Search results for: chicken protein hydrolysate

Authors: Paras Gupta, Rohit Goyal, Yamini Chauhan, Pyare Lal Sharma

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Background: Bombax ceiba Linn., commonly called as Semal, is used in various gastro-intestinal disturbances. It contains lupeol which inhibits PTP-1B, adipogenesis, TG synthesis and accumulation of lipids in adipocytes and adipokines whereas the flavonoids isolated from B. ceiba has FAS inhibitory activity. The present study was aimed to investigate ameliorative potential of Bombax ceiba to experimental obesity in Wistar rats, and its possible mechanism of action. Methods: Male Wistar albino rats weighing 180–220 g were employed in present study. Experimental obesity was induced by feeding high fat diet for 10 weeks. Methanolic extract of B. ceiba extract 100, 200 and 400 mg/kg and Gemfibrozil 50 mg/kg as standard drug were given orally from 7th to 10th week. Results: Induction with HFD for 10 weeks caused significant (p < 0.05) increase in % body wt, BMI, LEE indices; serum glucose, triglyceride, LDL, VLDL, cholesterol, free fatty acid, ALT, AST; tissue TBARS, nitrate/nitrite levels; different fat pads and relative liver weight; and significant decrease in food intake (g and kcal), serum HDL and tissue glutathione levels in HFD control rats. Treatment with B. ceiba extract and Gemfibrozil significantly attenuated these HFD induced changes, as compared to HFD control. The effect of B. ceiba 200 and 400 mg/kg was more pronounced in comparison to Gemfibrozil. Conclusion: On the basis of results obtained, it may be concluded that the methanolic extract of stem bark of Bombax ceiba has significant ameliorative potential against HFD induced obesity in rats, possibly through modulation of FAS and PTP-1B signaling due to the presence of flavonoids and lupeol.

Keywords: obesity, Bombax ceiba, free fatty acid, protein tyrosine phosphatase-1B, fatty acid synthase

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

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

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

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

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

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

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Authors: Hoa Thi Hoang, Stephan Sass, Michael U. Kumke

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Concanavalin A (conA) is a protein has been widely used in sensor system based on its specific binding to α-D-Glucose or α-D-Manose. For glucose sensor using conA, either fluoresence based techniques with intensity based or lifetime based are used. In this research, liposomes made from phospholipids were used as a biomimetic membrane system. In a first step, novel building blocks containing perylene labeled glucose units were added to the system and used to decorate the surface of the liposomes. Upon the binding between rhodamine labeled con A to the glucose units at the biomimetic membrane surface, a Förster resonance energy transfer system can be formed which combines unique fluorescence properties of perylene (e.g., high fluorescence quantum yield, no triplet formation) and its high hydrophobicity for efficient anchoring in membranes to form a novel probe for the investigation of sugar-driven binding reactions at biomimetic surfaces. Two glucose-labeled perylene derivatives were synthesized with different spacer length between the perylene and glucose unit in order to probe the binding of conA. The binding interaction was fully characterized by using high-end fluorescence techniques. Steady-state and time-resolved fluorescence techniques (e.g., fluorescence depolarization) in combination with single-molecule fluorescence spectroscopy techniques (fluorescence correlation spectroscopy, FCS) were used to monitor the interaction with conA. Base on the fluorescence depolarization, the rotational correlation times and the alteration in the diffusion coefficient (determined by FCS) the binding of the conA to the liposomes carrying the probe was studied. Moreover, single pair FRET experiments using pulsed interleaved excitation are used to characterize in detail the binding of conA to the liposome on a single molecule level avoiding averaging out effects.

Keywords: concanavalin A, FRET, sensor, biomimetic membrane

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Authors: Ali Kassem, Aly Taha, Abeer Hassan, Kazuhide Higuchi

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Introduction: Trans arterial chemoembolization (TACE) for hepatocellular carcinoma (HCC) is usually followed by hepatic dysfunction that limits its efficacy. L-carnitine is recently studied as hepatoprotective agent. Our aim is to evaluate the L-carnitine effects against the deterioration of liver functions after TACE. Method: 53 patients with intermediate stage HCC were assigned into two groups; L-carnitine group (26 patients) who received L-carnitine 300 mg tablet twice daily from 2 weeks before to 12 weeks after TACE and control group (27 patients) without L-carnitine therapy. 28 of studied patients received branched chain amino acids granules. Results: There were significant differences between L-carnitine Vs. control group in mean serum albumin change from baseline to 1 week and 4 weeks after TACE (p < 0.05). L-Carnitine maintained Child-Pugh score at 1 week after TACE and exhibited improvement at 4 weeks after TACE (p < 0.01 Vs 1 week after TACE). Control group has significant Child-Pugh score deterioration from baseline to 1 week after TACE (p < 0.05) and 12 weeks after TACE (p < 0.05). There were significant differences between L-carnitine and control groups in mean Child-Pugh score change from baseline to 4 weeks (p < 0.05) and 12 weeks after TACE (p < 0.05). L-carnitine displayed improvement in (PT) from baseline to 1 week, 4 w (p < 0.05) and 12 weeks after TACE. PT in control group declined less than baseline along all follow up intervals. Total bilirubin in L-carnitine group decreased at 1 week post TACE while in control group, it significantly increased at 1 week (p = 0.01). ALT and C-reactive protein elevation were suppressed at 1 week after TACE in Lcarnitine group. The hepatoprotective effects of L-carnitine were enhanced by concomitant use of branched chain amino acids. Conclusion: L-carnitine and BCAA combination therapy offer a novel supportive strategy after TACE in HCC patients.

Keywords: hepatocellular carcinoma, L-carnitine, liver functions , trans-arterial embolization

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Authors: S. Essa, H. Safar, R. Raghupathy

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Human cytomegalovirus (CMV) continues to be a source of severe complications to immunologically immature and immune-compromised hosts. Effective CMV vaccine that diminishes CMV disease in transplant patients and avoids congenital infection remains of high importance as no approved vaccines exist. Though the exact links of defense mechanisms are unidentified, viral-specific antibodies and Th1/Th2 cytokine responses have been involved in controlling viral infections. CMV envelope glycoprotein B (UL55/gB), the matrix proteins (UL83/pp65, UL99/pp28, UL32/pp150), and the assembly protein UL80a/pp38 are known to be targets of antiviral immune responses. In this study, mice were immunized with five HCMV antigens (UL32/pp150, UL80a/pp38, UL99/pp28, and UL83/pp65), and serum samples were collected and evaluated for eliciting viral-specific antibody responses. Moreover, Splenocytes were collected, stimulated, and assessed for cytokine responses. The results demonstrated a CMV-antigen-specific antibody response to pp38 and pp65 (E/C >2.0). The highest titers were detected with pp38 (average E/C 16.275) followed by pp65 (average E/C 7.72). Compared to control cells, splenocytes from PP38 antigen immunized mice gave a significantly higher concentration of GM-CSF, IFN-γ, IL-2 IL-4, IL-5, and IL-17A (P<0.05). Also, splenocytes from pp65 antigen immunized mice resulted in a significantly higher concentration of GM-CSF, IFN-γ, IL-2 IL-4, IL-10, IL-12, IL-17A, and TNF- α. The designation of target CMV peptides by identifying viral-specific antibodies and cytokine responses is vital for understanding the protective immune mechanisms during CMV infection and identifying appropriate viral antigens to develop novel vaccines.

Keywords: hepatitis C virus, peripheral blood mononuclear cells, neutrophils, cytokines

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Authors: B. E. Adeyanju, M. K. Bolade, V. N. Enijuigha

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This work examined the nutritional composition of maize-based snack (kango) fortified with kidney beans (Phaseolus vulgaris) and alligator pepper (Aframomum melegueta). The snack is essentially traditional food being consumed by all ages in the southwestern part of Nigeria. Three varieties of maize were obtained from the Institute of Agricultural Research and Training (IAR&T), Ibadan, Nigeria, namely: ART-98-SW06-W, Br 9943-DMR-SR-W and SUWAN-1-SR-Y. Flour blends were obtained using the Response Surface Methodology (RSM) which resulted in appropriate blending ratios of maize, kidney beans and alligator pepper. Kango was prepared by milling maize grain into flour; ingredients such as pepper, onion, salt and water were added to the maize flour, mixed together to make a slurry. The slurry was fried in hot groundnut oil at a temperature of 126°C for 8 minutes. The incorporation of kidney bean and alligator pepper in maize flour was observed to increase the water and oil absorption capacities of the resultant blends thereby giving 109.21 to 156.90 ml/mg and 110.68 to 136.67 ml/mg respectively for kango. The pasting properties of the maize flour blends were also enhanced due to the incorporation of kidney bean and alligator pepper. The peak viscosity of the flour blends ranged from 3.24 to 7.67 RVU. The incorporation of kidney bean and alligator pepper in the production of the snacks increased the protein contents from 9.63 to 16.37%. The mineral contents (sodium, potassium, calcium, magnesium, iron and zinc) of the snacks were equally increased due to the incorporation of kidney bean and alligator pepper. A general increase was observed for vitamin B1 (0.69- 1.25 mg/100g), B2 (0.09 - 0.46 mg/100g) and B3 (0.11 - 0.72 mg/100g) in the snacks due to the incorporation of kidney bean and alligator pepper. This research work showed that kango produced from the composited maize flour, kidney bean and alligator pepper had better functional properties and higher nutritional contents.

Keywords: functional properties, kango, nutritional composition, snack

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Authors: Magdalena B. Kaziuk, Waldemar Kosiba

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Background: In the XXI century we have to deal with the epidemic of obesity which is important risk factor for the cardiovascular and kidney diseases. Lipo proteins are directly involved in the atherosclerotic process. Non-high-density lipo protein (non-HDL) began following widespread recognition of its superiority over LDL as a measurement of vascular event risk. Non-HDL includes residual risk which persists in patients after achieved recommended level of LDL. Materials and Methods: The study covered 111 patients (52 females, 59 males, age 51,91±14 years), hospitalized on the intern department. Body composition was assessed using the bioimpendance method and anthropometric measurements. Physical activity data were collected during the interview. The nutritional status and the obesity type were determined with the Waist to Height Ratio and the Waist to Hip Ratio. A function of the kidney was evaluated by calculating the estimated glomerular filtration rate (eGFR) using MDRD formula. Non-HDL was calculated as a difference between concentration of the Total and HDL cholesterol. Results: 10% of patients were found to be underweight; 23.9 % had correct body weight; 15,08 % had overweight, while the remaining group had obesity: 51,02 %. People with the android shape have higher non-HDL cholesterol versus with the gynoid shape (p=0.003). The higher was non-HDL, the lower eGFR had studied subjects (p < 0.001). Significant correlation was found between high non-HDL and incorrect dietary habits in patients avoiding eating vegetables, fruits and having low physical activity (p < 0.005). Conclusions: Android type of figure raises the residual risk of the heart disease associated with higher levels of non-HDL. Increasing physical activity in these patients reduces the level of non-HDL. Non-HDL seems to be the best predictor among all cholesterol measures for the cardiovascular events and worsening eGFR.

Keywords: obesity, non-HDL cholesterol, glomerular filtration rate, lifestyle

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Authors: Zaide Montes Ortiz, Jorge Alberto Molina, Alejandro Reyes

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Insects are extremely successful organisms. A sophisticated olfactory system is in part responsible for their survival and reproduction. The detection of volatile organic compounds can positively or negatively affect many behaviors in insects. Compounds such as carbon dioxide (CO2), ammonium, indol, and lactic acid are essential for many species of mosquitoes like Anopheles gambiae in order to locate vertebrate hosts. For instance, in A. gambiae, the olfactory receptor AgOR2 is strongly activated by indol, which accounts for almost 30% of human sweat. On the other hand, in some insects of agricultural importance, the detection and identification of pheromone receptors (PRs) in lepidopteran species has become a promising field for integrated pest management. For example, with the disruption of the pheromone receptor, BmOR1, mediated by transcription activator-like effector nucleases (TALENs), the sensitivity to bombykol was completely removed affecting the pheromone-source searching behavior in male moths. Then, the detection and identification of olfactory receptors in the genomes of insects is fundamental to improve our understanding of the ecological interactions, and to provide alternatives in the integrated pests and vectors management. Hence, the objective of this study is to propose a bioinformatic workflow to enhance the detection and identification of potential olfactory receptors in genomes of relevant insects. Applying Hidden Markov models (Hmms) and different computational tools, potential candidates for pheromone receptors in Tuta absoluta were obtained, as well as potential carbon dioxide receptors in Rhodnius prolixus, the main vector of Chagas disease. This study showed the validity of a bioinformatic workflow with a potential to improve the identification of certain olfactory receptors in different orders of insects.

Keywords: bioinformatic workflow, insects, olfactory receptors, protein prediction

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Authors: Supat Chaiyakul, Direk Sukkasem, Patnachapa Natthapanpaisith

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Sinlek rice flour beverage or instant product is a dietary supplement for dysphagia, or difficulty swallowing. It is also consumed by individuals who need to consume supplements to maintain their calorific needs. This product provides protein, fat, iron, and a high concentration of carbohydrate from rice flour. However, the application of native flour is limited due to its high viscosity. Starch modification by controlling starch retrogradation was used in this study. The research studies the effects of rice flour concentration and retrogradation treatment on the physical properties of instant Sinlek brown rice. The native rice flour, gelatinized rice flour, and flour gels retrograded under 4 °C for 3 and 7 days were investigated. From the statistical results, significant differences between native and retrograded flour were observed. The concentration of rice flour was the main factor influencing the swelling power, solubility, and pasting properties. With the increase in rice flour content from 10 to 15%, swelling power, peak viscosity, trough, and final viscosity decreased; but, solubility, pasting temperature, peak time, breakdown, and setback increased. The peak time, pasting temperature, peak viscosity, trough, and final viscosity decreased as the storage period increased from 3 to 7 days. The retrograded rice flour powders had lower pasting temperature, peak viscosity, breakdown, and final viscosity than the gelatinized and native flour powders. Reduction of starch viscosity by gelatinization and controlling starch retrogradation could allow for increased quantities of rice flour in instant rice beverages. Also, the treatment could increase the energy and nutrient densities of rice beverages without affecting the viscosity of this product.

Keywords: instant rice, pasting properties, pregelatinization, retrogradation

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Authors: Coulibaly Fatoumata, Gragnon Biego, Aka N. David, Mbari K. Benjamin, Soro Y. René, Ndiaye Jean-louis

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Poultry is nowadays one of the most consumed sources of protein, and its livestock represents one of the few opportunities for savings, investment and protection against risk. It provides income for the most vulnerable sections of society, in particular, women (70%) and children who mainly practice this breeding. A study was conducted in the commune of Korhogo at the level of 52 poultry farms, the objective of which was to know the epidemiological situation of parasitism external and internal poultry in order to contribute to the improvement of the health status of modern poultry farms in the said commune. The method described by OIE (2005), consisting of using the standard formula (n = δ2*p*(1-p) *c /i2), made it possible to calculate the size of the sample. Then, samples of droppings and ectoparasites were taken from the affected farms. After analysis and identification, two (2) species of mallophagous lice, including Menopon gallinae (50%) and Menacanthus stramineus (33%) and a species of bug Cimex lectularius (17%) were highlighted. The laying hens were more infested than broilers. Regarding gastrointestinal parasites, different species (six) have been identified: Trichostrongylus tenuis (17%), Syngamus trachea (19%), Heterakis sp (10%), Ascaridia sp (17%), Raillietina sp (8%) and Eimeria sp (29%). In addition, coccidiosis (Eimeria sp) proved to be the dominant pathology representing 67% of pathologies in broiler farms and 33% in poultry farms. The presence of these parasitoses in these modern farms constitutes a constraint major contribution to productivity and their development In view of all these difficulties, proposals have been made in order to participate in the establishment of a good prophylaxis program (health and medical). In addition, the Ivorian government, with the support of veterinarians, must interfere more in the organization of the health monitoring of traditional chickens and poultry in general through supervision and training in order to preserve public health ( animal, human and environmental health).

Keywords: gastrointestinal parasites, ectoparasites, pathologies, poultry, korhogo.

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Authors: Marian Agbugui

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The West African Lung fishes are fishes rich in protein and serve as an important source of food supply for man. The kind of food ingested by this group of fishes is dependent on the alimentary canal as well as the fish’s digestive processes which provide suitable modifications for maximum utilization of food taken. A study of the alimentary canal of P. annectens will expose the best information on the anatomy and histology of the fish. Samples of P. annectens were dissected to reveal the liver, pancreas and entire gut wall. Digital pictures of the mouth, jaws and the Gastrointestinal Tract (GIT) were taken. The entire gut was identified, sectioned and micro graphed. P. annectens was observed to possess a terminal mouth that opens up to 10% of its total body length, an adaptive feature to enable the fish to swallow the whole of its pry. Its dentition is made up of incisors- scissor-like teeth which also help to firmly grip, seize and tear through the skin of prey before swallowing. A short, straight and longitudinal GIT was observed in P. annectens which is known to be common feature in lungfishes, though it is thought to be a primitive characteristic similar to the lamprey. The oesophagus is short and distensible similar to other predatory and carnivorous species. Food is temporarily stored in the stomach before it is passed down into the intestine. A pyloric aperture is seen at the end of the double folded pyloric valve which leads into an intestine that makes up 75% of the whole GIT. The intestine begins at the posterior end of the pyloric aperture and winds down in six coils through the whole length intestine and ends at the cloaca. From this study it is concluded that P. annectens possess a composite GIT with organs similar to other lung fishes; it is a detritor with carnivorous abilities.

Keywords: gastrointestinal tract, incisors scissor-like teeth, intestine, mucus, Protopterus annectens, serosa

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Authors: Murat Emre, Zeynep Tufekcioglu

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Objective: We describe a patient with late onset phenylketonuria which presented with rapidly progressive dementia and parkinsonism that were reversible after management. Background: Phenylketonuria is an autosomal recessive disorder due to mutations in the phenylalanine hydroxlase gene. It normally presents in childhood, in rare cases, however, it may have its onset in adulthood and may mimic other neurological disorders. Case description: A previously normal functioning, 59 year old man was admitted for blurred vision, cognitive impairment and gait difficulty which emerged over the past eight months. In neurological examination he had brisk reflexes, slow gait and left-dominant parkinsonism. Mini-mental state examination score was 25/30, neuropsychological testing revealed a dysexecutive syndrome with constructional apraxia and simultanagnosia. In cranial MRI there were bilateral diffuse hyper-intense lesions in parietal and occipital white matter with no significant atrophy. Electroencephalography showed diffuse slowing with predominance of teta waves. In cerebrospinal fluid examination protein level was slightly elevated (61mg/dL), oligoclonal bands were negative. Electromyography was normal. Routine laboratory examinations for rapidly progressive dementia and parkinsonism were also normal. Serum amino acid levels were determined to explore metabolic leukodystrophies and phenylalanine level was found to be highly elevated (1075 µmol/L) with normal tyrosine (61,20 µmol/L). His cognitive impairment and parkinsonian symptoms improved following three months of phenylalanine restricted diet. Conclusions: Late onset phenylketonuria is a rare, potentially reversible cause of rapidly progressive parkinsonism with dementia. It should be considered in the differential diagnosis of patients with suspicious features.

Keywords: dementia, neurology, Phenylketonuria, rapidly progressive parkinsonism

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Authors: Dafna Knani, Sarit S. Sivan

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Aggrecan, one of the main components of the intervertebral disc (IVD), belongs to the family of proteoglycans (PGs) that are composed of glycosaminoglycan (GAG) chains covalently attached to a core protein. Its primary function is to maintain tissue hydration and hence disc height under the high loads imposed by muscle activity and body weight. Significant PG loss is one of the first indications of disc degeneration. A possible solution to recover disc functions is by injecting a synthetic hydrogel into the joint cavity, hence mimicking the role of PGs. One of the hydrogels proposed is GAG-analogues, based on sulfate-containing polymers, which are responsible for hydration in disc tissue. In the present work, we used molecular dynamics (MD) to study the effect of the hydrogel crosslinking (type and degree) on the swelling behavior of the suggested GAG-analogue biomimetics by calculation of cohesive energy density (CED), solubility parameter, enthalpy of mixing (ΔEmix) and the interactions between the molecules at the pure form and as a mixture with water. The simulation results showed that hydrophobicity plays an important role in the swelling of the hydrogel, as indicated by the linear correlation observed between solubility parameter values of the copolymers and crosslinker weight ratio (w/w); this correlation was found useful in predicting the amount of PEGDA needed for the desirable hydration behavior of (CS)₄-peptide. Enthalpy of mixing calculations showed that all the GAG analogs, (CS)₄ and (CS)₄-peptide are water-soluble; radial distribution function analysis revealed that they form interactions with water molecules, which is important for the hydration process. To conclude, our simulation results, beyond supporting the experimental data, can be used as a useful predictive tool in the future development of biomaterials, such as disc replacement.

Keywords: molecular dynamics, proteoglycans, enthalpy of mixing, swelling

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Authors: Nurcan Cetinkaya

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The aim of the study was to estimate the microbial-N flow to small intestine based on daily urinary purine derivatives(PD) mainly xanthine, hypoxanthine, uric acid and allantoin excretion in Angora goats fed by grass hay and concentrate (Period I); barley straw and concentrate (Period II). Daily urine samples were collected during last 3 days of each period from 10 individually penned Angora bucks( LW 30-35 Kg, 2-3 years old) receiving ad libitum grass hay or barley straw and 300 g/d concentrate. Fresh water was always available. 4N H2SO4 was added to collected daily urine .samples to keep pH under 3 to avoid of uric acid precipitation. Diluted urine samples were stored at -20°C until analysis. Urine samples were analyzed for xanthine, hypoxanthine, uric acid, allantoin and creatinine by High-Performance Liquid Chromatographic Method (HPLC). Urine was diluted 1:15 in ratio with water and duplicate samples were prepared for HPLC analysis. Calculated mean levels (n=60) for urinary xanthine, hypoxanthine, uric acid, allantoin, total PD and creatinine excretion were 0.39±0.02 , 0.26±0.03, 0.59±0.06, 5.91±0.50, 7.15±0.57 and 3.75±0.40 mmol/L for Period I respectively; 0.35±0.03, 0.21±0.02, 0.55±0.05, 5.60±0.47, 6.71±0.46 and 3.73±0.41 mmol/L for Period II respectively.Mean values of Period I and II were significantly different (P< 0.05) except creatinine excretion. Estimated mean microbial-N supply to the small intestine for Period I and II in Angora goats were 5.72±0.46 and 5.41±0.61 g N/d respectively. The effects of grass hay and barley straw feeding on microbial-N supply to small intestine were found significantly different (P< 0.05). In conclusion, grass hay showed a better effect on the ruminal microbial protein synthesis compared to barley straw, therefore; grass hay is suggested as roughage source in Angora goat feeding.

Keywords: angora goat, HPLC method, microbial-N supply to small intestine, urinary purine derivatives

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Authors: Jin Young Kim, Kwon Kyoo Kang

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The SGR1 (STAYGREEN1) protein is a critical regulator of plant leaves in chlorophyll degradation and senescence. The functions and mechanisms of tomato SGR1 action are poorly understood and worthy of further investigation. To investigate the function of the SGR1 gene, we generated a SGR1-knockout (KO) null line via clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9-mediated gene editing and conducted RNA sequencing and gas chromatography tandem mass spectrometry (GC-MS/MS) analysis to identify the differentially expressed genes. The SlSGR1 (Solanum lycopersicum SGR1) knockout null line clearly showed a turbid brown color with significantly higher chlorophyll and carotenoid content compared to wild-type (WT) fruit. Differential gene expression analysis revealed 728 differentially expressed genes (DEGs) between WT and sgr1 #1-6 line, including 263 and 465 downregulated and upregulated genes, respectively, for which fold change was >2, and the adjusted p-value was <0.05. Most of the DEGs were related to photosynthesis and chloroplast function. In addition, the pigment, carotenoid changes in sgr1 #1-6 line was accumulated of key primary metabolites such as sucrose and its derivatives (fructose, galactinol, raffinose), glycolytic intermediates (glucose, G6P, Fru6P) and tricarboxylic acid cycle (TCA) intermediates (malate and fumarate). Taken together, the transcriptome and metabolite profiles of SGR1-KO lines presented here provide evidence for the mechanisms underlying the effects of SGR1 and molecular pathways involved in chlorophyll degradation and carotenoid biosynthesis.

Keywords: tomato, CRISPR/Cas9, null line, RNA-sequencing, metabolite profiling

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

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

Keywords: diet, lipid, obesity, T2DM

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Authors: Kadeejah Alsolami, Zainab Alrefay, Husaam Awad

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Obesity and vitamin D deficiency have both been related to cardiovascular disease. The present work aimed to investigate the possible protective effect of vitamin D on cardiac apoptosis in a rat model of dietary-induced obesity. Methods: 30 male Wistar rats included in this study. They were allocated into 4 groups: Control (n=5), animal were fed standard diet for 3 months: Control + vitamin D (VD) (n=5),animals were fed a standard diet with 400IU VD/kg for 3 months: hypercaloric diets group (n=10), animals were fed a high fat diet for 3 months: hypercaloric diet with VD group (n=10), animals were fed a high fat diet with 400IU VD/kg for 3 months. At the beginning of the experiment, the weight and length were measured to assess body mass index (BMI) and repeated every 45 days. Food intake and body weight were monitored throughout the study period. Then rats were sacrificed and heart tissues collected for Quantitative Real-time polymerase chain reaction (qRT-PCR). qRT-PCR used to detect different genetic markers of apoptosis (anti-apoptotic gene (BCL2), a pro-apoptotic gene(BAX), pro-apoptotic genes (FAS, FAS-L), tumour necrosis factor (TNF), mitogen-activated protein kinases (MAPK). Results: FAS and FAS-L gene expression were significantly upregulated in rats fed with high fat diet. And FAS-L gene expression was significantly upregulated in all groups on comparison with control. Whereas Bax gene expression was significantly downregulated in rats fed with high-fat diet supplied with vitamin D. TNF was significantly upregulated in rats fed with high-fat diet treated with vitamin D. MAPK was significantly upregulated in rats fed with high fat diet group, and in rats fed with high-fat diet supplied with vitamin D. Conclusion: The cardiac apoptotic pathways were more activated in rats fed with high-fat than lean rats. And vitamin D protect the heart from the cardiac mitochondrial-dependent apoptotic pathway.

Keywords: apoptosis, heart, obesity, Vitamin D

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Authors: Sekena H. Abd El-Aziem, Heba A. M. Abd El-Kader, Sally S. Alam, Othman E. Othman

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Growth hormone (GH) is an anabolic hormone synthesized and secreted by the somatotroph cells of the anterior lobe of the pituitary gland in a circadian and pulsatile manner, the pattern of which plays an important role in postnatal longitudinal growth and development, tissue growth, lactation, reproduction as well as protein, lipid and carbohydrate metabolism. The aim of this study was to detect the genetic polymorphism of GH gene in five camel breeds reared in Egypt; Sudany, Somali, Mowaled, Maghrabi and Falahy, using PCR-RFLP technique. Also this work aimed to identify the single nucleotide polymorphism between different genotypes detected in these camel breeds. The amplified fragment of camel GH at 613-bp was digested with the restriction enzyme MspI and the result revealed the presence of three different genotypes; CC, CT and TT in tested breeds and significant differences were recorded in the genotype frequencies between these camel breeds. The result showed that the Maghrabi breed that is classified as a dual purpose camels had higher frequency for allele C (0.75) than those in the other tested four breeds. The sequence analysis declared the presence of a SNP (C→T) at position 264 in the amplified fragment which is responsible for the destruction of the restriction site C^CGG and consequently the appearance of two different alleles C and T. The nucleotide sequences of camel GH alleles T and C were submitted to nucleotide sequences database NCBI/Bankit/GenBank and have accession numbers: KP143517 and KP143518, respectively. It is concluded that only one SNP C→T was detected in GH gene among the five tested camel breeds reared in Egypt and this nucleotide substitution can be used as a marker for the genetic biodiversity between camel breeds reared in Egypt. Also, due to the possible association between allele C and higher growth rate, we can used it in MAS for camels and enter the camels possess this allele in breeding program as a way for enhancement of growth trait in camel breeds reared in Egypt.

Keywords: camel breeds in Egypt, GH, PCR-RFLP, SNPs

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Authors: Nandini Nalika, Suhel Parvez

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Nanotechnology has emerged to play a vital role in developing all through the industrial world with an immense production of nanomaterials including nanoparticles (NPs). Many toxicological studies have confirmed that due to unique small size and physico-chemical properties of NPs (1-100nm), they can be potentially hazardous. Metallic NPs of small size have been shown to induce higher levels of cellular oxidative stress and can easily pass through the Blood Brain Barrier (BBB) and significantly accumulate in brain. With the wide applications of titanium dioxide nanoparticles (TNPs) in day-to-day life in form of cosmetics, paints, sterilisation and so on, there is growing concern regarding the deleterious effects of TNPs on central nervous system and mitochondria appear to be important cellular organelles targeted to the pro-oxidative effects of NPs and an important source that contribute significantly for the production of reactive oxygen species after some toxicity or an injury. The aim of our study was to elucidate the effect of TNPs in anatase form with different concentrations (5-50 µg/ml) following with various oxidative stress markers in isolated brain mitochondria as an in vitro model. Oxidative stress was determined by measuring the different oxidative stress markers like lipid peroxidation as well as the protein carbonyl content which was found to be significantly increased. Reduced glutathione content and major glutathione metabolizing enzymes were also modulated signifying the role of glutathione redox cycle in the pathophysiology of TNPs. The study also includes the mitochondrial enzymes (Complex 1, Complex II, complex IV, Complex V ) and the enzymes showed toxicity in a relatively short time due to the effect of TNPs. The study provide a range of concentration that were toxic to the neuronal cells and data pointing to a general toxicity in brain mitochondria by TNPs, therefore, it is in need to consider the proper utilization of NPs in the environment.

Keywords: mitochondria, nanoparticles, brain, in vitro

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Authors: Mohammad K. Parvez, Mohammed S. Al-Dosari

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Hepatitis E virus (HEV) is an emerging RNA virus that causes acute and chronic liver disease with a global mortality rate of about 2%. Despite milestone developments in understanding of HEV biology, there is still lack of a robust culture system or animal model. Therefore, in a novel approach, two recombinant-baculoviruses (vBac-ORF2 and vBac-ORF3) that could overexpress HEV ORF2 (structural/capsid) and ORF3 (nonstructural/regulatory) proteins, respectively were constructed. The established HEV-SAR55 (genotype 1) replicon that contained GFP gene, in place of ORF2/ORF3 sequences was in vitro transcribed, and GFP production in RNA transfected S10-3 cells was scored by FACS. Enhanced infectivity, if any, of nascent virions produced by exogenously-supplied ORF2 and viral RNA by co-expression of ORF3 was tested on naïve HepG2 cells. Co-transduction with vBac-ORF2/vBac-ORF3 (108 pfu/microL) produced high amounts of native ORF2/ORF3 in approximately 60% of S10-3 cells, determined by immunofluorescence microscopy and Western analysis. FACS analysis showed about 9% GFP positivity of S10-3 cells on day6 post-transfection (i.e, day5 post-transduction). Further, FACS scoring indicated that lysates from S10-3 cultures receiving the RNA plus vBac-ORF2 were capable of producing HEV particles with about 4% infectivity in HepG2 cells. However, lysates of cultures co-transduced with vBac-ORF3, were found to further enhance virion infectivity by approximately 17%. This supported a previously proposed role of ORF3 as a minor-structural protein in HEV virion assembly and infectivity. In conclusion, the present model for efficient genomic RNA packaging and production of infectious virions could be a valuable tool to study various aspects of HEV molecular biology, in vitro.

Keywords: chronic liver disease, hepatitis E virus, ORF2, ORF3, replicon

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Authors: Tettey Afi Pamela, Sotaro Fujii, Hidetoshi Saito, Kawaii Koichiro

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Introduction: Organisms employ adaptive mechanisms when faced with any stressor or risk of being wiped out. This has made it possible for them to survive in harsh environmental conditions such as increasing temperature, low pH, and anoxia. Some of the mechanisms they utilize include the expression of heat shock proteins, synthesis of cryoprotectants, and anhydrobiosis. Heat shock proteins (HSPs) have been widely studied to determine their involvement in stress tolerance among various organism, of which chironomid species have been no exception. We examined the survival and expression of genes encoding five (5) heat shock proteins (HSP70, HSP67, HSP60, HSP27, and HSP23) from Chironomus sulfurosus larvae reared from 1st instar at 25°C, 30°C, 35°C, and 40°C. Results: The highest survival rate was recorded at 30°C, followed by 25°C, then 35°C. Only a small percentage of C. sulfurosus survived at 40°C (14.5%). With regards to HSPs expression, some HSPs responded to an increase in high temperature. The relative expression levels were lowest at 30°C for HSP70, HSP60, HSP27, and HSP23. At 25°C and 40°C, HSP70, HSP67, HSP60, HSP27, and HSP23 had the highest expression. At 35°C, all had the lowest expression. Discussion: The expression of heat shock proteins varies from one species to another. We designated the genes HSP 70, HSP 67, HSP 60, HSP 27, and HSP 23 genes based on transcriptome analysis of C. sulfurosus. Our study can be termed as a long-heat shock study as C. sulfurosus was reared from the first instar to the fourth instar, and this might have led to a continuous induction of HSPs at 25°C. 40°C had the lowest survival but highest HSPs expression as C. sulfurosus larvae had to utilize HSPs for sustenance. These results and future high-throughput studies at both the transcriptome and proteome level will improve the information needed to predict the future geographic distribution of these species within the context of global warming.

Keywords: chironomid, heat shock proteins, high temperature, heat shock protein expression

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Authors: O. M. Alabi, F. A. Aderemi, M. O. Ayoola

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An aspect of biosecurity measures to ensure good welfare for chickens is rodents’ control. Rats and mice are rodents commonly found in poultry houses in most of the African countries. More than 20,000 species of rat have been identified in Africa among which are; Black house rats (Rattus rattus), East African mole rat (Tachyorcytes splendens), Naked mole rat (Heterocephalus glaber), Zambian mole rat (Fukomys mechowii), African grass rat (Arvicanthis niloticus), Nigerian mole rat (Cryptomys foxi), Target rat (Stochomys longicaudatus) and West African Shaggy rat (Dasymis rufulus). Apart from being destructive, rats and mice are voracious in that they compete with chickens for feed and water thereby causing economical losses to the farmer, they are also vectors to many pathogens of poultry diseases such as Salmonellosis, colibacillosis, ascaridiasis, coryza, pasteurellosis and mycoplasmosis. As bad as these rodents are to the poultry farmers, they are good sources of animal protein to local hunters and other farmers in most African countries. Rat is considered a delicacy in Nigeria and many other African countries hence the need to investigate into how the rats species will not go into extinction. Rodents are usually controlled by poultry farmers with the use of rodenticides which can either be anticoagulant or stomach poison, and with the use of baits. However, elimination of rats and mice is being considered as callous act against these species of animal and their natural existence as human food also. This paper therefore suggests that sanitation methods such as feed removal from rats and mice, controlling feed and water spillage, proper disposal of waste eggs, dead birds and garbage, keeping the surroundings of the poultry clean; rodent proofing by making it difficult for rodents to enter the poultry houses are some of the humane ways of controlling rodents in poultry production to avoid improving the welfare of a particular animal at the expense of the other.

Keywords: management, poultry, rodents, welfare

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Authors: Elif Cakir, Görkem Özülkü, Hatice Bekiroğlu, Muhammet Arici, Osman Sağdic

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It is important to be able to formulate cakes with a wide consumption mass with gluten-free and high nutritional value ingredients to increase the consumption possibilities of people with limited nutrition opportunities. Although people do not prefer Prunus spinosa (PS)because of its sour taste and its use in the food industry is limited on a local scale, the potential of using PS, which is a naturally rich source of many micronutrients and bioactive compounds, in glutenfree cake production has been investigated. In this study, the potential of using PS, a natural wild fruit, in the production of functional gluten-free cakes was investigated. It was aimed to evaluate the effects of freeze-dried and powdered PS-enriched rice flour cakes on tech functionality, nutrition and eating quality. In terms of physicochemical properties, PS raises increased the ash, protein, and moisture values of the cakes. PS with high phenolic content, phenolic component content, and radical reducing power made by ABTS, FRAP, and DPPH techniques were higher in all samples than control, and the highest 4% PS was determined in cakes. In terms of the glycemic index (GI), which is an important feature of diet products, it was determined that the GI in cakes decreased by 86.30±1.04.75.05±1.16 and 69.38±1.21, respectively, with the increase in PS ratio. Except for the 1%, PS added sample, the increase in PS caused a decrease in specific volume, % porosity and increase in hardness, including 4 days of storage. PS increase decreased the L* and b* values and increased a* value and redness of the cake. Sensory liking of the cake samples containing PS was scored significantly (p<0.05) higher of control.

Keywords: Prunus spinosa, gluten-free cake, antioxidant, phenolic, glycemic index

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Authors: Muñiz-Lino Marcos A.

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The ectomesenchymal tissues involved in tooth development and their remnants are the origin of different odontogenic lesions, including tumors and cysts of the jaws, with a wide range of clinical behaviors. A dentigerous cyst (DC) represents approximately 20% of all cases of odontogenic cysts, and it has been demonstrated that it can develop benign and malignant odontogenic tumors. DC is characterized by bone destruction of the area surrounding the crown of a tooth that has not erupted and contains liquid. The treatment of odontogenic tumors and cysts usually involves a partial or total removal of the jaw, causing important secondary co-morbidities. However, molecules implicated in DC pathogenesis, as well as in its development into odontogenic tumors, remain unknown. A cellular model may be useful to study these molecules, but that model has not been established yet. Here, we reported the establishment of a cell culture derived from a dentigerous cyst. This cell line was named DeCy-1. In spite of its ectomesenchymal morphology, DeCy-1 cells express epithelial markers such as cytokeratins 5, 6, and 8. Furthermore, these cells express the ODAM protein, which is present in odontogenesis and in dental follicles, indicating that DeCy-1 cells are derived from odontogenic epithelium. Analysis by electron microscopy of this cell line showed that it has a high vesicular activity, suggesting that DeCy-1 could secrete molecules that may be involved in DC pathogenesis. Thus, secreted proteins were analyzed by PAGE-SDS where we observed approximately 11 bands. In addition, the capacity of these secretions to degrade proteins was analyzed by gelatin substrate zymography. A degradation band of about 62 kDa was found in these assays. Western blot assays suggested that the matrix metalloproteinase 2 (MMP-2) is responsible for this protease activity. Thus, our results indicate that the establishment of a cell line derived from DC is a useful in vitro model to study the biology of this odontogenic lesion and its participation in the development of odontogenic tumors.

Keywords: dentigerous cyst, ameloblastoma, MMP-2, odontogenic tumors

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Authors: Flora Tajiki

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Infertility is a pervasive issue affecting women globally, profoundly impacting their psychological well-being and quality of life. This case study traces the six-year journey of a 35-year-old Iranian woman who married at 29 and grappled with infertility and premature menopause. Three years post-marriage, her initial pregnancy resulted in a miscarriage, followed by a year without successful conception. Diagnostic evaluations revealed small ovaries and a low AMH level of 0.19, leading to a diagnosis of premature menopause. Despite her primary therapist's recommendation to use a donated egg, she opted to pursue a comprehensive lifestyle modification approach through remote consultation with the author. The regimen encompassed regular physical activity, weight management, stress control, sleep regulation, meditation, abdominal massages, diaphragmatic breathing, and dietary changes toward a high-protein, low-carbohydrate diet. The recommended diet included nuts such as almonds and pistachios, as well as sprouted grains like wheat and mung beans. She was further advised to maintain adequate hydration and eliminate sugar, milk, preservatives, and processed foods. Regular follow-ups were conducted. Approximately three months into the regimen, the lifestyle overhaul led to a weight reduction from 99 kg to 89 kg and improved stress management. Remarkably, despite a continued decrease in AMH levels, she conceived naturally four months after adopting these lifestyle changes and is now the mother of a healthy 10-month-old girl. Similar cases under this healthy lifestyle approach, despite having low AMH levels, also resulted in successful pregnancies without IVF or hormone therapy. This underscores that AMH levels are not the sole determinant of fertility and that lifestyle modifications can play a significant role in overcoming infertility and achieving natural conception.

Keywords: egg donation, infertility, lifestyle Modification, natural conception, premature menopause

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Authors: Srishty Gulati, Anju Singh, Shrikant Kukreti

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The manifestation of structural polymorphism in DNA depends on the sequence and surrounding environment. Ample of folded DNA structures have been found in the cellular system out of which DNA hairpins are very common, however, are indispensable due to their role in the replication initiation sites, recombination, transcription regulation, and protein recognition. We enumerate this approach in our study, where the two base substitutions and change in temperature embark destabilization of DNA structure and misbalance the equilibrium between two structures of a sequence present at the overlapping region of the human COMT gene and MIR4761 gene. COMT and MIR4761 gene encodes for catechol-O-methyltransferase (COMT) enzyme and microRNAs (miRNAs), respectively. Environmental changes and errors during cell division lead to genetic abnormalities. The COMT gene entailed in dopamine regulation fosters neurological diseases like Parkinson's disease, schizophrenia, velocardiofacial syndrome, etc. A 19-mer deoxyoligonucleotide sequence 5'-AGGACAAGGTGTGCATGCC-3' (COMT19) is located at exon-4 on chromosome 22 and band q11.2 at the intersection of COMT and MIR4761 gene. Bioinformatics studies suggest that this sequence is conserved in humans and few other organisms and is involved in recognition of transcription factors in the vicinity of 3'-end. Non-denaturating gel electrophoresis and CD spectroscopy of COMT sequences indicate the formation of hairpin type DNA structures. Temperature-dependent CD studies revealed an unusual shift in the slipped DNA-Hairpin DNA equilibrium with the change in temperature. Also, UV-thermal melting techniques suggest that the two base substitutions on the complementary strand of COMT19 did not affect the structure but reduces the stability of duplex. This study gives insight about the possibility of existing structurally polymorphic transient states within DNA segments present at the intersection of COMT and MIR4761 gene.

Keywords: base-substitution, catechol-o-methyltransferase (COMT), hairpin-DNA, structural polymorphism

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Authors: Hui-Fang Lee, Hsuan-Jung Hsieh, Wen-Chi Lu, Meng-Chu Liu

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The purpose of the study was to examine abnormal BMI students of Chien Hsin University of Science and Technology, implement teaching aerobics course through elementary and advanced curriculum design, dietary education and three-day dietary record, analyze participant fitness improvement, an 10-week course as well as pre-test and post-test were carried out to evaluate the effect of the aerobics course on the fitness ability. The actual participate elementary and advanced courses each of 40 people, with low participation deduction course unfinished fitness testing, access to elementary curriculum valid samples 35 (87.5%) people, advanced courses valid samples 38(95%) people, 16 students participated in two consecutive courses. The fitness activities included sit-bending, one-minute sit-ups, standing long jump, and three minutes to board the stage. Analysis and comparison to the average three-day dietary record difference, an independent samples t-test was conducted to analyze the differences in the four activities between pre-test and post-test. The results showed that the elementary course had significant effects on females’ sit-bending and one minute sit-ups, the females also had high fat intake in three-day dietary record. The advanced course had significant effects on males’ sit-bending and on females’ BMI, sit-bending and standing long jump, males and females in three-day dietary record carbohydrate intake slightly low, slightly higher protein and fat intake. In conclusion, aerobics course teaching, dietary education and three-day, dietary record implementation can significantly enhance the physical fitness indicators, and continued to participate in advanced courses better. In the practice of sport should be the future course planning elementary and advanced courses, while introducing dietary education, achieve concrete results in improving physical fitness.

Keywords: physical fitness, aerobics course, dietary education, three-day dietary record

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Authors: Tatiana A. Kuznetsova, Maxim V. Vechersky, Natalia V. Kostina, Marat M. Umarov, Elena I. Naumova

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One of the main problems of nutrition of phytophagous animals is the insufficiency of protein in their forage. Usually, symbiotic microorganisms highly contribute both to carbohydrates and nitrogen compounds of the food. But it is not easy to utilize microbial biomass in the large intestine and caecum for the animals with hindgut fermentation. So that, some animals, as well hares, developed special mechanism of contribution of such biomass - obligate autocoprophagy, or reingestion. Hares have two types of feces - the hard and the soft. Hard feces are excreted at night, while hares are vigilance ("foraging period"), and the soft ones (caecotrophs) are produced and reingested in the day-time during hares "resting-period". We examine the role of microbial digestion in providing nitrogen nutrition of hare (Lepus europaeus). We determine the ability of nitrogen fixation in fornix and stomach body, small intestine, caecum and colon of hares' gastro-intestinal tract in two main period of hares activity - "resting-period" (day time) and "foraging period" (late-evening and very-early-morning). We use gas chromatography to measure levels of nitrogen fixing activity (acetylene reduction). Nitrogen fixing activity was detected in the contents of all analyzed parts of the gastrointestinal tract. Maximum values were recorded in the large intestine. Also daily dynamics of the process was detected. Thus, during hare “resting-period” (caecotrophs formation) N2-fixing activity was significantly higher than during “foraging period”, reaching 0,3 nmol C2H4/g*h. N2-fixing activity in the gastrointestinal tract can allocate to significant contribution of nitrogen fixers to microbial digestion in hare and confirms the importance of coprophagy as a nitrogen source in lagomorphs.

Keywords: coprophagy, gastrointestinal tract, lagomorphs, nitrogen fixation

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Authors: Alma Gómez-Loredo, José González-Valdez, Jorge Benavides, Marco Rito-Palomares

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Fucoxanthin is a carotenoid that exerts multiple beneficial effects on human health, including antioxidant, anti-cancer, antidiabetic and anti-obesity activity; making the development of a whole process for its production and recovery an important contribution. In this work, the lab-scale production and purification of fucoxanthin in Isocrhysis galbana have been studied. In batch cultures, low light intensities (13.5 μmol/m2s) and bubble agitation were the best conditions for production of the carotenoid with product yields of up to 0.143 mg/g. After fucoxanthin ethanolic extraction from biomass and hexane partition, further recovery and purification of the carotenoid has been accomplished by means of alcohol – salt Aqueous Two-Phase System (ATPS) extraction followed by an ultrafiltration (UF) step. An ATPS comprised of ethanol and potassium phosphate (Volume Ratio (VR) =3; Tie-line Length (TLL) 60% w/w) presented a fucoxanthin recovery yield of 76.24 ± 1.60% among the studied systems and was able to remove 64.89 ± 2.64% of the carotenoid and chlorophyll pollutants. For UF, the addition of ethanol to the original recovered ethanolic ATPS stream to a final relation of 74.15% (w/w) resulted in a reduction of approximately 16% of the protein contents, increasing product purity with a recovery yield of about 63% of the compound in the permeate stream. Considering the production, extraction and primary recovery (ATPS and UF) steps, around a 45% global fucoxanthin recovery should be expected. Although other purification technologies, such as Centrifugal Partition Chromatography are able to obtain fucoxanthin recoveries of up to 83%, the process developed in the present work does not require large volumes of solvents or expensive equipment. Moreover, it has a potential for scale up to commercial scale and represents a cost-effective strategy when compared to traditional separation techniques like chromatography.

Keywords: aqueous two-phase systems, fucoxanthin, Isochrysis galbana, microalgae, ultrafiltration

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