Search results for: lipid metabolism
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 164

Search results for: lipid metabolism

44 Effect of Real Wastewater on Biotransformation of 17α-ethynylestradiol by Ammonia-Oxidizing Bacteria in Nitrifying Activated Sludge

Authors: Natthawan Likitmongkonsakun, Tawan Limpiyakorn

Abstract:

17α-ethynylestradiol (EE2) is a synthetic estrogen used as a key ingredient in an oral contraceptives pill. EE2 is an endocrine disrupting compound, high in estrogenic potency. Although EE2 exhibits low degree of biodegradability with common microorganisms in wastewater treatment plants (WWTPs), this compound can be biotransformed by ammonia-oxidizing bacteria (AOB) via a co-metabolism mechanism in WWTPs. This study aimed to investigate the effect of real wastewater on biotransformation of EE2 by AOB. A preliminary experiment on the effect of nitrite and pH levels on abiotic transformation of EE2 suggested that the abiotic transformation occurred at only pH <6.8. Biotransformation of EE2 under the presence of municipal or industrial wastewater demonstrated that different types of wastewater affect EE2 biotransformation differently. Organic matters in wastewater were believed to deteriorate EE2 biotransformation via the competition effect. At a lower initial ammonium concentration, EE2 biotransformation can be retarded and the extent of the deterioration was COD-concentration dependent. However, when an initial ammonium concentration was elevated, thisphenomena disappeared. This is because when increasing the amount of the primary substrate, more AMO enzymes can be produced resulting in unlimited transformation of all compounds in the tests reducing the competitive effect of organic matters on EE2.

Keywords: 17α-ethynylestradiol, ammonia-oxidizing bacteria, nitrifying activated sludge, wastewater.

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43 Effects of Intrauterine and Extrauterine Exposure to 1800 MHz GSM-Like Radiofrequency Radiation on Liver Regulatory Enzymes Activities in Infant Female Rabbits

Authors: A. Tomruk, G. Guler, B. Tandogan, E. Ozgur, N.N. Ulusu, N. Seyhan

Abstract:

In the present study, we aimed to design the intrauterine and extrauterine exposure to 1800 MHz GSM-like RF radiation and investigate its possible bio-effects on infant female rabbits. Totally thirty-six New Zealand White female rabbits, onemonth old, were randomly divided into four groups which are composed of 9 rabbits; i. Group I [Intrauterine (IU) exposure(-); Extrauterine (EU) exposure (-)], Group II [IU exposure (-); EU exposure (+)], Group III [IU exposure(+);EU exposure(-)], Group IV [IU exposure (+);EU exposure(+)]. The master regulatory enzymes activities of pentose phosphate pathway (glucose-6-phosphate dehydrogenase, G-6PD; 6-phosphogluconate dehydrogenase, 6- PGDH) and glutathione-dependent metabolism (glutathione peroxidase, GSH-Px; glutathione reductase, GR; glutathione Stransferase, GST, thioredoxin reductase, TRx) were analyzed in liver tissues of young female rabbits. Decreased G-6PD, 6-PGD, GSH-Px, GR activities were found in Group III compared to Group I (p<0.05, Mann Whitney). Increased GSH-px and TRx activities were found in Group IV compared to Group I (p<0.05, Mann Whitney). It can be concluded that the intrauterine and extrauterine exposure to GSMlike RF radiation may influence the liver regulatory enzymes activities.

Keywords: Radiofrequency (RF), intrauterine (IU) andextrauterine (EU) exposure, infant female rabbits.

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42 Numbers and Biomass of Bacteria and Fungi Obtained by the Direct Microscopic Count Method

Authors: Ayuko Itsuki, Sachiyo Aburatani

Abstract:

The soil ecology of the organic and mineral soil layers of laurel-leaved and Cryptomeria japonica forest in the Kasuga-yama Hill Primeval Forest (Nara, Japan) was assessed. The number of bacteria obtained by the dilution plate count method was less than 0.05% of those counted by the direct microscopic count. We therefore found that forest soil contains large numbers of non-culturable bacteria compared with agricultural soils. The numbers of bacteria and fungi obtained by both the dilution plate count and the direct microscopic count were larger in the deeper horizons (F and H) of the organic layer than in the mineral soil layer. This suggests that active microbial metabolism takes place in the organic layer. The numbers of bacteria and the length of fungal hyphae obtained by the direct count method were greater in the H horizon than in the F horizon. The direct microscopic count revealed numerous non-culturable bacteria and fungi in the soil. The ratio of fungal to bacterial biomass was lower in the laurel-leaved forest soil. The fungal biomass was therefore relatively low in the laurel-leaved forest soil due to differences in forest vegetation.

Keywords: Bacterial number, Dilution plate count, Direct microscopic count, Forest soil.

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41 Evaluation of Hancornia speciosa Gomes Lyophilization at Different Stages of Maturation

Authors: D. C. Soares, J. T. S. Santos, D. G. Costa, A. K. S. Abud, T. P. Nunes, A. V. D. Figueiredo, A. M. de Oliveira Junior

Abstract:

Mangabeira (Hancornia speciosa Gomes), a native plant in Brazil, is found growing spontaneously in various regions of the country. The high perishability of tropical fruits such as mangaba, causes it to be necessary to use technologies that promote conservation, aiming to increase the shelf life of this fruit and add value. The objective of this study was to compare the mangabas lyophilization curves behaviors with different sizes and maturation stages. The fruits were freeze-dried for a period of approximately 45 hours at lyophilizer Liotop brand, model L -108. It has been considered large the fruits between 38 and 58 mm diameter and small, between 23 and 28 mm diameter and the two states of maturation, intermediate and mature. Large size mangabas drying curves in both states of maturation were linear behavior at all process, while the kinetic drying curves related to small fruits, independent of maturation state, had a typical behavior of drying, with all the well-defined steps. With these results it was noted that the time of lyophilization was suitable for small mangabas, a fact that did not happen with the larger one. This may indicate that the large mangabas require a longer time to freeze until reaches the equilibrium level, as it happens with the small fruits, going to have constant moisture at the end of the process. For both types of fruit were analyzed water activity, acidity, protein, lipid, and vitamin C before and after the process.

Keywords: Freeze dryer, mangaba, conservation, chemical characteristics.

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40 Ameliorating Effects of Silver Nanoparticles Synthesized Using Chlorophytum borivillianum against Gamma Radiation Induced Oxidative Stress in Testis of Swiss Albino Mice

Authors: Ruchi Vyas, Sanjay Singh, Rashmi Sisodia

Abstract:

Chlorophytum borivillianum root extract (CBE) was chosen as a reducing agent to fabricate silver nanoparticles with the aim of studying its radioprotective efficacy. The formation of synthesized nanoparticles was characterized by UV–visible analysis (UV–vis), Fourier transform infra-red (FT-IR), Transmission electron microscopy (TEM), Scanning electron microscope (SEM). TEM analysis showed particles size in the range of 20-30 nm. For this study, Swiss albino mice were selected from inbred colony and were divided into 4 groups: group I- control (irradiated-6 Gy), group II- normal (vehicle treated), group III- plant extract alone and group IV- CB-AgNPs (dose of 50 mg/kg body wt./day) administered orally for 7 consecutive days before irradiation to serve as experimental. CB-AgNPs pretreatment rendered significant increase in body weight and testes weight at various post irradiation intervals in comparison to irradiated group. Supplementation of CB-AgNPs reversed the adverse effects of gamma radiation on biochemical parameters as it notably ameliorated the elevation in lipid peroxidation and decline in glutathione concentration in testes. These observations indicate the radio-protective potential of CB-AgNPs in testicular constituents against gamma irradiation in mice.

Keywords: Chlorophytum borivillianum, gamma radiation, radioprotective, silver nanoparticles.

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39 Oil Extraction from Microalgae Dunalliela sp. by Polar and Non-Polar Solvents

Authors: A. Zonouzi, M. Auli, M. Javanmard Dakheli, M. A. Hejazi

Abstract:

Microalgae are tiny photosynthetic plants. Nowadays, microalgae are being used as nutrient-dense foods and sources of fine chemicals. They have significant amounts of lipid, carotenoids, vitamins, protein, minerals, chlorophyll, and pigments. Oil extraction from algae is a hotly debated topic currently because introducing an efficient method could decrease the process cost. This can determine the sustainability of algae-based foods. Scientific research works show that solvent extraction using chloroform/methanol (2:1) mixture is one of the efficient methods for oil extraction from algal cells, but both methanol and chloroform are toxic solvents, and therefore, the extracted oil will not be suitable for food application. In this paper, the effect of two food grade solvents (hexane and hexane/ isopropanol) on oil extraction yield from microalgae Dunaliella sp. was investigated and the results were compared with chloroform/methanol (2:1) extraction yield. It was observed that the oil extraction yield using hexane, hexane/isopropanol (3:2) and chloroform/methanol (2:1) mixture were 5.4, 13.93, and 17.5 (% w/w, dry basis), respectively. The fatty acid profile derived from GC illustrated that the palmitic (36.62%), oleic (18.62%), and stearic acids (19.08%) form the main portion of fatty acid composition of microalgae Dunalliela sp. oil. It was concluded that, the addition of isopropanol as polar solvent could increase the extraction yield significantly. Isopropanol solves cell wall phospholipids and enhances the release of intercellular lipids, which improves accessing of hexane to fatty acids.

Keywords: Fatty acid profile, Microalgae, Oil extraction, Polar solvent.

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38 Impact of Some Experimental Procedures on Behavioral Patterns and Physiological Traits of Rats

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

Abstract:

Welfare may be considered to be a subjective experience; it has a biological function that is related to the fitness and survival of the animal accordingly, researches have suggested that welfare is compromised when the animal's evolutionary fitness is reduced. This study was carried out to explain the effect of some managerial stressors as handling and restraint on behavioral patterns and biochemical parameters of rats. A total of 24 (12 males & 12 females) Sprague-Dawley rats (12 months & 150-180g) were allotted into 3 groups, handled group (4 male & 4 female), restrained group (4 male & 4 female) and control group (4 males & 4 females). The obtained results revealed that time spent feeding, drinking, movement and cage exploration frequencies increased significantly in handled rats than other groups, while lying time and licking increased significantly in restrained rats than handled and controls. Moreover, social behavior decreased in both stressed groups than control. Triglycerides were significantly increased in handled rats than other groups, while total lipid, total protein and globulin significantly increased in both treated groups than control. Corticosterone increased in restrained and handled rats than control ones. Moreover, there was an increment in packed cell volume significantly in restrained rats than others. These deducted that if we want to study the effect of stress on animal welfare it is necessary to study the effect of such stressors on animal’s behavior and physiological responses.

Keywords: Behavior, handling, restraint, rat, welfare.

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37 Effects of Different Dietary Crude Fiber Levels on the Growth Performance of Finishing Su-Shan Pigs

Authors: Li Bixia, Ren Shouwen, Fu Yanfeng, Tu Feng, Xiaoming Fang, Xueming Wang

Abstract:

The utilization of dietary crude fiber in different breed pigs is not the same. Su-shan pigs are a new breed formed by crossing Taihu pigs and Yorkshire pigs. In order to understand the resistance of Su-shan pigs to dietary crude fiber, 150 Su-shan pigs with 60 kg of average body weight and similar body conditions were allocated to three groups randomly, and there are 50 pigs in each group. The percentages of dietary crude fiber were 8.35%, 9.10%, and 11.39%, respectively. At the end of the experiment, 15 pigs randomly selected from each group were slaughtered. The results showed as follows: average daily gain of the 9.10% group was higher than that of the 8.35% group and the 11.39% group; there was a significant difference between the 9.10% group and the 8.35% group (p < 0.05. Levels of urea nitrogen, total cholesterol and high density lipoprotein in the 9.10% group were significantly higher than those in the 8.35% group and the 11.39% group (p < 0.05). Ratios of meat to fat in the 9.10% group and the 11.39% group were significantly higher than that in the 8.35% group (p < 0.05). Lean percentage of 9.10% group was higher than that of 8.35% group and 11.39% group, but there was no significant difference in three groups (p > 0.05). The weight of small intestine and large intestine in the 11.39% group was higher than that in the 8.35% group, and the 9.10% group and the difference reached a significant level (p < 0.05). In conclusion, increasing dietary crude fiber properly could reduce fat percentage, and improve the ratio of meat to fat of finishing Su-shan pigs. The digestion and metabolism of dietary crude fiber promoted the development of stomach and intestine of finishing Su-shan pig.

Keywords: Su-Shan pigs, dietary crude fiber, growth performance, serum biochemical indexes.

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36 Evaluation of Prevalence of the Types of Thyroid Disorders Using Ultrasound and Pathology of One Humped Camel in Iran (Camelus dromedarius)

Authors: M. Yadegari

Abstract:

The thyroid gland is the largest classic endocrine organ that effects many organs of the body and plays a significant role in the process of Metabolism in animals. The aim of this study was to investigate the prevalence of thyroid disorders diagnosed by ultrasound and microscopic Lesions of the thyroid during the slaughter of apparently healthy One Humped Camels (Camelus dromedarius) in Iran. Randomly, 520 male camels (With an age range of 4 to 8 years), were studied in 2012 to 2013. The Camels’ thyroid glands were evaluated by sonographic examination. In both longitudinal and transverse view and then tissue sections were provide and stained with H & E and finally examined by light microscopy. The results obtained indicated the following: hyperplastic goiter (21%), degenerative changes (12%), follicular cysts (8%), follicular atrophy (4%), nodular hyperplasia (3%), adenoma (1%), carcinoma (1%) and simple goiter colloid (1%). Ultrasound evaluation of thyroid gland in adenoma and carcinoma showed enlargement and irregular of the gland, decreased echogenicity, and the heterogeneous thyroid parenchyma. Also, in follicular cysts were observed in the enlarged gland with no echo structures of different sizes and decreased echogenicity as a local or general. In nodular hyperplasia, increase echogenicity and heterogeneous parenchymal were seen. These findings suggest the use of sonography and pathology as a screening test in the diagnosis of complications of thyroid disorders.

Keywords: One humped camel, pathology, sonography, thyroid gland.

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35 Higher Plants Ability to Assimilate Explosives

Authors: G. Khatisashvili, M. Gordeziani, G. Adamia, E. Kvesitadze, T. Sadunishvili, G. Kvesitadze

Abstract:

The ability of agricultural and decorative plants to absorb and detoxify TNT and RDX has been studied. All tested 8 plants, grown hydroponically, were able to absorb these explosives from water solutions: Alfalfa > Soybean > Chickpea> Chikling vetch >Ryegrass > Mung bean> China bean > Maize. Differently from TNT, RDX did not exhibit negative influence on seed germination and plant growth. Moreover, some plants, exposed to RDX containing solution were increased in their biomass by 20%. Study of the fate of absorbed [1-14ðí]-TNT revealed the label distribution in low and high-molecular mass compounds, both in roots and above ground parts of plants, prevailing in the later. Content of 14ðí in lowmolecular compounds in plant roots are much higher than in above ground parts. On the contrary, high-molecular compounds are more intensively labeled in aboveground parts of soybean. Most part (up to 70%) of metabolites of TNT, formed either by enzymatic reduction or oxidation, is found in high molecular insoluble conjugates. Activation of enzymes, responsible for reduction, oxidation and conjugation of TNT, such as nitroreductase, peroxidase, phenoloxidase and glutathione S-transferase has been demonstrated. Among these enzymes, only nitroreductase was shown to be induced in alfalfa, exposed to RDX. The increase in malate dehydrogenase activities in plants, exposed to both explosives, indicates intensification of Tricarboxylic Acid Cycle, that generates reduced equivalents of NAD(P)H, necessary for functioning of the nitroreductase. The hypothetic scheme of TNT metabolism in plants is proposed.

Keywords: Higher plants, TNT, RDX, transformation.

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34 The Role of Chemerin and Myostatin after Physical Activity

Authors: M. J. Pourvaghar, M. E. Bahram

Abstract:

Obesity and overweight is one of the most common metabolic disorders in industrialized countries and in developing countries. One consequence of pathological obesity is cardiovascular disease and metabolic syndrome. Chemerin is an adipocyne that plays a role in the regulation of the adipocyte function and the metabolism of glucose in the liver and musculoskeletal system. Most likely, chemerin is involved in obesity-related disorders such as type 2 diabetes and cardiovascular disease. Aerobic exercises reduce the level of chemerin and cause macrophage penetration into fat cells and inflammatory factors. Several efforts have been made to clarify the cellular and molecular mechanisms of hypertrophy and muscular atrophy. Myostatin, a new member of the TGF-β family, is a transforming growth factor β that its expression negatively regulates the growth of the skeletal muscle; and the increase of this hormone has been observed in conditions of muscular atrophy. While in response to muscle overload, its levels decrease after the atrophy period, TGF-β is the most important cytokine in the development of skeletal muscle. Myostatin plays an important role in muscle control, and animal and human studies show a negative role of myostatin in the growth of skeletal muscle. Separation of myostatin from Golgi begins on the ninth day of the onset period and continues until birth at all times of muscle growth. Higher levels of myostatin are found in obese people. Resistance training for 10 weeks could reduce levels of plasma myostatin.

Keywords: Chemerin, myostatin, obesity, physical activity.

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33 Mapping of Adrenal Gland Diseases Research in Middle East Countries: A Scientometric Analysis, 2007-2013

Authors: Zahra Emami, Mohammad Ebrahim Khamseh, Nahid Hashemi Madani, Iman Kermani

Abstract:

The aim of the study was to map scientific research on adrenal gland diseases in the Middle East countries through the Web of Science database using scientometric analysis. Data were analyzed with Excel software; and HistCite was used for mapping of the scientific texts. In this study, from a total of 268 retrieved records, 1125 authors from 328 institutions published their texts in 138 journals. Among 17 Middle East countries, Turkey ranked first with 164 documents (61.19%), Israel ranked second with 47 documents (15.53%) and Iran came in the third place with 26 documents. Most of the publications (185 documents, 69.2%) were articles. Among the universities of the Middle East, Istanbul University had the highest science production rate (9.7%). The Journal of Clinical Endocrinology & Metabolism had the highest TGCS (243 citations). In the scientific mapping, 7 clusters were formed based on TLCS (Total Local Citation Score) & TGCS (Total Global Citation Score). considering the study results, establishment of scientific connections and collaboration with other countries and use of publications on adrenal gland diseases from high ranking universities can help in the development of this field and promote the medical practice in this regard. Moreover, investigation of the formed clusters in relation to Congenital Hyperplasia and puberty related disorders can be research priorities for investigators.

Keywords: Mapping, scientific research, adrenal gland diseases, scientometric.

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32 The Expression of Lipoprotein Lipase Gene with Fat Accumulations and Serum Biochemical Levels in Betong (KU Line) and Broiler Chickens

Authors: W. Loongyai, N. Saengsawang, W. Danvilai, C. Kridtayopas, P. Sopannarath, C. Bunchasak

Abstract:

Betong chicken is a slow growing and a lean strain of chicken, while the rapid growth of broiler is accompanied by increased fat. We investigated the growth performance, fat accumulations, lipid serum biochemical levels and lipoprotein lipase (LPL) gene expression of female Betong (KU line) at the age of 4 and 6 weeks. A total of 80 female Betong chickens (KU line) and 80 female broiler chickens were reared under open system (each group had 4 replicates of 20 chicks per pen). The results showed that feed intake and average daily gain (ADG) of broiler chicken were significantly higher than Betong (KU line) (P < 0.01), while feed conversion ratio (FCR) of Betong (KU line) at week 6 were significantly lower than broiler chicken (P < 0.01) at 6 weeks. At 4 and 6 weeks, two birds per replicate were randomly selected and slaughtered. Carcass weight did not significantly differ between treatments; the percentage of abdominal fat and subcutaneous fat yield was higher in the broiler (P < 0.01) at 4 and 6 week. Total cholesterol and LDL level of broiler were higher than Betong (KU line) at 4 and 6 weeks (P < 0.05). Abdominal fat samples were collected for total RNA extraction. The cDNA was amplified using primers specific for LPL gene expression and analysed using real-time PCR. The results showed that the expression of LPL gene was not different when compared between Betong (KU line) and broiler chickens at the age of 4 and 6 weeks (P > 0.05). Our results indicated that broiler chickens had high growth rate and fat accumulation when compared with Betong (KU line) chickens, whereas LPL gene expression did not differ between breeds.

Keywords: Lipoprotein lipase gene, Betong (KU line), broiler, abdominal fat, gene expression.

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31 Oxidative and Hormonal Disruptions Underlie Bisphenol A - Induced Testicular Toxicity in Male Rabbits

Authors: Kadry M. Sadek, Tarek K. Abouzed, Mousa A. Ayoub

Abstract:

The presence of endocrine-disrupting compounds, such as bisphenol A (BPA), in the environment can cause serious health problems. However, there are controversial opinions. This study investigated the reproductive, metabolic, oxidative and immunologic-disrupting effects of bisphenol A in male rabbits. Rabbits were divided into five groups. The first four rabbit groups were administered oral BPA (1, 10, 50, or 100 mg/kg/day) for ten weeks. The fifth group was administered corn oil as the vehicle. BPA significantly decreased serum testosterone, estradiol and the free androgen index (FAI) and significantly increased sex hormone binding globulin (SHBG) compared with the placebo group. The higher doses of BPA showed a significant decrease in follicular stimulating hormone (FSH) and luteinizing hormone (LH). A significant increase in blood glucose levels was identified in the BPA groups. The non-significant difference in insulin levels is a novel finding. The cumulative testicular toxicity of BPA was clearly demonstrated by the dose-dependent decrease in absolute testes weight, primary measures of semen quality and a significant increase in testicular malonaldehyde (MDA). Moreover, BPA significantly decreased total antioxidant capacity (TAC) and significantly increased immunoglobulin G (IgG) at the highest concentration. Our results suggest that BPA, especially at higher doses, is associated with many adverse effects on metabolism, oxidative stress, immunity, sperm quality and markers of androgenic action.

Keywords: Bisphenol A, oxidative stress, rabbits, semen quality, steroidogenesis.

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30 Comparative Study of the Effects of Process Parameters on the Yield of Oil from Melon Seed (Cococynthis citrullus) and Coconut Fruit (Cocos nucifera)

Authors: Ndidi F. Amulu, Patrick E. Amulu, Gordian O. Mbah, Callistus N. Ude

Abstract:

Comparative analysis of the properties of melon seed, coconut fruit and their oil yield were evaluated in this work using standard analytical technique AOAC. The results of the analysis carried out revealed that the moisture contents of the samples studied are 11.15% (melon) and 7.59% (coconut). The crude lipid content are 46.10% (melon) and 55.15% (coconut).The treatment combinations used (leaching time, leaching temperature and solute: solvent ratio) showed significant difference (p < 0.05) in yield between the samples, with melon oil seed flour having a higher percentage range of oil yield (41.30 – 52.90%) and coconut (36.25 – 49.83%). The physical characterization of the extracted oil was also carried out. The values gotten for refractive index are 1.487 (melon seed oil) and 1.361 (coconut oil) and viscosities are 0.008 (melon seed oil) and 0.002 (coconut oil). The chemical analysis of the extracted oils shows acid value of 1.00mg NaOH/g oil (melon oil), 10.050mg NaOH/g oil (coconut oil) and saponification value of 187.00mg/KOH (melon oil) and 183.26mg/KOH (coconut oil). The iodine value of the melon oil gave 75.00mg I2/g and 81.00mg I2/g for coconut oil. A standard statistical package Minitab version 16.0 was used in the regression analysis and analysis of variance (ANOVA). The statistical software mentioned above was also used to optimize the leaching process. Both samples gave high oil yield at the same optimal conditions. The optimal conditions to obtain highest oil yield ≥ 52% (melon seed) and ≥ 48% (coconut seed) are solute - solvent ratio of 40g/ml, leaching time of 2hours and leaching temperature of 50oC. The two samples studied have potential of yielding oil with melon seed giving the higher yield.

Keywords: Coconut, melon, optimization, processing.

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29 Web–Based Tools and Databases for Micro-RNA Analysis: A Review

Authors: Sitansu Kumar Verma, Soni Yadav, Jitendra Singh, Shraddha, Ajay Kumar

Abstract:

MicroRNAs (miRNAs), a class of approximately 22 nucleotide long non coding RNAs which play critical role in different biological processes. The mature microRNA is usually 19–27 nucleotides long and is derived from a bigger precursor that folds into a flawed stem-loop structure. Mature micro RNAs are involved in many cellular processes that encompass development, proliferation, stress response, apoptosis, and fat metabolism by gene regulation. Resent finding reveals that certain viruses encode their own miRNA that processed by cellular RNAi machinery. In recent research indicate that cellular microRNA can target the genetic material of invading viruses. Cellular microRNA can be used in the virus life cycle; either to up regulate or down regulate viral gene expression Computational tools use in miRNA target prediction has been changing drastically in recent years. Many of the methods have been made available on the web and can be used by experimental researcher and scientist without expert knowledge of bioinformatics. With the development and ease of use of genomic technologies and computational tools in the field of microRNA biology has superior tremendously over the previous decade. This review attempts to give an overview over the genome wide approaches that have allow for the discovery of new miRNAs and development of new miRNA target prediction tools and databases.

Keywords: MicroRNAs, computational tools, gene regulation, databases, RNAi.

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28 Phyllantus niruri Protects against Fe2+ and SNP Induced Oxidative Damage in Mitochondrial Enriched Fractions of Rats Brain

Authors: Olusola Olalekan Elekofehinti, Isaac Gbadura Adanlawo, Joao Batista Teixeira Rocha

Abstract:

The potential neuroprotective effect of Phyllantus nuriri against Fe2+ and sodium nitroprusside (SNP) induced oxidative stress in mitochondria of rats brain was evaluated. Cellular viability was assessed by MTT reduction, reactive oxygen species (ROS) generation was measured using the probe 2,7-dichlorofluoresce indiacetate (DCFH-DA). Glutathione content was measured using dithionitrobenzoic acid (DTNB). Fe2+ (10μM) and SNP (5μM) significantly decreased mitochondrial activity, assessed by MTT reduction assay, in a dose-dependent manner, this occurred in parallel with increased glutathione oxidation, ROS production and lipid peroxidation end-products (thiobarbituric acid reactive substances, TBARS). The co-incubation with methanolic extract of Phyllantus nuriri (10-200 μg/ml) reduced the disruption of mitochondrial activity, gluthathione oxidation, ROS production as well as the increase in TBARS levels caused by both Fe2+ and SNP in a dose dependent manner. HPLC analysis of the extract revealed the presence of gallic acid (20.540.01), caffeic acid (7.930.02), rutin (25.310.05), quercetin (31.280.03) and kaemferol (14.360.01). This result suggests that these phytochemicals account for the protective actions of P. niruri against Fe2+ and SNP -induced oxidative stress. Our results show that P. nuriri consist important bioactive molecules in the search for an improved therapy against the deleterious effects of Fe2+, an intrinsic producer of reactive oxygen species (ROS), that leads to neuronal oxidative stress and neurodegeneration.

Keywords: Phyllantus niruri, mitochondria, antioxidant, oxidative stress, synaptosome.

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27 Microbubbles Enhanced Synthetic Phorbol Ester Degradation by Ozonolysis

Authors: Kuvshinov, D., Siswanto, A., Zimmerman, W. B.

Abstract:

A phorbol-12-myristate-13-acetate (TPA) is a synthetic analogue of phorbol ester (PE), a natural toxic compound of Euphorbiaceae plant. The oil extracted from plants of this family is useful source for primarily biofuel. However this oil might also be used as a foodstuff due to its significant nutrition content. The limitations for utilizing the oil as a foodstuff are mainly due to a toxicity of PE. Currently, a majority of PE detoxification processes are expensive as include multi steps alcohol extraction sequence.

Ozone is considered as a strong oxidative agent. It reacts with PE by attacking the carbon-carbon double bond of PE. This modification of PE molecular structure yields a non toxic ester with high lipid content.

This report presents data on development of simple and cheap PE detoxification process with water application as a buffer and ozone as reactive component. The core of this new technique is an application for a new microscale plasma unit to ozone production and the technology permits ozone injection to the water-TPA mixture in form of microbubbles.

The efficacy of a heterogeneous process depends on the diffusion coefficient which can be controlled by contact time and interfacial area. The low velocity of rising microbubbles and high surface to volume ratio allow efficient mass transfer to be achieved during the process. Direct injection of ozone is the most efficient way to process with such highly reactive and short lived chemical.

Data on the plasma unit behavior are presented and the influence of gas oscillation technology on the microbubble production mechanism has been discussed. Data on overall process efficacy for TPA degradation is shown.

Keywords: Microbubble, ozonolysis, synthetic phorbol ester.

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26 Evaluation of Electro-Flocculation for Biomass Production of Marine Microalgae Phaodactylum tricornutum

Authors: Luciana C. Ramos, Leandro J. Sousa, Antônio Ferreira da Silva, Valéria Gomes Oliveira Falcão, Suzana T. Cunha Lima

Abstract:

The commercial production of biodiesel using microalgae demands a high-energy input for harvesting biomass, making production economically unfeasible. Methods currently used involve mechanical, chemical, and biological procedures. In this work, a flocculation system is presented as a cost and energy effective process to increase biomass production of Phaeodactylum tricornutum. This diatom is the only species of the genus that present fast growth and lipid accumulation ability that are of great interest for biofuel production. The algae, selected from the Bank of Microalgae, Institute of Biology, Federal University of Bahia (Brazil), have been bred in tubular reactor with photoperiod of 12 h (clear/dark), providing luminance of about 35 μmol photons m-2s-1, and temperature of 22 °C. The medium used for growing cells was the Conway medium, with addition of silica. The seaweed growth curve was accompanied by cell count in Neubauer camera and by optical density in spectrophotometer, at 680 nm. The precipitation occurred at the end of the stationary phase of growth, 21 days after inoculation, using two methods: centrifugation at 5000 rpm for 5 min, and electro-flocculation at 19 EPD and 95 W. After precipitation, cells were frozen at -20 °C and, subsequently, lyophilized. Biomass obtained by electro-flocculation was approximately four times greater than the one achieved by centrifugation. The benefits of this method are that no addition of chemical flocculants is necessary and similar cultivation conditions can be used for the biodiesel production and pharmacological purposes. The results may contribute to improve biodiesel production costs using marine microalgae.

Keywords: Biomass, diatom, flocculation, microalgae.

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25 Chemical Characterization and Prebiotic Effect of Water-Soluble Polysaccharides from Zizyphus lotus Leaves

Authors: Zakaria Boual, Abdellah Kemassi, Toufik Chouana, Philippe Michaud, Mohammed Didi Ould El Hadj

Abstract:

In order to investigate the prebiotic potential of oligosaccharides prepared by chemical hydrolysis of water-soluble polysaccharides (WSP) from Zizyphus lotus leaves, the effect of oligosaccharides on bacterial growth was studied. The chemical composition of WSP was evaluated by colorimetric assays revealed the average values: 7.05±0.73% proteins and 86.21±0.74% carbohydrates, among them 64.81±0.42% is neutral sugar and the rest 16.25±1.62% is uronic acids. The characterization of monosaccharides was determined by high performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) was found to be composed of galactose (23.95%), glucose (21.30%), rhamnose (20.28%), arabinose (9.55%), and glucuronic acid (22.95%). The effects of oligosaccharides on the growth of lactic acid bacteria were compared with those of fructooligosaccharide (RP95). The oligosaccharides concentration was 1g/L of Man, Rogosa, Sharpe broth. Bacterial growth was assessed during 2, 4.5, 6.5, 9, 12, 16 and 24 h by measuring the optical density of the cultures at 600 nm (OD600) and pH values. During fermentation, pH in broth cultures decreased from 6.7 to 5.87±0.15. The enumeration of lactic acid bacteria indicated that oligosaccharides led to a significant increase in bacteria (P≤0.05) compared to the control. The fermentative metabolism appeared to be faster on RP95 than on oligosaccharides from Zizyphus lotus leaves. Both RP95 and oligosaccharides showed clear prebiotic effects, but had differences in fermentation kinetics because of to the different degree of polymerization. This study shows the prebiotic effectiveness of oligosaccharides, and provides proof for the selection of leaves of Zizyphus lotus for use as functional food ingredients.

Keywords: Zizyphus lotus, polysaccharides, characterization, prebiotic effects.

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24 A Review on the Importance of Nursing Approaches in Nutrition of Children with Cancer

Authors: Ş. Çiftcioğlu, E. Efe

Abstract:

In recent years, cancer has been at the top of diseases that cause death in children. Adequate and balanced nutrition plays an important role in the treatment of cancer. Cancer and cancer treatment is affecting food intake, absorption and metabolism, causing nutritional disorders. Appropriate nutrition is very important for the cancerous child to feel well before, during and after the treatment. There are various difficulties in feeding children with cancer. These are the cancer-related factors. Other factors are environmental and behavioral. As health professionals who spend more time with children in the hospital, nurses should be able to support the children on nutrition and help them to have balanced nutrition. This study aimed to evaluate the importance of nursing approaches in the nutrition of children with cancer. This article is planned as a review article by searching the literature on this field. Anorexia may develop due to psychogenic causes or chemotherapeutic agents or accompanying infections and nutrient uptake may be reduced.  In addition, stomatitis, mucositis, taste and odor changes in the mouth, the feeling of nausea, vomiting and diarrhea can also reduce oral intake and result in significant losses in the energy deficit. In assessing the nutritional status of children with cancer, determining weight loss and good nutrition is essential anamnesis of a child.  Some anthropometric measurements and biochemical tests should be used to evaluate the nutrition of the child. The nutritional status of pediatric cancer patients has been studied for a long time and malnutrition, in particular under nutrition, in this population has long been recognized. Yet, its management remains variable with many malnourished children going unrecognized and consequently untreated. Nutritional support is important to pediatric cancer patients and should be integrated into the overall treatment of these children.

Keywords: Cancer treatment, children, complication, nutrition, nursing approaches.

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23 Antioxidant Properties and Nutritive Values of Raw and Cooked Pool Barb (Puntius sophore) of Eastern Himalayas

Authors: Ch. Sarojnalini, Wahengbam Sarjubala Devi

Abstract:

Antioxidant properties and nutritive values of raw and cooked Pool barb, Puntius sophore (Hamilton-Buchanan) of Eastern Himalayas, India were determined. Antioxidant activity of the methanol extract of the raw, steamed, fried and curried Pool barb was evaluated by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging assay. In DPPH scavenging assay the IC50 value of the raw, steamed, fried and curried Pool barb was 1.66 micro-gram/ml, 16.09 micro-gram/ml, 8.99 micro-gram/ml, 0.59 micro-gram/ml whereas the IC50 of the reference ascorbic acid was 46.66miro-gram/ml. These results showed that the fish have high antioxidant activity. Protein content was found highest in raw (20.50±0.08%) and lowest in curried (18.66±0.13%). Moisture content in raw, fried and curried was 76.35±0.09, 46.27±0.14 and 57.46±0.24 respectively. Lipid content was recorded 2.46±0.14% in raw and 21.76±0.10% in curried. Ash content varied from 12.57±0.11 to 22.53±0.07%. The total amino acids varied from 36.79±0.02 and 288.43±0.12 mg/100g. Eleven essential mineral elements were found abundant in all the samples. The samples had considerable amount of Fe ranging from 152.17 to 320.39 milli-gram/100gram, Ca 902.06 to 1356.02 milli-gram/100gram, Zn 91.07 to 138.14 milli-gram/100gram, K 193.25 to 261.56 milli-gram/100gram, Mg 225.06 to 229.10 milli-gram/100gram. Ni was not detected in the curried fish. The Mg and K contents were significantly decreased in frying method; however the Fe, Cu, Ca, Co and Mn contents were increased significantly in all the cooked samples. The Mg and Na contents were significantly increased in curried sample and the Cr content was decreased significantly (p<0.05) in all the cooked samples.

Keywords: Antioxidant property, Pool barb, minerals, amino acids, proximate composition, cooking methods.

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22 The Effect of Saccharomyces cerevisiae Live Yeast Culture on Microbial Nitrogen Supply to Small Intestine in Male Kivircik Yearlings Fed with Different Forage-Concentrate Ratios

Authors: N. Cetinkaya, N. H. Ozdemir

Abstract:

The aim of the study was to investigate the effect of Saccharomyces cerevisiae (SC) live yeast culture on microbial protein supply to small intestine in Kivircik male yearlings when fed with different ratio of forage and concentrate diets. Four Kivircik male yearlings with permanent rumen canula were used in the experiment. The treatments were allocated to a 4x4 Latin square design. Diet I consisted of 70% alfalfa hay and 30% concentrate, Diet II consisted of 30% alfalfa hay and 70% concentrate, Diet I and II were supplemented with a SC. Daily urine was collected and stored at -20°C until analysis. Calorimetric methods were used for the determination of urinary allantoin and creatinine levels. The estimated microbial N supply to small intestine for Diets I, I+SC, II and II+SC were 2.51, 2.64, 2.95 and 3.43 g N/d respectively. Supplementation of Diets I and II with SC significantly affected the allantoin levels in μmol/W0.75 (p<0.05). Mean creatinine values in μmol/W0.75 and allantoin:creatinine ratios were not significantly different among diets. In conclusion, supplementation with SC live yeast culture had a significant effect on urinary allantoin excretion and microbial protein supply to small intestine in Kivircik yearlings fed with high concentrate Diet II (P<0.05). Hence urinary allantoin excretion may be used as a tool for estimating microbial protein supply in Kivircık yearlings. However, further studies are necessary to understand the metabolism of Saccharomyces cerevisiae live yeast culture with different forage:concentrate ratio in Kıvırcık Yearlings.

Keywords: Allantoin, creatinine, Kivircik yearling, microbial nitrogen, Saccharomyces cerevisiae.

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21 Role of Fish Hepatic Aldehyde Oxidase in Oxidative in vitro Metabolism of Phenanthridine Heterocyclic Aromatic Compound

Authors: Khaled S. Al Salhen

Abstract:

Aldehyde oxidase is molybdo-flavoenzyme involved in the oxidation of hundreds of endogenous and exogenous and N-heterocyclic compounds and environmental pollutants. Uncharged N-heterocyclic aromatic compounds such phenanthridine are commonly distributed pollutants in soil, air, sediments, surface water and groundwater, and in animal and plant tissues. Phenanthridine as uncharged N-heterocyclic aromatic compound was incubated with partially purified aldehyde oxidase from rainbow trout fish liver. Reversed-phase HLPC method was used to separate the oxidation products from phenanthridine and the metabolite was identified. The 6(5H)-phenanthridinone was identified the major metabolite by partially purified aldehyde oxidase from fish liver. Kinetic constant for the oxidation reactions were determined spectrophotometrically and showed that this substrate has a good affinity (Km = 78 ± 7.6µM) for hepatic aldehyde oxidase, will be a significant pathway. This study confirms that partially purified aldehyde oxidase from fish liver is indeed the enzyme responsible for the in vitro production 6(5H)-phenanthridinone metabolite as it is a major metabolite by mammalian aldehyde oxidase, coupled with a relatively high oxidation rate (0.77± 0.03 nmol/min/mg protein). In addition, the kinetic parameters of hepatic fish aldehyde oxidase towards the phenanthridine substrate indicate that in vitro biotransformation by hepatic fish aldehyde oxidase will be a significant pathway. This study confirms that partially purified aldehyde oxidase from fish liver is indeed the enzyme responsible for the in vitro production 6(5H)-phenanthridinone metabolite as it is a major metabolite by mammalian aldehyde oxidase.

Keywords: Aldehyde oxidase, Fish, Phenanthridine, Specificity.

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20 Influence of Pomegranate (Punica granatum L.) on Dimethoate Induced Hepatotoxicity in Rats

Authors: Abou-Baker Salim, A. A. K. Abou-Arab, Sherif R. Mohamed, T. A. Eldesouky

Abstract:

Pomegranate (Punica granatum L.) is an ancient fruit of great medical interest and rich source of antioxidants. Pesticides as dimethoate play a crucial role in the occurrence many diseases in plants, animal and human. Therefore the ability of Pomegranate (Punica granatum L.) to alleviate hepatotoxicity induced by organophosphate pesticide dimethoate was investigated. Albino male rats were divided randomly into 4 groups and kept at 7 animals per group in an environmentally controlled condition for 6 weeks. The first group was served as a control group (basal diet), the second group fed on basal diet supplemented with 5% freeze dried pomegranate seeds, the third group fed on 20 ppm dimethoate contaminated diet and the last group fed on dimethoate contaminated diet supplemented with 5% freeze dried pomegranate seeds. The results revealed that administration of dimethoate caused high significant increased in liver functions: alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) activities as well as lipid peroxide (malonaldhyde, MDA); on the other hand high significant decreased on glutathione (GSH), glutathione peroxidase (GPx), albumin and total protein were observed. However addition of 5% freeze dried pomegranate seeds significantly improved all previously mentioned parameters. These results indicate the dimethoate induced hepatotoxicity and highlight the protective effect of pomegranate seeds as a potential protective agent against dimethoate induced hepatotoxicity. This may be attributed to the powerful antioxidants (polyphenols, total phenols, and total flavonoids) which present in high levels in pomegranate as well as improving the immunity by activation of antioxidant enzymes GSH and GPx.

Keywords: Pomegranate, organophosphate pesticides, dimethoate, liver toxicity, free radicals, antioxidants.

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19 Mutation Analysis of the ATP7B Gene in 43 Vietnamese Wilson’s Disease Patients

Authors: Huong M. T. Nguyen, Hoa A. P. Nguyen, Mai P. T. Nguyen, Ngoc D. Ngo, Van T. Ta, Hai T. Le, Chi V. Phan

Abstract:

Wilson’s disease (WD) is an autosomal recessive disorder of the copper metabolism, which is caused by a mutation in the copper-transporting P-type ATPase (ATP7B). The mechanism of this disease is the failure of hepatic excretion of copper to bile, and leads to copper deposits in the liver and other organs. The ATP7B gene is located on the long arm of chromosome 13 (13q14.3). This study aimed to investigate the gene mutation in the Vietnamese patients with WD, and make a presymptomatic diagnosis for their familial members. Forty-three WD patients and their 65 siblings were identified as having ATP7B gene mutations. Genomic DNA was extracted from peripheral blood samples; 21 exons and exon-intron boundaries of the ATP7B gene were analyzed by direct sequencing. We recognized four mutations ([R723=; H724Tfs*34], V1042Cfs*79, D1027H, and IVS6+3A>G) in the sum of 20 detectable mutations, accounting for 87.2% of the total. Mutation S105* was determined to have a high rate (32.6%) in this study. The hotspot regions of ATP7B were found at exons 2, 16, and 8, and intron 14, in 39.6 %, 11.6 %, 9.3%, and 7 % of patients, respectively. Among nine homozygote/compound heterozygote siblings of the patients with WD, three individuals were determined as asymptomatic by screening mutations of the probands. They would begin treatment after diagnosis. In conclusion, 20 different mutations were detected in 43 WD patients. Of this number, four novel mutations were explored, including [R723=; H724Tfs*34], V1042Cfs*79, D1027H, and IVS6+3A>G. The mutation S105* is the most prevalent and has been considered as a biomarker that can be used in a rapid detection assay for diagnosis of WD patients. Exons 2, 8, and 16, and intron 14 should be screened initially for WD patients in Vietnam. Based on risk profile for WD, genetic testing for presymptomatic patients is also useful in diagnosis and treatment.

Keywords: ATP7B gene, mutation detection, presymptomatic diagnosis, Vietnamese Wilson’s disease.

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18 Comparative Correlation Investigation of Polynuclear Aromatic Hydrocarbons (PAHs) in Soils of Different Land Use: Sources Evaluation Perspective

Authors: O. Onoriode Emoyan, E. Eyitemi Akporhonor, Charles Otobrise

Abstract:

Polycyclic Aromatic Hydrocarbons (PAHs) are formed mainly because of incomplete combustion of organic materials during industrial, domestic activities or natural occurrence. Their toxicity and contamination of terrestrial and aquatic ecosystem have been established. However, with limited validity index, previous research has focused on PAHs isomer pair ratios of variable physicochemical properties in source identification. The objective of this investigation was to determine the empirical validity of Pearson Correlation Coefficient (PCC) and Cluster Analysis (CA) in PAHs source identification along soil samples of different land uses. Therefore, 16 PAHs grouped, as Endocrine Disruption Substances (EDSs) were determined in 10 sample stations in top and sub soils seasonally. PAHs was determined the use of Varian 300 gas chromatograph interfaced with flame ionization detector. Instruments and reagents used are of standard and chromatographic grades respectively. PCC and CA results showed that the classification of PAHs along pyrolitic and petrogenic organics used in source signature is about the predominance PAHs in environmental matrix. Therefore, the distribution of PAHs in the studied stations revealed the presence of trace quantities of the vast majority of the sixteen PAHs, which may ultimately inhabit the actual source signature authentication. Therefore, factors to be considered when evaluating possible sources of PAHs could be; type and extent of bacterial metabolism, transformation products/substrates, and environmental factors such as salinity, pH, oxygen concentration, nutrients, light intensity, temperature, co-substrates, and environmental medium are hereby recommended as factors to be considered when evaluating possible sources of PAHs.

Keywords: Comparative correlation, kinetically, polynuclear aromatic hydrocarbons, thermodynamically- favored PAHs, sources evaluation.

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17 An Open-Label Pilot Study of Efficacy and Safety of 2% Curcuma aeruginosa Roxb. Extract Cream in the Treatment of Mild to Moderate Facial Seborrheic Dermatitis

Authors: Kulaya Wimolwat, Panlop Chakravitthamrong, Neti Waranuch

Abstract:

Background: Seborrheic dermatitis is a common chronic skin condition affecting the face, scalp, chest, and trunk. The cause of seborrheic dermatitis is still unknown. Sebum production, lipid composition, hormone levels, and Malassezia species have been suggested as important factors in the development of seborrheic dermatitis. Curcuma aeruginosa Roxb. extract-containing cream with anti-inflammatory and anti-androgenic properties may be beneficial for treating mild to moderate facial seborrheic dermatitis. Objectives: We evaluated the efficacy and safety of 2% C. aeruginosa Roxb. extract-containing cream in the treatment of mild to moderate seborrheic dermatitis. Methods: This was a prospective, open-label, and non-comparative study. Ten adult patients clinically diagnosed with mild to moderate seborrheic dermatitis were enrolled in a four-week study. The 2% C. aeruginosa Roxb. cream was applied twice daily to a lesional area on the face for four weeks. The Scoring Index (SI) ranking system on days 14 and 28 was compared with that at baseline to determine the efficacy of treatment. The adverse events (burning sensation and erythema) were evaluated on days 14 and 28 to determine the safety of the treatment. Results: Significant improvement was observed in the reduction of the mean SI at day 14 (2.9) and 28 (1.4) compared to that at baseline (4.9). An adverse reaction was observed on day 14 (mild erythema 20% and mild burning sensation 10%) and was resolved by the end of the study. Conclusion: This open-label pilot study has shown that there was a significant improvement in the severity in these seborrheic patients and most reported they were satisfied with it. Reported adverse events were all mild.

Keywords: Anti-androgenic, antifungals, anti-inflammatory, Curcuma aeruginosa, seborrheic dermatitis, efficacy, safety.

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16 Apoptosis Pathway Targeted by Thymoquinone in MCF7 Breast Cancer Cell Line

Authors: M. Marjaneh, M. Y. Narazah, H. Shahrul

Abstract:

Array-based gene expression analysis is a powerful tool to profile expression of genes and to generate information on therapeutic effects of new anti-cancer compounds. Anti-apoptotic effect of thymoquinone was studied in MCF7 breast cancer cell line using gene expression profiling with cDNA microarray. The purity and yield of RNA samples were determined using RNeasyPlus Mini kit. The Agilent RNA 6000 NanoLabChip kit evaluated the quantity of the RNA samples. AffinityScript RT oligo-dT promoter primer was used to generate cDNA strands. T7 RNA polymerase was used to convert cDNA to cRNA. The cRNA samples and human universal reference RNA were labelled with Cy-3-CTP and Cy-5-CTP, respectively. Feature Extraction and GeneSpring softwares analysed the data. The single experiment analysis revealed involvement of 64 pathways with up-regulated genes and 78 pathways with downregulated genes. The MAPK and p38-MAPK pathways were inhibited due to the up-regulation of PTPRR gene. The inhibition of p38-MAPK suggested up-regulation of TGF-ß pathway. Inhibition of p38-MAPK caused up-regulation of TP53 and down-regulation of Bcl2 genes indicating involvement of intrinsic apoptotic pathway. Down-regulation of CARD16 gene as an adaptor molecule regulated CASP1 and suggested necrosis-like programmed cell death and involvement of caspase in apoptosis. Furthermore, down-regulation of GPCR, EGF-EGFR signalling pathways suggested reduction of ER. Involvement of AhR pathway which control cytochrome P450 and glucuronidation pathways showed metabolism of Thymoquinone. The findings showed differential expression of several genes in apoptosis pathways with thymoquinone treatment in estrogen receptor-positive breast cancer cells.

Keywords: CARD16, CASP10, cDNA microarray, PTPRR, Thymoquinone.

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15 In vitro Effects of Salvia officinalis on Bovine Spermatozoa

Authors: Eva Tvrdá, Boris Botman, Marek Halenár, Tomáš Slanina, Norbert Lukáč

Abstract:

In vitro storage and processing of animal semen represents a risk factor to spermatozoa vitality, potentially leading to reduced fertility. A variety of substances isolated from natural sources may exhibit protective or antioxidant properties on the spermatozoon, thus extending the lifespan of stored ejaculates. This study compared the ability of different concentrations of the Salvia officinalis extract on the motility, mitochondrial activity, viability and reactive oxygen species (ROS) production by bovine spermatozoa during different time periods (0, 2, 6 and 24 h) of in vitro culture. Spermatozoa motility was assessed using the Computer-assisted sperm analysis (CASA) system. Cell viability was examined using the metabolic activity MTT assay, the eosin-nigrosin staining technique was used to evaluate the sperm viability and ROS generation was quantified using luminometry. The CASA analysis revealed that the motility in the experimental groups supplemented with 0.5-2 µg/mL Salvia extract was significantly lower in comparison with the control (P<0.05; Time 24 h). At the same time, a long-term exposure of spermatozoa to concentrations ranging between 0.05 µg/mL and 2 µg/mL had a negative impact on the mitochondrial metabolism (P<0.05; Time 24 h). The viability staining revealed that 0.001-1 µg/mL Salvia extract had no effects on bovine male gametes, however 2 µg/mL Salvia had a persisting negative effect on spermatozoa (P<0.05). Furthermore 0.05-2 µg/mL Salvia exhibited an immediate ROS-promoting effect on the sperm culture (P>0.05; Time 0 h and 2 h), which remained significant throughout the entire in vitro culture (P<0.05; Time 24 h). Our results point out to the necessity to examine specific effects the biomolecules present in Salvia officinalis may have individually or collectively on the in vitro sperm vitality and oxidative profile.

Keywords: Bulls, CASA, MTT test, reactive oxygen species, sage, Salvia officinalis, spermatozoa.

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