cartilage degradation were evident histologically during the development of OA. Mechanical hyperalgesia was most severe at week 1, then persisted thereafter. It was associated with stress coping strategies. Similar to the pain behavioral results, levels of IL-1β and TNF-α in synovial lavage fluid were significantly elevated in the OA group at weeks 1 and 4, respectively. Sema4D expression in cartilage and the synovial membrane was also enhanced in the OA group and was correlated with pain and pro-inflammatory cytokines. The marker of neurogenesis, PGP9.5, was also enhanced during the development of OA. Discussion: OA induced mechanical hyperalgesia, which might be through upregulating IL-1β/TNF-α-mediated Sema4D expressions. If anti-Sema4D treatment could reduce OA-induced mechanical hyperalgesia and prevent the subsequent progression of OA needs to be further investigated. Significance: OA can induce mechanical hyperalgesia through upregulation of IL-1β/TNF-α-mediated Sema4D and PGP9.5 expressions. And the upregulation of Sema4D may indicate the severity or active status of OA and OA-induced pain.

Keywords: traumatic osteoarthritis, mechanical hyperalgesia, Sema4D, inflammatory cytokines

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204 Differential Expression of Biomarkers in Cancer Stem Cells and Side Populations in Breast Cancer Cell Lines

Authors: Dipali Dhawan

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Cancerous epithelial cells are confined to a primary site by the continued expression of adhesion molecules and the intact basal lamina. However, as the cancer progresses some cells are believed to undergo an epithelial-mesenchymal transition (EMT) event, leading to increased motility, invasion and, ultimately, metastasis of the cells from the primary tumour to secondary sites within the body. These disseminated cancer cells need the ability to self-renew, as stem cells do, in order to establish and maintain a heterogeneous metastatic tumour mass. Identification of the specific subpopulation of cancer stem cells amenable to the process of metastasis is highly desirable. In this study, we have isolated and characterized cancer stem cells from luminal and basal breast cancer cell lines (MDA-MB-231, MDA-MB-453, MDA-MB-468, MCF7 and T47D) on the basis of cell surface markers CD44 and CD24; as well as Side Populations (SP) using Hoechst 33342 dye efflux. The isolated populations were analysed for epithelial and mesenchymal markers like E-cadherin, N-cadherin, Sfrp1 and Vimentin by Western blotting and Immunocytochemistry. MDA-MB-231 cell lines contain a major population of CD44+CD24- cells whereas MCF7, T47D and MDA-MB-231 cell lines show a side population. We observed higher expression of N-cadherin in MCF-7 SP cells as compared to MCF-7NSP (Non-side population) cells suggesting that the SP cells are mesenchymal like cells and hence express increased N-cadherin with stem cell-like properties. There was an expression of Sfrp1 in the MCF7- NSP cells as compared to no expression in MCF7-SP cells, which suggests that the Wnt pathway is expressed in the MCF7-SP cells. The mesenchymal marker Vimentin was expressed only in MDA-MB-231 cells. Hence, understanding the breast cancer heterogeneity would enable a better understanding of the disease progression and therapeutic targeting.

Keywords: cancer stem cells, epithelial to mesenchymal transition, biomarkers, breast cancer

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203 Utility of CK7, CK20 and CDX-2 as a Potential Panel in Differentiating Primary Ovarian Surface Epithelial Tumors from Metastatic Adenocarcinoma to the Ovary

Authors: Ghada Esheba, Ghadeer Aldoobi, Salwa Almalk, Abrar Alshareef, Eman Al-khairi, Eman Yaseen

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Background: In Saudi Arabia, ovarian cancer ranked seventh among female population and is the most common female genital tract malignancy after endometrial cancer. A slight increase in the incidence of ovarian cancer was observed from 2001–2008. Makkah, Riyadh, and the eastern region of Saudi Arabia had the highest incidence rate ratio for the number of ovarian cancer cases (1). Differentiating metastatic adenocarcinomas from primary ovarian carcinomas, especially those of endometrioid and mucinous type is clinically significant and a challenge for clinicians and pathologists, yet the distinction has important therapeutic and prognostic implications. Aim: To clarify the most important histopathological criteria to differentiate between primary ovarian surface epithelial tumors especially mucinous and endometrioid subtypes, and metastatic adenocarcinoma and to evaluate the value of a panel of antibodies consisting of CK7, CK20, and CDX-2 in the distinction between primary ovarian surface epithelial tumors and metastatic adenocarcinoma. Material and methods: This study was carried out on 26 cases of primary ovarian surface epithelial neoplasms and 14 cases of metastatic ovarian adenocarcinoma. All cases were studied immunohistochemically using CK7, CK20, and CDX-2. Results: All cases of primary ovarian adenocarcinoma were positive for CK7. 25% and 58% of mucinous borderline mucinous tumor and mucinous carcinoma respectively were positive for CK20. Only 42% of mucinous carcinoma were positive for CDX-2. All cases of endometrioid carcinomas were negative for both CK20 and CDX-2. All cases of metastatic adenocarcinoma from the colon were negative for CK7 and positive for CK20 and CDX-2. Conclusions: CK7 is an important positive marker for primary ovarian tumors, while CK20 and CDX-2 are useful markers for colorectal carcinoma metastatic to the ovary. Caution should be taken as primary ovarian mucinous tumors may stain positive for CK20, CDX-2, or both, however, they usually exhibit a focal pattern of reactivity.

Keywords: adenoma, endometrioid, malignancy, ovarian

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