Search results for: underutilised species

Authors: S. Al-Khalasi, O. Mahgoub, H. Yaakub

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This study evaluated the use of raw or processed Prosopis juliflora (Meskit) pods as a major ingredient in a formulated ration to provide an alternative non-conventional concentrate for livestock feeding in Oman. Dry Meskit pods were reduced to lengths of 0.5- 1.0 cm to ensure thorough mixing into three diets. Meskit pods were subjected to two types of treatments; roasting and soaking. They were roasted at 150оC for 30 minutes using a locally-made roasting device (40 kg barrel container rotated by electric motor and heated by flame gas cooker). Chopped pods were soaked in tap water for 24 hours and dried for 2 days under the sun with frequent turning. The Meskit-pod-based diets (MPBD) were formulated and pelleted from 500 g/kg ground Meskit pods, 240 g/kg wheat bran, 200 g/kg barley grain, 50 g/kg local dried sardines and 10 g/kg of salt. Twenty four 10 months-old intact Omani male lambs with average body weight of 27.3 kg (± 0.5 kg) were used in a feeding trial for 84 days. They were divided (on body weight basis) and allocated to four diet combination groups. These were: Rhodes grass hay (RGH) plus a general ruminant concentrate (GRC); RGH plus raw Meskit pods (RMP) based concentrate; RGH plus roasted Meskit pods (ROMP) based concentrate; RGH plus soaked Meskit pods (SMP) based concentrate Daily feed intakes and bi-weekly body weights were recorded. MPBD had higher contents of crude protein (CP), acid detergent fibre (ADF) and neutral detergent fibre (NDF) than the GRC. Animals fed various types of MPBD did not show signs of ill health. There was a significant effect of feeding ROMP on the performance of Omani sheep compared to RMP and SMP. The ROMP fed animals had similar performance to those fed the GRC in terms of feed intake, body weight gain and feed conversion ratio (FCR).This study indicated that roasted Meskit pods based diet may be used instead of the commercial concentrate for feeding Omani sheep without adverse effects on performance. It offers a cheap alternative source of protein and energy for feeding Omani sheep. Also, it might help in solving the spread impact of Meskit trees, maintain the ecosystem and helping in preserving the local tree species.

Keywords: growth, Meskit, Omani sheep, Prosopis juliflora

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Authors: Aktar-Uz-Zaman, M. Tuhina-Khatun, Mohamed Hanafi Musa

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Three rust diseases: leaf (brown) rust caused by Puccinia triticina Eriks, stripe (yellow) rust caused by Puccinia striiformis West, and stem (black) rust caused by Puccinia graminis f. sp. tritici are economically important diseases of wheat in world wide. Yield loss due to leaf rust is 40% in susceptible cultivars. Yield losses caused by the stem rust pathogens in the mid of 20 century reached 20-30% in Eastern and Central Europe and the most virulent stem rust race Ug99 emerged first in Uganda and after that in Kenya, Ethiopia, Yemen, in the Middle East and South Asia. Yield losses were estimated up to 100%, whereas, up to 80% have been reported in Kenya during 1999. In case of stripe rust, severity level has been recorded 60% - 70% as compared to 100% severity of susceptible check in disease screening nurseries in Kenya. Improvement of resistant varieties or cultivars is the sustainable, economical and environmentally friendly approaches for increasing the global wheat production to suppress the rust diseases. More than 68 leaf rust, 49 stripe rust and 53 stem rust resistance genes have been identified in the global wheat cultivars or varieties using different molecular breeding approaches. Among these, Lr1, Lr9, Lr10, Lr19, Lr21, Lr24, Lr25, Lr28, Lr29, Lr34, Lr35, Lr37, Lr39, Lr47, Lr51, Lr3bg, Lr18, Lr40, Lr46, and Lr50 leaf rust resistance genes have been identified by using molecular, enzymatic and microsatellite markers from African, Asian, European cultivars of hexaploid wheat (Triticum aestivum), durum wheat and diploid wheat species. These genes are located on 20, of the 21 chromosomes of hexaploid wheat. Similarly, Sr1, Sr2, Sr24, and Sr3, Sr31 stem rust resistance genes have been recognized from wheat cultivars of Pakistan, India, Kenya, and Uganda etc. A race of P. striiformis (stripe rust) Yr9, Yr18, and Yr29 was first observed in East Africa, Italy, Pakistan and India wheat cultivars. These stripe rust resistance genes are located on chromosomes 1BL, 4BL, 6AL, 3BS and 6BL in bread wheat cultivars. All these identified resistant genes could be used for notable improvement of susceptible wheat cultivars in the future.

Keywords: hexaploid wheat, resistance genes, rust disease, triticum aestivum

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Authors: Sara Taghdisi, Mehrosadat Mirmohammadi, Mostafa Mokhtarian, Mohammad Hossein Pazandeh

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Leishmaniasis is one of the 10 most important diseases of the World Health Organization with health problems in more than 90 countries. Over one billion people are at risk of these diseases on almost every continent. The present human study was performed to evaluate the therapeutic effect of cotton plant on cutaneous leishmaniasis leision. firstly, the cotton seeds were cleaned and grinded to smaller particles. In the second step, the seeds were oiled by cold press method. In order to separate bioactive compound, after saponification of the oil, its gossypol was hydrolyzed and crystalized. finally, the therapeutic effect of Cottonseed Oil on cutaneous leishmaniasis was investigated. In the current project, Gossypol was extracted with a liquid-liquid extraction method in 120 minutes in the presence of Phosphoric acid from the cotton seed oil of Golestan beach varieties, then got crystallized in darkness using Acetic acid and isolated as Gossypol Acetic acid. The efficiency of the extracted crystal was obtained at 1.28±0.12. the cotton plant could be efficient in the treatment of Cutaneous leishmaniasis. This double-blind randomized controlled clinical trial was performed on 88 cases of leishmaniasis wounds. Patients were randomly divided into two groups of 44 cases. two groups received conventional treatment. In addition to the usual treatment (glucantime), the first group received cottonseed oil and the control group received placebo. The results of the present study showed that the surface of lesion before the intervention and in the first to fourth weeks after the intervention was not significantly different between the two groups (P-value> 0.05). But the surface of lesion in the Intervention group in the eighth and twelfth weeks was lower than the control group (P-value <0.05). This study showed that the improvement of leishmaniasis lesion using topical cotton plant mark in the eighth and twelfth weeks after the intervention was significantly more than the control group. Considering the most common chemical drugs for Cutaneous leishmaniasis treatment are sodium stibogluconate, and meglumine antimonate, which not only have relatively many side effects, but also some species of the Leishmania genus have become resistant to them. Therefore, a plant base bioactive compound such as cottonseed oil can be useful whit fewer side effects.

Keywords: cottonseed oil, crystallization, gossypol, leishmaniasis

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Authors: Luís R. Silva, Ana C. Gonçalves, Catarina Bento, Fábio Jesus, Branca M. Silva

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Prunus avium Linnaeus, more known as sweet cherry, is one of the most appreciated fruit worldwide. Most of these quantities are produced in Fundão region, being Saco the cultivar most produced. Saco is very rich in bioactive compounds, especially phenolics, and presents great antioxidant capacity. The purpose of the present study was to investigate the chemical profile and biological potential, concerning antioxidant, anti-diabetic activity and protective effects towards erythrocytes by Saco sweet cherry collected from Fundão region (Portugal). The hydroethanolic extracts were prepared and passed through a C18 solid-phase extraction column. The phenolic profile analyzed by LC-DAD method allowed to the identification of 22 phenolic compounds, being 16 non-phenolics and 6 anthocyanins. In respect to non-coloured phenolics, 3-O-caffeoylquinic and ρ-coumaroylquinic acids were the main ones. Concerning to anthocyanins, cyanidin-3-O-rutinoside was found in higher amounts. Relatively to biological potential, Saco showed great antioxidant potential, through DPPH and NO radical assays, with IC50 =16.24 ± 0.46 µg/mL and IC50 = 176.69 ± 3.35 µg/mL for DPPH and NO, respectively. These results were similar to those obtained for ascorbic acid control (IC50 = 16.92 ± 0.69 and IC50 = 162.66 ± 1.31 μg/mL for DPPH and NO, respectively). In respect to antidiabetic potential, Saco revealed capacity to inhibit α-glucosidase in a dose-dependent manner (IC50 = 10.79 ± 0.40 µg/mL), being much active than positive control acarbose (IC50 = 306.66 ± 0.84 μg/mL). Additionally, Saco extracts revealed protective effects against ROO•-mediated toxicity generated by AAPH in human blood erythrocytes, inhibiting hemoglobin oxidation (IC50 = 38.57 ± 0.96 μg/mL) and hemolysis (IC50 = 73.03 ± 1.48 μg/mL), in a concentration-dependent manner. However, Saco extracts were less effective than quercetin control (IC50 = 3.10 μg/mL and IC50 = 0.7 μg/mL for inhibition of hemoglobin oxidation and hemolysis, respectively). The results obtained showed that Saco is an excellent source of phenolic compounds. These ones are natural antioxidant substances, which easily capture reactive species. This work presents new insights regarding sweet cherry antioxidant properties which may be useful for the future development of new therapeutic strategies for preventing or attenuating oxidative-related disorders.

Keywords: antioxidant capacity, health benefits, phenolic compounds, saco

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Authors: Dhritilekha Deka, Deepak Patwa, Ravi K., Archana M. Nair

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Bioaccumulation of heavy metal contaminants due to intense anthropogenic interference degrades the environment and ecosystem functions. Conventional physicochemical methods involve energy-intensive and costly methodologies. Phytoremediation, on the other hand, provides an efficient nature-based strategy for the reclamation of heavy metal-contaminated sites. However, the slow process and adaptation to high-concentration contaminant sequestration often limit the efficiency of the method. This necessitates natural amendments such as biochar to improve phytoextraction and stabilize the green cover. Biochar is a highly porous structure with high carbon sequestration potential and containing negatively charged functional groups that provide binding sites for the positively charged metals. This study aims to develop and determine the synergy between sugarcane bagasse biochar content and phytoremediation. A 60-day pot experiment using perennial tussock vetiver grass (Chrysopogon zizanioides) was conducted for different biochar contents of 1%, 2%, and 4% for the removal of cadmium and zinc. A concentration of 500 ppm is maintained for the amended and unamended control (CK) samples. The survival rates of the plants, biomass production, and leaf area index were measured for the plant growth characteristics. Results indicate a visible change in the plant growth and the heavy metal concentration with the biochar content. The bioconcentration factor (BCF) in the plant improved significantly for the 4% biochar content by 57% in comparison to the control CK treatment in Cd-treated soils. The Zn soils indicated the highest reduction in the metal concentration by 50% in the 2% amended samples and an increase in the BCF in all the amended samples. The translocation from the rhizosphere to the shoots was low but not dependent on the amendment content and varied for each contaminant type. The root-to-shoot ratio indicates higher values compared to the control samples. The enhanced tolerance capacities can be attributed to the nutrients released by the biochar in the soil. The study reveals the high potential of biochar as a phytoremediation amendment, but its effect is dependent on the soil and heavy metal and accumulator species.

Keywords: phytoextraction, biochar, heavy metals, chrysopogon zizanioides, bioaccumulation factor

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Authors: Juan Facetti, Rocky McBride, Karina Loup

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The Chaco Region of Paraguay is a key biodiverse area for the conservation of jaguars (Panthera onca), the largest feline of the Americas. It comprises five eco-regions, which holds important but decreasing populations of this species. The last decades, the expansion of soybean over the Atlantic Forest, forced the translocation of cattle-ranches towards the Chaco. Few studies of Jaguar's population densities in the American hemisphere were done until now. In the region, the specie is listed as vulnerable or threatened and more information is needed to implement any conservation policy. Among the factors that threaten the populations are land-use change, habitat fragmentation, prey depletion and illegal hunting. Two largest eco-regions were studied: the Wet Chaco and the Dry Chaco. From 2002 more than 20 jaguars were captured and fitted with GPS-collar. Data collected from 11 GPS-collars were processed, transformed numerically and finally converted into maps for analyzing. 8.092 locations were determined for four adult females (AF) and one adult male (AM) in the Wet Chaco, and one AF, one juvenile male (JM) and four AM in the Dry Chaco, during 1,867 days. GIS and kernel methodology were used to calculate daily distance of movement, home range-HR (95% isopleth), and core area (considered as 50% isopleth). In the Wet Chaco HR were 56 Km2 and 238 km2 for females and males respectively; while in the Dry Chaco HR were 685 Km2 and 844.5 km2 for females and males respectively, and 172 Km2 for a juvenile. Core areas of individual activity for each jaguar, were on average 11.5 Km2 and 33.55 km2 for AF and AM respectively in the Wet Chaco, while in the Dry Chaco were larger: 115 km2 for five AM and 225 Km2 for an AF and 32.4 Km2 for a JM. In both ecoregions, only one relevant overlap of HR of adults was reported. During the reproduction season, the HR (95% K) of one AM overlapped 49.83% with that of one AF. At the Wet Chaco, the maximum daily distance moved by an AF was 14.5 Km and 11.6 Km for the AM, while the Maximum Mean Daily Moved (MMDM) distance was 5.6 km for an AF and 3.1 km for an AM. At the Dry Chaco, the maximum daily distance for an AF was 61.7Km., 50.9Km for the AM and 6.6 Km for the JM, while the MMDM distance was 13.2 km for an AM and 8.4 km for an AF. This study confirmed that, as the invasion to jaguar habitat increased, it resulted in fragmented landscapes that influence spacing patterns of jaguars. Males used largest HR that of the smaller females and males covers largest distances that of the females. There appeared to be important spatial segregation between not only females but also males. It is likely that the larger areas used by males are partly caused by the sexual dimorphism in body size that entails differences in prey requirements. These could explain the larger distances travelled daily by males.

Keywords: Chaco ecoregions, Jaguar, home range, Panthera onca, Paraguay

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Authors: Emmanuel Billy

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This work is in the CABRISS project (H2020 projects) which aims at developing innovative cost-effective methods for the extraction of materials from the different sources of PV waste: Si based panels, thin film panels or Si water diluted slurries. Aluminum, silicon, indium, and silver will especially be extracted from these wastes in order to constitute materials feedstock which can be used later in a closed-loop process. The extraction of metals from silicon solar cells is often an energy-intensive process. It requires either smelting or leaching at elevated temperature, or the use of large quantities of strong acids or bases that require energy to produce. The energy input equates to a significant cost and an associated CO2 footprint, both of which it would be desirable to reduce. Thus there is a need to develop more energy-efficient and environmentally-compatible processes. Thus, ‘ionometallurgy’ could offer a new set of environmentally-benign process for metallurgy. This work demonstrates that ionic liquids provide one such method since they can be used to dissolve and recover silver. The overall process associates leaching, recovery and the possibility to re-use the solution in closed-loop process. This study aims to evaluate and compare different ionic liquids to leach and recover silver. An electrochemical analysis is first implemented to define the best system for the Ag dissolution. Effects of temperature, concentration and oxidizing agent are evaluated by this approach. Further, a comparative study between conventional approach (nitric acid, thiourea) and the ionic liquids (Cu and Al) focused on the leaching efficiency is conducted. A specific attention has been paid to the selection of the Ionic Liquids. Electrolytes composed of chelating anions are used to facilitate the lixiviation (Cl, Br, I,), avoid problems dealing with solubility issues of metallic species and of classical additional ligands. This approach reduces the cost of the process and facilitates the re-use of the leaching medium. To define the most suitable ionic liquids, electrochemical experiments have been carried out to evaluate the oxidation potential of silver include in the crystalline solar cells. Then, chemical dissolution of metals for crystalline solar cells have been performed for the most promising ionic liquids. After the chemical dissolution, electrodeposition has been performed to recover silver under a metallic form.

Keywords: electrodeposition, ionometallurgy, leaching, recycling, silver

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Authors: Berina Pilipović, Saša Pilipović, Maja Pašić-Kulenović

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Inflammation is the body's response to impaired homeostasis caused by infection, injury or trauma resulting in systemic and local effects. Inflammation causes the body's response to injury and is characterized by a series of events including inflammatory response, response to pain receptors and the recovery process. Inflammation can be acute and chronic. The inflammatory response is described in three different phases. Free radical is an atom or molecule that has the unpaired electron and is therefore generally very reactive chemical species. Biologically important example of reaction with free radicals is called Lipid peroxidation (LP). Lipid peroxidation reactions occur in biological membranes, and if at the outset is not stopped with the action of antioxidants, it will bring damage to the membrane, which results in partial or complete loss of their physiological functions. Calcium antagonists and beta-adrenergic receptor antagonists are known drugs, and for many years and widely used in the treatment of cardiovascular diseases. Some of these compounds also show antioxidant activity. The mechanism of antioxidant activities of calcium antagonists and beta-blockers is unknown, since their structure varies widely. This research investigated the possible local anti-inflammatory activity of ointments containing 1% carvedilol in the white petrolatum USP. Ear inflammation was induced by 3% croton oil acetone solution, in quantity of 10 µl on both mouse ears. Albino Swiss mouse (n = 8) are treated with 2.5 mg/ear ointment, and control group was treated on the same way as previous with hydrocortisone 1% ointment (2.5 mg/ear). The other ear of the same animal was used as control one. Ointments were administered once per day, on the left ear. After treatment, ears were observed for three days. After three days, we measured mass (mg) of 6 mm ear punch of treated and controlled ears. The results of testing anti-inflammatory effects of ointments with carvedilol in the mouse ear model show stronger observed effect than ointment with 1% hydrocortisone in the same basis. Identical results were confirmed by the difference between the mass of 6 mm ears punch. The results were also confirmed by histological examination. Ointments with carvedilol showed significant reduction of the inflammation process caused by croton oil on the mouse inflammation model.

Keywords: antioxidant, carvedilol, inflammation, mouse ear

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Authors: Olubukayo-Opeyemi Oyetayo, Oscar Mendez-Lucio, Andreas Bender, Hans Kiefer

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The thermal melting curve of a protein provides information on its conformational stability and could provide cues on its aggregation behavior. Naturally-occurring osmolytes have been shown to improve the thermal stability of most proteins in a concentration-dependent manner. They are therefore commonly employed as additives in therapeutic protein purification and formulation. A number of intertwined and seemingly conflicting mechanisms have been put forward to explain the observed stabilizing effects, the most prominent being the preferential exclusion mechanism. We attempted to probe and summarize molecular mechanisms for thermal stabilization of a monoclonal antibody (mAb) by developing quantitative structure-activity relationships using a rationally-selected library of 120 osmolyte-like compounds in the polyhydric alcohols, amino acids and methylamines classes. Thermal stabilization potencies were experimentally determined by thermal shift assays based on differential scanning fluorimetry. The cross-validated QSAR model was developed by partial least squares regression using descriptors generated from Molecular Operating Environment software. Careful evaluation of the results with the use of variable importance in projection parameter (VIP) and regression coefficients guided the selection of the most relevant descriptors influencing mAb thermal stability. For the mAb studied and at pH 7, the thermal stabilization effects of tested compounds correlated positively with their fractional polar surface area and inversely with their fractional hydrophobic surface area. We cannot claim that the observed trends are universal for osmolyte-protein interactions because of protein-specific effects, however this approach should guide the quick selection of (de)stabilizing compounds for a protein from a chemical library. Further work with a large variety of proteins and at different pH values would help the derivation of a solid explanation as to the nature of favorable osmolyte-protein interactions for improved thermal stability. This approach may be beneficial in the design of novel protein stabilizers with optimal property values, especially when the influence of solution conditions like the pH and buffer species and the protein properties are factored in.

Keywords: thermal stability, monoclonal antibodies, quantitative structure-activity relationships, osmolytes

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Authors: Amr Saeb, Khalid Al-Rubeaan, Mohamed Abouelhoda, Manojkumar Selvaraju, Hamsa Tayeb

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Background: P. mirabilis is a common uropathogenic bacterium that can cause major complications in patients with long-standing indwelling catheters or patients with urinary tract anomalies. In addition, P. mirabilis is a common cause of chronic osteomyelitis in diabetic foot ulcer (DFU) patients. Methodology: P. mirabilis SCDR1 was isolated from a diabetic ulcer patient. We examined P. mirabilis SCDR1 levels of resistance against nano-silver colloids, the commercial nano-silver and silver containing bandages and commonly used antibiotics. We utilized next generation sequencing techniques (NGS), bioinformatics, phylogenetic analysis and pathogenomics in the identification and characterization of the infectious pathogen. Results: P. mirabilis SCDR1 is a multi-drug resistant isolate that also showed high levels of resistance against nano-silver colloids, nano-silver chitosan composite and the commercially available nano-silver and silver bandages. The P. mirabilis-SCDR1 genome size is 3,815,621 bp with G+C content of 38.44%. P. mirabilis-SCDR1 genome contains a total of 3,533 genes, 3,414 coding DNA sequence genes, 11, 10, 18 rRNAs (5S, 16S, and 23S), and 76 tRNAs. Our isolate contains all the required pathogenicity and virulence factors to establish a successful infection. P. mirabilis SCDR1 isolate is a potential virulent pathogen that despite its original isolation site, wound, it can establish kidney infection and its associated complications. P. mirabilis SCDR1 contains several mechanisms for antibiotics and metals resistance including, biofilm formation, swarming mobility, efflux systems, and enzymatic detoxification. Conclusion: P. mirabilis SCDR1 is the spontaneous nano-silver resistant bacterial strain. P. mirabilis SCDR1 strain contains all reported pathogenic and virulence factors characteristic for the species. In addition, it possesses several mechanisms that may lead to the observed nano-silver resistance.

Keywords: Proteus mirabilis, multi-drug resistance, silver nanoparticles, resistance, next generation sequencing techniques, genome analysis, bioinformatics, phylogeny, pathogenomics, diabetic foot ulcer, xenobiotics, multidrug resistance efflux, biofilm formation, swarming mobility, resistome, glutathione S-transferase, copper/silver efflux system, altruism

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Authors: Alina Beraudi, Simona Catalani, Dalila De Pasquale, Eva Bianconi, Umberto Santoro, Susanna Stea, Pietro Apostoli

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Recently the European Community, in line with the international scientific community such as with the Consensus Statement, has determined to stop the use of metal-on-metal big head stemmed hip prosthesis. Among the factors accounted as responsible for the high failure rates of these hip implants are the release and accumulation of metal ions. Many studies have correlated the presence of these ions, besides other factors, with the induction of oxidative stress response. In our study on 12 subjects, we observed the patient specific capability to eliminate metal ions after revision surgery. While for cobalt all the patients were able to completely excrete cobalt ions within 5-7 months after metal-on-metal bearing removal, for chromium ions it didn’t happen. If on the one hand the toxicokinetic differences between the two types of ions are confirmed by toxicological and occupational studies, on the other hand, this peculiar way of exposition represents a novel and important point of view. Thus, two different approaches were performed to better understand the subject specific capability to transport metal ions (albumin study) and to manage the response to them (heme-oxygenase-1 study): - a mutational screening of ALBUMIN gene was conducted in 30 MoM prosthetic patients resulting in the absence of nucleotidic changes compared with the ALB reference sequence. To this study was also added the analysis of expression of modified albumin protein; - a gene and protein expression study on 44 patients of heme-oxygenase-1, that is one of the most important antioxidant enzyme induced by metallic ions, was performed. This study resulted in no statistically significant differences in the expression of the gene and protein heme-oxygenase-1 between prosthetic and non-prosthetic patients, as well as between patients with high and low ions levels. Our results show that the protein studied (albumin and heme-oxygenase-1) seem to be not involved in determining chromium and cobalt ions level. On the other hand, achromium and cobalt elimination rates are different, but similar in all patients analyzed, suggesting that this process could be not patient-related. We support the importance of researching more about ions transport within the organism once released by hip prosthesis, about the chemical species involved, the districts where they are contained and the mechanisms of elimination, not excluding the existence of a subjective susceptibility to these metals ions.

Keywords: chromium, cobalt, hip prosthesis, individual susceptibility

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Authors: U. K. Hulihalli, Shantveerayya

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Buckwheat (Fagopyrum esculentum Moench) is an annual crop belongs to family Poligonaceae. The cultivated buckwheat species are notable for their exceptional nutritive values. It is an important source of carbohydrates, fibre, macro, and microelements such as K, Ca, Mg, Na and Mn, Zn, Se, and Cu. It also contains rutin, flavonoids, riboflavin, pyridoxine and many amino acids which have beneficial effects on human health, including lowering both blood lipid and sugar levels. Rutin, quercetin and some other polyphenols are potent carcinogens against colon and other cancers. Buckwheat has significant nutritive value and plenty of uses. Cultivation of buckwheat in Sothern part of India is very meager. Hence, a study was planned with an objective to know the performance of buckwheat genotypes to different planting geometries and fertility levels. The field experiment was conducted at Main Agriculture Research Station, University of Agriculture Sciences, Dharwad, India, during 2017 Kharif. The experiment was laid-out in split-plot design with three replications having three planting geometries as main plots, two genotypes as sub plots and three fertility levels as sub-sub plot treatments. The soil of the experimental site was vertisol. The standard procedures are followed to record the observations. The planting geometry of 30*10 cm was recorded significantly higher seed yield (893 kg/ha⁻¹), stover yield (1507 kg ha⁻¹), clusters plant⁻¹ (7.4), seeds clusters⁻¹ (7.9) and 1000 seed weight (26.1 g) as compared to 40*10 cm and 20*10 cm planting geometries. Between the genotypes, significantly higher seed yield (943 kg ha⁻¹) and harvest index (45.1) was observed with genotype IC-79147 as compared to PRB-1 genotype (687 kg ha⁻¹ and 34.2, respectively). However, the genotype PRB-1 recorded significantly higher stover yield (1344 kg ha⁻¹) as compared to genotype IC-79147 (1173 kg ha⁻¹). The genotype IC-79147 was recorded significantly higher clusters plant⁻¹ (7.1), seeds clusters⁻¹ (7.9) and 1000 seed weight (24.5 g) as compared PRB-1 (5.4, 5.8 and 22.3 g, respectively). Among the fertility levels tried, the fertility level of 60:30 NP kg ha⁻¹ recorded significantly higher seed yield (845 kg ha^-1) and stover yield (1359 kg ha⁻¹) as compared to 40:20 NP kg ha^-1 (808 and 1259 kg ha⁻¹ respectively) and 20:10 NP kg ha^-1 (793 and 1144 kg ha⁻¹ respectively). Within the treatment combinations, IC 79147 genotype having 30*10 cm planting geometry with 60:30 NP kg ha⁻¹ recorded significantly higher seed yield (1070 kg ha⁻¹), clusters plant⁻¹ (10.3), seeds clusters⁻¹ (9.9) and 1000 seed weight (27.3 g) compared to other treatment combinations.

Keywords: buckwheat, planting geometry, genotypes, fertility levels

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Authors: Ruchi Pathania, Ahmad Ahmad, Shireesh Srivastava

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Cyanobacteria is a promising microbe that can capture and convert atmospheric CO₂ and light into valuable industrial bio-products like biofuels, biodegradable plastics, etc. Among their most attractive traits are faster autotrophic growth, whole year cultivation using non-arable land, high photosynthetic activity, much greater biomass and productivity and easy for genetic manipulations. Cyanobacteria store carbon in the form of glycogen which can be hydrolyzed to release glucose and fermented to form bioethanol or other valuable products. Marine cyanobacterial species are especially attractive for countries with scarcity of freshwater. We recently identified a marine native cyanobacterium Synechococcus sp. BDU 130192 which has good growth rate and high level of polyglucans accumulation compared to Synechococcus PCC 7002. In this study, firstly we sequenced the whole genome and the sequences were annotated using the RAST server. Genome scale metabolic model (GSMM) was reconstructed through COBRA toolbox. GSMM is a computational representation of the metabolic reactions and metabolites of the target strain. GSMMs construction through the application of Flux Balance Analysis (FBA), which uses external nutrient uptake rates and estimate steady state intracellular and extracellular reaction fluxes, including maximization of cell growth. The model, which we have named isyn942, includes 942 reactions and 913 metabolites having 831 metabolic, 78 transport and 33 exchange reactions. The phylogenetic tree obtained by BLAST search revealed that the strain was a close relative of Synechococcus PCC 7002. The flux balance analysis (FBA) was applied on the model iSyn942 to predict the theoretical yields (mol product produced/mol CO₂ consumed) for native and non-native products like acetone, butanol, etc. under phototrophic condition by applying metabolic engineering strategies. The reported strain can be a viable strain for biotechnological applications, and the model will be helpful to researchers interested in understanding the metabolism as well as to design metabolic engineering strategies for enhanced production of various bioproducts.

Keywords: cyanobacteria, flux balance analysis, genome scale metabolic model, metabolic engineering

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Authors: Chloe Richards, Adrian Delgado Ollero, Yan Delaure, Fiona Regan

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Growing concern about the natural environment has accelerated the search for non-toxic, but at the same time, economically reasonable, antifouling materials. Bioinspired surfaces, due to their nano and micro topographical antifouling capabilities, provide a hopeful approach to the design of novel antifouling surfaces. Biological organisms are known to have highly evolved and complex topographies, demonstrating antifouling potential, i.e. shark skin. Previous studies have examined the antifouling ability of topographic patterns, textures and roughness scales found on natural organisms. One of the mechanisms used to explain the adhesion of cells to a substrate is called attachment point theory. Here, the fouling organism experiences increased attachment where there are multiple attachment points and reduced attachment, where the number of attachment points are decreased. In this study, an attempt to characterize the microtopography of the common brill fish, Scophthalmus rhombus, was undertaken. Scophthalmus rhombus is a small flatfish of the family Scophthalmidae, inhabiting regions from Norway to the Mediterranean and the Black Sea. They reside in shallow sandy and muddy coastal areas at depths of around 70 – 80 meters. Six engineered surfaces (inspired by the Brill fish scale) produced by a 2-photon polymerization (2PP) process were evaluated for their potential as an antifouling solution for incorporation onto tidal energy blades. The micro-textures were analyzed for their AF potential under both static and dynamic laboratory conditions using two laboratory grown diatom species, Amphora coffeaeformis and Nitzschia ovalis. The incorporation of a surface topography was observed to cause a disruption in the growth of A. coffeaeformis and N. ovalis cells on the surface in comparison to control surfaces. This work has demonstrated the importance of understanding cell-surface interaction, in particular, topography for the design of novel antifouling technology. The study concluded that biofouling can be controlled by physical modification, and has contributed significant knowledge to the use of a successful novel bioinspired AF technology, based on Brill, for the first time.

Keywords: attachment point theory, biofouling, Scophthalmus rhombus, topography

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Authors: Madhusudana S. N., Reeta Mani, Ashwini S. Satishchandra P., Netravati, Udhani V., Fiaz A., Karande S.

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Rabies is an acute encephalitis which is considered 100% fatal despite occasional reports of survivors. However, in recent times more cases of human rabies survivors are being reported. In the last 5 years, there are six laboratories confirmed human rabies survivors in India alone. All cases were children below 15 years and all contracted the disease by dog bites. All of them also had received the full or partial course of rabies vaccination and 4 out of 6 had also received rabies immunoglobulin. All cases were treated in intensive care units in hospitals at Bangalore, Mumbai, Chandigarh, Lucknow and Goa. We report here the results of immunological and virological studies conducted at our laboratory on these patients. The clinical samples that were obtained from these patients were Serum, CSF, nuchal skin biopsy and saliva. Serum and CSF samples were subjected to standard RFFIT for estimation of rabies neutralizing antibodies. Skin biopsy, CSF and saliva were processed by TaqMan real-time PCR for detection of viral RNA. CSF, saliva and skin homogenates were also processed for virus isolation by inoculation of suckling mice. The PBMCs isolated from fresh blood was subjected to ELISPOT assay to determine the type of immune response (Th1/Th2). Both CSF and serum were also investigated for selected cytokines by Luminex assay. The level of antibodies to virus G protein and N protein were determined by ELISA. All survivors had very high titers of RVNA in serum and CSF 100 fold higher than non-survivors and vaccine controls. A five-fold rise in titer could be demonstrated in 4 out of 6 patients. All survivors had a significant increase in antibodies to G protein in both CSF and serum when compared to non-survivors. There was a profound and robust Th1 response in all survivors indicating that interferon gamma could play an important factor in virus clearance. We could isolate viral RNA in only one patient four years after he had developed symptoms. The partial N gene sequencing revealed 99% homology to species I strain prevalent in India. Levels of selected cytokines in CSF and serum did not reveal any difference between survivors and non-survivors. To conclude, survival from rabies is mediated by virus-specific immune responses of the host and clearance of rabies virus from CNS may involve the participation of both Th2 and Th1 immune responses.

Keywords: rabies, rabies treatment, rabies survivors, immune reponse in rabies encephalitis

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Authors: Jasmal A. Syamsu, Muhammad Yusuf, Hikmah M. Ali, Mawardi A. Asja, Zulkharnaim

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This study was conducted and aimed in identifying and mapping the pasture as an ecological base of beef cattle. A survey was carried out during a period of April to June 2016, in Suppa, Mattirobulu, the district of Pinrang, South Sulawesi province. The mapping process of grazing area was conducted in several stages; inputting and tracking of data points into Google Earth Pro (version 7.1.4.1529), affirmation and confirmation of tracking line visualized by satellite with a variety of records at the point, a certain point and tracking input data into ArcMap Application (ArcGIS version 10.1), data processing DEM/SRTM (S04E119) with respect to the location of the grazing areas, creation of a contour map (a distance of 5 m) and mapping tilt (slope) of land and land cover map-making. Analysis of land cover, particularly the state of the vegetation was done through the identification procedure NDVI (Normalized Differences Vegetation Index). This procedure was performed by making use of the Landsat-8. The results showed that the topography of the grazing areas of hills and some sloping surfaces and flat with elevation vary from 74 to 145 above sea level (asl), while the requirements for growing superior grass and legume is an altitude of up to 143-159 asl. Slope varied between 0 - > 40% and was dominated by a slope of 0-15%, according to the slope/topography pasture maximum of 15%. The range of NDVI values for pasture image analysis results was between 0.1 and 0.27. Characteristics of vegetation cover of pasture land in the category of vegetation density were low, 70% of the land was the land for cattle grazing, while the remaining approximately 30% was a grove and forest included plant water where the place for shelter of the cattle during the heat and drinking water supply. There are seven types of graminae and 5 types of legume that was dominant in the region. Proportionally, graminae class dominated up 75.6% and legume crops up to 22.1% and the remaining 2.3% was another plant trees that grow in the region. The dominant weed species in the region were Cromolaenaodorata and Lantana camara, besides that there were 6 types of floor plant that did not include as forage fodder.

Keywords: pastoral, ecology, mapping, beef cattle

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Authors: D. Hammad, D. P. Tonge

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Rheumatoid arthritis (RA) is a disabling and common autoimmune disease during which the body's immune system attacks healthy tissues. This results in complicated and long-lasting actions being carried out by the immune system, which typically only occurs when the immune system encounters a foreign object. In the case of RA, the disease affects millions of people and causes joint inflammation, ultimately leading to the destruction of cartilage and bone. Interestingly, the disease mechanism still remains unclear. It is likely that RA occurs as a result of a complex interplay of genetic and environmental factors including an imbalance in the microorganism population inside our body. The human microbiome or microbiota is an extensive community of microorganisms in and on the bodies of animals, which comprises bacteria, fungi, viruses, and protozoa. Recently, the development of molecular techniques to characterize entire bacterial communities has renewed interest in the involvement of the microbiome in the development and progression of RA. We believe that an imbalance in some of the specific bacterial species in the gut, mouth and other sites may lead to atopobiosis; the translocation of these organisms into the blood, and that this may lead to changes in immune system status. The aim of this study was, therefore, to characterize the microbiome of RA serum samples in comparison to healthy control subjects using 16S rRNA gene amplification and sequencing. Serum samples were obtained from healthy control volunteers and from patients with RA both prior to, and following treatment. The bacterial community present in each sample was identified utilizing V4 region 16S rRNA amplification and sequencing. Bacterial identification, to the lowest taxonomic rank, was performed using a range of bioinformatics tools. Significantly, the proportions of the Lachnospiraceae, Ruminococcaceae, and Halmonadaceae families were significantly increased in the serum of RA patients compared with healthy control serum. Furthermore, the abundance of Bacteroides and Lachnospiraceae nk4a136_group, Lachnospiraceae_UGC-001, RuminococcaceaeUCG-014, Rumnococcus-1, and Shewanella was also raised in the serum of RA patients relative to healthy control serum. These data support the notion of a blood microbiome and reveal RA-associated changes that may have significant implications for biomarker development and may present much-needed opportunities for novel therapeutic development.

Keywords: blood microbiome, gut and oral bacteria, Rheumatoid arthritis, 16S rRNA gene sequencing

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Authors: Suganya Chandrababu, Dhundy Bastola

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Like in the gut, the subgingival microbiota plays a crucial role in oral hygiene, health, and cariogenic diseases. Human activities like diet, antibiotics, and periodontal treatments alter the bacterial communities, metabolism, and functions in the oral cavity, leading to a dysbiotic state and changes in the plaques of orthodontic patients. Fixed periodontal appliances hinder oral hygiene and cause changes in the dental plaques influencing the subgingival microbiota. However, the microbial species’ diversity and complexity pose a great challenge in understanding the taxa’s community distribution patterns and their role in oral health. In this research, we analyze the subgingival microbial samples from individuals with fixed dental appliances (metal/clear) using an in silico approach. We employ exploratory hypothesis-driven multivariate and regression analysis to shed light on the microbial community and its functional fluctuations due to dental appliances used and identify risks associated with complex disease phenotypes. Our findings confirm the changes in oral microbiota composition due to the presence and type of fixed orthodontal devices. We identified seven main periodontic pathogens, including Bacteroidetes, Actinobacteria, Proteobacteria, Fusobacteria, and Firmicutes, whose abundances were significantly altered due to the presence and type of fixed appliances used. In the case of metal braces, the abundances of Bacteroidetes, Proteobacteria, Fusobacteria, Candidatus saccharibacteria, and Spirochaetes significantly increased, while the abundance of Firmicutes and Actinobacteria decreased. However, in individuals With clear braces, the abundance of Bacteroidetes and Candidatus saccharibacteria increased. The highest abundance value (P-value=0.004 < 0.05) was observed with Bacteroidetes in individuals with the metal appliance, which is associated with gingivitis, periodontitis, endodontic infections, and odontogenic abscesses. Overall, the bacterial abundances decrease with clear type and increase with metal type of braces. Regression analysis further validated the multivariate analysis of variance (MANOVA) results, supporting the hypothesis that the presence and type of the fixed oral appliances significantly alter the bacterial abundance and composition.

Keywords: oral microbiota, statistical analysis, fixed or-thodontal appliances, bacterial abundance, multivariate analysis, regression analysis

Procedia PDF Downloads 166

Authors: Débora P. Jaeschke, Eduardo A. Merlo, Rosane Rech, Giovana D. Mercali, Ligia D. F. Marczak

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Carotenoids are high value added pigments that can be alternatively extracted from some microalgae species. However, the application of carotenoids synthetized by microalgae is still limited due to the utilization of organic toxic solvents. In this context, studies involving alternative extraction methods have been conducted with more sustainable solvents to replace and reduce the solvent volume and the extraction time. The aim of the present work was to evaluate the extraction time of carotenoids from the microalgae Heterochlorella luteoviridis using moderate electric field (MEF) as a pre-treatment to the extraction. The extraction methodology consisted of a pre-treatment in the presence of MEF (180 V) and ethanol (25 %, v/v) for 10 min, followed by a diffusive step performed for 50 min using a higher ethanol concentration (75 %, v/v). The extraction experiments were conducted at 30 °C and, to keep the temperature at this value, it was used an extraction cell with a water jacket that was connected to a water bath. Also, to enable the evaluation of MEF effect on the extraction, control experiments were performed using the same cell and conditions without voltage application. During the extraction experiments, samples were withdrawn at 1, 5 and 10 min of the pre-treatment and at 1, 5, 30, 40 and 50 min of the diffusive step. Samples were, then, centrifuged and carotenoids analyses were performed in the supernatant. Furthermore, an exhaustive extraction with ethyl acetate and methanol was performed, and the carotenoids content found for this analyses was considered as the total carotenoids content of the microalgae. The results showed that the application of MEF as a pre-treatment to the extraction influenced the extraction yield and the extraction time during the diffusive step; after the MEF pre-treatment and 50 min of the diffusive step, it was possible to extract up to 60 % of the total carotenoids content. Also, results found for carotenoids concentration of the extracts withdrawn at 5 and 30 min of the diffusive step did not presented statistical difference, meaning that carotenoids diffusion occurs mainly in the very beginning of the extraction. On the other hand, the results for control experiments showed that carotenoids diffusion occurs mostly during 30 min of the diffusive step, which evidenced MEF effect on the extraction time. Moreover, carotenoids concentration on samples withdrawn during the pre-treatment (1, 5 and 10 min) were below the quantification limit of the analyses, indicating that the extraction occurred in the diffusive step, when ethanol (75 %, v/v) was added to the medium. It is possible that MEF promoted cell membrane permeabilization and, when ethanol (75 %) was added, carotenoids interacted with the solvent and the diffusion occurred easily. Based on the results, it is possible to infer that MEF promoted the decrease of carotenoids extraction time due to the increasing of the permeability of the cell membrane which facilitates the diffusion from the cell to the medium.

Keywords: moderate electric field (MEF), pigments, microalgae, ethanol

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Authors: Sheng-Yu Lee

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Aim: Opioid use disorder is often characterized by repetitive drug-seeking and drug-taking behaviors with severe public health consequences. Animal model showed that opioid-induced perturbations in the gut microbiota causally relate to neuroinflammation, deficits in reward responding, and opioid tolerance, possibly due to changes in gut microbiota. Therefore, we propose that the dysbiosis of gut microbiota can be associated with pathogenesis of opioid dependence. In this current study, we explored the differences in gut microbiota between patients and normal controls and in patients before and after initiation of methadone treatment program for 12 weeks. Methods: Patients with opioid use disorder between 20 and 65 years were recruited from the methadone maintenance outpatient clinic in 2 medical centers in the Southern Taiwan. Healthy controls without any family history of major psychiatric disorders (schizophrenia, bipolar disorder and major depressive disorder) were recruited from the community. After initial screening, 15 patients with opioid use disorder joined the study for initial evaluation (Week 0), 12 of them completed the 12-week follow-up while receiving methadone treatment and ceased heroin use (Week 12). Fecal samples were collected from the patients at baseline and the end of 12th week. A one-time fecal sample was collected from the healthy controls. The microbiota of fecal samples were investigated using 16S rRNA V3V4 amplicon sequencing, followed by bioinformatics and statistical analyses. Results: We found no significant differences in species diversity in opioid dependent patients between Week 0 and Week 12, nor compared between patients at both points and controls. For beta diversity, using principal component analysis, we found no significant differences between patients at Week 0 and Week 12, however, both patient groups showed significant differences compared to control (P=0.011). Furthermore, the linear discriminant analysis effect size (LEfSe) analysis was used to identify differentially enriched bacteria between opioid use patients and healthy controls. Compared to controls, the relative abundance of Lactobacillaceae Lactobacillus (L. Lactobacillus), Megasphaera Megasphaerahexanoica (M. Megasphaerahexanoica) and Caecibacter Caecibactermassiliensis (C Caecibactermassiliensis) were increased in patients at Week 0, while Coriobacteriales Atopobiaceae (C. Atopobiaceae), Acidaminococcus Acidaminococcusintestini (A. Acidaminococcusintestini) and Tractidigestivibacter Tractidigestivibacterscatoligenes (T. Tractidigestivibacterscatoligenes) were increased in patients at Week 12. Conclusion: In conclusion, we suggest that the gut microbiome community maybe linked to opioid use disorder, such differences may not be altered even after 12-week of cessation of opioid use.

Keywords: opioid use disorder, gut microbiota, methadone treatment, follow up study

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Authors: Bouchekoua Myriam, Aloui Dorsaf, Trabelsi Sonia

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Introduction Food hygiene in hospitals is important, particularly among patients who could be more vulnerable than healthy subjects to microbiological and nutritional risks. The consumption of contaminated food may be responsible for foodborne diseases, which can be severe among hospitalized patients, especially those immunocompromised. The aim of our study was to assess the state of hygiene in the internal catering department of a Tunisian hospital. Methodology and major results: A prospective study was conducted for one year in the Parasitology-Mycology laboratory of Charles Nicolle Hospital. Samples were taken from the kitchen staff, worktops, and cooking utensils used in the internal catering department. Thirty one employees have benefited from stool exams and scotch tape in order to evaluate the degree of infestation of parasites. 35% of stool exams were positive. Protozoa were the only parasites detected. Blastocystis sp was the species mostly found in nine food handlers. Its role as a human pathogen is still controversial. Pathogenic protozoa were detected in two food handlers (Giardia intestinalis in one person and Dientamoeba fragilis in the other one. Non-pathogenic protozoa were found in two cases; among them, only one had digestive symptoms without a statistically significant association with the carriage of intestinal parasites. Moreover, samples were performed from the hands of the staff in order to search for a fungal carriage. Thus, 25 employees (81%) were colonized by fungi, including molds. Besides, mycological examination among food handlers with a suspected dermatomycosis for diagnostic confirmation concluded foot onychomycosis in 32% of cases and interdigital intertrigo in 26%. Only one person had hand onychomycosis. Among the 17 samples taken from worktops and kitchen utensils, fungal contamination was detected in 13 sites. Hot and cold equipment were the most contaminated. Molds were mainly identified as belonging to five different genera. Cladosporium sp was predominant. Conclusion: In the view of the importance of intestinal parasites among food handlers, the intensity of fungi hand carriage among these employees, and the high level of fungal contamination in worktops and kitchen utensils, a reinforcement of hygiene measures is more than essential in order to minimize the alimentary contamination-risk.

Keywords: hospital kitchen, environment, intestinal parasitosis, fungal carriage, fungal contamination

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Authors: William E. Bauta, Jennifer Rebeles, Reggie Jacob

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Porphyrins are noteworthy in biomedical science for their cancer tissue accumulation and photophysical properties. The preferential accumulation of some porphyrins in cancerous tissue has been known for many years. This, combined with their characteristic photophysical and photochemical properties, including their strong fluorescence and their ability to generate reactive oxygen species in vivo upon laser irradiation, has led to much research into the application of porphyrins as cancer diagnostic and therapeutic agents. Porphyrins have been used as dyes to detect cancer cells both in vivo and, less commonly, in vitro. In one example, human sputum samples from lung cancer patients and patients without the disease were dissociated and stained with the porphyrin TCPP (5,10,15,20-tetrakis-(4-carboxyphenyl)-porphine). Cells were analyzed by flow cytometry. Cancer samples were identified by their higher TCPP fluorescence intensity relative to the no-cancer controls. However, quantitative analysis of fluorescence in cell suspensions stained with multiple fluorophores requires particles stained with each of the individual fluorophores as controls. Fluorescent control particles must be compatible in size with flow cytometer fluidics and have favorable hydrodynamic properties in suspension. They must also display fluorescence comparable to the cells of interest and be stable upon storage amine-functionalized spherical polystyrene beads in the 5 to 20-micron diameter range that was reacted with TCPP and EDC in aqueous pH six buffer overnight to form amide bonds. Beads were isolated by centrifugation and tested by flow cytometry. The 10-micron amine-functionalized beads displayed the best combination of fluorescence intensity and hydrodynamic properties, such as lack of clumping and remaining in suspension during the experiment. These beads were further optimized by varying the stoichiometry of EDC and TCPP relative to the amine. The reaction was accompanied by the formation of a TCPP-related particulate, which was removed, after bead centrifugation, using a microfiltration process. The resultant TCPP-functionalized beads were compatible with flow cytometry conditions and displayed a fluorescence comparable to that of stained cells, which allowed their use as fluorescence standards. The beads were stable in refrigerated storage in the dark for more than eight months. This work demonstrates the first preparation of porphyrin-functionalized flow cytometry control beads.

Keywords: tetraaryl porphyrin, polystyrene beads, flow cytometry, peptide coupling

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Authors: Alija Uzunovic, Sasa Pilipovic, Aida Sapcanin, Zahida Ademovic, Berina Pilipović

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Capsaicin is a naturally occurring alkaloid extracted from capsicum fruit extracts of different of Capsicum species. It has been employed topically to treat many diseases such as rheumatoid arthritis, osteoarthritis, cancer pain and nerve pain in diabetes. The high degree of pre-systemic metabolism of intragastrical capsaicin and the short half-life of capsaicin by intravenous administration made topical application of capsaicin advantageous. In this study, we have evaluated differences in the dissolution characteristics of capsaicin patch 11 mg (purchased from market) at different dissolution rotation speed. The proposed patch area is 308 cm2 (22 cm x 14 cm; it contains 36 µg of capsaicin per square centimeter of adhesive). USP Apparatus 5 (Paddle Over Disc) is used for transdermal patch testing. The dissolution study was conducted using USP apparatus 5 (n=6), ERWEKA DT800 dissolution tester (paddle-type) with addition of a disc. The fabricated patch of 308 cm2 is to be cut into 9 cm2 was placed against a disc (delivery side up) retained with the stainless-steel screen and exposed to 500 mL of phosphate buffer solution pH 7.4. All dissolution studies were carried out at 32 ± 0.5 °C and different rotation speed (50± 5; 100± 5 and 150± 5 rpm). 5 ml aliquots of samples were withdrawn at various time intervals (1, 4, 8 and 12 hours) and replaced with 5 ml of dissolution medium. Withdrawn were appropriately diluted and analyzed by reversed-phase liquid chromatography (RP-LC). A Reversed Phase Liquid Chromatography (RP-LC) method has been developed, optimized and validated for the separation and quantitation of capsaicin in a transdermal patch. The method uses a ProntoSIL 120-3-C18 AQ 125 x 4,0 mm (3 μm) column maintained at 600C. The mobile phase consisted of acetonitrile: water (50:50 v/v), the flow rate of 0.9 mL/min, the injection volume 10 μL and the detection wavelength 222 nm. The used RP-LC method is simple, sensitive and accurate and can be applied for fast (total chromatographic run time was 4.0 minutes) and simultaneous analysis of capsaicin and dihydrocapsaicin in a transdermal patch. According to the results obtained in this study, we can conclude that the relative difference of dissolution rate of capsaicin after 12 hours was elevated by increase of dissolution rotation speed (100 rpm vs 50 rpm: 84.9± 11.3% and 150 rpm vs 100 rpm: 39.8± 8.3%). Although several apparatus and procedures (USP apparatus 5, 6, 7 and a paddle over extraction cell method) have been used to study in vitro release characteristics of transdermal patches, USP Apparatus 5 (Paddle Over Disc) could be considered as a discriminatory test. would be able to point out the differences in the dissolution rate of capsaicin at different rotation speed.

Keywords: capsaicin, in vitro, patch, RP-LC, transdermal

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Authors: Joulié Aurélien, Jourdain Elsa, Bailly Xavier, Gasqui Patrick, Yang Elise, Leblond Agnès, Rousset Elodie, Sidi-Boumedine Karim

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Q fever is a worldwide zoonosis caused by the gram-negative obligate intracellular bacterium Coxiella burnetii. Following the recent outbreaks in the Netherlands, a hyper virulent clone was found to be the cause of severe human cases of Q fever. In livestock, Q fever clinical manifestations are mainly abortions. Although the abortion rates differ between ruminant species, C. burnetii’s virulence remains understudied, especially in enzootic areas. In this study, the infectious potential of three C. burnetii isolates collected from French farms of small ruminants were compared to the reference strain Nine Mile (in phase II and in an intermediate phase) using an in vivo (CD1 mice) model. Mice were challenged with 105 live bacteria discriminated by propidium monoazide-qPCR targeting the icd-gene. After footpad inoculation, spleen and popliteal lymph node were harvested at 10 days post-inoculation (p.i). The strain invasiveness in spleen and popliteal nodes was assessed by qPCR assays targeting the icd-gene. Preliminary results showed that the avirulent strains (in phase 2) failed to pass the popliteal barrier and then to colonize the spleen. This model allowed a significant differentiation between strain’s invasiveness on biological host and therefore identifying distinct virulence profiles. In view of these results, we plan to go further by testing fifteen additional C. burnetii isolates from French farms of sheep, goat and cattle by using the above-mentioned in vivo model. All 15 strains display distant MLVA (multiple-locus variable-number of tandem repeat analysis) genotypic profiles. Five of the fifteen isolates will bee also tested in vitro on ovine and bovine macrophage cells. Cells and supernatants will be harvested at day1, day2, day3 and day6 p.i to assess in vitro multiplication kinetics of strains. In conclusion, our findings might help the implementation of surveillance of virulent strains and ultimately allow adapting prophylaxis measures in livestock farms.

Keywords: Q fever, invasiveness, ruminant, virulence

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Authors: Jeremy Johnson, Craig Nitschke, Luke Kelly

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Fire regimes, climate and topographic gradients interact to influence ecosystem structure and function across fire-prone, montane landscapes worldwide. Biota have developed a range of adaptations to historic fire regime thresholds, which allow them to persist in these environments. In south-eastern Australia, a signal of fire regime changes is emerging across these landscapes, and anthropogenic climate change is likely to be one of the main drivers of an increase in burnt area and more frequent wildfire over the last 25 years. This shift has the potential to modify vegetation structure and composition at broad scales, which may lead to landscape patterns to which biota are not adapted, increasing the likelihood of local extirpation of some mammal species. This study aimed to address concerns related to the influence of abrupt changes in fire regimes on mammal populations in montane landscapes. It first examined the impact of climate, topography, and vegetation on fire patterns and then explored the consequences of these changes on mammal populations and their habitats. Field studies were undertaken across diverse vegetation, fire severity and fire frequency gradients, utilising camera trapping and passive acoustic monitoring methodologies and the collection of fine-scale vegetation data. Results show that drought is a primary contributor to fire regime shifts at the landscape scale, while topographic factors have a variable influence on wildfire occurrence at finer scales. Frequent, high severity wildfire influenced forest structure and composition at broad spatial scales, and at fine scales, it reduced occurrence of hollow-bearing trees and promoted coarse woody debris. Mammals responded differently to shifts in forest structure and composition depending on their habitat requirements. This study highlights the complex interplay between fire regimes, environmental gradients, and biotic adaptations across temporal and spatial scales. It emphasizes the importance of understanding complex interactions to effectively manage fire-prone ecosystems in the face of climate change.

Keywords: fire, ecology, biodiversity, landscape ecology

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Authors: A. Díaz-Roa, P. I. Silva Junior, F. J. Bello

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Sarconesiopsis magellanica (Diptera: Calliphoridae) is a medically important necrophagous fly which is used for establishing the post-mortem interval. Dipterous maggots release diverse proteins and peptides contained in larval excretion and secretion (ES) products playing a key role in digestion. The most important mechanism for combating infection using larval therapy depends on larval ES. These larvae are protected against infection by a diverse spectrum of antimicrobial peptides (AMPs), one already known like lucifensin. Special interest in these peptides has also been aroused regarding understanding their role in wound healing since they degrade necrotic tissue and kill different bacteria during larval therapy. The action of larvae on wounds occurs through 3 mechanisms of action: removal of necrotic tissue, stimulation of granulation tissue, and antibacterial action of larval ES. Some components of the ES include calcium, urea, allantoin ammonium bicarbonate and reducing the viability of Gram positive and Gram negative bacteria. The Lucilia sericata fly larvae have been the most used, however, we need to evaluate new species that could potentially be similar or more effective than fly above. This study was thus aimed at identifying and characterizing S. magellanica AMPs contained in ES products for the first time and compared them with the common fly used L. sericata. These products were obtained from third-instar larvae taken from a previously established colony. For the first analysis, ES fractions were separate by Sep-Pak C18 disposable columns (first step). The material obtained was fractionated by RP-HPLC by using Júpiter C18 semi-preparative column. The products were then lyophilized and their antimicrobial activity was characterized by incubation with different bacterial strains. The first chromatographic analysis of ES from L. sericata gives 6 fractions with antimicrobial activity against Gram-positive bacteria Micrococus luteus, and 3 fractions with activity against Gram-negative bacteria Pseudomonae aeruginosa while the one from S. magellanica gaves 1 fraction against M. luteus and 4 against P. aeruginosa. Maybe one of these fractions could correspond to the peptide already known from L. sericata. These results show the first work for supporting further experiments aimed at validating S. magellanica use in larval therapy. We still need to search if we find some new molecules, by making mass spectrometry and ‘de novo sequencing’. Further studies are necessary to identify and characterize them to better understand their functioning.

Keywords: antimicrobial peptides, larval therapy, Lucilia sericata, Sarconesiopsis magellanica

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Authors: Chiu-Wen Chang, I-Yun Chang, Wei-Ting Chen, Hui-Ping Ho, Chao-Ying Pan, Joh-Jong Huang

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Dengue is a ‘community disease’ or ‘environmental disease’, as long as the environment exist suitable container (including natural and artificial) for mosquito breeding, once the virus invade will lead to the dengue epidemic. Surveillance of vector density is critical to effective infectious disease control and play an important role in monitoring the dynamics of mosquitoes in community, such as mosquito species, density, distribution area. The objective of this study was to examine the relationship in vector density survey (Breteau index, Adult index, House index, Container index, and Larvae index) form 2014 to 2016 in Kaohsiung City and evaluate the effects of introducing the Breeding Elimination and Appraisal Team (hereinafter referred to as BEAT) as an intervention measure on eliminating dengue vector breeding site started from May 2016. BEAT were performed on people who were suspected of contracting dengue fever, a surrounding area measuring 50 meters by 50 meters was demarcated as the emergency prevention and treatment zone. BEAT would perform weekly vector mosquito inspections and vector mosquito inspections in regions with a high Gravitrap index and assign a risk assessment index to each region. These indices as well as the prevention and treatment results were immediately reported to epidemic prevention-related units every week. The results indicated that, vector indices from 2014 to 2016 showed no statistically significant differences in the Breteau index, adult index, and house index (p > 0.05) but statistically significant differences in the container index and larvae index (p <0.05). After executing the integrated elimination work, container index and larvae index are statistically significant different from 2014 to 2016 in the (p < 0.05). A post hoc test indicated that the container index of 2014 (M = 12.793) was significantly higher than that of 2016 (M = 7.631), and that the larvae index of 2015 (M = 34.065) was significantly lower than that of 2014 (M = 66.867). The results revealed that effective vector density surveillance could highlight the focus breeding site and then implement the immediate control action (BEAT), which successfully decreased the vector density and the risk of dengue epidemic.

Keywords: Breteau index, dengue control, monitoring method, vector density

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Authors: Hanbyul Kim, Hye-Jin Kin, Taehun Park, Woo Jun Sul, Susun An

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Skin is the largest organ of the human body and can provide the diverse habitat for various microorganisms. The ecology of the skin surface selects distinctive sets of microorganisms and is influenced by both endogenous intrinsic factors and exogenous environmental factors. The diversity of the bacterial community in the skin also depends on multiple host factors: gender, age, health status, location. Among them, age-related changes in skin structure and function are attributable to combinations of endogenous intrinsic factors and exogenous environmental factors. Skin aging is characterized by a decrease in sweat, sebum and the immune functions thus resulting in significant alterations in skin surface physiology including pH, lipid composition, and sebum secretion. The present study gives a comprehensive clue on the variation of skin microbiota and the correlations between ages by analyzing and comparing the metagenome of skin microbiome using Next Generation Sequencing method. Skin bacterial diversity and composition were characterized and compared between two different age groups: younger (20 – 30y) and older (60 - 70y) Xian, Chinese women. A total of 73 healthy women meet two conditions: (I) living in Xian, China; (II) maintaining healthy skin status during the period of this study. Based on Ribosomal Database Project (RDP) database, skin samples of 73 participants were enclosed with ten most abundant genera: Chryseobacterium, Propionibacterium, Enhydrobacter, Staphylococcus and so on. Although these genera are the most predominant genus overall, each genus showed different proportion in each group. The most dominant genus, Chryseobacterium was more present relatively in Young group than in an old group. Similarly, Propionibacterium and Enhydrobacter occupied a higher proportion of skin bacterial composition of the young group. Staphylococcus, in contrast, inhabited more in the old group. The beta diversity that represents the ratio between regional and local species diversity showed significantly different between two age groups. Likewise, The Principal Coordinate Analysis (PCoA) values representing each phylogenetic distance in the two-dimensional framework using the OTU (Operational taxonomic unit) values of the samples also showed differences between the two groups. Thus, our data suggested that the composition and diversification of skin microbiomes in adult women were largely affected by chronological and physiological skin aging.

Keywords: next generation sequencing, age, Xian, skin microbiome

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Authors: Ekta, G. K. Darbha

Abstract:

Nanotechnology offers the potential of simultaneously increasing efficiency as compared to their bulk material as well as reducing harmful environmental impacts of pesticides in field of agriculture. The term nanopesticide covers different pesticides that are cumulative of several surfactants, polymers, metal ions, etc. of nanometer size ranges from 1-1000 nm and exhibit abnormal behavior (high efficacy and high specific surface area) of nanomaterials. Commercial formulations of pesticides used by farmers nowadays cannot be used effectively due to a number of problems associated with them. For example, more than 90% of applied formulations are either lost in the environment or unable to reach the target area required for effective pest control. Around 20−30% of pesticides are lost through emissions. A number of factors (application methods, physicochemical properties of the formulations, and environmental conditions) can influence the extent of loss during application. It is known that among various formulations, polymer-based formulations show the greatest potential due to their greater efficacy, slow release and protection against premature degradation of active ingredient as compared to other commercial formulations. However, the nanoformulations can have a significant effect on the fate of active ingredient as well as may release some new ingredients by reacting with existing soil contaminants. Environmental fate of these newly generated species is still not explored very well which is essential to field scale experiments and hence a lot to be explored in the field of environmental fate, nanotoxicology, transport properties and stability of such formulations. In our preliminary work, we have synthesized polymer based nanoformulation of commercially used weedicide atrazine. Atrazine belongs to triazine class of herbicide, which is used in the effective control of seed germinated dicot weeds and grasses. It functions by binding to the plastoquinone-binding protein in PS-II. Plant death results from starvation and oxidative damage caused by breakdown in electron transport system. The stability of the suspension of nanoformulation containing herbicide has been evaluated by considering different parameters like polydispersity index, particle diameter, zeta-potential under different environmental relevance condition such as pH range 4-10, temperature range from 25°C to 65°C and stability of encapsulation also have been studied for different amount of added polymer. Morphological characterization has been done by using SEM.

Keywords: atrazine, nanoformulation, nanopesticide, nanotoxicology

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Authors: Maxence Queyrel, Edi Prifti, Alexandre Templier, Jean-Daniel Zucker

Abstract:

Analysis of the human microbiome using metagenomic sequencing data has demonstrated high ability in discriminating various human diseases. Raw metagenomic sequencing data require multiple complex and computationally heavy bioinformatics steps prior to data analysis. Such data contain millions of short sequences read from the fragmented DNA sequences and stored as fastq files. Conventional processing pipelines consist in multiple steps including quality control, filtering, alignment of sequences against genomic catalogs (genes, species, taxonomic levels, functional pathways, etc.). These pipelines are complex to use, time consuming and rely on a large number of parameters that often provide variability and impact the estimation of the microbiome elements. Training Deep Neural Networks directly from raw sequencing data is a promising approach to bypass some of the challenges associated with mainstream bioinformatics pipelines. Most of these methods use the concept of word and sentence embeddings that create a meaningful and numerical representation of DNA sequences, while extracting features and reducing the dimensionality of the data. In this paper we present an end-to-end approach that classifies patients into disease groups directly from raw metagenomic reads: metagenome2vec. This approach is composed of four steps (i) generating a vocabulary of k-mers and learning their numerical embeddings; (ii) learning DNA sequence (read) embeddings; (iii) identifying the genome from which the sequence is most likely to come and (iv) training a multiple instance learning classifier which predicts the phenotype based on the vector representation of the raw data. An attention mechanism is applied in the network so that the model can be interpreted, assigning a weight to the influence of the prediction for each genome. Using two public real-life data-sets as well a simulated one, we demonstrated that this original approach reaches high performance, comparable with the state-of-the-art methods applied directly on processed data though mainstream bioinformatics workflows. These results are encouraging for this proof of concept work. We believe that with further dedication, the DNN models have the potential to surpass mainstream bioinformatics workflows in disease classification tasks.

Keywords: deep learning, disease prediction, end-to-end machine learning, metagenomics, multiple instance learning, precision medicine

Procedia PDF Downloads 101