Search results for: animal protein (AP)

Authors: Naveen Kumar B. T., Anuj Tyagi, Prabjeet Singh, Shanthanagouda A. H., Sumeet Rai

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Inland saline water resources are gaining the importance in expanding the aquaculture activities to mitigate the nutritional and food security issues of the world. For profitable and sustainable aquaculture practices, scientific farming, biosecurity measure, and best fish health management should be the integral part of developmental activities. Keeping in line with global awareness and trends, the Indian government has taken an innovative step to conduct disease surveillance and awareness programme for aquatic disease through network project. This ‘National Surveillance Programme for Aquatic Animal Diseases (NSPAAD)’ is being implemented in collaboration of national institutes and state agriculture universities with funding support from National Fisheries Development Board (NFDB), Govt. of India. Guru Angad Dev Veterinary and Animal Sciences University (GADVASU), Ludhiana, an NSPAAD collaborator, has been actively engaged in disease surveillance in the Indian state of Punjab. Shrimp farming in inland saline areas of Punjab is expanding at a tremendous pace under the guidance of GADVASU along with the support of State Fisheries Department. Under this national disease surveillance programme, we reported Enterocytozoon hepatopenaei (EHP) infection in the Litopenaeus vannamei cultured in the inland saline waters. Polymerase chain reaction (PCR) based diagnosis was carried out using the OIE (World Organisation for Animal Health) protocol. It was observed that out of 20 shrimp farms, two farms were 1st step PCR positive and two more farms were nested PCR positive. All the EHP positive ponds had shown the white faeces along with mortalities at very low rate. Therefore, implementation of biosecurity and continuous surveillance and monitoring program for finfish and shellfish aquaculture are in need of the hour to prevent and control the large-scale disease outbreaks and subsequent economic losses.

Keywords: disease, EHP, inland saline water, shrimp culture

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Authors: H. Li, L. Ji, J. Su

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Forward osmosis is an emerging technology for separation and has great potential in the concentration of liquid products such as protein, pharmaceutical, and natural products. In pharmacy industry, one of the very tough talks is to concentrate the product in a gentle way since some of the key components may lose bioactivity when exposed to heating or pressurization. Therefore, forward osmosis (FO), which uses inherently existed osmosis pressure instead of externally applied hydraulic pressure, is attractive for pharmaceutical enrichments in a much efficient and energy-saving way. Recently, coordination complexes have been explored as the new class of draw solutes in FO processes due to their bulky configuration and excellent performance in terms of high water flux and low reverse solute flux. Among these coordination complexes, ferric citrate complex with lots of hydrophilic groups and ionic species which make them good solubility and high osmotic pressure in aqueous solution, as well as its low toxicity, has received much attention. However, the chemistry of ferric complexation by citrate is complicated, and disagreement prevails in the literature, especially for the structure of the ferric citrate. In this study, we investigated the chemical reaction with various molar ratio of iron and citrate. It was observed that the ferric citrate complex (Fe-CA2) with molar ratio of 1:1 for iron and citrate formed at the beginning of the reaction, then Fecit would convert to ferric citrate complex at the molar ratio of 1:2 with the proper excess of citrate in the base solution. The structures of the ferric citrate complexes synthesized were systematically characterized by X-ray diffraction (XRD), UV-vis spectroscopy, X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FT-IR) and Thermogravimetric analysis (TGA). Fe-CA2 solutions exhibit osmotic pressures more than twice of that for NaCl solutions at the same concentrations. Higher osmotic pressure means higher driving force, and this is preferable for the FO process. Fe-CA2 and NaCl draw solutions were prepared with the same osmotic pressure and used in FO process for BSA protein concentration. Within 180 min, BSA concentration was enriched from 0.2 to 0.27 L using Fe-CA draw solutions. However, it was only increased from 0.20 to 0.22 g/L using NaCl draw solutions. A reverse flux of 11 g/m²h was observed for NaCl draw solutes while it was only 0.1 g/m²h for Fe-CA2 draw solutes. It is safe to conclude that Fe-CA2 is much better than NaCl as draw solute and it is suitable for the enrichment of liquid product.

Keywords: draw solutes, ferric citrate complex, forward osmosis, protein enrichment

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Authors: Laura Ulitzky, Dougbeh-Chris Nyan, Manuel M. Lafer, Erica Silberstein, Nicoleta Cehan, Deborah R. Taylor

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Hepatitis C virus (HCV)-associated hepatocellular carcinoma, fibrosis and cirrhosis are more frequent in men and postmenopausal women than in premenopausal women and women receiving hormone replacement therapy, suggesting that β-estradiol (estrogen) plays an innate role in preventing viral infection and liver disease. Estrogen classically acts through nuclear estrogen receptors or, alternatively, through the membrane-bound G-protein-coupled estrogen receptor (GPR30 or GPER). We observed a marked decrease in detectable virus when HCV-infected human hepatoma cells were treated with estrogen. The effect was mimicked by both Tamoxifen (Tam) and G1, a GPR30-specific agonist, and was reversed by the GPR30-specific antagonist, G15. Through GPR30, estrogen-mediated the down-regulation of occludin; a tight junction protein and HCV receptor, by promoting activation of matrix metalloproteinases (MMPs). Activated MMP-9 was secreted in response to estrogen, cleaving occludin in the extracellular Domain D, the motif required for HCV entry and spread. This pathway gives new insight into a novel innate immune pathway and the disparate host-virus responses to HCV demonstrated by the two sexes. Moreover, these data suggest that hormone replacement therapy may have beneficial antiviral properties for HCV-infected postmenopausal women and show promise for new antiviral treatments for both men and women.

Keywords: HCV, estrogen, occludin, MMPs

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Authors: Salwa Karboune, Amanda Waglay

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Recent studies indicate that health-promoting functional ingredients and nutraceuticals can help support and improve the overall public health, which is timely given the aging of the population and the increasing cost of health care. The development of novel ‘natural’ functional ingredients is increasingly challenging. Biocatalysis offers powerful approaches to achieve this goal. Our recent research has been focusing on the development of innovative biocatalytic approaches towards the isolation of protein isolates from potato by-products and the generation of peptides. Potato is a vegetable whose high-quality proteins are underestimated. In addition to their high proportion in the essential amino acids, potato proteins possess angiotensin-converting enzyme-inhibitory potency, an ability to reduce plasma triglycerides associated with a reduced risk of atherosclerosis, and stimulate the release of the appetite regulating hormone CCK. Potato proteins have long been considered not economically feasible due to the low protein content (27% dry matter) found in tuber (Solanum tuberosum). However, potatoes rank the second largest protein supplying crop grown per hectare following wheat. Potato proteins include patatin (40-45 kDa), protease inhibitors (5-25 kDa), and various high MW proteins. Non-destructive techniques for the extraction of proteins from potato pulp and for the generation of peptides are needed in order to minimize functional losses and enhance quality. A promising approach for isolating the potato proteins was developed, which involves the use of multi-enzymatic systems containing selected glycosyl hydrolase enzymes that synergistically work to open the plant cell wall network. This enzymatic approach is advantageous due to: (1) the use of milder reaction conditions, (2) the high selectivity and specificity of enzymes, (3) the low cost and (4) the ability to market natural ingredients. Another major benefit to this enzymatic approach is the elimination of a costly purification step; indeed, these multi-enzymatic systems have the ability to isolate proteins, while fractionating them due to their specificity and selectivity with minimal proteolytic activities. The isolated proteins were used for the enzymatic generation of active peptides. In addition, they were applied into a reduced gluten cookie formulation as consumers are putting a high demand for easy ready to eat snack foods, with high nutritional quality and limited to no gluten incorporation. The addition of potato protein significantly improved the textural hardness of reduced gluten cookies, more comparable to wheat flour alone. The presentation will focus on our recent ‘proof-of principle’ results illustrating the feasibility and the efficiency of new biocatalytic processes for the production of innovative functional food ingredients, from potato by-products, whose potential health benefits are increasingly being recognized.

Keywords: biocatalytic approaches, functional ingredients, potato proteins, peptides

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Authors: Athar Mahmood, Asad Ali

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Management considerations including harvesting time and nitrogen application considerably influence the biomass yield, quality and biogas production. Therefore, a field study was conducted to determine the effect of various harvesting times and nitrogen rates on the biomass yield, quality and biogas yield of maize crop. This experiment was consisted of various harvesting times i.e., harvesting after 45, 55 and 65 days of sowing (DAS) and nitrogen rates i.e., 0, 100, 150 and 200 kg ha-1 respectively. The data indicated that maximum plant height, leaf area, dry matter (DM) yield, protein, acid detergent fiber, neutral detergent fiber, crude fiber contents and biogas yield were recorded 65 days after sowing while lowest was recorded 45 days after sowing. In contrary to that significantly higher chlorophyll contents were observed at 45 DAS. In case of nitrogen rates maximum plant height, leaf area, and DM yield, protein contents, ash contents, acid detergent fiber, neutral detergent fiber, crude fiber contents and chlorophyll contents were determined with nitrogen at the rate of 200 kg ha-1, while minimum was observed when no N was applied. Therefore, harvesting 65 DAS and N application @ 200 kg ha-1 can be suitable for getting the higher biomass and biogas production.

Keywords: chemical composition, fiber contents, biogas, nitrogen, harvesting time

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Authors: Jannika Dombrowski, Sabine Ambros, Ulrich Kulozik

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Due to the increasing popularity of healthy products, probiotics are still of rising importance in food manufacturing. With the aim to amplify the field of probiotic application to non-chilled products, the cultures have to be preserved by drying. Microwave drying has proved to be a suitable technique to achieve relatively high survival rates, resulting from drying at gentle temperatures, among others. However, diffusion limitation due to compaction of cell suspension during drying can prolong drying times as well as deteriorate product properties (grindability, rehydration performance). Therefore, we aimed to embed probiotics in an aerated matrix of whey proteins (surfactants) and di-/polysaccharides (foam stabilization, probiotic protection) during drying. As a result of the manifold increased inner surface of the cell suspension, drying performance was enhanced significantly as compared to non-foamed suspensions. This work comprises investigations on suitable foam matrices, being stable under vacuum (variation of protein concentration, type and concentration of di-/polysaccharide) as well as development of an applicable microwave drying process in terms of microwave power, chamber pressure and maximum product temperatures. Performed analyses included foam characteristics (overrun, drainage, firmness, bubble sizes), and properties of the dried cultures (survival, activity). In addition, efficiency of the drying process was evaluated.

Keywords: foam structure, microwave drying, polysaccharides, probiotics

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Authors: Waranya Imprasittichai, Patsarawadee Paojinda, 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. In this study, we constructed gene fusions of Plasmodium falciparum OMPDC-OPRT (1,836 bp) in pTrcHisA vector and expressed as an 6xHis-tag bifunctional protein in three Escherichia coli strains (BL21, Rosetta, TOP10) at 18 °C, 25 °C and 37 °C. The recombinant bifunctional protein was partially purified by Ni-Nitrilotriacetic acid-affinity chromatography. Specific activities of OPRT and OMPDC domains in the bifunctional enzyme expressed in E. coli TOP10 cells were approximately 3-4-fold higher than those in BL21 cells. There were no enzymatic activities when the construct vector expressed in Rosetta cells. Maximal expression of the fused gene was observed at 18 °C and the bifunctional enzyme had specific activities of OPRT and OMPDC domains in a ratio of 1:2. These results provide greater yields and better catalytic activities of the bifunctional OMPDC-OPRT enzyme for further purification and kinetic study.

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

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Authors: Raj K. Gupta, X. Gary Tan, Andrzej Przekwas

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Bomb blasts from Improvised Explosive Devices (IEDs) account for vast majority of terrorist attacks worldwide. Injuries caused by IEDs result from a combination of the primary blast wave, penetrating fragments, and human body accelerations and impacts. This paper presents a multiscale computational model of coupled blast physics, whole human body biodynamics and injury biomechanics of sensitive organs. The disparity of the involved space- and time-scales is used to conduct sequential modeling of an IED explosion event, CFD simulation of blast loads on the human body and FEM modeling of body biodynamics and injury biomechanics. The paper presents simulation results for blast-induced brain injury coupling macro-scale brain biomechanics and micro-scale response of sensitive neuro-axonal structures. Validation results on animal models and physical surrogates are discussed. Results of our model can be used to 'replicate' filed blast loadings in laboratory controlled experiments using animal models and in vitro neuro-cultures.

Keywords: blast waves, improvised explosive devices, injury biomechanics, mathematical models, traumatic brain injury

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Authors: Redie Kidane Ghebrehawariat, Betiel Habte Hadgu, Filmon Berhane Kahsay, Rim Berhane Fisehaye, Samuel Haile Kahsay, Saron Yemane Yosief, Selemawit Mosazghi Gilazghi

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A cross-sectional study was conducted from 22 February to 9 April 2022 on small, medium, and large holder dairy farms to determine the bovine mastitis prevalence and associated risk factors in the Anseba region, Eritrea. A total of 34 dairy farms and 193 dairy cows were randomly selected. Dairy cows were physically examined for any change on the udder and milk; a California mastitis test was performed to check sub-clinical mastitis; a closed-ended semi-structured questionnaire composed of 28 variables/risk factors (21 management risk factors and 7 animal-level risk factors) was used to determine the risk factors responsible for clinical and sub-clinical mastitis in the dairy cows. The overall cow-level prevalence of mastitis was 147 (76.2%). The animal level prevalence rate of clinical and sub-clinical mastitis was found to be 22 (11.4%) and 125 (64.8%), respectively, while herd level prevalence both for clinical and subclinical mastitis was found to be 14 (41.2%) and 26 (76.5%) respectively. Based on the already set P-value, which is <0.05, a number of risk factors were found to have a significant relationship with the occurrence of clinical and sub-clinical mastitis. Generally, animal risk factors such as animal age, parity, injury on the udder or teat, and previous history of mastitis presence of injury on the udder and lactation stage were risk factors with a significant relationship with the occurrence of clinical and sub-clinical mastitis. On the other hand, management risk factors with a significant relationship to the occurrence of clinical and sub-clinical mastitis were herd size, failure to milk mastitic cow, at last, educational level, floor type, failure to use a towel, using one towel for more than one cow and failure to practice mastitis test. From a total of 772 quarters, 280 (36.3%) were found positive for sub-clinical mastitis using the California mastitis test; of these, 70 (9%) were weakly positive, 90 (11.7%) were distinct positive, and 120 (15.5%) were strongly positive. Furthermore, 13 (1.7%) quarters were blocked. Quarter level prevalence was right front 80 (41.5%), left front 64 (33.3%), right hind 69 (35.8%) and left hind 67 (34.7%). The study has shown that mastitis is a major problem for dairy farms and the findings suggested that mastitis is one of the limiting factors in increasing milk production. Subclinical mastitis was found to be a devastating problem, and it occurred in all three breeds of lactating dairy cattle. Therefore, farmers should work hard to avoid the above-mentioned risk factors to minimize the infection of their dairy cattle by mastitis and thereby increase their profit. On the other hand, the Ministry of Agriculture, through the extension unit, should work in close contact with the farmers to increase awareness of the economic importance of the disease and associated risk factors.

Keywords: mastitis, prevalence, dairy cattle, Anseba, Eritrea

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Authors: Ezzati Givi M., Ezzatigivi N., Eimani H.

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Trigeminal Neuralgia (TN) consists of painful attacks often triggered with general activities, which cause impairment and disability. The first line of treatment consists of pharmacotherapy. However, the occurrence of many side-effects limits its application. Acute pain relief is crucial for titrating oral drugs and making time for neurosurgical intervention. This study aimed to examine the long-term anesthetic effect of QX-314 and capsaicin in trigeminal neuralgia using an animal model. TN was stimulated by surgical constriction of the infraorbital nerve in rats. After seven days, anesthesia infiltration was done, and the duration of mechanical allodynia was compared. Thirty-five male Wistar rats were randomly divided into seven groups as follows: control (normal saline); lidocaine (2%); QX314 (30 mM); lidocaine (2%)+QX314 (15 mM); lidocaine (2%)+QX314 (22 mM); lidocaine (2%)+QX314 (30 mM); and lidocaine (2%)+QX314 (30 mM) +capsaicin (1μg). QX314 in combination with lidocaine significantly increased the duration of anesthesia, which was dose-dependent. The combination of lidocaine+QX314+capsaicin could significantly increase the duration of anesthesia in trigeminal neuralgia. In the present study, we demonstrated that the combination of QX-314 with lidocaine and capsaicin produced a long-lasting, reversible local anesthesia and was superior to lidocaine alone in the fields of the duration of trigeminal neuropathic pain blockage.

Keywords: trigeminal neuralgia, capsaicin, lidocaine, long-lasting

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Authors: H. Sakamoto, J. Tochimoto, S. Kurosawa, M. Suzuki, S. Oguri

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Salt stress adversely affects plant growth at various stages of development including seed germination, seedling establishment, vegetative growth and finally reproduction. Because of their immobile nature, plants have evolved mechanisms to sense and respond to salt stress. Seed dormancy is an adaptive trait that enables seed germination to coincide with favorable environmental conditions. We identified a novel locus of Arabidopsis, designated SHG1 (salt hypersensitive germination 1), whose disruption leads to reduced germination rate under moderate salt stress conditions. SHG1 encodes a transmembrane protein with an ankyrin repeat motif that has been implicated in diverse cellular processes such as signal transduction. The SGH1-disrupted Arabidopsis mutant died at the cotyledon stage when sown on salt-containing medium, although wild type plants could form true leaves under the same conditions. On the other hand, this mutant showed similar phenotypes to wild type plants when sown on medium without salt and transferred to salt-containing medium at the vegetative stage. These results suggested that SHG1 played indispensable role in the seed germination and seedling establishment under moderate salt stress conditions. SHG1 may be involved in the release of seed dormancy.

Keywords: germination, ankyrin repeat, arabidopsis, salt tolerance

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Authors: A. F. C. Nassar, D. K. Tessler, L. Okuda, C. Del Fava, D. P. Chiebao, A. H. C. N. Romaldini, A. P. Alvim, M. J. Sanchez-Vazquez, M. S. Rosa, J. C. Pompei, R. Harakava, M. C. S. Araujo, G. H. F. Marques, E. M. Pituco

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Glanders is a zoonotic disease of Equidae caused by the bacterium Burkholderia mallei presented in acute or chronic clinical forms with inflammatory nodules in the respiratory tract, lymphangitis and caseous lymph nodes. There is not a treatment with veterinary drugs to this life-threatening disease; thus, its occurrence must be notified to official animal health services and any infected animal must be eliminated. This study aims to detect B. mallei from horses euthanized in outbreaks of glanders in Brazil, providing a better understanding of the bacterial characteristics and determine a proper protocol for isolation. The work was carried out with the collaboration of the Ministry of Agriculture and the Sao Paulo State Animal Health Department, while its procedures were approved by the Committee of Ethics in Animal Experimentation from the Instituto Biologico (CETEA n°156/2017). To the present time, 16 horses from farms with outbreaks of glanders detected by complement fixation test (CFT) serology method were analyzed. During the necropsy, samples of possibly affected organs (lymph nodes, lungs, heart, liver, spleen, kidneys and trachea) were collected for bacterial isolation, molecular tests and pathology. Isolation was performed using two enriched mediums, a potato infusion agar with 5% sheep blood, 4% glycerol and antibiotics (penicilin100U/ mL), and another with the same ingredients except the antibiotic. A PCR protocol was modified for this study using primers design to identify a region of the Flip gen of B. mallei. Thru isolation, 12.5% (2/16) animals were confirmed positive using only the enriched medium with antibiotic and confirmed by PCR: from mediastinal and submandibular lymph nodes and lungs in one animal and from mediastinal lymph node in the other. The detection of the bacterium using PCR showed positivity of 100% (16/16) horses from 144 samples of organs. Pathology macroscopic lesions observed were catarrhal nasal discharge, fetlock ulcers, emaciation, lymphangitis in limbs, suppurative lymphangitis, lymph node enlargement, star shaped liver, and spleen scars, adherence of the renal capsule, pulmonary hemorrhage, and miliary nodules. Microscopic lesions were suppurative bronchopneumonia with microabscesses and Langhans giant cells in lungs; lymph nodes with abscesses and intense lymphoid reaction; hemosiderosis and abscesses in spleen. Positive samples on PCR will be sequenced later and analyzed comparing with previous records in the literature. A throughout description of the recent acute cases of glanders occurring in Brazil and characterization of the bacterium related will contribute to advances in the knowledge of the pathogenicity, clinical symptoms, and epidemiology of this zoonotic disease. Acknowledgment: This project is sponsored by FAPESP.

Keywords: equines, bacterial isolation, zoonosis, PCR, pathology

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Authors: Teklay Gebrecherkos, Mahmud Abdulkader, Tobias Rinke De Wit, Britta C. Urban, Feyissa Chala, Yazezew Kebede, Dawit Welday

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Background: C-reactive protein (CRP) levels are a reliable surrogate for interleukin-6 bioactivity that plays a pivotal role in the pathogenesis of cytokine storm associated with severe COVID-19. There is a lack of data on the role of CRP as a determinant of COVID-19 severity status in the African context. Methods: We determined the longitudinal kinetics of CRP levels on 78 RT-PCR-confirmed COVID-19 patients (49 non-severe and 29 severe cases) and 50 PCR-negative controls. Results: COVID-19 patients had overall significantly elevated CRP at baseline when compared to PCR-negative controls [median 11.1 (IQR: 2.0-127.8) mg/L vs. 0.9 (IQR: 0.5-1.9) mg/L; p=0.0004)]. Moreover, severe COVID-19 patients had significantly higher median CRP levels than non-severe cases [166.1 (IQR: 48.6-332.5) mg/L vs. 2.4 (IQR: 1.2-7.6) mg/L; p<0.00001)]. In addition, persistently elevated levels of CRP were exhibited among those with comorbidities and higher age groups. Area under receiver operating characteristic curve (AUC) analysis of CRP levels distinguished PCR-confirmed COVID-19 patients from the ones with PCR-negative non-COVID-19 individuals, with an AUC value of 0.77 (95% CI: 0.68-0.84; p=0.001). Moreover, it clearly distinguished severe from non-severe COVID-19 patients, with an AUC value of 0.83 (95% CI: 0.73-0.91). After adjusting for age and the presence of comorbidities, CRP levels above 30 mg/L were significantly associated with an increased risk of developing severe COVID-19 (adjusted relative risk 3.99 (95%CI: 1.35-11.82; p=0.013). Conclusions: Determining CRP levels in COVID-19 patients in African settings may provide a simple, prompt, and inexpensive assessment of the severity status at baseline and monitoring of treatment outcomes.

Keywords: CRP, COVID-19, SARS-CoV-2, biomarker

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Authors: Hady Atef, Zinab Helmy, Heba Abdeen, Mostafa Fadel

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Background and purpose: After the success of the first trials of this experiment which were done on rabbits, a new study were conducted on dogs to ensure the past results; in a step forward to use low-level LASER therapy in the treatment of congenital septal defects in infants. The aim of this study was to investigate the effect of low-level LASER irradiation on congenital septal defects in dogs. Subjects and Methodology: six male dogs who have congenital septal defects in their hearts -with age ranged 6-10 months- enrolled in this study for one and half months. They were assigned into two groups: Group (A): The study group consisted of 3 canine hearts who received routine animal care associated with LASER irradiation. Group (B): The control group consisted of 3 canine hearts who received only routine animal care. Sizes of the septal defects were measured for both groups at the beginning and after the end of the study. Results: There was a significant decrease in the size of the diameter of the congenital septal defect with the study group (percentage of improvement was 42.19%) when compared with control group. Conclusion: It was concluded that low-level LASER therapy can be considered as a promising therapy for congenital heart defects in animals and to be examined on children with similar congenital lesions after then.

Keywords: laser, congenital septal defects, dogs, infants

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Authors: Rabah Iratni, Husain El Hasasna, Khawlah Athamneh, Halima Al Sameri, Nehla Benhalilou, Asma Al Rashedi

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Background: Cancer therapies have witnessed great advances in the recent past, however, cancer continues to be a leading cause of death, with colorectal cancer being the fourth cause of cancer-related deaths. Colorectal cancer affects both sexes equally with poor survival rate once it metastasizes. Phytochemicals, which are plant derived compounds, have been on a steady rise as anti-cancer drugs due to the accumulation of evidences that support their potential. Here, we investigated the anticancer effect of Rhus coriaria on colon cancer cells. Material and Method: Human colon cancer HT-29 cell line was used. Protein expression and protein phosphorylation were examined using Western blotting. Transcription activity was measure using Quantitative RT-PCR. Human tumoral clonogenic assay was used to assess cell survival. Senescence was assessed by the senescence-associated beta-galactosidase assay. Results: Rhus coriaria extract (RCE) was found to significantly inhibit the viability and colony growth of human HT-29 colon cancer cells. RCE induced senescence and cell cycle arrest at G1 phase. These changes were concomitant with upregulation of p21, p16, downregulation of cyclin D1, p27, c-myc and expression of Senescence-associated-β-Galactosidase activity. Moreover, RCE induced non-canonical beclin-1independent autophagy and subsequent apoptotic cell death through activation of activation caspase 8 and caspase 7. The blocking of autophagy by 3-methyladenine (3-MA) or chloroquine (CQ) reduced RCE-induced cell death. Further, RCE induced DNA damage, reduced mutant p53 protein level and downregulated phospho-AKT and phospho-mTOR, events that preceded autophagy. Mechanistically, we found that RCE inhibited the AKT and mTOR pathway, a regulator of autophagy, by promoting the proteasome-dependent degradation of both AKT and mTOR proteins. Conclusion: Our findings provide strong evidence that Rhus coriaria possesses strong anti-colon cancer activity through induction of senescence and autophagic cell death, making it a promising alternative or adjunct therapeutic candidate against colon cancer.

Keywords: autophagy, proteasome degradation, senescence, mTOR, apoptosis, Beclin-1

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Authors: Ahsan Abdullah, Ahmed A. S. Bakshwain

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Livestock is one of the fastest-growing sectors in agriculture. If carefully managed, have potential opportunities for economic growth, food sovereignty and food security. In this study we mainly analyse and compare long-term i.e. for year 2030 climate variability impact on predicted productivity of meat i.e. beef, mutton and poultry for the Kingdom of Saudi Arabia w.r.t three factors i.e. i) climatic-change vulnerability ii) CO2 fertilization and iii) water scarcity and compare the results with two countries of the region i.e. Iraq and Yemen. We do the analysis using data from diverse sources, which was extracted, transformed and integrated before usage. The collective impact of the three factors had an overall negative effect on the production of meat for all the three countries, with adverse impact on Iraq. High similarity was found between CO2 fertilization (effecting animal fodder) and water scarcity i.e. higher than that between production of beef and mutton for the three countries considered. Overall, the three factors do not seem to be favorable for the three Middle-East countries considered. This points to possibility of a vegetarian year 2030 based on dependency on indigenous live-stock population.

Keywords: prediction, animal-source foods, pastures, CO2 fertilization, climatic-change vulnerability, water scarcity

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Authors: Muhammad Jawad Saleem, Faisal Hafeez, Muhammad Arshad, Afifa Naeem, Ayesha Iftekhar

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Bacillus thuringiensis is a gram-positive spore-forming bacterium that belongs to the Bacillus cereus group of Bacilli and it produces ICP (insecticidal crystal protein) Cry toxins or Cysts toxins. Spores are produced as parasporal crystalline inclusions bodies (also known as endotoxins) at the onset of sporulation during the stationary growth phase. During vegetative growth that does not form crystals and is called vegetative insecticidal proteins (VIP) and secreted an insecticidal protein (SIP). Bacillus thuringiensis (Bt) is important for pest management either in the form of insecticides or through incorporated in the gene of the crop. Bioassays were conducted on the F2 generation of 1st instar larvae of H. armigera by the diet incorporation method to determine the susceptibility to Bt Cry toxins (Cry1Ac, Cry2Ab, Cry2A). The median lethal concentration (LC₅₀) of Cry1Ac, Cry2Ab, Cry2A ranged from 0.11 to 1.06 µg/ml and moult inhibitory concentration (MIC₅₀) of Cry1Ac, Cry2Ab, Cry2A ranged from 0.05 to 0.25 µg/ml. Cry1Ac was found most toxic to 1st instar larvae of H. armigera as compared to other Bt Cry toxins (Cry1Ac, Cry2Ab, Cry2A). The experimental results are important to policy-makers and technology providers to develop strategies for the exploitation of transgenic Bt cotton varieties as a component of integrated pest management.

Keywords: Bt toxin, Cry1Ac, Cry2Ab, Cry2A, susceptibility, Helicoverpa armigera

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Authors: Shujun Xie, Shirong Zhang, Shenglin Ma

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Background: Chronic inflammation might lead to many malignancies, and inadequate resolution could play a crucial role in tumor invasion, progression, and metastases. A randomised, double-blind, placebo-controlled trial shows that IL-1β inhibition with canakinumab could reduce incident lung cancer and lung cancer mortality in patients with atherosclerosis. The process and secretion of proinflammatory cytokine IL-1β are controlled by the inflammasome. Here we showed the correlation of the innate immune system and afatinib, a tyrosine kinase inhibitor targeting epidermal growth factor receptor (EGFR) in non-small cell lung cancer. Methods: Murine Bone marrow derived macrophages (BMDMs), peritoneal macrophages (PMs) and THP-1 were used to check the effect of afatinib on the activation of NLRP3 inflammasome. The assembly of NLRP3 inflammasome was check by co-immunoprecipitation of NLRP3 and apoptosis-associated speck-like protein containing CARD (ASC), disuccinimidyl suberate (DSS)-cross link of ASC. Lipopolysaccharide (LPS)-induced sepsis and Alum-induced peritonitis were conducted to confirm that afatinib could inhibit the activation of NLRP3 in vivo. Peripheral blood mononuclear cells (PBMCs) from non-small cell lung cancer (NSCLC) patients before or after taking afatinib were used to check that afatinib inhibits inflammation in NSCLC therapy. Results: Our data showed that afatinib could inhibit the secretion of IL-1β in a dose-dependent manner in macrophage. Moreover, afatinib could inhibit the maturation of IL-1β and caspase-1 without affecting the precursors of IL-1β and caspase-1. Next, we found that afatinib could block the assembly of NLRP3 inflammasome and the ASC speck by blocking the interaction of the sensor protein NLRP3 and the adaptor protein ASC. We also found that afatinib was able to alleviate the LPS-induced sepsis in vivo. Conclusion: Our study found that afatinib could inhibit the activation of NLRP3 inflammasome in macrophage, providing new evidence that afatinib could target the innate immune system to control chronic inflammation. These investigations will provide significant experimental evidence in afatinib as therapeutic drug for non-small cell lung cancer or other tumors and NLRP3-related diseases and will explore new targets for afatinib.

Keywords: inflammasome, afatinib, inflammation, tyrosine kinase inhibitor

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Authors: Reihane Mehrparvar

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Human age could exceed further by altering gene expression through food intaking, although as a consequence of recent century eating patterns, human life-span getting shorter by emerging irregulating in autophagy mechanism, insulin, leptin, gut microbiota which are important etiological factors of type-2 diabetes, obesity, infertility, cancer, metabolic and autoimmune diseases. However, restricted calorie intake and vigorous exercise might be beneficial for losing weight and metabolic regulation in a short period but could not be implementable in the long term as a way of life. Therefore, the lack of a dietary program that is compatible with the genes of the body is essential. Sweet and high-glycemic-index (HGI) foods were associated with type-2 diabetes and cancer morbidity. The neuropsychological perspective characterizes the inclination of sweet and HGI-food consumption as addictive behavior; hence this process engages preference of gut microbiota, neural node, and dopaminergic functions. Moreover, meal composition is not the only factor that affects body hemostasis. In this narrative review, it is believed to attempt to investigate how the body responded to different food intakes and represent an accurate model based on current evidence. Eating frequently and ingesting unassimilable protein and carbohydrates may not be compatible with human genes and could cause impairments in the self-renovation mechanism. This trajectory indicates our body is more adapted to starvation and eating animal meat and marrow. Here has been recommended a model that takes into account three important factors: frequent eating, meal composition, and circadian rhythm, which may offer a promising intervention for obesity, inflammation, cardiovascular, autoimmune disorder, type-2 diabetes, insulin resistance, infertility, and cancer through intensifying autophagy-mechanism and eliminate medical costs.

Keywords: metabolic disease, anti-aging, type-2 diabetes, autophagy

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Authors: Rueyhung Weng, Chia-En Huang, Jung-Chi-Liao

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The transition zone (TZ) serves as a diffusion barrier to regulate the ins and outs of the proteins recruited to the primary cilia. TCTN2 is one of the TZ proteins and its mutation causes Joubert syndrome, a serious multi-organ disease. Despite its important medical relevance, the functions of TCTN2 remain elusive. Here we created a TCTN2 gene deleted retinal pigment epithelial cells (RPE1) using CRISPR/Cas9-based genome editing technique and used this knockout line to reveal roles of TCTN2. TCTN2 knockout RPE1 cells displayed a significantly reduced ciliogenesis or a shortened primary cilium length in the cilium-remaining population. Intraflagellar transport protein IFT88 aberrantly accumulated at the tip of TCTN2 deficient cells. Guanine nucleotide exchange factor Arl13B was mostly absent from the ciliary compartment, with a small population localizing at the ciliary tip. The deficient TZ was corroborated with the mislocalization of two other TZ proteins TMEM67 and MKS1. In addition, TCTN2 deficiency induced TZ impairment led to the suppression of Sonic hedgehog signaling in response to Smoothened (Smo) agonist. Together, depletion of TCTN2 destabilizes other TZ proteins and considerably alters the localization of key transport and signaling-associated proteins, including IFT88, Arl13B, and Smo.

Keywords: CRISPR/Cas9, primary cilia, Sonic hedgehog signaling, transition zone

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Authors: Thauane Silva, Gabrielle do Vale, Andre Ferreira, Marilda Siqueira, Thiago Moreno L. Souza, Milene D. Miranda

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The exposure of A(H1N1)pdm09-infected epithelial cells (HeLa) to HIV-1 viral particles, or its gp120, enhanced interferon-induced transmembrane protein (IFITM3) content, a viral restriction factor (RF), resulting in a decrease in influenza replication. The gp120 binds to CCR5 (R5) or CXCR4 (X4) cell receptors during HIV-1 infection. Then, it is possible that the endogenous ligands of these receptors also modulate the expression of IFITM3 and other cellular factors that restrict influenza virus replication. Thus, the aim of this study is to analyze the role of cellular receptors R5 and X4 in modulating RFs in order to inhibit the replication of the influenza virus. A549 cells were treated with 2x effective dose (ED50) of endogenous R5 or X4 receptor agonists, CCL3 (20 ng/ml), CCL4 (10 ng/ml), CCL5 (10 ng/ml) and CXCL12 (100 ng/mL) or exogenous agonists, gp120 Bal-R5, gp120 IIIB-X4 and its mutants (5 µg/mL). The interferon α (10 ng/mL) and oseltamivir (60 nM) were used as a control. After 24 h post agonists exposure, the cells were infected with virus influenza A(H3N2) at 2 MOI (multiplicity of infection) for 1 h. Then, 24 h post infection, the supernatant was harvested and, the viral titre was evaluated by qRT-PCR. To evaluate IFITM3 and SAM and HD domain containing deoxynucleoside triphosphate triphosphohydrolase 1 (SAMHD1) protein levels, A549 were exposed to agonists for 24 h, and the monolayer was lysed with Laemmli buffer for western blot (WB) assay or fixed for indirect immunofluorescence (IFI) assay. In addition to this, we analyzed other RFs modulation in A549, after 24 h post agonists exposure by customized RT² Profiler Polymerase Chain Reaction Array. We also performed a functional assay in which SAMHD1-knocked-down, by single-stranded RNA (siRNA), A549 cells were infected with A(H3N2). In addition, the cells were treated with guanosine to assess the regulatory role of dNTPs by SAMHD1. We found that R5 and X4 agonists inhibited influenza replication in 54 ± 9%. We observed a four-fold increase in SAMHD1 transcripts by RFs mRNA quantification panel. After 24 h post agonists exposure, we did not observe an increase in IFITM3 protein levels through WB or IFI assays, but we observed an upregulation up to three-fold in the protein content of SAMHD1, in A549 exposed to agonists. Besides this, influenza replication enhanced in 20% in cell cultures that SAMDH1 was knockdown. Guanosine treatment in cells exposed to R5 ligands further inhibited influenza virus replication, suggesting that the inhibitory mechanism may involve the activation of the SAMHD1 deoxynucleotide triphosphohydrolase activity. Thus, our data show for the first time a direct relationship of SAMHD1 and inhibition of influenza replication, and provides perspectives for new studies on the signaling modulation, through cellular receptors, to induce proteins of great importance in the control of relevant infections for public health.

Keywords: chemokine receptors, gp120, influenza, virus restriction factors

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Authors: Dibyendu Das, Santanu Kumar Pal

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Milk protein lactoferrin (Lf) exhibits potent antibacterial activity due to its interaction with Gram-negative bacterial cell membrane component, lipopolysaccharide (LPS). This paper represents fabrication of new Liquid crystals (LCs) based biosensors to explore the interaction between Lf and LPS. LPS self-assembled at aqueous/LCs interface and orients interfacial nematic 4-cyano-4’- pentylbiphenyl (5CB) LCs in a homeotropic fashion (exhibiting dark optical image under polarized optical microscope). Interestingly, on the exposure of Lf on LPS decorated aqueous/LCs interface, an optical image of LCs changed from dark to bright indicating an ordering alteration of interfacial LCs from homeotropic to tilted/planar state. The ordering transition reflects strong binding between Lf and interfacial LPS that, in turn, perturbs the orientation of LCs. With the help of epifluorescence microscopy, we further affirmed the interfacial LPS-Lf binding event by imaging the presence of FITC tagged Lf at the LPS laden aqueous/LCs interface. Finally, we have investigated the conformational behavior of Lf in solution as well as in the presence of LPS using Circular Dichroism (CD) spectroscopy and further reconfirmed with Vibrational Circular Dichroism (VCD) spectroscopy where we found that Lf undergoes alpha-helix to random coil-like structure in the presence of LPS. As a whole the entire results described in this paper establish a robust approach to envisage the interaction between LPS and Lf through the ordering transitions of LCs at aqueous/LCs interface.

Keywords: endotoxin, interface, lactoferrin, lipopolysaccharide

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Authors: Ankan Roy, Niharika, Samir Kumar Patra

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Anomalous nexus of complex topological assemblies and spatiotemporal epigenetic choreography at chromosomal territory may forms the most sophisticated regulatory layer of gene expression in cancer. Colon cancer is one of the leading malignant neoplasms of the lower gastrointestinal tract worldwide. There is still a paucity of information about the complex molecular mechanisms of colonic cancerogenesis. Bioinformatics prediction and analysis helps to identify essential genes and significant pathways for monitoring and conquering this deadly disease. The present study investigates and explores potential hub genes as biomarkers and effective therapeutic targets for colon cancer treatment. Colon cancer patient sample containing gene expression profile datasets, such as GSE44076, GSE20916, and GSE37364 were downloaded from Gene Expression Omnibus (GEO) database and thoroughly screened using the GEO2R tool and Funrich software to find out common 2 differentially expressed genes (DEGs). Other approaches, including Gene Ontology (GO) and KEGG pathway analysis, Protein-Protein Interaction (PPI) network construction and hub gene investigation, Overall Survival (OS) analysis, gene correlation analysis, methylation pattern analysis, and hub gene-Transcription factors regulatory network construction, were performed and validated using various bioinformatics tool. Initially, we identified 166 DEGs, including 68 up-regulated and 98 down-regulated genes. Up-regulated genes are mainly associated with the Cytokine-cytokine receptor interaction, IL17 signaling pathway, ECM-receptor interaction, Focal adhesion and PI3K-Akt pathway. Downregulated genes are enriched in metabolic pathways, retinol metabolism, Steroid hormone biosynthesis, and bile secretion. From the protein-protein interaction network, thirty hub genes with high connectivity are selected using the MCODE and cytoHubba plugin. Survival analysis, expression validation, correlation analysis, and methylation pattern analysis were further verified using TCGA data. Finally, we predicted COL1A1, COL1A2, COL4A1, SPP1, SPARC, and THBS2 as potential master regulators in colonic cancerogenesis. Moreover, our experimental data highlights that disruption of lipid raft and RAS/MAPK signaling cascade affects this gene hub at mRNA level. We identified COL1A1, COL1A2, COL4A1, SPP1, SPARC, and THBS2 as determinant hub genes in colon cancer progression. They can be considered as biomarkers for diagnosis and promising therapeutic targets in colon cancer treatment. Additionally, our experimental data advertise that signaling pathway act as connecting link between membrane hub and gene hub.

Keywords: hub genes, colon cancer, DNA methylation, epigenetic engineering, bioinformatic predictions

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Authors: Muhammad Arsalan Mahmoo, Allah Rakha, Muhammad Sohail

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The supplementation of wheat flour with high lysine legume flours has positive effects on the nutritional value of bread. In present study, germinated and terminated soya flour blends were prepared and supplemented in bread in variable proportions (10 % and 20 % of each) to check its impact on quality and sensory attributes of bread. The results showed that there was a significant increase in protein, ash and crude fat contents due to increase in the level of germinated and ungerminated soya flour. However, the moisture and crude fiber contents of composite flours containing germinated and ungerminated soya flour decreased with increased level of supplementation. Mean values for physical analysis (loaf volume, specific volume, weight loss and force for texture) were significantly higher in breads prepared with germinated soya bean flour.The scores assigned to sensory parameters of breads like volume, color of crust, symmetry, color of crumb, texture, taste and aroma decreased significantly by increasing the level of germinated and ungerminated soya flour in wheat flour while color of crust and taste slightly improved. The scores given to overall acceptability of bread prepared from composite flour supplemented with 10 % germinated soya flour.

Keywords: composite bread, protein energy malnutrition, supplementation, amino acid profile, grain legumes

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Authors: Mamadou Sileye Niang

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The study is a contribution to the development of a feed for juvenile tilapia Oreochromis niloticus, from local raw materials in order to reduce the cost of feeding farmed tilapia in Senegal. Three feeds were formulated from local raw materials. The basic composition of the tested feeds is as follows: A1 (peanut meal, rice bran, millet bran, maize meal and no fish meal); A2 (peanut meal, rice bran, millet bran, maize meal and 10% fish meal) and A3 (peanut meal, rice bran, millet bran, maize meal and 25% fish meal). All feeds contain 31% protein. The trial compared three batches, in 2 replicates, with different diets. The initial weight of the juveniles was 0.37± 0.5g. The daily ration was distributed at 9 am and 4 pm. After 90 days of the experiment, the final mean weights were 2.45 ± 0.5g; 2.75±0.5g; and 4.67 ± 0.5g for A1, A2, and A3, respectively. A performance test, of which the objective was to compare growth parameters, was conducted. The results of the growth parameters of juveniles fed A3 were significantly higher (p < 0.05) than those fed A1 and A2. The weight growth study shows similar growth during the first month. However, from this date onwards, juveniles fed A3 show a faster growth, which is maintained throughout the experiment. On the other hand, the Protein Efficiency Coefficient and the Survival Rate showed no significant difference. The zootechnical parameters are not significantly different (p > 0.05) between the two tanks for the same feed treatment.

Keywords: nutrition, feed, fingerlings, Oreochromis, local raw materials, feed cost

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Authors: Harithas Aruchchunan

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Silage production is an essential aspect of dairy farming, used to provide high-quality feed to ruminants. However, dairy farmers in Northern Province Sri Lanka are facing multiple challenges that compromise the quality and quantity of silage produced. To tackle these challenges, promoting silage feeding has become an essential component of sustainable dairy farming practices. In this study, silage bale samples were collected from a newly started silage baling factory in Jaffna, Northern province and their quality was analysed at the Veterinary Research Institute laboratory in Kandy in March 2023. The results show the nutritional composition of three Napier grass cultivars: Super Napier, CO6, and Indian Red Napier (BH18). The main parameters analysed were dry matter, pH, lactic acid, soluble carbohydrate, ammonia nitrogen, ash, crude protein, NDF, and ADF. The results indicate that Super Napier and CO6 have higher crude protein content and lower ADF levels, making them suitable for producing high-quality silage. The pH levels of all three cultivars were safe, and the ammonia nitrogen levels were considered appropriate. However, laboratory results indicate that the quality of silage bales produced can be further enhanced. Dairy farmers should be encouraged to adopt these cultivars to achieve better yields as they are high in protein and are better suited to Northern Province's soil and climate. Therefore, it is vital to educate small-scale fodder producers, who supply the raw material to silage factories, on the best practices of cultivating these new cultivars. To improve silage bale production and quality in Northern Province Sri Lanka, we recommend increasing public awareness about silage feeding, providing education and training to dairy farmers and small-scale fodder producers on modern silage production techniques and improving the availability of raw materials for silage production. Additionally, Napier grass cultivars need to be promoted among dairy farmers for better production and quality of silage bales. Failing to improve the quality and quantity of silage bale production could not only lead to the decline of dairy farming in Northern Province Sri Lanka but also the negative impact on the economy

Keywords: silage bales, dairy farming, economic crisis, Sri Lanka

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Authors: Anjana Goel, Ashok Kumar Bhatia

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Ocimum sanctum, a well known medicinal plant is used for various alignments in Ayurvedic medicines. It was found to be effective in treating the humans suffering from different viral infections like chicken pox, small pox, measles and influenza. In addition, curative effect of the plant in malignant patients was also reported. In the present study, leaves of this plant were screened against animal viruses i.e. Bovine Herpes Virus-type-1 (BHV-1), Foot and Mouth disease virus (FMDV) and Newcastle Disease Virus (NDV). BHV-1 and FMDV were screened in MDBK and BHK cell lines respectively using cytopathic inhibition test. While NDV was propagated in chick embryo fibroblast culture and tested by haemagglutination inhibition test. Maximum non toxic dose of aqueous extract of Ocimum sanctum leaves was calculated by MTT assay in all the cell cultures and nontoxic doses were used for antiviral activity against viruses. 98.4% and 85.3% protection were recorded against NDV and BHV-1 respectively. However, Ocimum sanctum extract failed to show any inhibitory effect on the cytopathic effect caused by FMD virus. It can be concluded that Ocimum sanctum is a very effective remedy for curing viral infections in animals also.

Keywords: bovine herpes virus-type-1, foot and mouth disease virus, newcastle disease virus, Ocimum sanctum

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Authors: Elif Tugce Aksun Tumerkan

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Fluorescent proteins (FPs) have become very popular since green fluorescent protein discovered from crystal jellyfish. It is known that Anthozoa species have a wide range of chromophore organisms, and the initial crystal structure for non-fluorescent chromophores obtained from the reef-building coral has been determined. There are also differently coloured pigments in non-bioluminescent Anthozoa zooxanthellate and azooxanthellate which are frequently members of the GFP-like protein family. The development of fluorescent proteins (FPs) and their applications is an outstanding example of basic science leading to practical biotechnological and medical applications. Fluorescent proteins have several applications in science and are used as important indicators in molecular biology and cell-based research. With rising interest in cell biology, FPs have used as biosensor indicators and probes in pharmacology and cell biology. Using fluorescent proteins in genetically encoded metabolite sensors has many advantages than chemical probes for metabolites such as easily introduced into any cell or organism in any sub-cellular localization and giving chance to fixing to fluoresce of different colours or characteristics. There are different factors effects to signalling mechanism when they used as a biosensor. While there are wide ranges of research have been done on the significance and applications of fluorescent proteins, the cell signalling response of FPs and target cell are less well understood. In this study, it was aimed to clarify the response of adaptive mechanisms of coral species such as pH, temperature and symbiotic relationship and target cells properties on the signalling capacity. Corals are a rich natural source of fluorescent proteins that change with environmental conditions such as light, heat stress and injury. Adaptation mechanism of coral species to these types of environmental variations is important factor due to FPs properties have affected by this mechanism. Since fluorescent proteins obtained from nature, their own ecological property like the symbiotic relationship is observed very commonly in coral species and living conditions have the impact on FPs efficiency. Target cell properties also have an effect on signalling and visualization. The dynamicity of detector that used for reading fluorescence and the level of background fluorescence are key parameters for the quality of the fluorescent signal. Among the factors, it can be concluded that coral species adaptive characteristics have the strongest effect on FPs signalling capacity.

Keywords: biosensor, cell biology, environmental conditions, fluorescent protein, sea anemone

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Authors: Muhammad Ali Siddiquee, Md. Alauddin, Md. Omar Faruque, Zakir Hossain Howlader, Mohammad Asaduzzaman

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Rice bran (RB) and rice bran oil (RBO) are explored as prominent food components worldwide. In this study, fermented rice bran (FRB) was produced by employing edible gram-positive bacteria (Lactobacillus acidophilus, Lactobacillus bulgaricus, and Bifidobacterium bifidum) at 125 x 10⁵ spore g⁻¹ of rice bran, and investigated to evaluate nutritional quality. The crude rice bran oil (CRBO) was extracted from RB, and its quality was also investigated compared to market-available rice bran oil (MRBO) in Bangladesh. We found that fermentation of rice bran with lactic acid bacteria increased total proteins (29.52%), fat (5.38%), ash (48.47%), crude fiber (38.96%), and moisture (61.04%) and reduced the carbohydrate content (36.61%). We also found that essential amino acids (methionine, tryptophan, threonine, valine, leucine, lysine, histidine, and phenylalanine) and non-essential amino acids (alanine, aspartate, glycine, glutamine, proline, serine, and tyrosine) were increased in FRB except methionine and proline. Moreover, total phenolic content, tannin content, flavonoid content, and antioxidant activity were increased in FRB. The RBO analysis showed that γ-oryzanol content (10.00mg/g) was found in CRBO compared to MRBO (ranging from 7.40 to 12.70 mg/g) and Vitamin-E content 0.20% was found higher in CRBO compared to MRBO (ranging 0.097 to 0.12%). The total saturated (25.16%) and total unsaturated fatty acids (74.44%) were found in CRBO, whereas MRBO contained total saturated (22.08 to 24.13%) and total unsaturated fatty acids (71.91 to 83.29%), respectively. The physiochemical parameters were found satisfactory in all samples except acid value and peroxide value higher in CRBO. Finally, animal experiments showed that FRB and CRBO reduce the body weight, glucose, and lipid profile in high-fat diet-induced animal models. Thus, FRB and RBO could be value-added food supplements for human health.

Keywords: fermented rice bran, crude rice bran oil, amino acids, proximate composition, gamma-oryzanol, fatty acids, heavy metals, physiochemical parameters

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Authors: Alan Luo, Hunter N. B. Moseley

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Many macromolecular structure entries in the Protein Data Bank (PDB) have a range of regional (localized) quality issues, be it derived from x-ray crystallography, Nuclear Magnetic Resonance (NMR) spectroscopy, or other experimental approaches. However, most PDB entries are judged by global quality metrics like R-factor, R-free, and resolution for x-ray crystallography or backbone phi-psi distribution statistics and average restraint violations for NMR. Regional quality is often ignored when PDB entries are re-used for a variety of structurally based analyses. The binding of ligands, especially ligands involved in energy metabolism, is of particular interest in many structurally focused protein studies. Using a regional quality metric that provides chemically interpretable information from electron density maps, a significant number of outliers in regional structural quality was detected across x-ray crystallographic PDB entries for proteins bound to biochemically critical ligands. In this study, a series of analyses was performed to evaluate both specific and general potential factors that could promote these outliers. In particular, these potential factors were the minimum distance to a metal ion, the minimum distance to a crystal contact, and the isotropic atomic b-factor. To evaluate these potential factors, Fisher’s exact tests were performed, using regional quality criteria of outlier (top 1%, 2.5%, 5%, or 10%) versus non-outlier compared to a potential factor metric above versus below a certain outlier cutoff. The results revealed a consistent general effect from region-specific normalized b-factors but no specific effect from metal ion contact distances and only a very weak effect from crystal contact distance as compared to the b-factor results. These findings indicate that no single specific potential factor explains a majority of the outlier ligand-bound regions, implying that human error is likely as important as these other factors. Thus, all factors, including human error, should be considered when regions of low structural quality are detected. Also, the downstream re-use of protein structures for studying ligand-bound conformations should screen the regional quality of the binding sites. Doing so prevents misinterpretation due to the presence of structural uncertainty or flaws in regions of interest.

Keywords: biomacromolecular structure, coenzyme, electron density discrepancy analysis, x-ray crystallography

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