Search results for: wild rats

Authors: Mekonen Hailu, Liu Liping

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Cage aquaculture is highly preferred due to higher production per unit volume of water, lower costs of investment, and simpler routine farm management procedures compared to pond systems. In the 1980s, cage culture was first used on a trial basis in sub-Saharan Africa. Over the past 20 years, a small number of prosperous freshwater cage culture operations have started to emerge in Egypt, Rwanda, Kenya, Uganda, Tanzania, Ghana, Malawi, Zambia and Zimbabwe. Brackish and marine cage culture also offers a lot of potential, although this subsector hasn't seen any significant commercial growth to date. In 2019, 263 cage aquaculture installations on the African inland waters on 18 water bodies within eight countries with an estimated 20,114 cages were reported. The lakes Victoria, Kariba, Volta, and River Volta, which together account for 82.9% of all cage aquaculture installations regarded as sub-Saharan Africa's principal cage aquaculture regions (Fig 1). Except few small-scale trials with North African catfish (Clarias gariepinus), almost all farms in Sub-Saharan Africa and Egypt grow Nile tilapia (Oreochromis niloticus). More than 247,398 tonnes of fish are produced yearly from ten African countries through cage aquaculture. The expansion of cage culture in Africa provides job opportunities for both skilled and unskilled workers, nutritious food and foreign currency. The escaping non-native strains of tilapia in Lake Volta and the occurrence of a risky Tilapia lake virus (syncytial hepatitis), which has the potential to wipe out entire populations in both wild and farmed Nile tilapia on Lake Victoria, are threats coming with the expansion of cage aquaculture in Africa. In addition, the installations of 138 cage aquacultures were found in contrary to best cage culture practices. To sustain cage aquaculture development and maintain harmony with other water uses, developers must strictly abide by best practices. Hence, the exclusion of protected areas and small lakes (average depth 5 m or less) should be done, as well an Environmental Impact Assessment should be conducted before establishing the cage farms.

Keywords: Africa, cage aquaculture, production, threats

Procedia PDF Downloads 33

Authors: Liaqat Ali, Hubert E. Blum, Türkân Sakinc

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Enterococcus faecium is an emerging multidrug-resistant nosocomial pathogen increased dramatically worldwide and causing bacteremia, endocarditis, urinary tract and surgical site infections in immunocomprised patients. The capsular polysaccharides that contribute to pathogenesis through evasion of the host innate immune system are also involved in hindering leukocyte killing of enterococci. The gene cluster (enterococcal polysaccharide antigen) of E. faecalis encoding homologues of many genes involved in polysaccharide biosynthesis. We identified two putative loci with 22 kb and 19 kb which contained 11 genes encoding for glycosyltransferases (GTFs); this was confirmed by using genome comparison of already sequenced strains that has no homology to known capsule genes and the epa-locus. The polysaccharide-conjugate vaccines have rapidly emerged as a suitable strategy to combat different pathogenic bacteria, therefore, we investigated a polysaccharide biosynthesis CapD protein in E. faecium contains 336 amino acids and had putative function for N-linked glycosylation. The deletion/knock-out capD mutant was constructed and complemented by homologues recombination method and confirmed by using PCR and sequencing. For further characterization and functional analysis, in-vitro cell culture and in-vivo a mouse infection models were used. Our ΔcapD mutant shows a strong hydrophobicity and all strains exhibited biofilm production. Subsequently, the opsonic activity was tested in an opsonophagocytic assay which shows increased in mutant compared complemented and wild type strains but more than two fold decreased in colonization and adherence was seen on surface of uroepithelial cells. However, a significant higher bacterial colonialization was observed in capD mutant during animal bacteremia infection. Unlike other polysaccharides biosynthesis proteins, CapD does not seems to be a major virulence factor in enterococci but further experiments and attention is needed to clarify its function, exact mechanism and involvement in pathogenesis of enteroccocal nosocomial infections eventually to develop a vaccine/ or targeted therapy.

Keywords: E. faecium, pathogenesis, polysaccharides, biofilm formation

Procedia PDF Downloads 298

Authors: Yan Lu, Song Hong, Janakiraman Udaiyappan, Aarti Nagayach, Quoc-Viet A. Duong, Masao Morita, Shun Saito, Yuichi Kobayashi, Yuhai, Zhao, Hongying Peng, Nicholas B. Pham, Walter J Lukiw, Christopher A. Vuong, Nicolas G. Bazan

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Aims: Neurodegeneration and behavior dysfunction occurs in patients with Alzheimer's Disease (AD), and as the disease progresses many patients develop cognitive impairment. 5XFAD mouse model of AD is widely used to study AD pathogenesis and treatment. This study aimed to investigate the effect of maresin like 1 (MaR-L1) treatment in AD pathology using 5XFAD mice. Methods: We tested 12-month-old male 5XFAD mice and wild type control mice treated with MaR-L1 in a battery of behavioral tasks. We performed open field test, beam walking test, clasping test, inverted grid test, acetone test, marble burring test, elevated plus maze test, cross maze test and novel object recognition test. We also studied neuronal loss, amyloid β burden, and inflammation in the brains of 5XFAD mice using immunohistology and Western blotting. Results: MaR-L1 treatment to the 5XFAD mice showed improved cognitive function of 5XFAD mice. MaR-L1 showed decreased anxiety behavior in open field test and marble burring test, increased muscular strength in the beam walking test, clasping test and inverted grid test. Cognitive function was improved in MaR-L1 treated 5XFAD mice in the novel object recognition test. MaR-L1 prevented neuronal loss and aberrant inflammation. Conclusion: Our finding suggests that behavioral abnormalities were normalized by the administration of MaR-L1 and the neuroprotective role of MaR-L1 in the AD. It also indicates that MaR-L1 treatment is able to prevent and or ameliorate neuronal loss and aberrant inflammation. Further experiments to validate the results are warranted using other AD models in the future.

Keywords: Alzheimer's disease, motor and cognitive behavior, 5XFAD mice, Maresin Like 1, microglial cell, astrocyte, neurodegeneration, inflammation, resolution of inflammation

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Authors: Nolitha L. Skenjana, Bongani P. Kubheka, Maxwell A. Poswal

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The Eastern Cape Province of South Africa is characterized by subsistence farming, which is mostly distributed in the rural areas of the province. It is estimated that cereal crops such as maize and sorghum, and vegetables such as cabbage are grown in more than 400.000 rural households, with maize being the most dominant crop. However, compared to commercial agriculture, small-scale farmers receive minimal support from research and development, limited technology transfer on the latest production practices and systems and have poor production infrastructure and equipment. Similarly, there is limited farmers' appreciation on best practices in insect pest management and control. The paper presents findings from the primary literature and personal observations on insect pest management practices of small-scale farmers in the province. Inferences from literature and personal experiences in the production areas have led to a number of deductions regarding the level of exposure and extent of vulnerability. Farmers' pest management practices, which included not controlling at all though there is a pest problem, resulted in their crop stands to be more vulnerable to pest attacks. This became more evident with the recent brown locust, African armyworm, and Fall armyworm outbreaks, and with the incidences of opportunistic phytophagous insects previously collected on wild hosts only, found causing serious damages on crops. In most of these occurrences, damage to crops resulted in low or no yield. Improvements on farmers' reaction and response to pest problems were only observed in areas where focused awareness campaigns and trainings on specific pests and their management techniques were done. This then calls for a concerted effort from all role players in the sphere of small-scale crop production, to train and equip farmers with relevant skills, and provide them with information on affordable and climate-smart strategies and technologies in order to create a state of preparedness. This is necessary for the prevention of substantial crop losses that may exacerbate food insecurity in the province.

Keywords: Eastern Cape Province, small-scale farmers, insect pest management, vulnerability

Procedia PDF Downloads 118

Authors: Asif Bilal, Fouzia Tanvir, Sibtain Ahmad

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Estrogen receptor alpha, also known as estrogen receptor-1, is highly involved in risk of mammary carcinoma. The objectives of this study were to identify non-synonymous SNPs of estrogen receptor and their association with breast cancer and to identify the chemotherapeutic responses of phytochemicals against it via in-silico study design. For this purpose, different online tools. to identify pathogenic SNPs the tools were SIFT, Polyphen, Polyphen-2, fuNTRp, SNAP2, for finding disease associated SNPs the tools SNP&GO, PhD-SNP, PredictSNP, MAPP, SNAP, MetaSNP, PANTHER, and to check protein stability Mu-Pro, I-Mutant, and CONSURF were used. Post-translational modifications (PTMs) were detected by Musitedeep, Protein secondary structure by SOPMA, protein to protein interaction by STRING, molecular docking by PyRx. Seven SNPs having rsIDs (rs760766066, rs779180038, rs956399300, rs773683317, rs397509428, rs755020320, and rs1131692059) showing mutations on I229T, R243C, Y246H, P336R, Q375H, R394S, and R394H, respectively found to be completely deleterious. The PTMs found were 96 times Glycosylation; 30 times Ubiquitination, a single time Acetylation; and no Hydroxylation and Phosphorylation were found. The protein secondary structure consisted of Alpha helix (Hh) is (28%), Extended strand (Ee) is (21%), Beta turn (Tt) is 7.89% and Random coil (Cc) is (44.11%). Protein-protein interaction analysis revealed that it has strong interaction with Myeloperoxidase, Xanthine dehydrogenase, carboxylesterase 1, Glutathione S-transferase Mu 1, and with estrogen receptors. For molecular docking we used Asiaticoside, Ilekudinuside, Robustoflavone, Irinoticane, Withanolides, and 9-amin0-5 as ligands that extract from phytochemicals and docked with this protein. We found that there was great interaction (from -8.6 to -9.7) of these ligands of phytochemicals at ESR1 wild and two mutants (I229T and R394S). It is concluded that these SNPs found in ESR1 are involved in breast cancer and given phytochemicals are highly helpful against breast cancer as chemotherapeutic agents. Further in vitro and in vivo analysis should be performed to conduct these interactions.

Keywords: breast cancer, ESR1, phytochemicals, molecular docking

Procedia PDF Downloads 45

Authors: Himanshu Kushwah, Nidhi Sandal, Meenakshi K. Chauhan, Gaurav Mittal

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Excessive bleeding is the primary factor of avoidable death in both civilian trauma centers as well as the military battlefield. Hundreds of Indian troops die every year due to blood loss caused by combat-related injuries. These deaths are avoidable and can be prevented to a large extent by making available a suitable hemostatic dressing in an emergency medical kit. In this study, natural gums were evaluated as potential hemostatic agents in combination with calcium gluconate. The study compares the hemostatic activity of Gum Tragacanth (GT), Guar Gum (GG), Xanthan Gum (XG) and Gum Acacia (GA) by carrying out different in-vitro and in-vivo studies. In-vitro studies were performed using the Lee-White method and Eustrek method, which includes the visual and microscopic analysis of blood clotting. MTT assay was also performed using human lymphocytes to check the cytotoxicity of the gums. The in-vivo studies were performed in Sprague Dawley rats using tail bleeding assay to evaluate the hemostatic efficacy of the gums and compared with a commercially available hemostatic sponge, Surgispon. Erythrocyte agglutination test was also performed to check the interaction between blood cells and the natural gums. Other parameters like blood loss, adherence strength of the developed hemostatic dressing material incorporating these gums, re-bleeding, and survival of the animals were also studied. The data obtained from the MTT assay showed that Guar gum, Gum Tragacanth, and Gum Acacia were not significantly cytotoxic, but substantial cytotoxicity was observed in Xanthan gum samples at high concentrations. Also, Xanthan gum took the least time with its minimum concentration to achieve hemostasis, (approximately 50 seconds at 3mg concentration). Gum Tragacanth also showed efficient hemostasis at a concentration of 35mg at the same time, but the other two gums tested were not able to clot the blood in significantly less time. A sponge dressing made of Tragacanth gum was found to be more efficient in achieving hemostasis and showed better practical applicability among all the gums studied and also when compared to the commercially available product, Surgispon, thus making it a potentially better alternative.

Keywords: cytotoxicity, hemostasis, natural gums, sponge

Procedia PDF Downloads 126

Authors: Rania F. Ahmed, Sally A. El Awdan, Gehad A. Abdel Jaleel, Dalia O. Saleh, Omar A. H. Ahmed-Farid

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The metabolic syndrome is an important public health concern often related to obesity, improper diet, and sedentary lifestyles and can predispose individuals to the development of many dangerous health conditions, disability and early death. This research aimed to investigate the efficacy of resveratrol (RSV) to reverse the neuro-complications associated with metabolic syndrome experimentally-induced in rats using an eight weeks high fat, high fructose diet (HFHF) model. The corresponding drug treatments were administered orally during the last 10 days of the diet. Behavioural tests namely the open field test (OFT) and the forced swimming test (FST) were conducted. Brain levels of monoamines viz. serotonin, norepinephrine and dopamine as well as their metabolites were assessed. 8-hydroxyguanosine (8-OHDG) as an indicative of DNA-fragmentation, nitric oxide (NOx) and tumor necrosis factor-α (TNF- α) were estimated. Finally, brain antioxidant parameters namely malondialdehyde (MDA), reduced and oxidized glutathione (GSH, GSSG) were evaluated. HFHF-induced metabolic syndrome resulted in decreased activity in the OFT and increased immobility duration in the FST. Furthermore, HFHF-induced metabolic syndrome lead to a significant increase in brain monoamines turn over as well as elevation in 8-OHDG, NOx, TNF- α, MDA and GSSG; and reduction in GSH. Ten days daily treatment with RSV (20 and 40 mg/kg p.o) dose dependently increased activity in the OFT and decreased immobility duration in the FST. Moreover, RSV normalized brain monoamines contents, reduced 8-OHDG, NOx, TNF- α, MDA and GSSG; and elevated GSH. In conclusion, we can say that RSV showed neuro-protective properties against HFHF-induced metabolic syndrome represented by monoamines preservation, prevention of neurodegeneration, anti-inflammatory and antioxidant potentials and could be recommended as a beneficial daily dietary supplement to treat the neuronal side effects associated with HFHF-induced metabolic syndrome.

Keywords: antioxidants, DNA-fragmentation, forced swimming test, HFHF-induced metabolic syndrome, monoamines, nitric oxide (NOx), open field, resveratrol, tumor necrosis factor-α (TNF- α), 8-hydroxyguanosine (8-OHDG)

Procedia PDF Downloads 254

Authors: Eryati Darwin, Eka Fithra Elfi, Indria Hafizah

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Background: Physical exercise induces a pattern of hormonal and immunological responses that prevent endothelial dysfunction by maintaining the availability of nitric oxide (NO). Regular and moderate exercise stimulates NO release, that can be considered as protective factor of cardiovascular diseases, while strenuous exercise induces increased levels in a number of pro-inflammatory and anti-inflammatory cytokines. Pro-inflammatory cytokines tumor necrosis factor-α (TNF-α) triggers endothelial activation which results in an increased vascular permeability. Nuclear gene factor kappa B (NF-κB) activates biological effect of TNF-α. Aim of Study: To determine the effect of physical exercise on the endothelial and skeletal muscle, we measured the level of NF-κB on rats’ serum, macrophages, and myocytes after strenuous physical exercise. Methods: 30 male Rattus norvegicus in the age of eight weeks were randomly divided into five groups (each containing six), and there were treated groups (T) and control group (C). The treated groups obtain strenuous physical exercise by ran on treadmill at 32 m/minutes for 1 hour or until exhaustion. Blood samples, myocytes of gastrocnemius muscle, and intraperitoneal macrophages were collected sequentially. There were investigated immediately, 2 hours, 6 hours, and 24 hours (T1, T2, T3, and T4) after sacrifice. The levels of NF-κB were measured by ELISA methods. Results: From our study, we found that the levels of NF-κB on myocytes in treated group from which its specimen was taken immediately (T1), 2 hours after treadmill (T2), and 6 hours after treadmill (T3) were significantly higher than control group (p<0.05), while the group from which its specimen was taken 24 hours after treadmill, was no significantly different (p>0.05). Also on macrophages, NF-κB in treated groups T1, T2, and T3 was significantly higher than control group (p<0.05), but there was no difference between T4 and control group (p>0.05). The level of serum NF-κB was not significantly different between treatment group as well as compared to control group (p>0.05). Serum NF-κB was significantly higher than the level on macrophages and myocytes (p<0.05). Conclusion: This study demonstrated that strenuous physical exercise stimulates the activation of NF-κB that plays a role in vascular inflammation and muscular damage, and may be recovered after resting period.

Keywords: endothelial function, inflammation, NFkB, physical exercise

Procedia PDF Downloads 239

Authors: Sunghee Nam

Abstract:

Entomopathogenic fungi is a Cordyceps species that is isolated from dead silkworm and cicada. Fungi on cicadas were described in old Chinese medicinal books and from ancient times, vegetable wasps and plant worms were widely known to have active substance and have been studied for pharmacological use. Among many fungi belonging to the genus Cordyceps, Cordyceps sinensis have been demonstrated to yield natural products possessing various biological activities and many bioactive components. Generally, It is commonly used to replenish the kidney and soothe the lung, and for the treatment of fatigue. Due to their commercial and economic importance, the demand for Cordyceps has been rapidly increased. However, a supply of Cordyceps specimen could not meet the increasing demand because of their sole dependence on field collection and habitat destruction. Because it is difficult to obtain many insect hosts in nature and the edibility of host insect needs to be verified in a pharmacological aspect. Recently, this setback was overcome that P. tenuipes was able to be cultivated in a large scale using silkworm as host. Pharmacological effects of P. tenuipes cultured on silkworm such as strengthening immune function, anti-fatigue, anti-tumor activity and controlling liver etc. have been proved. They are widely commercialized. In this study, we attempted to establish a method for stable growth inhibition of P. tenuipes on silkworm hosts and an optimal condition for synnemata formation. To determine optimum culturing conditions, temperature and light conditions were varied. The length and number of synnemata was highest at 25℃ temperature and 100~300 lux illumination. On an average, the synnemata of wild P. tenuipes measures 70 ㎜ in length and 20 in number; those of the cultured strain were relatively shorter and more in number. The number of synnemata may have increased as a result of inoculating the host with highly concentrated conidia, while the length may have decreased due to limited nutrition per individual. It is not able that changes in light illumination cause morphological variations in the synnemata. However, regulation of only light and temperature could not produce stromata like perithecia, asci, and ascospores.

Keywords: optimization of culture conditions of paecilomyces tenuipes, entomopathogenic fungi optimization of culture conditions of paecilomyces tenuipes, entomopathogenic fungi silkworm larva, bombyx mori

Procedia PDF Downloads 234

Authors: Jyh-Ping Chen, Chia-Lin Sheu

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In this study, we propose to use electrospinning to fabricate porous nanofibrous membranes as postsurgical anti-adhesion barriers and to improve the properties of current post-surgical anti-adhesion products. We propose to combine FDA-approved biomaterials with anti-adhesion properties, polycaprolactone (PCL), polyethylene glycol (PEG), hyaluronic acid (HA) with silver nanoparticles (Ag) and ibuprofen (IBU), to produce anti-adhesion barrier nanofibrous membranes. For this purpose, PEG/PCL/Ag/HA/IBU core-shell nanofibers were prepared. The shell layer contains PEG + PCL to provide mechanical supports and Ag was added to the outer PEG-PCL shell layer during electrospinning to endow the nanofibrous membrane with anti-bacterial properties. The core contains HA to exert anti-adhesion and IBU to exert anti-inflammation effects, respectively. The nanofibrous structure of the membranes can reduce cell penetration while allowing nutrient and waste transports to prevent postsurgical adhesion. Nanofibers with different core/shell thickness ratio were prepared. The nanofibrous membranes were first characterized for their physico-chemical properties in detail, followed by in vitro cell culture studies for cell attachment and proliferation. The HA released from the core region showed extended release up to 21 days for prolonged anti-adhesion effects. The attachment of adhesion-forming fibroblasts is reduced using the nanofibrous membrane from DNA assays and confocal microscopic observation of adhesion protein vinculin expression. The Ag released from the shell showed burst release to prevent E Coli and S. aureus infection immediately and prevent bacterial resistance to Ag. Minimum cytotoxicity was observed from Ag and IBU when fibroblasts were culture with the extraction medium of the nanofibrous membranes. The peritendinous anti-adhesion model in rabbits and the peritoneal anti-adhesion model in rats were used to test the efficacy of the anti-adhesion barriers as determined by gross observation, histology, and biomechanical tests. Within all membranes, the PEG/PCL/Ag/HA/IBU core-shell nanofibers showed the best reduction in cell attachment and proliferation when tested with fibroblasts in vitro. The PEG/PCL/Ag/HA/IBU nanofibrous membranes also showed significant improvement in preventing both peritendinous and peritoneal adhesions when compared with other groups and a commercial adhesion barrier film.

Keywords: anti-adhesion, electrospinning, hyaluronic acid, ibuprofen, nanofibers

Procedia PDF Downloads 157

Authors: M. R. Aher, R. B. Laware, G. S. Asane, B. S. Kuchekar

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Objective: Studies have shown that a new combination therapy with Artemisinin derivatives and curcumin is unique, with potential advantages over known ACTs. In present study an attempt was made to prepare microparticulate drug delivery system of Curcumin-Zn complex and evaluate it in combination with artemether for antimalarial activity. Material and method: Curcumin Zn complex was prepared and encapsulated using sodium alginate. Microparticles thus obtained are further coated with various enteric polymers at different coating thickness to control the release. Microparticles are evaluated for encapsulation efficiency, drug loading and in vitro drug release. Roentgenographic Studies was conducted in rabbits with BaSO 4 tagged formulation. Optimized formulation was screened for antimalarial activity using P. berghei-infected mice survival test and % paracetemia inhibition, alone (three oral dose of 5mg/day) and in combination with arthemether (i.p. 500, 1000 and 1500µg). Curcumin-Zn(II) was estimated in serum after oral administration to rats by using spectroflurometry. Result: Microparticles coated with Cellulose acetate phthalate showed most satisfactory and controlled release with 479 min time for 60% drug release. X-ray images taken at different time intervals confirmed the retention of formulation in GI tract. Estimation of curcumin in serum by spectroflurometry showed that drug concentration is maintained in the blood for longer time with tmax of 6 hours. The survival time (40 days post treatment) of mice infected with P. berghei was compared to survival after treatment with either Curcumin-Zn(II) microparticles artemether combination, curcumin-Zn complex and artemether. Oral administration of Curcumin-Zn(II)-artemether prolonged the survival of P.berghei-infected mice. All the mice treated with Curcumin-Zn(II) microparticles (5mg/day) artemether (1000µg) survived for more than 40 days and recovered with no detectable parasitemia. Administration of Curcumin-Zn(II) artemether combination reduced the parasitemia in mice by more than 90% compared to that in control mice for the first 3 days after treatment. Conclusion: Antimalarial activity of the curcumin Zn-artemether combination was more pronounced than mono therapy. A single dose of 1000µg of artemether in curcumin-Zn combination gives complete protection in P. berghei-infected mice. This may reduce the chances of drug resistance in malaria management.

Keywords: formulation, microparticulate drug delivery, antimalarial, pharmaceutics

Procedia PDF Downloads 372

Authors: L. Musak, J. Valachova, T. Vasicko, O. Osina

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Introduction: Stainless steel is resistant to electrochemical corrosion by passivation. Welders are greatly exposed to welding fumes of toxic metals, which added to this steel. The content of chromium (Cr) in steel was above 11.5%, Ni and Mo from 2 to 6.5%. The aim of the study was the evaluation of occupational exposure to Cr, chromosome analysis and valuation of individual susceptibility polymorphism of gene CCND1 c.870 G>A. Materials and Methods: The exposed group was consisted from 117 welders of stainless steels. The average age was 38.43 years and average exposure time 7.14 years. Smokers represented 40.17%. The control group consisted of 123 non-exposed workers with an average age of 39.74 years and time employment 16.67 years. Smokers accounted for 22.76%. Analysis of Cr in blood and urine was performed by atomic absorption spectrophotometry (AAS Varian SpectraAA 30P) with electrothermal decomposition of the sample in the graphite furnace. For the evaluation of chromosomal aberrations (CA) was used cytogenetic analysis of peripheral blood lymphocytes, gene polymorphism was determined by PCR-RFLP reaction using appropriate primers and restriction enzymes. For statistical analysis was used the Mann-Whitney U-test. Results: The mean Cr level in exposed group was 0.095 mmol/l (0.019 min-max 0.504). No value does exceed the average normal value. The average value Cr in urine was 7.9 mmol/mol creatinine (min 0.026 to max 19.26). The total number of CA was 1.86% in compared to 1.70% controls. (CTA-type 0.90% vs 0.80% and CSA-type 0.96% vs 0.90%). In the number of total CA was observed statistical difference between smokers and non-smokers of exposed group (S-1.57% vs. NS-2.04%, P<0.05). In CCND1 gene polymorphisms was observed the increasing of the total CA with wild-type allele (WT) via heterozygous to the VAR genotype (1.44%<1.82%<2.13%). There was observed a statistically higher incidence of CTA-type aberrations in variant genotypes between exposed and control groups (1.22% vs. 0.59%, P<0.05). Discussion and conclusions: The work place is usually higher source of exposure to harmful factors. Workers need consistently and checked frequently health control. In assessing the risk of adverse effects of metals is important to consider their persistence, behavior and bioavailability. Prolonged exposure to carcinogens may not manifest symptoms of poisoning, but delayed effects may occur, which resulted in a higher incidence of malignant tumors.

Keywords: genotoxicity, chromium, stainless steels, welders

Procedia PDF Downloads 344

Authors: Dugeree Otgongerel, Kyong Jin Cho, Yu-Han Kim, Sangmee Ahn Jo

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Excessive activation of glutamate receptor is thought to be involved in retinal ganglion cell (RGC) death after ischemia- reperfusion damage. Glutamate carboxypeptidase II (GCPII) is an enzyme responsible for the synthesis of glutamate. Several studies showed that inhibition of GCPII prevents or reduces cellular damage in brain diseases. Thus, in this study, we examined the expression of GCPII in rat retina and the role of GCPII in acute high IOP ischemia-reperfusion damage of eye by using a GCPII inhibitor, 2-(phosphonomethyl) pentanedioic acid (2-PMPA). Animal model of ischemia-reperfusion was induced by raising the intraocular pressure for 60 min and followed by reperfusion for 3 days. Rats were randomly divided into four groups: either intra-vitreous injection of 2-PMPA (11 or 110 ng per eye) or PBS after ischemia-reperfusion, 2-PMPA treatment without ischemia-reperfusion and sham-operated normal control. GCPII immunoreactivity in normal rat retina was detected weakly in retinal nerve fiber layer (RNFL) and retinal ganglionic cell layer (RGL) and also inner plexiform layer (IPL) and outer plexiform layer (OPL) strongly where are co-stained with an anti-GFAP antibody, suggesting that GCPII is expressed mostly in Muller and astrocytes. Immunostaining with anti-BRN antibody showed that ischemia- reperfusion caused RGC death (31.5 %) and decreased retinal thickness in all layers of damaged retina, but the treatment of 2-PMPA twice at 0 and 48 hour after reperfusion blocked these retinal damages. GCPII level in RNFL layer was enhanced after ischemia-reperfusion but was blocked by PMPA treatment. This result was confirmed by western blot analysis showing that the level of GCPII protein after ischemia- reperfusion increased by 2.2- fold compared to control, but this increase was blocked almost completely by 110 ng PMPA treatment. Interestingly, GFAP immunoreactivity in the retina after ischemia- reperfusion followed by treatment with PMPA showed similar pattern to GCPII, increase after ischemia-reperfusion but reduction to the normal level by PMPA treatment. Our data demonstrate that increase of GCPII protein level after ischemia-reperfusion injury is likely to cause glial activation and/or retinal cell death which are mediated by glutamate, and GCPII inhibitors may be useful in treatment of retinal disorders in which glutamate excitotoxicity is pathogenic.

Keywords: glutamate carboxypepptidase II, glutamate excitotoxicity, ischemia-reperfusion, retinal ganglion cell

Procedia PDF Downloads 323

Authors: Mateusz Hermyt, Pawel Kaczmarek, Weronika Rupik

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The egg tooth is a crucial structure during hatching of lizards and snakes. In contrast to birds, turtles, crocodiles, and monotremes, egg tooth of squamate reptiles is a true tooth sharing common features of structure and development with all the other teeth of vertebrates. The egg tooth; however, due to its function, exhibits structural differences in relation to regular teeth. External morphology seems to be important in the context of phylogenetic relationships within Squamata but up to date, there is scarce information concerning structure and development of the egg tooth at the submicroscopical level. In presented studies detailed analysis of the egg tooth development in grass snake has been performed with the usage of light (including fluorescent), transmission and scanning electron microscopy. Grass snake embryo’s heads have been used in our studies. Grass snake is common snake species occurring in most of Europe including Poland. The grass snake is characterized by the presence of single unpaired egg tooth (as in most squamates) in contrast to geckos and dibamids possessing paired egg teeth. Studies show changes occurring on the external morphology, tissue and cellular levels of differentiating egg tooth. The egg tooth during its development changes its curvature. Initially, faces directly downward and in the course of its differentiation, it gradually changes to rostro-ventral orientation. Additionally, it forms conical dentinal protrusions on the sides. Histological analysis showed that egg tooth development occurs in similar steps in relation to regular teeth. It undergoes initiation, bud, cap and bell morphological stages. Analyses focused on describing morphological changes in hard tissues (mainly dentin and predentin) of egg tooth and in cells which enamel organ consists of. It included: outer enamel epithelium, stratum intermedium, inner enamel epithelium, odontoblasts, and cells of dental pulp. All specimens used in the study were captured according to the Polish regulations concerning the protection of wild species. Permission was granted by the Local Ethics Commission in Katowice (41/2010; 87/2015) and the Regional Directorate for Environmental Protection in Katowice (WPN.6401.257.2015.DC).

Keywords: hatching, organogenesis, reptile, Squamata

Procedia PDF Downloads 158

Authors: Adnan A. Kadi, Nasser S. Al-Shakliah, Haitham Al-Rabiah

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Zorifertinib is a novel, potent, oral, a small molecule used to treat non-small cell lung cancer (NSCLC). zorifertinib is an Epidermal Growth Factor Receptor (EGFR) inhibitor and has good blood–brain barrier permeability for (NSCLC) patients with EGFR mutations. zorifertinibis currently at phase II/III clinical trials. The current research reports the characterization and identification of in vitro, in vivo and reactive intermediates of zorifertinib. Prediction of susceptible sites of metabolism and reactivity pathways (cyanide and GSH) of zorifertinib were performed by the Xenosite web predictor tool. In-vitro metabolites of zorifertinib were performed by incubation with rat liver microsomes (RLMs) and isolated perfused rat liver hepatocytes. Extraction of zorifertinib and it's in vitro metabolites from the incubation mixtures were done by protein precipitation. In vivo metabolism was done by giving a single oral dose of zorifertinib(10 mg/Kg) to Sprague Dawely rats in metabolic cages by using oral gavage. Urine was gathered and filtered at specific time intervals (0, 6, 12, 18, 24, 48, 72,96and 120 hr) from zorifertinib dosing. A similar volume of ACN was added to each collected urine sample. Both layers (organic and aqueous) were injected into liquid chromatography ion trap mass spectrometry(LC-IT-MS) to detect vivozorifertinib metabolites. N-methyl piperizine ring and quinazoline group of zorifertinib undergoe metabolism forming iminium and electro deficient conjugated system respectively, which are very reactive toward nucleophilic macromolecules. Incubation of zorifertinib with RLMs in the presence of 1.0 mM KCN and 1.0 Mm glutathione were made to check reactive metabolites as it is often responsible for toxicities associated with this drug. For in vitro metabolites there were nine in vitro phase I metabolites, four in vitro phase II metabolites, eleven reactive metabolites(three cyano adducts, five GSH conjugates metabolites, and three methoxy metabolites of zorifertinib were detected by LC-IT-MS. For in vivo metabolites, there were eight in vivo phase I, tenin vivo phase II metabolitesofzorifertinib were detected by LC-IT-MS. In vitro and in vivo phase I metabolic pathways wereN- demthylation, O-demethylation, hydroxylation, reduction, defluorination, and dechlorination. In vivo phase II metabolic reaction was direct conjugation of zorifertinib with glucuronic acid and sulphate.

Keywords: in vivo metabolites, in vitro metabolites, cyano adducts, GSH conjugate

Procedia PDF Downloads 174

Authors: Tiisetso Mpai, Sanjay K. Jaiswal, Felix D. Dakora

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The South African Cape fynbos biome is a global biodiversity hotspot. This biome contains a diversity of endemic shrub legumes, including Polhillia, Wiborgia, and Wiborgiella species, which are important for ecotourism as well as for improving soil fertility status. This is due to their proven N₂-fixing abilities when in association with compatible soil bacteria. In fact, Polhillia, Wiborgia, and Wiborgiella species have been reported to derive over 61% of their needed nitrogen through biological nitrogen fixation and to exhibit acid and alkaline phosphatase activity in their rhizospheres. Thus, their interactions with soil microbes may explain their survival mechanisms under the continued summer droughts and acidic, nutrient-poor soils in this region. However, information regarding their rhizosphere microbiome is still unavailable, yet it is important for Fynbos biodiversity management. Therefore, the aim of this study was to assess the microbial community structures associated with rhizosphere soils of Polhillia pallens, Polhillia brevicalyx, Wiborgia obcordata, Wiborgia sericea, and Wiborgiella sessilifolia growing at different locations of the South African Cape fynbos, during the wet and dry seasons. The hypothesis is that the microbial communities in these legume rhizospheres are the same type and are not affected by the growing season due to the restricted habitat of these wild fynbos legumes. To obtain the results, DNA was extracted from 0.5 g of each rhizosphere soil using PowerSoil™ DNA Isolation Kit, and sequences were obtained using the 16S rDNA Miseq Illumina technology. The results showed that in both seasons, bacteria were the most abundant microbial taxa in the rhizosphere soils of all five legume species, with Actinobacteria showing the highest number of sequences (about 30%). However, over 19.91% of the inhabitants in all five legume rhizospheres were unclassified. In terms of genera, Mycobacterium and Conexibacter were common in rhizosphere soils of all legumes in both seasons except for W. obcordata soils sampled during the dry season, which had Dehalogenimonas as the major inhabitant (6.08%). In conclusion, plant species and season were found to be the main drivers of microbial community structure in Cape fynbos, with the wet season being more dominant in shaping microbial diversity relative to the dry season. Wiborgia obcordata had a greater influence on microbial community structure than the other four legume species.

Keywords: 16S rDNA, Cape fynbos, endemic legumes, microbiome, rhizosphere

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Authors: Goksen Capar, Levent Dogankaya

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According to the circular economy model, resource productivity should be maximized and wastes should be reduced. Since earth’s natural resources are continuously depleted, resource recovery has gained great interest in recent years. As part of our research study on the recovery and reuse of silk wastes, this paper focuses on the utilization of silkworm pupae as fishmeal replacement, which would replace the original fishmeal raw material, namely the fish itself. This, in turn, would contribute to sustainable management of wild fish resources. Silk fibre is secreted by the silkworm Bombyx mori in order to construct a 'room' for itself during its transformation process from pupae to an adult moth. When the cocoons are boiled in hot water, silk fibre becomes loose and the silk yarn is produced by combining thin silk fibres. The remaining wastes are 1) sericin protein, which is dissolved in water, 2) remaining part of cocoon, including the dead body of B. mori pupae. In this study, an eight weeks trial was carried out to determine the growth performance of common carp juveniles fed with waste silkworm pupae meal (SWPM) as a replacement for fishmeal (FM). Four isonitrogenous diets (40% CP) were prepared replacing 0%, 33%, 50%, and 100% of the dietary FM with non-defatted silkworm pupae meal as a dietary protein source for experiments in C. carpio. Triplicate groups comprising of 20 fish (0.92±0.29 g) were fed twice/day with one of the four diets. Over a period of 8 weeks, results showed that the diet containing 50% of its protein from SWPM had significantly higher (p ≤ 0.05) growth rates in all groups. The increasing levels of SWPM were resulted in a decrease in growth performance and significantly lower growth (p ≤ 0.05) was observed with diets having 100% SWPM. The study demonstrates that it is practical to replace 50% of the FM protein with SWPM with a significantly better utilization of the diet but higher SWPM levels are not recommended for juvenile carp. Further experiments are under study to have more detailed results on the possible effects of this alternative diet on the growth performance of juvenile carp.

Keywords: Bombyx mori, Cyprinus carpio, fish meal, silk, waste pupae

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Authors: Ivna Mororó, Lise P. Labéjof, Stephanie Ribeiro, Kely Almeida

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Lipid droplets (LDs) are normally presented in greater or lesser number in the cytoplasm of almost all eukaryotic and some prokaryotic cells. They are independent organelles composed of a lipid ester core and a surface phospholipid monolayer. As a lipid storage form, they provide an available source of energy for the cell. Recently it was demonstrated that they play an important role in other many cellular processes. Among the many unresolved questions about them, it is not even known how LDs is formed, how lipids are recruited to LDs and how they interact with the other organelles. Excess fat in the organism is pathological and often associated with the development of some genetic, hormonal or behavioral diseases. The formation and accumulation of lipid droplets in the cytoplasm can be increased by exogenous physical or chemical agents. It is well known that ionizing radiation affects lipid metabolism resulting in increased lipogenesis in cells, but the details of this process are unknown. To better understand the mode of formation of LDs in liver cells, we investigate their ultrastructural morphology after irradiation. For that, Wistar rats were exposed to whole body gamma radiation from 60-cobalt at various single doses. Samples of the livers were processed for analysis under a conventional transmission electron microscope. We found that when compared to controls, morphological changes in liver cells were evident at the higher doses of radiation used. It was detected a great number of lipid droplets of different sizes and homogeneous content and some of them merged each other. In some cells, it was observed diffused LDs, not limited by a monolayer of phospholipids. This finding suggests that the phospholipid monolayer of the LDs was disrupted by ionizing radiation exposure that promotes lipid peroxydation of endo membranes. Thus the absence of the phospholipid monolayer may prevent the realization of some cellular activities as follow: - lipid exocytosis which requires the merging of LDs membrane with the plasma membrane; - the interaction of LDs with other membrane-bound organelles such as the endoplasmic reticulum (ER), the golgi and mitochondria and; - lipolysis of lipid esters contained in the LDs which requires the presence of enzymes located in membrane-bound organelles as ER. All these impediments can contribute to lipid accumulation in the cytoplasm and the development of diseases such as liver steatosis, cirrhosis and cancer.

Keywords: radiobiology, hepatocytes, lipid metabolism, transmission electron microscopy

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Authors: Nisha Singh, Munish Puri, Collin Barrow, Deepak Tuli, Anshu S. Mathur

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The biological conversion of lignocellulosic biomass to ethanol is a promising strategy to solve the present global crisis of exhausting fossil fuels. The existing bioethanol production technologies have cost constraints due to the involvement of mandate pretreatment and extensive enzyme production steps. A unique process configuration known as consolidated bioprocessing (CBP) is believed to be a potential cost-effective process due to its efficient integration of enzyme production, saccharification, and fermentation into one step. Due to several favorable reasons like single step conversion, no need of adding exogenous enzymes and facilitated product recovery, CBP has gained the attention of researchers worldwide. However, there are several technical and economic barriers which need to be overcome for making consolidated bioprocessing a commercially viable process. Finding a natural candidate CBP organism is critically important and thermophilic anaerobes are preferred microorganisms. The thermophilic anaerobes that can represent CBP mainly belong to genus Clostridium, Caldicellulosiruptor, Thermoanaerobacter, Thermoanaero bacterium, and Geobacillus etc. Amongst them, Clostridium thermocellum has received increased attention as a high utility CBP candidate due to its highest growth rate on crystalline cellulose, the presence of highly efficient cellulosome system and ability to produce ethanol directly from cellulose. Recently with the availability of genetic and molecular tools aiding the metabolic engineering of Clostridium thermocellum have further facilitated the viability of commercial CBP process. With this view, we have specifically screened cellulolytic and xylanolytic thermophilic anaerobic ethanol producing bacteria, from unexplored hot spring/s in India. One of the isolates is a potential CBP organism identified as a new strain of Clostridium thermocellum. This strain has shown superior avicel and xylan degradation under unoptimized conditions compared to reported wild type strains of Clostridium thermocellum and produced more than 50 mM ethanol in 72 hours from 1 % avicel at 60°C. Besides, this strain shows good ethanol tolerance and growth on both hexose and pentose sugars. Hence, with further optimization this new strain could be developed as a potential CBP microbe.

Keywords: Clostridium thermocellum, consolidated bioprocessing, ethanol, thermophilic anaerobes

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Authors: Milda Nainyte, Viktoras Masevicius

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Biological methylation is a methyl group transfer from S-adenosyl-L-methionine (AdoMet) onto N-, C-, O- or S-nucleophiles in DNA, RNA, proteins or small biomolecules. The reaction is catalyzed by enzymes called AdoMet-dependent methyltransferases (MTases), which represent more than 3 % of the proteins in the cell. As a general mechanism, the methyl group from AdoMet replaces a hydrogen atom of nucleophilic center producing methylated DNA and S-adenosyl-L-homocysteine (AdoHcy). Recently, DNA methyltransferases have been used for the sequence-specific, covalent labeling of biopolymers. Two types of MTase catalyzed labeling of biopolymers are known, referred as two-step and one-step. During two-step labeling, an alkylating fragment is transferred onto DNA in a sequence-specific manner and then the reporter group, such as biotin, is attached for selective visualization using suitable chemistries of coupling. This approach of labeling is quite difficult and the chemical hitching does not always proceed at 100 %, but in the second step the variety of reporter groups can be selected and that gives the flexibility for this labeling method. In the one-step labeling, AdoMet analog is designed with the reporter group already attached to the functional group. Thus, the one-step labeling method would be more comfortable tool for labeling of biopolymers in order to prevent additional chemical reactions and selection of reaction conditions. Also, time costs would be reduced. However, effective AdoMet analog appropriate for one-step labeling of biopolymers and containing cleavable bond, required for reduction of PCR interferation, is still not known. To expand the practical utility of this important enzymatic reaction, cofactors with activated sulfonium-bound side-chains have been produced and can serve as surrogate cofactors for a variety of wild-type and mutant DNA and RNA MTases enabling covalent attachment of these chains to their target sites in DNA, RNA or proteins (the approach named methyltransferase-directed Transfer of Activated Groups, mTAG). Compounds containing hex-2-yn-1-yl moiety has proved to be efficient alkylating agents for labeling of DNA. Herein we describe synthetic procedures for the preparation of N-biotinoyl-N’-(pent-4-ynoyl)cystamine starting from the coupling of cystamine with pentynoic acid and finally attaching the biotin as a reporter group. The synthesis of the first AdoMet based cofactor containing a cleavable reporter group and appropriate for one-step labeling was developed.

Keywords: adoMet analogs, DNA alkylation, cofactor, methyltransferases

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Authors: Manqhai Kraai, Adrian M. Shrader, Peter F. Scogings

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For animals living in herds, competition between group members increases as herd size increases. The intensity of this competition is likely greater across poor quality landscapes and during the dry season. In contrast to wild herbivores, herd size in domestic livestock is determined by their owners. This then raises the question, how do domestic livestock, like goats, reduce competition for food within these defined herds? To explore this question, large-scale foraging behaviour of both small (12 to 28 individuals) and large (42 to 83 individuals) herds of free-ranging goats were recorded in Tugela Ferry, KwaZulu-Natal, South Africa. The study was conducted on three different landscapes that varied in both food quality and availability, during the wet and dry seasons of 2013-2014. The goats were housed in kraals overnight and let out in the mornings to forage unattended. Thus, foraging decisions were made by the goats and not by herders. The large-scale foraging behaviours focussed on included, (i) total distance travelled by goats while foraging, (ii) distance travelled before starting to feed, (iii) travel speed, and (iv) feeding duration. This was done using Garmin Foretrex 401 GPS devices harnessed to two goats per herd. Irrespective of season, there was no difference in the total distance travelled by the different sized herds across the different quality landscapes. However, both small and large herds started feeding farther from the kraal in the dry compared to the wet season. Despite this, there was no significant seasonal difference in total amount of time the herds spent feeding across the different landscapes. Finally, both small and large herds increased their travel speed across all the landscapes in the dry season, but large herds travelled faster than small herds. This increase was likely to maximise the time that large herds could spend feeding in good areas. Ultimately, these results indicate that both small and large herds were affected by declines in food quality and quantity during the dry season. However, as large herds made greater behavioural adjustments compared to smaller herds (i.e., feeding farther away from the kraal and travelling faster), it appeared that they were more affected by the seasonal increases in intra-herd competition.

Keywords: distance, feeding duration, food availability, food quality, travel speed

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Authors: Mark Morgan, Christine Li, Shuangyu Xu, Jenny McCarty

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The National Wild and Scenic Rivers System was created by the U.S. Congress in 1968 (Public Law 90-542; 16 U.S.C. 1271 et seq.) to preserve outstanding natural, cultural, and recreational values of some U.S. rivers in a free-flowing condition for the enjoyment of present and future generations. This Act is notable for safeguarding the special character of these rivers while supporting management action that encourages public participation for co-creating river protection goals and strategies. This is not an easy task. To meet the challenges of modern ecosystem management, federal resource agencies must address many legal, environmental, economic, political, and social issues. The U.S. Forest Service manages a 44-mile section of the Eleven Point National Scenic River (EPR) in southern Missouri, mainly for outdoor recreation purposes. About half of the acreage is in private lands, while the remainder flows through the Mark Twain National Forest. Private land along the river is managed by scenic easements to ensure protection of scenic values and natural resources, without public access. A portion of the EPR lies adjacent to a 16,500-acre tract known as the Irish Wilderness. The spring-fed river has steep bluffs, deep pools, clear water, and a slow current, making it an ideal setting for outdoor enthusiasts. A 10-month visitor study was conducted at five access points along the EPR during 2019 so the US Forest Service could update their river management plan. A mail-back survey was administered to 560 on-site visitors, yielding a response rate of 53%. Although different types of visitors use the EPR, boating and fishing were the predominant forms of outdoor recreation. Some river use was from locals, but other visitors came from farther away. Formulating unbiased policies for outdoor recreation is difficult because managers must assign relative values to recreational activities and travel distance. Because policymaking is a subjective process, management decisions can affect user groups in different ways (i.e., boaters vs. fishers; proximate vs. distal visitors), as seen through a GIS analysis.

Keywords: activity selection, forest recreation, policy, travel distance

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Authors: Gema Gómez-Casado, Alba Rodríguez-Muñoz, Virginia Mela-Rivas, Pallavi Kompella, Francisco José Tinahones-Madueña, Isabel Moreno-Indias, Almudena Ortega-Gómez

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Obesity is a high-priority health problem worldwide due to its high prevalence. The proportion of obese and overweight subjects in industrialized countries exceeds half of the population in most cases. Beyond the metabolic problem, obesity boosts inflammation levels in the organism. The gut microbiota, considered an organ by itself, controls a high variety of processes at a systemic level. In fact, the microbiota interacts closely with the immune system, being crucial in determining the maturation state of neutrophils, key effectors of the innate immune response. It is known that changes in the diet exert strong effects on the variety and activity of the gut microbiota. The effect that those changes have on the axis microbiota-immune response is an unexplored field. In this study, 10 patients with obesity (weight 114,3 ± 14,5Kg, BMI 40,47±3,66) followed a Mediterranean-hypocaloric diet for 3 months, reducing their initial weight by 12,71 ± 3%. A transplant of microbiota from these patients before and after the diet was performed into wild type “germ-free” mice (n=10/group), treated with antibiotics. Six weeks after the transplant, mice were euthanized, and the presence of cells from the innate immune system were analysed in different organs (bone marrow, blood, spleen, visceral adipose tissue, and intestine) by flow cytometry. No differences were observed in the number of myeloid cells in bone marrow, blood, spleen, or visceral adipose tissue of mice transplanted with patient’s microbiota before and after following the Mediterranean diet. However, the intestine of mice that received post-diet microbiota presented a marked decrease in the number of neutrophils (whose presence is associated with tissue inflammation), as well as macrophages. In line with these findings, intestine monocytes from mice with post-diet microbiota showed a less inflammatory profile (lower Ly6Gˡᵒʷ proportion of cells). These results point toward a decrease in the inflammatory state of the intestinal tissue, derived from changes in the gut microbiota, which occurred after a 3-month Mediterranean diet.

Keywords: obesity, nutrition, Mediterranean diet, gut microbiota, immune system

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Authors: Naira Sahakyan, Margarit Petrosyan, Armen Trchounian

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One of the tasks in contemporary biotechnology, pharmacology and other fields of human activities is to obtain biologically active substances from plants. They are very essential in the treatment of many diseases due to their actually high therapeutic value without visible side effects. However, sometimes the possibility of obtaining the metabolites is limited due to the reduction of wild-growing plants. That is why the plant cell cultures are of great interest as alternative sources of biologically active substances. Besides, during the monitored cultivation, it is possible to obtain substances that are not synthesized by plants in nature. Isolated culture of Ajuga genevensis with high growth activity and ability of regeneration was obtained using MS nutrient medium. The agar-diffusion method showed that aqueous extracts of callus culture revealed high antimicrobial activity towards various gram-positive (Bacillus subtilis A1WT; B. mesentericus WDCM 1873; Staphylococcus aureus WDCM 5233; Staph. citreus WT) and gram-negative (Escherichia coli WKPM M-17; Salmonella typhimurium TA 100) microorganisms. The broth dilution method revealed that the minimal and half maximal inhibitory concentration values against E. coli corresponded to the 70 μg/mL and 140 μg/mL concentration of the extract respectively. According to the photochemiluminescent analysis, callus tissue extracts of leaf and root origin showed higher antioxidant activity than the same quantity of A. genevensis intact plant extract. A. genevensis intact plant and callus culture extracts showed no cytotoxic effect on K-562 suspension cell line of human chronic myeloid leukemia. The GC-MS analysis showed deep differences between the qualitative and quantitative composition of callus culture and intact plant extracts. Hexacosane (11.17%); n-hexadecanoic acid (9.33%); and 2-methoxy-4-vinylphenol (4.28%) were the main components of intact plant extracts. 10-Methylnonadecane (57.0%); methoxyacetic acid, 2-tetradecyl ester (17.75%) and 1-Bromopentadecane (14.55%) were the main components of A. genevensis callus culture extracts. Obtained data indicate that callus culture of A. genevensis can be used as an alternative source of biologically active substances.

Keywords: Ajuga genevensis, antibacterial activity, antioxidant activity, callus cultures

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Authors: Qianqian He, Naian Liu, Xiaodong Xie, Linhe Zhang, Yang Zhang, Weidong Yan

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In the wild, the vegetation layer with exceptionally complex fuel composition and heterogeneous spatial distribution strongly affects the rate of fire spread (ROS) and fire intensity. Clarifying the influence of fuel bed structure on fire spread behavior is of great significance to wildland fire management and prediction. The packing ratio is one of the key physical parameters describing the property of the fuel bed. There is a threshold value of the packing ratio for ROS, but little is known about the controlling mechanism. In this study, to address this deficiency, a series of fire spread experiments were performed across a discrete fuel bed composed of some regularly arranged laser-cut cardboards, with constant wind speed and different packing ratios (0.0125-0.0375). The experiment aims to explore the relative importance of the internal and surface heat transfer with packing ratio. The dependence of the measured ROS on the packing ratio was almost consistent with the previous researches. The data of the radiative and total heat fluxes show that the internal heat transfer and surface heat transfer are both enhanced with increasing packing ratio (referred to as ‘Stage 1’). The trend agrees well with the variation of the flame length. The results extracted from the video show that the flame length markedly increases with increasing packing ratio in Stage 1. Combustion intensity is suggested to be increased, which, in turn, enhances the heat radiation. The heat flux data shows that the surface heat transfer appears to be more important than the internal heat transfer (fuel preheating inside the fuel bed) in Stage 1. On the contrary, the internal heat transfer dominates the fuel preheating mechanism when the packing ratio further increases (referred to as ‘Stage 2’) because the surface heat flux keeps almost stable with the packing ratio in Stage 2. As for the heat convection, the flow velocity was measured using Pitot tubes both inside and on the upper surface of the fuel bed during the fire spread. Based on the gas velocity distribution ahead of the flame front, it is found that the airflow inside the fuel bed is restricted in Stage 2, which can reduce the internal heat convection in theory. However, the analysis indicates not the influence of inside flow on convection and combustion, but the decreased internal radiation of per unit fuel is responsible for the decrease of ROS.

Keywords: discrete fuel bed, fire spread, packing ratio, wildfire

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Authors: Pravina Gurjar, Bothiraja Pour, Vijay Kumbhar, Ganesh Dama

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Andrographolide (AG), a bioactive phytoconstituent, has a wider range of pharmacological action. However, due to the intestinal degradation, shows low oral bioavailability. The aim of the present work was to develop Floating In-situ gelling Gastro retentive System (FISGS) for AG in order to enhance its site specific absorption and minimize pH dependent hydrolysis in alkaline environment. Further to increase its therapeutic efficacy for peptic ulcer disease caused by H. pyroli. Gellan based floating in situ gelling system of AG were prepared by using sodium citrate and calcium carbonate. The 32 factorial designs was used to study the effect of gellan and calcium carbonate concentration (independent variables) on dependent variable such as viscosity, floating lag time and drug release. Developed system was evaluated for drug content, floating lag time, viscosity, and drug release studies. Drug content, viscosity, and floating lag time was found to be 81-99%, 67-117 Cps, and 3-5 sec, respectively. The obtained system showed good in vitro floating ability for more than 12 h using 0.1 N HCl as dissolution medium with initial burst release followed by the controlled zero order drug release up to 24 hrs. In vivo testing of FISGS of AG to rats demonstrated significant antiulcer activity that were evaluated by various parameters like pH, volume, total acidity, millimole equivalent of H+ ions/30 min, and protein content of gastric content. The densities of all the formulation batches were found to be near about 0.9 and floating duration above 12 hr. It was observed that with the increase in conc. of gellan there was increase in the viscosity of formulation but all formulations were in optimum range. The drug content of optimized batch was found to be 99.23. In histopathology study of stomach, the villi at the mucosal surface, the intercellular junction, the intestinal lumen were intact; no destruction of the epithelium, and submucosal gland in formulation treated and control group animals as compared to pure drug AG and standard ranitidine. Gellan-based in situ gastro retentive floating system could be advantageous in terms of increased bioavailability of AG to maintain an effective drug conc. in gastric fluid as well as in serum for longer period of time.

Keywords: andrographolide, floating drug delivery, in situ gelling system, gastroretentive system

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Authors: Bharat Rawat, Shekhar Rajbhandari, Yadav Bhatta, Jay Prakash Jaiswal, Shivaji Bikram Silwal, Rajiv Shrestha, Shova Sunuwar

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Introduction: The honey produced by the bees fed on Rhobdodendron species containing grayanotoxin is known as mad honey. Grayanotoxin is found in honey obtained from the nectar of Rhododendron species growing on the mountains of the Black Sea region of Turkey and also in Japan, Nepal, Brazil, parts of North America, and Europe. Although the incidence of grayanotoxin poisoning is rare, there is concern that the number of cases per year will rise with the increasing demand for organic products. Mad honey intoxication might present with mild symptoms of cardiovascular, gastrointestinal and neurological systems or might also present with a life-threatening form with AV block and cardiovascular collapse. In this article, we describe the summary of five cases, which came to our hospital with mad honey related cardiac complications. Findings: In last one year, five cases presented in the emergency department with sudden onset of Loss of consciousness, dizziness, shortness of breath. They felt difficulty after the consumption of 1-3 teaspoonful of wild honey. The honey was brought from most of the rural parts of Nepal like khotang. Some of them also came with vomiting, dizziness, and loose stool. On examination, most of them had severe bradycardia and low blood pressure. No abnormalities were detected on systemic examinations. In one patient, ECG and cardiac enzymes showed features of the acute coronary syndrome, but his treadmill test done few days later was normal. All patients were managed with inj. Atropine, I/V normal saline, and other supportive measures and discharged in a stable condition within one or two days. Conclusions: Rhododendrons is the national flower of Nepal. The specific species of rhododendron found in Nepal which contains the toxin is not known. Bees feeding on these rhododendrons are known to transfer the grayanotoxin to the honey they produce. Most symptoms are mild and resolve themselves without medical intervention. Signs and symptoms of grayanotoxin poisoning rarely last more than 24 hours and are usually not fatal. Some signs of mad honey poisoning include Bradycardia, Cardiac arrhythmia, Hypotension, Nausea and Vomiting. They respond to close monitoring and appropriate supportive treatment. Normally, patients recover completely with no residual damage to the heart or its conduction system.

Keywords: rhobdodendron, honey, grayanotoxin, bradycardia

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Authors: Bedilu Tafesse

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The diversity and ecological analysis of vascular epiphytes was studied in Gera Forest in southwestern Ethiopia at altitudes between 1600 and 2400 m.a.s.l. A total area of 4.5 ha was surveyed in coffee and non-coffee forest vegetation. Fifty sampling plots, each 30 m x 30 m (900 m2), were used for the purpose of data collection. A total of 59 species of vascular epiphytes were recorded, of which 34 (59%) were holo epiphytes, two (4%) were hemi epiphytes and 22 (37%) species were accidental vascular epiphytes. To study the altitudinal distribution of vascular epiphytes, altitudes were classified into higher >2000, middle 1800-2000 and lower 1600-1800 m.a.s.l. According to Shannon-Wiener Index (H/= 3.411) of alpha diversity the epiphyte community in the study area is medium. There was a statistically significant difference between host bark type and epiphyte richness as determined by one-way ANOVA p = 0.001 < 0.05. The post-hoc test shows that there is significant difference of vascular epiphytes richness between smooth bark with rough, flack and corky bark (P =0.001< 0.05), as well as rough and cork bark (p =0.43 <0.05). However, between rough and flack bark (p = 0.753 > 0.05) and between flack and corky bark (p = 0.854 > 0.05) no significant difference of epiphyte abundance was observed. Rough bark had 38%, corky 26%, flack 25%, and only 11% vascular epiphytes abundance occurred on smooth bark. The regression correlation test, (R2 = 0.773, p = 0.0001 < 0.05), showed that the number of species of vascular epiphytes and host DBH size are positively correlated. The regression correlation test (R2 = 0.28, p = 0.0001 < 0.05), showed that the number of species and host tree height positively correlated. The host tree preference of vascular epiphytes was recorded for only Vittaria volkensii species hosted on Syzygium guineense trees. The result of similarity analysis indicated that Gera Forest showed the highest vascular epiphytic similarity (0.35) with Yayu Forest and shared the least vascular epiphytic similarity (0.295) with Harenna Forest. It was concluded that horizontal stems and branches, large and rough, flack and corky bark type trees are more suitable for vascular epiphytes seedling attachments and growth. Conservation and protection of these phorophytes are important for the survival of vascular epiphytes and increase their ecological importance.

Keywords: accidental epiphytes, hemiepiphyte, holoepiphyte, phorophyte

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Authors: Gurfateh Singh, Mu Khan, Razia Khanam, Govind Mohan

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Objective: The present study has been designed to investigate the beneficial role of Fenofibrate & Clofibrate in attenuated the cardioprotective effect of ischemic preconditioning (IPC) in hyperlipidemic rat hearts. Materials & Methods: Experimental hyperlipidemia was produced by feeding high fat diet to rats for a period of 28 days. Isolated langendorff’s perfused normal and hyperlipidemic rat hearts were subjected to global ischemia for 30 min followed by reperfusion for 120 min. The myocardial infarct size was assessed macroscopically using triphenyltetrazolium chloride staining. Coronary effluent was analyzed for lactate dehydrogenase (LDH) and creatine kinase-MB release to assess the extent of cardiac injury. Moreover, the oxidative stress in heart was assessed by measuring thiobarbituric acid reactive substance, superoxide anion generation and reduced form of glutathione. Results: The ischemia-reperfusion (I/R) has been noted to induce oxidative stress by increasing TBARS, superoxide anion generation and decreasing reduced form of glutathione in normal and hyperlipidemic rat hearts. Moreover, I/R produced myocardial injury, which was assessed in terms of increase in myocardial infarct size, LDH and CK-MB release in coronary effluent and decrease in coronary flow rate in normal and hyperlipidemic rat hearts. In addition, the hyperlipidemic rat hearts showed enhanced I/R-induced myocardial injury with high degree of oxidative stress as compared with normal rat hearts subjected to I/R. Four episodes of IPC (5 min each) afforded cardioprotection against I/R-induced myocardial injury in normal rat hearts as assessed in terms of improvement in coronary flow rate and reduction in myocardial infarct size, LDH, CK-MB and oxidative stress. On the other hand, IPC mediated myocardial protection against I/R-injury was abolished in hyperlipidemic rat hearts. However, Treatment with Fenofibrate (100 mg/kg/day, i.p.), Clofibrate (300mg/kg/day, i.p.) as a agonists of PPAR-α have not affected the cardioprotective effect of IPC in normal rat hearts, but its treatment markedly restored the cardioprotective potentials of IPC in hyperlipidemic rat hearts. Conclusion: It is noted that the high degree of oxidative stress produced in hyperlipidemic rat heart during reperfusion and consequent down regulation of PPAR-α may be responsible to abolish the cardioprotective potentials of IPC.

Keywords: Hyperlipidemia, ischemia-reperfusion injury, ischemic preconditioning, PPAR-α

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Authors: Viviane A. O. Silva, Angela M. Costa, Glaucia N. M. Hajj, Ana Preto, Aline Tansini, Martin Roffé, Peter Jordan, Rui M. Reis

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Autophagy is a recycling and degradative system suggested to be a major cell death pathway in cancer cells. Autophagy pathway is interconnected with the endocytosis pathways sharing the same ultimate lysosomal destination. Lysosomes are crucial regulators of cell homeostasis, responsible to downregulate receptor signalling and turnover. It seems highly likely that derailed endocytosis can make major contributions to several hallmarks of cancer. WNK2, a member of the WNK (with-no-lysine [K]) subfamily of protein kinases, had been found downregulated by its promoter hypermethylation, and has been proposed to act as a specific tumour-suppressor gene in brain tumors. Although some contradictory studies indicated WNK2 as an autophagy modulator, its role in cancer cell death is largely unknown. There is also growing evidence for additional roles of WNK kinases in vesicular traffic. Aim: To evaluate the role of WNK2 in autophagy and endocytosis on glioma context. Methods: Wild-type (wt) A172 cells (WNK2 promoter-methylated), and A172 transfected either with an empty vector (Ev) or with a WNK2 expression vector, were used to assess the cellular basal capacities to promote autophagy, through western blot and flow-cytometry analysis. Additionally, we evaluated the effect of WNK2 on general endocytosis trafficking routes by immunofluorescence. Results: The re-expression of ectopic WNK2 did not interfere with autophagy-related protein light chain 3 (LC3-II) expression levels as well as did not promote mTOR signaling pathway alteration when compared with Ev or wt A172 cells. However, the restoration of WNK2 resulted in a marked increase (8 to 92,4%) of Acidic Vesicular Organelles formation (AVOs). Moreover, our results also suggest that WNK2 cells promotes delay in uptake and internalization rate of cholera toxin B and transferrin ligands. Conclusions: The restoration of WNK2 interferes in vesicular traffic during endocytosis pathway and increase AVOs formation. This results also suggest the role of WNK2 in growth factor receptor turnover related to cell growth and homeostasis and associates one more time, WNK2 silencing contribution in genesis of gliomas.

Keywords: autophagy, endocytosis, glioma, WNK2

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