Search results for: acyl-coa binding protein (ACBP)

Authors: Ali Bayram, Burak Uz, Ilhan Dolasik, Remzi Yiğiter

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The GABARAP (GABAA-receptor-associated protein) family consists of GABARAP, GABARAPL1 (GABARAP-like 1) and GABARAPL2 (GABARAP-like 2). GABARAPL1, like GABARAP, was described to interact with both GABAA receptor and tubulin, and to be involved in intracellular GABAA receptor trafficking and promoting tubulin polymerization. In addition, GABARAPL1 is thought to be involved in various physiological (autophagosome closure, regulation of circadian rhythms) and/or pathological mechanisms (cancer, neurodegeneration). Alzheimer’s disease (AD) is a progressive neuro degenerative disorder characterized with impaired cognitive functions. Disruption of the GABAergic neuro transmission as well as cholinergic and glutamatergic interactions, may also be involved in the pathogenesis of AD. GABARAPL1 presents a regulated tissue expression and is the most expressed gene among the GABARAP family members in the central nervous system. We, herein, conducted a study to investigate the GABARAPL1 mRNA expression levels in patients with AD. 50 patients with AD and 49 control patients were enrolled to the present study. Messenger RNA expression levels of GABARAPL1 were detected by real-time polymerase chain reaction. GABARAPL1 mRNA expression in AD / control patients was 0,495 (95% confidence interval: 0,404-0,607), p= 0,00000002646. Reduced activity of GABARAPL1 gene might play a role, at least partly, in the pathophysiology of AD.

Keywords: Alzheimer’s disease, GABARAPL1, mRNA expression, RT-PCR

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Authors: E. Irankhah, M. Zarif, E. Mazrooei Rad, K. Ghandehari

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Alzheimer's prevalence is on the rise, and the disease comes with problems like cessation of treatment, high cost of treatment, and the lack of early detection methods. The pathology of this disease causes the formation of protein deposits in the brain of patients called plaque amyloid. Generally, the diagnosis of this disease is done by performing tests such as a cerebrospinal fluid, CT scan, MRI, and spinal cord fluid testing, or mental testing tests and eye tracing tests. In this paper, we tried to use the Medial Temporal Atrophy (MTA) method and the Leave One Out (LOO) cycle to extract the statistical properties of the three Fz, Pz, and Cz channels of ERP signals for early diagnosis of this disease. In the process of CT scan images, the accuracy of the results is 81% for the healthy person and 88% for the severe patient. After the process of ERP signaling, the accuracy of the results for a healthy person in the delta band in the Cz channel is 81% and in the alpha band the Pz channel is 90%. In the results obtained from the signal processing, the results of the severe patient in the delta band of the Cz channel were 89% and in the alpha band Pz channel 92%.

Keywords: Alzheimer's disease, image and signal processing, LOO cycle, medial temporal atrophy

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Authors: Novia Dwi Prabandari, Diah Ayu Asmarani

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Soluble fiber can accelerate the metabolism of cholesterol. Pectin and gum has been used in the form of substance additive for material stabilizer and emulsifier. Pectin supplementation in laying hens can decimate the cholesterol content in egg yolk and muscle. Therefore, this laying hens’ feed is regular feed chickens enriched with soluble fiber (Pectin, Xanthan gum, and Guar gum) to produce eggs and muscle with lower cholesterol than usual.The ingredients are mixed in the ratio of concentrate 45%, corn flour 25%, soybean meal 20%, and extract of soluble fiber 10%. Once all the ingredients are mixed and then evaporated with temperature < 80 °C. Then put in the grinding machine resulting in a circular shape with holes 2-3 mm in diameter, after it dried up the water content in the feed is less than 14%. Eggs from laying hen with soluble fiber fortification feed intake will have lower cholesterol levels in eggs than regular feed. So even with the cholesterol content in the muscle, it is because chicken feed fortified with soluble fiber will accelerate the metabolism of cholesterol and cause cholesterol deposits in the chicken less. The use of this kind of laying hens feed is produce eggs with high protein content can be consumed more for people who have hypercholesterolemia.

Keywords: pectin, xanthan gum, guar gum, laying hen, cholesterol

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Authors: Mir Hussain Nawaz, Lyudmila Nedyalkova, Haizhong Zhu, Wael M. Rabeh

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In this study, we aim to biochemically and structurally characterize the interactions of human HK2 with the mitochondria in addition to the role of its N-terminal domain in catalysis and stability of the full-length enzyme. Here, we solved the crystal structure of human HK2 in complex with glucose and glucose-6-phosphate (PDB code: 2NZT), where it is a homodimer with catalytically active N- and C-terminal domains linked by a seven-turn α-helix. Different from the inactive N-terminal domains of isozymes 1 and 3, the N- domain of HK2 not only capable to catalyze a reaction but it is responsible for the thermodynamic stabilizes of the full-length enzyme. Deletion of first α-helix of the N-domain that binds to the mitochondria altered the stability and catalytic activity of the full-length HK2. In addition, we found the linker helix between the N- and C-terminal domains to play an important role in controlling the catalytic activity of the N-terminal domain. HK2 is a major step in the regulation of glucose metabolism in cancer making it an ideal target for the development of new anticancer therapeutics. Characterizing the structural and molecular mechanisms of human HK2 and its role in cancer metabolism will accelerate the design and development of new cancer therapeutics that are safe and cancer specific.

Keywords: cancer metabolism, enzymology, drug discovery, protein stability

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Authors: Haralabos C. karantonis, Afroditi Michalaki

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High intakes of carbohydrates and fats have been associated with an increased risk of chronic diseases due to the role of postprandial oxidative stress. This pilot nutritional intervention aimed to examine the acute effect of consuming two different types of breadsticks prepared from durum wheat flour mixtures differing in total phenolic content on postprandial inflammatory and oxidant responses in healthy volunteers. A cross-over, controlled, and single-blind clinical trial was designed, and two isocaloric high-fat and high-carbohydrate meals were tested. Serum total, High Density Lipoprotein (HDL)- and Low Density Lipoprotein (LDL)-cholesterol, triglycerides, glucose, C-reactive protein (CRP), uric acid, plasma total antioxidant capacity, and antiplatelet activity were determined in fasting and 30, 60, and 120 min after consumption. The results showed a better postprandial HDL-cholesterol and total antioxidant activity response in the intervention group. The choice of durum wheat flours with higher phenolic content and antioxidant activity is presented as promising for human health, and clinical studies will expand to draw safer conclusions.

Keywords: antioxidant, antiplatelet, durum wheat, nutritional value

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Authors: Xianbao Sun, Jinlong Zhao, Shudong He, Jing Li

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Volutharpa ampullacea perryi is a high-protein marine shellfish. However, few data are available on the effects of boiling temperatures and time on quality of the meat. In this study, colour, texture and sensory characteristics of Volutharpa ampullacea perryi meat during the boiling cooking processes (75-100 °C, 5-60 min) were investigated by colors analysis, texture profile analysis (TPA), scanning electron microscope (SEM) and sensory evaluation. The ratio of cooking loss gradually increased with the increase of temperature and time. The colour of meat became lighter and more yellower from 85 °C to 95 °C in a short time (5-20 min), but it became brown after a 30 min treatment. TPA results showed that the Volutharpa ampullacea perryi meat were more firm and less cohesive after a higher temperature (95-100 °C) treatment even in a short period (5-15 min). Based on the SEM analysis, it was easily found that the myofibrils structure was destroyed at a higher temperature (85-100 °C). Sensory data revealed that the meat cooked at 85-90 °C in 10-20 min showed higher scores in overall acceptance, as well as color, hardness and taste. Based on these results, it could be constructed that Volutharpa ampullacea perryi meat should be heated on a suitable condition (such as 85 °C 15 min or 90 °C 10 min) in the boiling cooking to be ensure a better acceptability.

Keywords: Volutharpa ampullacea perryi meat, boiling cooking, colour, sensory, texture

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Authors: L. Bouzina, A. Bensafi, M. Duval, C. Mathis, M. Rawiso

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We study the stability and structural properties of mixtures of model nanoparticles and non-adsorbing polymers in the 'protein limit', where the size of polymers exceeds the particle size substantially. We have synthesized in institute (Charles Sadron Strasbourg) model nanoparticles by coating fullerene C60 molecules with low molecular weight polystyrene (PS) chains (6 PS chains with a degree of polymerization close to 25 and 50 are grafted on each fullerene C60 molecule. We will present a Small Angle Neutron scattering (SANS) study of Tetrahydrofuran (THF) solutions involving long polystyrene (PS) chains and fullerene (C60) nanoparticles. Long PS chains and C60 nanoparticles with different arm lengths were synthesized either hydrogenated or deuteriated. They were characterized through Size Exclusion Chromatography (SEC) and Quasielastic Light Scattering (QLS). In this way, the solubility of the C60 nanoparticles in the usual good solvents of PS was controlled. SANS experiments were performed by use of the contrast variation method in order to measure the partial scattering functions related to both components. They allow us to obtain information about the dispersion state of the C60 nanoparticles as well as the average conformation of the long PS chains. Specifically, they show that the addition of long polymer chains leads to the existence of an additional attractive interaction in between soft nanoparticles.

Keywords: fulleren nanoparticles, polymer, small angle neutron scattering, solubility

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Authors: Mona Hamdy, Iman Shaheen, Hadeel Seif Eldin, Basma Ali, Omnia Abdeldayem

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Summary: Bone involvement of sickle cell disease (SCD) patients varies from acute clinical manifestations of painful vaso-occlusive crises or osteomyelitis to more chronic affection of bone mineral density (BMD) and debilitating osteonecrosis and osteoporosis. Secreted klotho protein is involved in calcium (Ca) reabsorption in the kidney. This study aimed to measure serum klotho levels in children with SCD to determine the possibility of using it as a marker of low BMD in children with SCD in correlation with a dual-energy radiograph absorptiometry scan. This study included 60 sickle disease patients and 30 age-matched and sex-matched control participants without SCD. A highly statistically significant difference was found between patients with normal BMD and those with low BMD, with serum Ca and klotho levels being lower in the latter group. Klotho serum level correlated positively with both serum Ca and BMD. Serum klotho level showed 94.9% sensitivity and 95.2% specificity in the detection of low BMD. Both serum Ca and klotho serum levels may be useful markers for detection of low BMD related to SCD with high sensitivity and specificity; however, klotho may be a better indicator as it is less affected by the nutritional and endocrinal status of patients or by intake of Ca supplements.

Keywords: sickle cell disease, BMD, osteoporosis, DEXA, klotho

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Authors: Mohammed A. Alfayyadh, Halla A. AlAbdulhadi, Mahdi H. Almubarak

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Purpose: Eales’ disease is an idiopathic vasculitis that affects the peripheral retina. It is characterized by recurrent vitreous hemorrhage as a complication of retinal neovascularization. It is more prevalent in India and affects young males. Here we present a patient with neovascular glaucoma as a rare first presentation of Eales’ disease. Observations: This is a 24-year-old Indian gentleman, who complained of a sudden decrease in vision in the left eye over less than 24 hours, along with frontal headache and eye pain for the last three weeks. Ocular examination revealed peripheral retinal ischemia in the right eye, very high intraocular pressure, rubeosis iridis, vitreous hemorrhage and extensive retinal ischemia in the left eye, vascular sheathing and neovascularization in both eyes. Purified protein derivative skin test was positive. The patient was managed with anti-glaucoma, intravitreal anti-vascular endothelial growth factor and laser photocoagulation. Systemic steroids and anti-tuberculous therapy were also initiated. Conclusions: Neovascular glaucoma is an infrequent complication of Eales’ disease. However, the lack of early detection of the disease in the early stages might lead to such serious complication.

Keywords: case report, Eales’ disease, mycobacterium tuberculosis, neovascular glaucoma

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Authors: E. Oskoueian, E. Maroufyan, Y. M. Goh, E. Ramezani-Fard, M. Ebrahimi

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The fish meat plays an important role in the human health as it contains high quality protein. The tilapia fish considered as the third largest group of farmed fish. The oxidative deterioration of fish meat may occur during the cooking process. The proper cooking process and using natural antioxidant to prevent oxidation and enhance the quality of the tilapia fish fillet is necessary. Hence, this research was carried out to evaluate the potential of clove essential oil to prevent lipid peroxidation and enhance the antioxidant activity of tilapia fish fillet cooked using microwave and griller. The results showed that cooking using microwave significantly (p < 0.05) increased the lipid peroxidation and decreased the DPPH and ferric reducing activity power of the fish fillet as compared to grilling. The fortification of fish fillet using clove essential oil prevented from lipid peroxidation and enhanced the antioxidant activity of the fish fillet significantly (p < 0.05). Consequently, fortification of tilapia fish fillet using clove essential oil followed by cooking using griller to have high quality cooked fish meat is recommended.

Keywords: antioxidant activity, fillet, fish, fortification, lipid peroxidation

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Authors: Patsarawadee Paojinda, Waranya Imprasittichai, Sudaratana R. Krungkrai, Nirianne Marie Q. Palacpac, Toshihiro Horii, Jerapan Krungkrai

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Fusion of the last two enzymes in the pyrimidine biosynthetic pathway in the inversed order by having COOH-terminal orotate phosphoribosyltransferase (OPRT) and NH2-terminal orotidine 5'-monophosphate decarboxylase (OMPDC), as OMPDC-OPRT, are described in many organisms. Here, we produced gene fusions of Plasmodium falciparum OMPDC-OPRT and expressed the bifunctional protein in Escherichia coli. The enzyme was purified to homogeneity using affinity and anion-exchange chromatography, exhibited enzymatic activities and functioned as a dimer. The activities, although unstable, can be stabilized by its substrate and product during purification and long-term storage. Furthermore, the enzyme expressed a perfect catalytic efficiency (kcat/Km). The kcat was selectively enhanced up to 3 orders of magnitude, while the Km was not much affected and remained at low µM levels when compared to the monofunctional enzymes. The fusion of the two enzymes, creating a “super-enzyme” with perfect catalytic power and more flexibility, reflects cryptic relationship of enzymatic reactivaties and metabolic functions on molecular evolution.

Keywords: bifunctional enzyme, orotate phosphoribosyltransferase, orotidine 5'-monophosphate decarboxylase, plasmodium falciparum

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Authors: A. Albaridi Najla

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Flaxseed (Linum usitatissimum) is well known to have beneficial effect on health. The seeds are rich in protein, α-linolenic fatty acid and dietary fiber. Bakery products are important part of our daily meals. Functional food recently received considerable attention among consumers. The increase in bread daily consumption leads to the production of breads with functional ingredients such as flaxseed The aim of this Study was to improve the nutritional value of bread by adding flaxseed flour and assessing the effect of adding 0, 5, 10 and 15% flaxseed on whole wheat bread rheological and sensorial properties. The total consumer's acceptability of the flaxseed bread was assessed. Dough characteristics were determined using Farinograph (C.W. Brabender® Instruments, Inc). The result shows no change was observed in water absorption between the stander dough (without flaxseed) and the bread with flaxseed (67%). An Increase in the peak time and dough stickiness was observed with the increase in flaxseed level. Further, breads were evaluated for sensory parameters, colour and texture. High flaxseed level increased the bread crumb softness. Bread with 5% flaxseed was optimized for total sensory evaluation. Overall, flaxseed bread produced in this study was highly acceptable for daily consumption as a functional foods with a potentially health benefits.

Keywords: bread, flaxseed, rheological properties, whole-wheat bread

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Authors: Vandana Singh, Subash Singh

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Introduction: Oral squamous cell carcinoma is the most common malignant tumor of the oral cavity. Normally the death of cell and the growth are active processes and depend not only on external factors but also on the expression of genes like Bcl-2, which activate and inhibit apoptosis. The term Bcl-2 is an acronym for B-cell lymphoma/ leukemia -2 genes. Objectives: An attempt was made to evaluate Bcl-2 oncoprotein expression in patients with oral precancer and cancer and to assess possible correlation between Bcl-2 oncoprotein expression and clinicopathological features of oral precancer and cancer. Material and Methods: This is a selective prospective clinical and immunohistochemical study. Clinicopathological examination is correlated with immunohistochemical findings. The immunolocalization of Bcl-2 protein is performed using the labeled streptavidin biotin (LSAB) method. To visualize the reaction, 3, 3-diaminobenzidine (DAB) is used. Results: Bcl-2 expression was positive in 11 [36.66 %, low Bcl-2 expression 3 (10.00 %), moderate Bcl-2 expression 7 (23.33 %), and high Bcl-2 expression 1 (3.33 %)] oral cancer cases and in 14 [87.50 %, low expression 8 (50 %), moderate expression 6 (37.50 %)] precancer cases. Conclusion: On the basis of the results of our study we conclude that positive Bcl-2 expression may be an indicator of poor prognosis in oral cancer and precancer. Relevance: It has been reported that there is deregulation of Bcl-2 expression during progression from oral epithelial dysplasia to squamous cell carcinoma. It can be used for revealing progression of epithelial dysplasia to malignancy and as a prognostic marker in oral precancer and cancer.

Keywords: BcL-2, immunohistochemistry, oral cancer, oral precancer

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Authors: Hae Jeong Park, Joo-Ho Chung

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In this study, the effect of essential oil from Chamaecyparis obtuse (EOCO) on early life stress using maternal separation (MS) rats was investigated. Anxiety-related behaviors were examined in MS rats using the elevated plus-maze (EPM) test. The changes of gene expressions by EOCO in the hippocampus of MS rats were analyzed using a microarray method. Rats in the MS groups were separated from their respective mothers from postnatal day (pnd) 14 to 28. Rats in the EOCO-treated groups were exposed to EOCO for 1 h or 2 h by inhalation from pnd 21 to 28. The EOCO-treated MS rats showed decreased anxiety-related behaviors compared to the MS rats in the EPM test. In the microarray analysis, EOCO downregulated the expressions of cytokine genes such as Ccl2, Il6, Cxcl10, Ccl19, and Il1rl in the hippocampus of MS rats, and it was also confirmed through RT-PCR. In particular, the expressions of Ccl2 and Il6 were predominantly decreased by EOCO in the hippocampus of MS rats. Interestingly, their protein expressions were also reduced by EOCO in MS rats. These results indicate that EOCO decreases MS-induced anxiety-related behaviors, and modulate cytokines, particularly Ccl2 and Il6, in the hippocampus of MS rats.

Keywords: anxiety-related behavior, Chamaecyparis obtuse, cytokine gene, early-life stress, maternal separation

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Authors: Sagarjyoti Roy

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Dengue is an arboviral infection, belonging to family flaviviridae, comprising of four serotypes; DENV1, DENV2, DENV3 and DENV4. All four serotypes are capable of causing full-spectrum of clinical features, ranging from self-limiting fever to severe dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). Complications may affect any organ system, including those involving gastrointestinal system and serositis. We report a case, of a 28 years, non-alcoholic male, presenting with a 7 day history of fever and malaise followed by abdominal pain and distension, from 4th day of fever. He was admitted in medicine department of RG KAR medical college hospital. Dengue fever was confirmed by NS1 and dengue IgM positivity. Platelet count was 30,000/cc (1.5- 4 lac/cc) and haematocrit was 52% (38- 50% for men). Clinicoradiological findings revealed bilateral pleural effusion, ascites and splenomegaly. Ascitic fluid was hemorrhagic in nature, with a high protein and RBC content. Liver function tests revealed mild transaminitis with normal coagulation profile. Patient was managed conservatively. A diagnosis of dengue fever complicated by serositis and spontaneous haemoperitoneum was made. The symptoms subsided after a hospital stay of 10 days. The case highlights haemorrhage into peritoneal cavity as a possible complication of dengue fever. Although a definite explanation requires more detailed studies, platelet or endothelial cell dysfunction might be contributory.

Keywords: ascites, dengue, haemoperitoneum, serositis

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Authors: Tamer El-Messery, Beraat Ozcelik

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This study aimed to produce functional yogurt supplemented with microencapsulated krill oil as a source of omega 3, which is known, to maintain the normal brain function, reduce the risk of cancer, and preventing cardiovascular disease. Krill oil was mixed with black cumin oil (1:1) in order to increase its oxidative stability. β-caroteine (10 mg/100 ml) was used as a standard antioxidant. Maltodextrin (MD) was mixed with whey protein concentrate (WPC) and gum Arabic (GA) at the ratio of 8:2:0.5 ratios and used for microencapsulation of single or mixed oils. The microcapsules were dried by freeze and spray drying in order to maximize encapsulation efficiency and minimize lipid oxidation. The feed emulsions used for particle production were characterized for stability, viscosity and particle size, zeta potential, and oxidative stability. The oxidative stability for mixed krill oil and black cumin oil was the highest. The highest encapsulation efficiency was obtained using spray drying, which also showed the highest oxidative stability. The addition of encapsulated krill and black cumin oils (1:1) powder in yogurt manufacture reduced slightly effects on the development of acidity, textural parameters, and water holding capacity of yogurt as compared to control.

Keywords: Krill oil, black cumin oil, micro-encapsulation, oxidative stability, functional yogurt

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Authors: Indranil Chakraborty, Kalyan Mandal

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Three new α-hydroxy carboxylate group functionalized MnFe₂O₄ nanoparticles (NPs) have been developed to explore the microscopic origin of ligand modified fluorescence and magnetic properties of nearly monodispersed MnFe₂O₄ NPs. The surface functionalization has been carried out with three small organic ligands (tartrate, malate, and citrate) having different number of α-hydroxy carboxylate functional group along with steric effect. Detailed study unveils that α-hydroxy carboxylate moiety of the ligands plays key role to generate intrinsic fluorescence in functionalized MnFe₂O₄ NPs through the activation of ligand to metal charge transfer transitions, associated with ligand-Mn²⁺/Fe³⁺ interactions along with d-d transition corresponding to d-orbital energy level splitting of Fe³⁺ ions on NP surface. Further, MnFe₂O₄ NPs show a maximum 140.88% increase in coercivity and 97.95% decrease in magnetization compared to its bare one upon functionalization. The ligands that induce smallest crystal field splitting of d-orbital energy level of transition metal ions are found to result in strongest ferromagnetic activation of the NPs. Finally, our developed tartrate functionalized MnFe₂O₄ (T-MnFe₂O₄) NPs have been utilized for studying DNA binding interaction and nuclease activity for stimulating their beneficial activities toward diverse biomedical applications. The spectroscopic measurements indicate that T-MnFe₂O₄ NPs bind calf thymus DNA by intercalative mode. The ability of T-MnFe₂O₄ NPs to induce DNA cleavage was studied by gel electrophoresis technique where the complex is found to promote the cleavage of pBR322 plasmid DNA from the super coiled form I to linear coiled form II and nicked coiled form III with good efficiency. This may be taken into account for designing new biomolecular detection agents and anti-cancer drug which can open up a new door toward diverse non-invasive biomedical applications.

Keywords: MnFe₂O₄ nanoparticle, α-hydroxy carboxylic acid, comparative fluorescence, magnetism study, DNA interaction, nuclease activity

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Authors: Z. M. Iqbal, M. Abdullah, K. Javed, M. A. Jabbar, A. Haque, M. Saadullah, F. Shahzad

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The current study was conducted to set the appropriate concentrate level for Nili-Ravi buffalo heifers. Twenty seven buffalo heifers were randomly divided into three different groups A, B and C having nine animals in each group. All the heifers were given free access to chopped green fodder and fresh water. In addition, heifers of group A, B and C were given concentrate at the rate of 0.5%, 1% and 1.5% of their body weight. The average daily dry matter intake was 2.69, 3.06 and 3.83 kg with average daily gain of 456.09, 398.56 and 515.87 gm in group A, B and C, respectively. The feed conversion ratio of heifers of these groups was 5.89, 7.74 and 7.52, respectively. There was non-significant (P>0.05) difference in the body measurements (height at wither, body length and heart girth), final body condition and scoring and blood serum (glucose, total protein and cholesterol) of heifers of all the three groups. The results of current study shows that there is non-significant (P>0.05) difference in the growth rate of Nili-Ravi heifers at varying levels of concentrate so, it is cost effective to raise 6-8 month calves by offering concentrate at the rate of 0.5% body weight along with free access of green fodder.

Keywords: concentrate level, buffalo heifer, body measurement, green fodder

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Authors: Sadaf Ilyas

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Recombinant cytokines have been employed successfully as potential therapeutic agent. Some cytokine therapies are already used as a part of clinical practice, ranging from early exploratory trials to well established therapies that have already received approval. Interleukin 15 is a pleiotropic cytokine having multiple roles in peripheral innate and adaptive immune cell function. It regulates the activation, proliferation and maturation of NK cells, T-cells, monocytes/macrophages and granulocytes, and the interactions between them thus acting as a bridge between innate and adaptive immune responses. Unraveling the biology of IL-15 has revealed some interesting surprises that may point toward some of the first therapeutic applications for this cytokine. In this study, the human interleukin 15 gene was isolated, amplified and ligated to a TA vector which was then transfected to a bacterial host, E. coli Top10F’. The sequence of cloned gene was confirmed and it showed 100% homology with the reported sequence. The confirmed gene was then subcloned in pET Expression system to study the IPTG induced expression of IL-15 gene. Positive expression was obtained for number of clones that showed 15 kd band of IL-15 in SDS-PAGE analysis, indicating the successful strain development that can be studied further to assess the potential therapeutic intervention of this cytokine in relevance to human diseases.

Keywords: Interleukin 15, pET expression system, immune therapy, protein purification

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Authors: Babatunde Stephen Oladeji, Gloria Asuquo Edet

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The proximate, functional, pasting, and sensory properties of semolina from blends of cassava, maize, and rice were investigated. Cassava, maize, and rice were milled and sieved to pass through a 1000 µm sieve, then blended in the following ratios to produce five samples; FS₁ (40:30:30), FS₂ (20:50:30), FS₃ (25:25:50), FS₄ (34:33:33) and FS₅ (60:20:20) for cassava, maize, and rice, respectively. A market sample of wheat semolina labeled as FSc served as the control. The proximate composition, functional properties, pasting profile, and sensory characteristics of the blends were determined using standard analytical methods. The protein content of the samples ranged from 5.66% to 6.15%, with sample FS₂ having the highest value and being significantly different (p ≤ 0.05). The bulk density of the formulated samples ranged from 0.60 and 0.62 g/ml. The control (FSc) had a higher bulk density of 0.71 g/ml. The water absorption capacity of both the formulated and control samples ranged from 0.67% to 2.02%, with FS₃ having the highest value and FSc having the lowest value (0.67%). The peak viscosity of the samples ranged from 60.83-169.42 RVU, and the final viscosity of semolina samples ranged from 131.17 to 235.42 RVU. FS₅ had the highest overall acceptability score (7.46), but there was no significant difference (p ≤ 0.05) from other samples except for FS₂ (6.54) and FS₃ (6.29). This study establishes that high-quality and consumer-acceptable semolina that is comparable to the market sample could be produced from blends of cassava, maize, and rice.

Keywords: semolina, gluten, celiac disease, wheat allergies

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Authors: Hafiz Khuram Wasim Aslam, Muhammad Saeed, Azam Shakeel, Muhammad Inam Ur Raheem, Moazzam Rafiq Khan, Muhammad Atif Randhawa

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Inulin is significant ingredient used in food industry that functions technologically as a fat replacer often without compromising taste and texture. In this study inulin was extracted from the chicory roots and the effect of inulin addition as a fat replacer on the physiochemical, microbiological and sensory properties of non-fat yogurt was investigated. The supplementation of chicory inulin reduced the magnitude of firmness in comparison with non-inulin ¬supplemented non-fat yoghurt. Higher values of acidity were observed due to the more microbial fermentation in the inulin containing yogurt as compared to non-inulin yogurt and were in the range of 0.56 to 0.75 during storage days. Syneresis in control sample increased from 43.9% to 47.9% during the storage study. However inulin addition at different treatment enhanced syneresis from 44.5% to 47.6%. Inulin addition at various concentrations caused an increase in the TPC due to its probiotic effect. No effects of inuline addition on fat and protein contents were observed. Non-fat yoghurt supplemented with inulin demonstrated sensory behavior better than that of the control yoghurt. The most important effect of the addition of inulin to non-fat yoghurt is an increase in the sensory attributes appearance, body and texture, taste and mouth feel, overall acceptability. On an average, yoghurt supplemented with 1 to 2% inulin was better in overall acceptance as compared to control yoghurt.

Keywords: inulin, fat replacer, yoghurt, sensory evaluation, low fat

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Authors: Banu Aytül Ekmekçi

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Salinity stress has negative effects on agricultural yield throughout the world, affecting production whether it is for subsistence or economic gain. This study investigates the inductive role of vitamin C and its application mode in mitigating the detrimental effects of irrigation with diluted (10, 20 and 30 %) NaCl + water on carthamus tinctorius plants. The results show that 10% of salt water exhibited insignificant changes, while the higher levels impaired growth by reducing seed germination, dry weights of shoot and root, water status and chlorophyll contents. However, irrigation with salt water enhanced carotenoids and antioxidant enzyme activities. The detrimental effects of salt water were ameliorated by application of 100 ppm ascorbic acid (vitamin C). The inductive role of vitamin was associated with the improvement of seed germination, growth, plant water status, carotenoids, endogenous ascorbic acid and antioxidant enzyme activities. Moreover, vitamin C alone or in combination with 30% NaCl water increased the intensity of protein bands as well as synthesized additional new proteins with molecular weights of 205, 87, 84, 65 and 45 kDa. This could increase tolerance mechanisms of treated plants towards water salinity.

Keywords: salinity, stress, vitamin c, antioxidant, NaCl, enzyme

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Authors: Onyinyechi Lilian Uche

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Limiting arbitrariness and political power in governance is expressed in the concept of constitutionalism. Constitutionalism acknowledges the necessity for government but insists upon a limitation being placed upon its powers. It is therefore clear that the essence of constitutionalism is obviation of arbitrariness in governance and maximisation of liberty with adequate and expedient restraint on government. The doctrine of separation of powers accompanied by a system of checks and balances in Nigeria like many other African countries is marked by elements of ‘personal government’ and this has raised questions about whether the apparent separation of powers provided for in the Nigerian Constitution is not just a euphemism for the hegemony of the executive over the other two arms of government; the legislature and the judiciary. Another question raised in the article is whether the doctrine is merely an abstract philosophical inheritance that lacks both content and relevance to the realities of the country and region today? The current happenings in Nigeria and most African countries such as the flagrant disregard of court orders by the Executive, indicate clearly that the concept constitutionalism ordinarily goes beyond mere form and strikes at the substance of a constitution. It, therefore, involves a consideration of whether there are provisions in the constitution which limit arbitrariness in the exercise of political powers by providing checks and balances upon such exercise. These questions underscore the need for Africa to craft its own understanding of the separation of powers between the arms of government in furtherance of good governance as it has been seen that it is possible to have a constitution in place which may just be a mere statement of unenforceable ‘rights’ or may be bereft of provisions guaranteeing liberty or adequate and necessary restraint on exercise of government. This paper seeks to expatiate on the importance of the nexus between constitutionalism and democratic process and a juxtaposition of practices between Nigeria and South Africa. The article notes that an abstract analysis of constitutionalism without recourse to the democratic process is meaningless and also analyses the structure of government of some selected African countries. These are examined the extent to which the doctrine operates within the arms of government and concludes that it should not just be regarded as a general constitutional principle but made rigid or perhaps effective and binding through law and institutional reforms.

Keywords: checks and balances, constitutionalism, democratic process, separation of power

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Authors: Aslihan Cetinbas Genc, Meral Unal

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This study presents the structural and cytochemical alterations of stigma at the stages of pre-anthesis, anthesis and post-anthesis in Vibirnum tinus. Capitate stigma continues with a closed style. The receptive surface of stigma is composed of unicellular papillae which are short and flattened at pre-anthesis stage. The papillae in this stage have dense cytoplasm with small vacuoles and a centrally located nucleus. With the start of anthesis, the stigma widens, papillae lengthen and become cylindrical. At anthesis stage, vacuoles enlarge, and nucleus moves to the base of the cell. At post-anthesis stage, the boundaries of the papillae become less noticeable. As proved by Periodic Acid Schiff procedure, the cytoplasm of papillae is rich in insoluble polysaccharides at all stages of development but it becomes remarkable at post-anthesis, particularly at the sub-papillar area. Although there is no significant difference in the content of protein in all stages of the development, it is more abundant at post-anthesis stage, as in Coomassie Brillant Blue stained sections. The surface of papillae is covered by a cuticle which becomes thicker at post-anthesis, and it gives positive reaction with Sudan Black B and Auramine O. The cuticle is covered by a pellicle stained by Coomassie Brillant Blue, indicating dry type of stigma.

Keywords: develeopmental features, histochemistry, stigma, Vibirnum tinus

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Authors: Sheila M. Wicks, Gail Mahady, Udesh Patel, Joanna Michel, Armando Caceres

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Introduction: The genus piperaceae contains approximately 1000 species of herbs scrubs small trees and hanging vines distributed in both hemispheres. During our ethno medical work in Guatemala of the 27 plant families documented for us e by the Qeqchi Maya for reproductive disorders the most prominent were the Piperaceae (15%) and Menispermiaceae. Our Previous work showed that extracts from form Piper and Cissampelos species bound to both and progesterone and the estrogen receptors. In this work active extracts from Piper aeruginosibaccum Trelease, P auritum, P tuerckheimii and Cissampels tropaeolifolia were tested in functionalized cell based assays including a SEAP reporter gene and by qPCR of ER-responsive gene expression in MCF-7cells. In the reporter gene assay P aeruginosibaccum was estrogenic and enhanced E2 EFFECTS IN MCF-7 CELLS. P. tuerckheimi was not estrogenic alone but significantly enhanced the effects of E2 on SEAP reporter gene expression. Both altered mRNA expression of E2 responsive genes in MCF-7. Methods: this is collaborative project between University of Illinois at Chicago and University of San Carlos Guatemala City. 144 spices of plants were collected in Guatemala of which 57 used to treat a variety of women's reproductive health. The Genus Piperaraceae contains approximately 1000 species of herbs scrubs and small trees. Active extracts of the plants were tested in functionalized in cell-based bioassays including SEAP reporter genes. Results demonstrated altered mRNA expression of E2 responsive genes in MC-7 cells plants were collected in Guatemala of which 57 used. Conclusion of the 5 plants tested all were shown to contain components of binding to estrogenic receptor to a greater or lesser degree. These effects support the use of QEqchi Maya women in Guatemala for reproductive.

Keywords: reporter genes, MC7, guatemala piperaceae, reproductive health

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Authors: Anna Pajdak, Mateusz Kudasik, Aleksandra Gajda, Katarzyna Kozieł

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Shales are a type of fine-grained sedimentary rocks, which are composed of small grains of several to several dozen μm in size and consist of a variable mixture of clay minerals, quartz, feldspars, carbonates, sulphides, amorphous material and organic matter. The study involved an analysis of the basic physical properties of shale rocks from several research wells in Poland. The structural, sorption and seepage parameters of these rocks were determined. The total porosity of granular rock samples reached several percent, including the share of closed pores up to half a percent. The volume and distribution of pores, which are of significant importance in the context of the mechanisms of methane binding to the rock matrix and methods of stimulating its desorption and the possibility of CO₂ storage, were determined. The BET surface area of the samples ranged from a few to a dozen or so m²/g, and the share of micropores was dominant. In order to determine the interaction of rocks with gases, the sorption capacity in relation to CO₂ and CH₄ was determined at a pressure of 0-1.4 MPa. Sorption capacities, sorption isotherms and diffusion coefficients were also determined. Studies of competitive sorption of CO₂/CH₄ on shales showed a preference for CO₂ sorption over CH₄, and the selectivity of CO₂/CH₄ sorption decreased with increasing pressure. In addition to the pore structure, the adsorption capacity of gases in shale rocks is significantly influenced by the carbon content in their organic matter. The sorbed gas can constitute from 20% to 80% of the total gas contained in the shales. With the increasing depth of shale gas occurrence, the share of free gas to sorbed gas increases, among others, due to the increase in temperature and surrounding pressure. Determining the share of free gas to sorbed gas in shale, depending on the depth of its deposition, is one of the key elements of recognizing the gas/sorption exchange processes of CO₂/CH₄, which are the basis of CO₂-ESGR technology. The main objective of the work was to identify the correlation between different forms of gas occurrence in rocks and the parameters describing the pore space of shales.

Keywords: shale, CH₄, CO₂, shale gas, CO₂ -ESGR, pores structure

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Authors: S. Lekhavat, U. Srimongkoluk, P. Ratanachamnong, G. Laungsopapun

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Quercetin was encapsulated with whey protein and high methoxyl pectin by spray drying technique. Feed solution, consisting of 0.1875 0.125 and 0.0625 % w/w quercetin, respectively, was prepared and then sprays at outlet temperature of 70, 80 and 90 °C. Quercetin contents either in feed solution or in spray dried powder were determined by HPLC technique. Physicochemical properties such as viscosity and total soluble solid of feed solution as well as moisture content and water activity of spray dried powder were examined. Particle morphology was imaged using scanning electron microscope. The results showed that feed solution has total soluble solid and viscosity in range of 1.73-5.60 ºBrix and 2.58-8.15 cP, in that order. After spray drying, the moisture content and water activity value of powder are in range of 0.58-2.72 % and 0.18-0.31, respectively. Quercetin content in dried sample increased along with outlet drying temperature but decreased when total soluble solid increased. It was shown that particles are likely to shrivel when spray drying at high temperature. The suggested conditions for encapsulation of quercetin are feed solution with 0.0625 % (w/w) quercetin and spray drying at drying outlet temperature of 90°C.

Keywords: drying temperature, particle morphology, spray drying, quercetin

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Authors: Marina Stramarkou, Sofia Papadaki, Maria Kaloupi, Ioannis Batzakas

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Biofouling is a complex problem of the aquaculture industry, as it reduces the efficiency of the equipment and causes significant losses of cultured organisms. Nowadays, the current antifouling methods are proved to be labor intensive, have limited lifetime and use toxic substances that result in fish mortality. Several species of marine algae produce a wide variety of biogenic compounds with antibacterial and antifouling properties, which are effective in the prevention and control of biofouling and can be incorporated in antifouling coatings. In the present work, Fucus spiralis, a species of macro algae, and Chlorella vulgaris, a well-known species of microalgae, were used for the isolation and recovery of bioactive compounds, belonging to groups of fatty acids, lipopeptides and amides. The recovery of the compounds was achieved through the application of the ultrasound- assisted extraction, an environmentally friendly method, using green, non-toxic solvents. Moreover, the coating of the antifouling agents was done by innovative encapsulation and coating methods, such as electro-hydrodynamic process. For the encapsulation of the bioactive compounds natural matrices were used, such as polysaccharides and proteins. Water extracts that were incorporated in protein matrices were considered the most efficient antifouling coating.

Keywords: algae, electrospinning, fatty acids, ultrasound-assisted extraction

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Authors: Maciej Sobczak, Julita Pietrzak, Tomasz Płoszaj, Agnieszka Robaszkiewicz

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Brg1, a member of SWI/SNF complex, plays a role in chromatin remodeling, therefore, regulates expression of many genes. Brg1 is an ATPase of SWI/SNF complex, thus its activity requires ATP. Through its bromodomain recognizes acetylated histone residues and evicts them, thus promoting transcriptionally active state of chromatin. One of the enzymes that is responsible for acetylation of histone residues is Ep300. It was previously shown in the literature that cooperation of Brg1 and Ep300 occurs at the promoter regions that have binding sites for E2F-family transcription factors as well as CpG islands. According to literature, approximately 20% of human cancer possess mutation in Brg1 or any other crucial SWI/SNF subunit. That phenomenon makes Brg1-Ep300 a very promising target for anti-cancer therapy. Therefore in our study, we investigated if physical interaction between Brg1 and Ep300 exists and what impact those two proteins have on key for breast cancer cells processes such as DNA damage repair and cell proliferation. Bioinformatical analysis pointed out, that genes involved in cell proliferation and DNA damage repair are overexpressed in MCF7 and MDA-MB-231 cells. Moreover, promoter regions of these genes are highly acetylated, which suggests high transcriptional activity of those sites. Notably, many of those gene possess within their promoters an E2F, Brg1 motives, as well as CpG islands and acetylated histones. Our data show that Brg1 physically interacts with Ep300, and together they regulate expression of genes involved in DNA damage repair and cell proliferation. Upon inhibiting Brg1 or Ep300, expression of vital for cancer cell survival genes such as CDK2/4, BRCA1/2, PCNA, and XRCC1 is decreased in MDA-MB-231 and MCF7 cells. Moreover, inhibition or silencing of either Brg1 or Ep300 leads to cell cycle arrest in G1. After inhibition of BRG1 or Ep300 on tested gene promoters, the repressor complex including Rb, HDAC1, and EZH2 is formed, which inhibits gene expression. These results highlight potentially significant target for targeted anticancer therapy to be introduced as a supportive therapy.

Keywords: brg1, ep300, breast cancer, epigenetics

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Authors: Kuladip Jana

Abstract:

Benzo(a)pyrene [B(a)P] is an environmental toxicant present mostly in cigarette smoke and car exhaust, is an aryl hydrocarbon receptor (AhR) ligand that exerts its toxic effects on both male and female reproductive systems. In this study, the effect of B(a)P at different doses (0.1, 0.25, 0.5, 1 and 5 mg /kg body weight) was studied on male reproductive system of rat. A significant decrease in cauda epididymal sperm count and motility along with the presence of sperm head abnormalities and altered epididymal and testicular histology were documented following B(a)P treatment. B(a)P treatment resulted apoptotic sperm cells as observed by TUNEL and Annexin V-PI assay with increased ROS, altered sperm mitochondrial membrane potential (ΔΨm) with a simultaneous decrease in the activity of antioxidant enzymes and GSH status. TUNEL positive apoptotic cells also observed in testis as well as isolated germ and Leydig cells following B(a)P exposure. Western Blot analysis revealed the activation of p38MAPK, cytosolic translocation of cytochrome-c, up-regulation of Bax and inducible nitric oxide synthase (iNOS) with cleavage of PARP and down-regulation of BCl2 in testis upon B(a)P treatment. The protein and mRNA levels of testicular key steroidogenesis regulatory proteins like StAR, cytochrome P450 IIA1 (CYPIIA1), 3β HSD, 17β HSD showed a significant decrease in a dose dependent manner while an increase in the expression of cytochrome P450 1A1 (CYP1A1), Aryl hydrocarbon Receptor (AhR), active caspase- 9 and caspase- 3 following B(a)P exposure. We conclude that exposure of benzo(a)pyrene caused testicular gamatogenic and steroidogenic disorders by induction of oxidative stress, inhibition of StAR and other steroidogenic enzymes along with activation of p38MAPK and initiated caspase-3 mediated germ and Leydig cell apoptosis.The possible protective role of naturally occurring phytochemicals against B(a)P induced testicular toxicity needs immediate consideration. Curcumin and resveratrol separately were found to protect against B(a)P induced germ cell apoptosis, and their combinatorial effect was more significant. Our present study in isolated testicular germ cell population from adult male Wistar rats, highlighted their synergistic protective effect against B(a)P induced germ cell apoptosis. Curcumin-resveratrol co-treatment decreased the expression of pro-apoptotic proteins like cleaved caspase 3,8,9, cleaved PARP, Apaf1, FasL, tBid. Curcumin-resveratrol co-treatment decreased Bax/Bcl2 ratio, mitochondria to cytosolic translocation of cytochrome c and activated the survival protein Akt. Curcumin-resveratrol decreased the expression of p53 dependent apoptotic genes like Fas, FasL, Bax, Bcl2, Apaf1.Curcumin-resveratrol co-treatment thus prevented B(a)P induced germ cell apoptosis. B(a)P induced testicular ROS generation and oxidative stress were significantly ameliorated with curcumin and resveratrol. Curcumin-resveratrol co-treatment prevented B(a)P induced nuclear translocation of AhR and CYP1A1 production. The combinatorial treatment significantly inhibited B(a)P induced ERK 1/2, p38 MAPK and JNK 1/2 activation. B(a)P treatment increased the expression of p53 and its phosphorylation (p53 ser 15). Curcumin-resveratrol co-treatment significantly decreased p53 level and its phosphorylation (p53 ser 15). The study concludes that curcumin-resveratrol synergistically modulated MAPKs and p53, prevented oxidative stress, regulated the expression of pro and anti-apoptotic proteins as well as the proteins involved in B(a)P metabolism thus protected germ cells from B(a)P induced apoptosis.

Keywords: benzo(a)pyrene, germ cell, apoptosis, oxidative stress, resveratrol, curcumin

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