Search results for: plasma protein electrophoresis

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: Mir Md Abdus Salam, Muhammad Mohsin, Pertti Pulkkinen, Paavo Pelkonen, Ari Pappinen

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Soils contaminated with metals, e.g., copper (Cu), zinc (Zn) and nickel (Ni) are one of the main global environmental problems. Zn is an important element for plant growth, but excess levels may become a threat to plant survival. Soils polluted with metals may also pose risks and hazards to human health. Afforestation based on short rotation Salix crops may be a good solution for the reduction of metals toxicity levels in the soil and in ecosystem restoration of severely polluted sites. In a greenhouse experiment, plant growth and zinc (Zn) uptake by four Salix cultivars grown in Zn contaminated soils collected from a mining area in Finland were tested to assess their suitability for phytoextraction. The sequential extraction technique and inductively coupled plasma‒mass spectrometry (ICP–MS) were used to determine the extractable metals and evaluate the fraction of metals in the soil that could be potentially available for plant uptake. The cultivars displayed resistance to heavily polluted soils throughout the whole experiment. After uptake, the total mean Zn concentrations ranged from 776 to 1823 mg kg⁻¹. The average uptake percentage of Zn across all cultivars and treatments ranged from 97 to 223%. Lime and wood ash addition showed a significant effect on plant dry biomass growth and metal uptake percentage of Zn in most of the cultivars. The results revealed that Salix cultivars have the potential to accumulate and take up significant amounts of Zn. Ecological restoration of polluted soils could be environmentally favorable in conjunction with economically profitable practices, such as forestry and bioenergy production. As such, the utilization of Salix for phytoextraction and bioenergy purposes is of considerable interest.

Keywords: lime, phytoextraction, Salix, wood ash, zinc

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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: Hyun Joon Chang, Jae In Kim, Sungsoo Na

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Kinesin-1 (hereafter called kinesin) is a molecular motor protein that moves cargos toward the end of microtubules using the energy of adenosine triphosphate (ATP) hydrolysis. When kinesin is inactive, its tail autoinhibits the motor chain in order to prevent from reacting with the ATP by cross-linking of the tail domain to the motor domains at two positions. However, the morphological study of kinesin during autoinhibition is yet remained obscured. In this study, we report the effect of the binding site of the tail domain using the normal mode analysis of the elastic network model on kinesin in the tail-free form and tail-bind form. Considering the relationship between the connectivity of conventional network model with respect to the cutoff length and the functionality of the binding site of the tail, we revaluated the network model to observe the key role of the tail domain in its structural aspect. Contingent on the existence of the tail domain, the results suggest the morphological stability of the motor domain. Furthermore, employing the results from normal mode analysis, we have determined the strain energy of the neck linker, an essential portion of the motor domain for ATP hydrolysis. The results of the neck linker also converge to the same indication, i.e. the morphological analysis of the motor domain.

Keywords: elastic network model, Kinesin-1, autoinhibition

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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

Procedia PDF Downloads 107

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: Sara Arafeh, Kovriguine Evguine

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Cytochromes P450 are enzymes that require a supply of electrons to catalyze the synthesis of steroid hormones, fatty acids, and prostaglandin hormone. Cytochrome P450 Oxidoreductase (CYPOR), a membrane bound enzyme, provides these electrons in its open conformation. CYPOR has two cytosolic domains (FAD domain and FMN domain) and an N-terminal in the membrane. In its open conformation, electrons flow from NADPH, FAD, and finally to FMN where cytochrome P450 picks up these electrons. In the closed conformation, cytochrome P450 does not bind to the FMN domain to take the electrons. It was found that when the cytosolic domains are isolated, CYPOR could not bind to cytochrome P450. This suggested that the membrane environment is important for CYPOR function. This project takes the initiative to better understand the dynamics of CYPOR in its full length. Here, we determine the distance between specific sites in the FAD and FMN binding domains in CYPOR by Forster Resonance Energy Transfer (FRET) and Ultrafast TA spectroscopy with and without NADPH. The approach to determine these distances will rely on labeling these sites with red and infrared fluorophores. Mimic membrane attachment is done by inserting CYPOR in lipid nanodiscs. By determining the distances between the donor-acceptor sites in these domains, we can observe the open/closed conformations upon reducing CYPOR in the presence and absence of cytochrome P450. Such study is important to better understand CYPOR mechanism of action in various endosomal membranes including hepatic CYPOR which is vital in plasma cholesterol homeostasis. By investigating the conformational cycles of CYPOR, we can synthesize drugs that would be more efficient in affecting the steroid hormonal levels and metabolism of toxins catalyzed by Cytochrome P450.

Keywords: conformational cycle of CYPOR, cytochrome P450, cytochrome P450 oxidoreductase, FAD domain, FMN domain, FRET, Ultrafast TA Spectroscopy

Procedia PDF Downloads 279

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

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

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

Procedia PDF Downloads 294

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: Fang Liu, Jia Jia Yao, Guan Lin Wu, Ren Jie Liu, Zhuang Guo

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Leveraging plasma-assisted molecular beam epitaxy (PA-MBE) on c-plane free-standing GaN substrates, this work demonstrates high-performance AlN/GaN double-barrier resonant tunneling diodes (RTDs) featuring stable and repeatable negative differential resistance (NDR) characteristics at room temperature. By scaling down the barrier thickness of AlN and the lateral mesa size of collector, a record peak current density of 1551 kA/cm2 is achieved, accompanied by a peak-to-valley current ratio (PVCR) of 1.24. This can be attributed to the reduced resonant tunneling time under thinner AlN barrier and the suppressed external incoherent valley current by reducing the dislocation number contained in the RTD device with the smaller size of collector. Statistical analysis of the NDR performance of RTD devices with different AlN barrier thicknesses reveals that, as the AlN barrier thickness decreases from 1.5 nm to 1.25 nm, the average peak current density increases from 145.7 kA/cm2 to 1215.1 kA/cm2, while the average PVCR decreases from 1.45 to 1.1, and the peak voltage drops from 6.89 V to 5.49 V. The peak current density obtained in this work represents the highest value reported for nitride-based RTDs to date, while maintaining a high PVCR value simultaneously. This illustrates that an ultra-scaled RTD based on a vertical quantum-well structure and lateral collector size is a valuable approach for the development of nitride-based RTDs with excellent NDR characteristics, revealing their great potential applications in high-frequency oscillation sources and high-speed switch circuits.

Keywords: GaN resonant tunneling diode, peak current density, peak-to-valley current ratio, negative differential resistance

Procedia PDF Downloads 63

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: Abdullah Ijaz Hussain

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The purpose of this study was to explore the cardioprotective and anti-inflammatory effects of polyphenol-rich extracts from cucurbitaceae family members, including Cucurbita pepo, C. moschata, and C. maxima, on rat models. The initial crude extracts from these cucurbits were further separated into hexane, chloroform, ethyl acetate, butanol, and aqueous ethanol fractions, labeled as HEF, CHF, EAF, BUF, and AEF, respectively. Of these, AEF yielded the highest amount, followed by BUF, HEF, EAF, and CHF in descending order. Notably, EAF contained the greatest concentration of total phenolics, flavonoids, and flavonols. In terms of antioxidant activity, EAF demonstrated the most potent DPPH radical scavenging capability, succeeded by CHF, BUF, AEF, and HEF. EAF also exhibited the strongest reducing potential among the fractions. RP-HPLC analysis identified various phenolic acids and flavonoids across the cucurbita fractions, including ferulic acid, vanillic acid, p-coumeric acid, gallic acid, p-hydroxybenzoic acid, chlorogenic acid, catechin, rutin, quercetin, myricetin, and kaempferol. Doses of 250 and 500 mg/kg body weight of cucurbita fractions were administered orally to male WKY rats daily for 21 days. The rats' body weight, heart rate, and blood pressure were monitored bi-weekly. Oxidative status assessments were conducted using plasma samples to measure levels of malondialdehyde (MDA), superoxide dismutase (SOD), reduced glutathione (GSH), nitric oxide (NO), and total antioxidant capacity (TAC). At the study's conclusion, surgical assessments, including blood pressure, pulse wave velocity (PWV), and echocardiograms (ECG) were performed. The findings indicated that EAF from cucurbita significantly enhanced antihypertensive and antioxidant activities in the SHR rat group.

Keywords: polyphenols, chlorogenic acid, antihypertensive activity, oxidative stress, lcms

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Authors: Maria L. G. Lourenco, Viviane Y. Hibaru, Keylla H. N. P. Pereira, Fabiana F. Souza, Joao C. P. Ferreira, Simone B. Chiacchio, Luiz H. A. Machado

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Neonatal conjunctivitis, or ophthalmia, is an infection of the conjunctiva or cornea before opening the eyelids. It is believed that immunodeficiency contributes to the development of the condition. This study aims at reporting a case of ophthalmia neonatorum in a dog, in addition to its diagnosis and treatment. A litter of five pug neonates was admitted to the Sao Paulo State University (UNESP) Veterinary Hospital, Botucatu, Sao Paulo, Brazil, with complaints of ocular secretion. The neonates were five days old. The clinical examination revealed that three newborns presented swelling in the ocular region and a purulent secretion in the medial corner of the eye that was exerting pressure on the ocular globes, which are compatible with the description of this disease. The diagnosis was made based on the clinical signs and bacterial culture of the secretion, which revealed the presence of bacteria belonging to the genus Staphylococcus sp. The laboratory assays did not reveal any alterations. The treatment was instituted gently, opening the eyelids early and cleaning the purulent ocular secretion with saline solution. An ophthalmic ointment with retinol, amino acids, methionine, and chloramphenicol (Epitezan®) was prescribed four times a day for seven days. Blood plasma (2 mL/100 g) was administered subcutaneously because bacterial infections in neonates may represent a failure in the transference of passive immunity. A more thorough cleaning of the environment was also recommended. Neonatal conjunctivitis has a simple diagnosis and treatment. If not treated early, it can evolve to adherence of the eyelids to the cornea, ulceration, and perforation of the cornea. Therefore, the prognosis is favorable as long as the condition is diagnosed early, and the treatment is instituted quickly.

Keywords: ophthalmia neonatorum, neonatal infection, puppy, newborn

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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: 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: Kuladip Jana

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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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Authors: Emmanuelle Maria, Pierre Crançon, Gaëtane Lespes

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Colloids are ubiquitous in the environment and are known to play a major role in enhancing the transport of trace elements, thus being an important vector for contaminants dispersion. Colloids study and characterization are necessary to improve our understanding of the fate of pollutants in the environment. However, in stream water and groundwater, colloids are often very poorly concentrated. It is therefore necessary to pre-concentrate colloids in order to get enough material for analysis, while preserving their initial structure. Many techniques are used to extract and/or pre-concentrate the colloidal phase from bulk aqueous phase, but yet there is neither reference method nor estimation of the impact of these different techniques on the colloids structure, as well as the bias introduced by the separation method. In the present work, we have tested and compared several methods of colloidal phase extraction/pre-concentration, and their impact on colloids properties, particularly their size distribution and their elementary composition. Ultrafiltration methods (frontal, tangential and centrifugal) have been considered since they are widely used for the extraction of colloids in natural waters. To compare these methods, a ‘synthetic groundwater’ was used as a reference. The size distribution (obtained by Field-Flow Fractionation (FFF)) and the chemical composition of the colloidal phase (obtained by Inductively Coupled Plasma Mass Spectrometry (ICPMS) and Total Organic Carbon analysis (TOC)) were chosen as comparison factors. In this way, it is possible to estimate the pre-concentration impact on the colloidal phase preservation. It appears that some of these methods preserve in a more efficient manner the colloidal phase composition while others are easier/faster to use. The choice of the extraction/pre-concentration method is therefore a compromise between efficiency (including speed and ease of use) and impact on the structural and chemical composition of the colloidal phase. In perspective, the use of these methods should enhance the consideration of colloidal phase in the transport of pollutants in environmental assessment studies and forensics.

Keywords: chemical composition, colloids, extraction, preconcentration methods, size distribution

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Authors: Ghada Badr, Arwa Alturki

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The biological function of an RNA molecule depends on its structure. The objective of the alignment is finding the homology between two or more RNA secondary structures. Knowing the common functionalities between two RNA structures allows a better understanding and a discovery of other relationships between them. Besides, identifying non-coding RNAs -that is not translated into a protein- is a popular application in which RNA structural alignment is the first step A few methods for RNA structure-to-structure alignment have been developed. Most of these methods are partial structure-to-structure, sequence-to-structure, or structure-to-sequence alignment. Less attention is given in the literature to the use of efficient RNA structure representation and the structure-to-structure alignment methods are lacking. In this paper, we introduce an O(N2) Component-based Pairwise RNA Structure Alignment (CompPSA) algorithm, where structures are given as a component-based representation and where N is the maximum number of components in the two structures. The proposed algorithm compares the two RNA secondary structures based on their weighted component features rather than on their base-pair details. Extensive experiments are conducted illustrating the efficiency of the CompPSA algorithm when compared to other approaches and on different real and simulated datasets. The CompPSA algorithm shows an accurate similarity measure between components. The algorithm gives the flexibility for the user to align the two RNA structures based on their weighted features (position, full length, and/or stem length). Moreover, the algorithm proves scalability and efficiency in time and memory performance.

Keywords: alignment, RNA secondary structure, pairwise, component-based, data mining

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Authors: Navneet Sharma, Himanshu Ojha, Dharam Pal Pathak, Rakesh Kumar Sharma

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Radio-nuclides decontamination is an important task because any extra second of deposition leads to deleterious health effects. We had developed and characterise nanoemulsion of p-tertbutylcalix[4]arens using phase inversion temperature (PIT) method and evaluate its decontamination efficacy (DE). The solubility of the drug was determined in various oils and surfactants. Nanoemulsion developed with an HLB value of 11 and different ratios of the surfactants 10% (7:3, w/w), oil (20%, w/w), and double distilled water (70%) were selected. Formulation was characterised by multi-photon spectroscopy and parameters like viscosity, droplet size distribution, zeta potential and stability were optimised. In vitro and Ex vivo decontamination efficacy (DE) was evaluated against Technetium-99m, Iodine-131, and Thallium-201 as radio-contaminants applied over skin of Sprague-Dawley rat and human tissue equivalent model. Contaminants were removed using formulation soaked in cotton swabs at different time intervals and whole body imaging and static counts were recorded using SPECT gamma camera before and after decontamination attempt. Data were analysed using one-way analysis of variance (ANOVA) and was found to be significant (p <0.05). DE of the nanoemulsion loaded with p-tertbutylcalix[4]arens was compared with placebo and recorded to be 88±5%, 90±3% and 89±3% for 99mTc, 131I and 201Tl respectively. Ex-vivo complexation study of p-tertbutylcalix[4]arene nanoemulsion with surrogate nuclides of radioactive thallium and Iodine, were performed on rat skin mounted on Franz diffusion cell using high-resolution sector field inductively coupled plasma mass spectroscopy (HR-SF-ICPMS). More than 90% complexation of the formulation with these nuclides was observed. Results demonstrate that the prepared nanoemulsion formulation was found efficacious for the decontamination of radionuclides from a large contaminated population.

Keywords: p-tertbutylcalix[4]arens, skin decontamination, radiological emergencies, nanoemulsion, iodine-131, thallium-201

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Authors: A. Aly Salwa, H. Diekmann, S. Hafiz Ragaa, DG Abo Elhassan

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This study was aimed to evaluate the effect of Milbond (HSCAS) on aflatoxin M1 in artificially contaminated cows milk. Chemisorption compounds used in this experiment were MIlBond, hydrated sodium calcium aluminosilicate (HSCAS). Raw cow milk were artificially exposed to aflatoxin M1 in a concentration of 100 ppb) with addition of Nilbond at 0.5, 1, 2 and 3 % at room temperature for 30 minutes. Aflatoxin M1 was decreased more than 95% by HSCAS at 2%. Milk composition consist of protein, fat, lactose, solid non fat and total solid were affected by addition of some adsorbents were not significantly affected (p 0.05). Tthis method did not involve degrading the toxin, milk may be free from toxin degradation products and is safe for consumption. In addition, the added material may be easily separated from milk after the substance adsorbs the toxin. Thus, this method should be developed by further researches for determining effects of these compounds on functional properties of milk. The ability of hydrated sodium calcium aluminosilicate to prevent or reduce the level of aflatoxin MI residues in milk is critically needed. This finding has important implications, because milk is ultimately consumed by humans and animals, and the reduction of aflatoxin contamination in the milk could have an important impact on their health.

Keywords: aflatoxin M1, Hydrated sodium calcium aluminium silicate, detoxification, raw cow milk

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Authors: Supabhorn Yimthiang, Wiyada Khanwian

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Cadmium is a heavy metal that is important in the environment because it is highly toxic. The incidence and severity of type 2 diabetes mellitus are known to be associated with cadmium. The purpose of this study was to investigate the cadmium levels in patients with type 2 diabetes mellitus at diabetes mellitus clinic, Thasala hospital, Nakhon Si Thummarat, Thailand. The study population was composed of forty five subjects. Among them, twenty two were diabetic patients and twenty three were apparently healthy non-diabetic individual subjects. After an overnight fasting, blood and morning urine samples were collected from each subject to determine fasting blood sugar and cadmium levels in urine, respectively. Systolic and diastolic blood pressure values were measured by aneroid sphygmomanometer. Study approval was taken from the human subject ethics committee of Walailak University. Verbal and written informed consent was taken from all participants. In the study samples, there were 31.8% males and 68.2% females with mean age of 47+10.53 years. The geometric mean of urine cadmium was significantly higher in diabetic patients (1.015 + 0.79 µg/g creatinine) when compared with the healthy subjects (0.395 + 0.53 µg/g creatinine) (P<0.05). This result also showed that urine cadmium excretion in diabetic patients was higher than in healthy subjects by 2.6 times. Moreover, fasting blood sugar (153+47.86 μg/dl) and systolic blood pressure (183.26+17.15 mmHg) of diabetic patients was significantly different when compared with healthy subjects (79+5.38 μg/dl and 112.78+11.32 mmHg, respectively) (P<0.05). Meanwhile, the concentration of cadmium in urine showed positive correlation with fasting plasma glucose (r=0.616) and systolic blood pressure (r=0.487). This preliminary study showed that cadmium might play an important role in the development and pathogenesis of diabetes mellitus in general population. However, these findings require confirmation through additional epidemiological and biological research.

Keywords: blood pressure, cadmium, fasting blood sugar, type 2 diabetes mellitus

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Authors: Zemmouri Hassiba, Lounici Hakim, Mameri Nabil

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Dried crushed seeds of Moringa oleifera contain an effective soluble protein; a natural cationic polyelectrolyte which causes coagulation. The present study aims to investigate the performance of Moringa oleifera seed extract as natural coagulant in clarification of secondary wastewater treatment highly charged in colloidal. A series of Jar tests was undertaken using raw wastewater providing from secondary decanter of Reghaia municipal wastewater treatment plant (MWWTP) located in East of Algiers, Algeria. Coagulation flocculation performance of Moringa oleifera was evaluated through supernatant residual turbidity. Various influence parameters namely Moringa oleifera dosage and pH have been considered. Tests on Reghaia wastewater, having 129 NTU of initial turbidity, showed a removal of 69.45% of residual turbidity with only 1.5 mg/l of Moringa oleifera. This sufficient removal capability encourages the use of this bioflocculant for treatment of turbid waters. Based on this result, the coagulant seed extract of Moringa oleifera is better suited to clarify municipal wastewater by removing turbidity. Indeed, Moringa oleifera which is a natural resource available locally (South of Algeria) coupled to the non-toxicity, biocompatibility and biodegradability, may be a very interesting alternative to the conventional coagulants used so far.

Keywords: coagulation flocculation, colloids, moringa oleifera, secondary wastewater

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Authors: Arun Kumar Singh, Rupesh M. Das, Shailendra Saini

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The present work is an attempt to evaluate the sub-auroral ionospheric behavior under changing space weather conditions especially during high solar activity year 2014. In view of this, the GPS TEC along with Ionosonde data over Indian permanent scientific base 'Maitri', Antarctica (70°46′00″ S, 11°43′56″ E) has been utilized. The results suggested that the nature of ionospheric responses to the geomagnetic disturbances mainly depended upon the status of high latitudinal electro-dynamic processes along with the season of occurrence. Fortunately, in this study, both negative and positive ionospheric impact to the geomagnetic disturbances has been observed in a single year but in different seasons. The study reveals that the combination of equator-ward plasma transportation along with ionospheric compositional changes causes a negative ionospheric impact during summer and equinox seasons. However, the combination of pole-ward contraction of the oval region along with particle precipitation may lead to exhibiting positive ionospheric response during the winter season. Other than this, some Ionosonde based new experimental evidence also provided clear evidence of particle precipitation deep up to the low altitudinal ionospheric heights, i.e., up to E-layer by the sudden and strong appearance of E-layer at 100 km altitudes. The sudden appearance of E-layer along with a decrease in F-layer electron density suggested the dominance of NO⁺ over O⁺ at a considered region under geomagnetic disturbed condition. The strengthening of E-layer is responsible for modification of auroral electrojet and field-aligned current system. The present study provided a good scientific insight on sub-auroral ionospheric to the changing space weather condition.

Keywords: high latitude ionosphere, space weather, geomagnetic storms, sub-storm

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Authors: Hai Xu, Yiwei Wang, Xi Bao, Bihua Deng, Pengcheng Li, Yu Lu

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T7 phage could be used as a perfect vector for peptides expression and haptens presentation. T7-3GnRH recombinant phage was constructed by inserting three copies of Gonadotrophin Releasing Hormone (GnRH) gene into the multiple cloning site of T7 Select 415-1b phage genome. The positive T7-3GnRH phage was selected by using polymerase chain reaction amplification, and the p10B-3GnRH fusion protein was verified by SDS-PAGE and Western-blotting assay. T7-3GnRH vaccine was made and immunized with 10¹⁰ pfu in 0.2 ml per dose in mice. Blood samples were collected at an interval in weeks, and anti-GnRH antibody and testosterone concentrations were detected by ELISA and radioimmunoassay, respectively. The results show that T7-3GnRH phage particles confer a high immunogenicity to the GnRH-derived epitope. Moreover, the T7-3GnRH vaccine induced higher level of anti-GnRH antibody than ImproVac^®. However, the testosterone concentrations in both immunized groups were at a similar level, and the testis developments were significantly inhibited compared to controls. These findings demonstrated that the anti-GnRH antibody could neutralize the endogenous GnRH to down regulate testosterone level and limit testis development, highlighting the potential value of T7-3GnRH in the immunocastration vaccine research.

Keywords: Gonadotrophin Releasing Hormone (GnRH), Immunocastration, T7 phage, Phage vaccine

Procedia PDF Downloads 287