Search results for: nutrient restriction

Authors: Radheshyam Yadav, Ramakrishna Wusirika

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Plant Growth Promoting Bacteria (PGPB) and Arbuscular Mycorrhizal (AM) Fungi are ubiquitous in soil and often very critical for crop yield and agriculture sustainability, and this has motivated the agricultural practices to support and promote PGPB and AM Fungi in agriculture. PGPB can be involved in a range of processes that affect Nitrogen (N) and Phosphorus (P) transformations in soil and thus influence nutrient availability and uptake to the plants. A field study with two wheat cultivars, HD-3086, and HD-2967 was performed in Malwa region, Bathinda of Punjab, India, to evaluate the effect of native and non-native PGPB alone and in combination with AM fungi as an inoculant on wheat grain yield, nutrient uptake and soil health parameters (dehydrogenase, urease, β‐glucosidase). Our results showed that despite an early insignificant increase in shoot length, plants treated with PGPB (Bacillus sp.) and AM Fungi led to a significant increase in shoot growth at maturity, aboveground biomass, nitrogen (45% - 40%) and phosphorus (40% - 34%) content in wheat grains relative to untreated control plants. Similarly, enhanced grain yield and nutrients uptake i.e. copper (27.15% - 36.25%) iron (43% - 53%) and zinc (44% - 47%) was recorded in PGPB and AM Fungi treated plants relative to untreated control. Overall, inoculation with native PGPB alone and in combination with AM Fungi provided benefits to enhance grain yield, wheat biofortification, and improved soil fertility, despite this effect varied depending on different PGPB isolates and wheat cultivars. These field study results provide evidence of the benefits of agricultural practices involving native PGPB and AM Fungi to the plants. These native strains and AM Fungi increased accumulations of copper, iron, and zinc in wheat grains, enhanced grain yield, and soil fertility.

Keywords: AM Fungi, biofortification, PGPB, soil microbial enzymes

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Authors: Sophie Oakes-Rogers, Di Bailey, Karen Slade

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Female offenders are a uniquely vulnerable group, who are at high risk of suicide. Whilst the prevention of self-harm and suicide remains a key global priority, we need to better understand the relationship between these challenging behaviours that constitute a pressing problem, particularly in environments designed to prioritise safety and security. Method choice is unlikely to be random, and is instead influenced by a range of cultural, social, psychological and environmental factors, which change over time and between countries. A key aspect of self-harm and suicide in women receiving forensic care is the lack of free access to methods. At a time where self-harm and suicide rates continue to rise internationally, understanding the role of these influencing factors and the impact of current suicide prevention strategies on the use of near-lethal methods is crucial. This poster presentation will present findings from 25 interviews and 3 focus groups, which enlisted a Participatory Action Research approach to explore the differences between self-harming and suicidal behavior. A key element of this research was using the lived experiences of women receiving forensic care from one forensic pathway in the UK, and the staffs who care for them, to discuss the role of near-lethal self-harm (NLSH). The findings and suggestions from the lived accounts of the women and staff will inform a draft assessment tool, which better assesses the risk of suicide based on the lethality of methods. This tool will be the first of its kind, which specifically captures the needs of women receiving forensic services. Preliminary findings indicate women engage in NLSH for two key reasons and is determined by their history of self-harm. Women who have a history of superficial non-life threatening self-harm appear to engage in NLSH in response to a significant life event such as family bereavement or sentencing. For these women, suicide appears to be a realistic option to overcome their distress. This, however, differs from women who appear to have a lifetime history of NLSH, who engage in such behavior in a bid to overcome the grief and shame associated with historical abuse. NLSH in these women reflects a lifetime of suicidality and indicates they pose the greatest risk of completed suicide. Findings also indicate differences in method selection between forensic provisions. Restriction of means appears to play a role in method selection, and findings suggest it causes method substitution. Implications will be discussed relating to the screening of female forensic patients and improvements to the current suicide prevention strategies.

Keywords: forensic mental health, method substitution, restriction of means, suicide

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Authors: Hai-Hong Gu, Yan-Jun Ai, Zheng Zhou

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Vanadium and titanium are rare metals with superior properties and are important resources in aerospace, aviation, and military. The vanadium/titanium magnetite are mostly ultra-lean ores, and a large number of tailings has been produced in the exploitation process. The tailings are characterized by loose structure, poor nutrient, complex composition and high trace metal contents. Returning green manure has been shown to not only increase plant biomass and soil nutrients but also change the bioavailability of trace metals and the microbial community structure. Fungi play an important role in decomposing organic matter and increasing soil fertility, and the application of organic matter also affects the community structure of fungi. The effects of green manure plants, alfalfa (Medicago sativa L.), returned to the tailings in situ on community structure of fungi, nutrients and bioavailability of trace metals in vanadium/titanium magnetite tailings were investigated in a pot experiment. The results showed that the fungal community diversity and richness were increase after alfalfa green manure returned in situ. The dominant phyla of the fungal community were Ascomycota, Basidiomycota and Ciliophora, especially, the phyla Ciliophora was rare in ordinary soil, but had been found to be the dominant phyla in tailings. Meanwhile, the nutrient properties and various trace metals may shape the microbial communities by affecting the abundance of fungi. It was found that the plant growth was stimulated and the available N and organic C were significantly improved in the vanadium/titanium magnetite tailing with the long-term returning of alfalfa green manure. Moreover, the DTPA-TEA extractable Cd and Zn concentrations in the vanadium/titanium magnetite tailing were reduced by 7.72%~23.8% and 8.02%~24.4%, respectively, compared with those in the non-returning treatment. The above results suggest that the returning of alfalfa green manure could be a potential approach to improve fungal community structure and restore mine tailing ecosystem.

Keywords: fungal community, green manure returning, vanadium/titanium magnet tailings, trace metals

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Authors: Saeid Doaei

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The various studies have examined the relationship between FTO gene expression and macronutrients levels. In order to obtain better viewpoint from this interactions, all of the existing studies were reviewed systematically. All published papers have been obtained and reviewed using standard and sensitive keywords from databases such as CINAHL, Embase, PubMed, PsycInfo, and the Cochrane, from 1990 to 2016. The results indicated that all of 6 studies that met the inclusion criteria (from a total of 428 published article) found FTO gene expression changes at short-term follow-ups. Four of six studies found an increased FTO gene expression after calorie restriction, while two of them indicated decreased FTO gene expression. The effect of protein, carbohydrate and fat were separately assessed and suggested by all of six studies. In conclusion, the level of FTO gene expression in hypothalamus is related to macronutrients levels. Future research should evaluate the long-term impact of dietary interventions.

Keywords: obesity, gene expression, FTO, macronutrients

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Authors: Chansiri Singhtaun

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This paper proposes a mathematical model and examines the performance of an exact algorithm for a location–transportation problems in humanitarian relief. The model determines the number and location of distribution centers in a relief network, the amount of relief supplies to be stocked at each distribution center and the vehicles to take the supplies to meet the needs of disaster victims under capacity restriction, transportation and budgetary constraints. The computational experiments are conducted on the various sizes of problems that are generated. Branch and bound algorithm is applied for these problems. The results show that this algorithm can solve problem sizes of up to three candidate locations with five demand points and one candidate location with up to twenty demand points without premature termination.

Keywords: disaster response, facility location, humanitarian relief, transportation

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Authors: Savannah Rugg, Carlo Moreno, Pushpa Soti, Alexis Racelis

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Situated in deep South Texas, the Lower Rio Grande Valley (LRGV) is considered one the most productive agricultural regions in the southern US. With the highest concentration of organic farms in the state (Hidalgo county), the LRGV has a strong potential to be leaders in sustainable agriculture. Finding management practices that comply with organic certification and increase the health of the agroecosytem and the farmers working the land is increasingly pertinent. Cover cropping, or the intentional planting of non-cash crop vegetation, can serve multiple functions in an agroecosystem by decreasing environmental pollutants that originate from the agroecosystem, reducing inputs needed for crop production, and potentially decreasing on-farm costs for farmers—overall increasing the sustainability of the farm. Use of cover crops on otherwise fallow lands have shown to enhance ecosystem services such as: attracting native beneficial insects (pollinators), increase nutrient availability in topsoil, prevent nutrient leaching, increase soil organic matter, and reduces soil erosion. In this study, four cover crops (Lablab, Sudan Grass, Sunn Hemp, and Pearl Millet) were analyzed in the subtropical region of south Texas to see how their multiple functions enhance ecosystem services. The four cover crops were assessed to see their potential to harbor native insects, their potential to increase soil nitrogen, to increase soil organic matter, and to suppress weeds. The preliminary results suggest that these subtropical varieties of cover crops have potential to enhance ecosystem services on agricultural land in the RGV by increasing soil organic matter (in all varieties), increasing nitrogen in topsoil (Lablab, Sunn Hemp), and reducing weeds (Sudan Grass).

Keywords: cover crops, ecosystem services, subtropical agriculture, sustainable agriculture

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Authors: Y. B. Sharma, K. B. Biswas

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This research project examines a 5000 cal yr BP sediment core record to reveal the consequences of human impact and climate variability on the tropical dry forests of the Mahi river basin, western India. To date there has been little research to assess the impact of climate variability and human impact on the vegetation dynamics of this region. There has also been little work to link changes in vegetation cover to documented changes in the basin hydrology over the past 100 years – although it is assumed that the two are closely linked. The key objective of this research project therefore is to understand the driving mechanisms responsible for the abrupt changes in the Mahi river basin as detailed in historical documentation and its impact on water resource management. The Mahi river basin is located in western India (22° 11’-24° 35’ N 72° 46’-74° 52’ E). Mahi river arises in the Malwa Plateau, Madhya Pradesh near Moripara and flows through the uplands and alluvial plain of Rajasthan and Gujarat provinces before draining into the Gulf of Cambay. Palaeoecological procedures (sedimentology, geochemical analysis, C&N isotopes and fossil pollen evidences) have been applied on sedimentary sequences collected from lakes in the Mahi basin. These techniques then facilitate to reconstruct the soil erosion, nutrient cycling, vegetation changes and climatic variability over the last 5000 years. Historical documentation detailing changes in demography, climate and landscape use over the past 100 years in this region will also be collated to compare with the most recent palaeoecological records. The results of the research work provide a detailed record of vegetation change, soil erosion, changes in aridity, and rainfall patterns in the region over the past 5000 years. This research therefore aims to determine the drivers of change and natural variability in the basin. Such information is essential for its current and future management including restoration.

Keywords: human impact, climate variability, vegetation cover, hydrology, water resource management, Mahi river basin, sedimentology, geochemistry, fossil pollen, nutrient cycling, vegetation changes, palaeoecology, aridity, rainfall, drivers of change

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Authors: Revathi S. Rajan, Pratibha Malik, Nupur Garg, Smitha Avula, Kamini A. Rao

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This study aimed to analyse the pregnancy outcomes in patients with TPO positivity after appropriate L-Thyroxin supplementation with close surveillance. All pregnant women attending the antenatal clinic at Milann-The Fertility Center, Bangalore, India- from Aug 2013 to Oct 2014 whose booking TSH was more than 2.5 mIU/L were included along with those pregnant women with prior hypothyroidism who were TPO positive. Those with TPO positive status were vigorously managed with appropriate thyroxin supplementation and the doses were readjusted every 3 to 4 weeks until delivery. Women with recurrent pregnancy loss were also tested for TPO positivity and if tested positive, were monitored serially with TSH and fT4 levels every 3 to 4 weeks and appropriately supplemented with thyroxin when the levels fluctuated. The testing was done after an informed consent in all these women. The statistical software namely SAS 9.2, SPSS 15.0, Stata 10.1, MedCalc 9.0.1, Systat 12.0 and R environment ver.2.11.1 were used for the analysis of the data. 460 pregnant women were screened for thyroid dysfunction at booking of which 52% were hypothyroid. Majority of them (31.08%) were subclinically hypothyroid and the remaining were overt. 25% of the total no. of patients screened were TPO positive. The various pregnancy complications that were observed in the TPO positive women were gestational glucose intolerance [60%], threatened abortion [21%], midtrimester abortion [4.3%], premature rupture of membranes [4.3%], cervical funneling [4.3%] and fetal growth restriction [3.5%]. 95.6% of the patients who followed up till the end delivered beyond 30 weeks. 42.6% of these patients had previous history of recurrent abortions or adverse obstetric outcome and 21.7% of the delivered babies required NICU admission. Obstetric outcomes in our study in terms of midtrimester abortions, placental abruption, and preterm delivery improved for the better after close monitoring of the thyroid hormone [TSH and fT4] levels every 3 to 4 weeks with appropriate dose adjustment throughout pregnancy. Euthyroid women with TPO positive status enrolled in the study incidentally were those with recurrent abortions/infertility and required thyroxin supplements due to elevated Thyroid hormone (TSH, fT4) levels during the course of their pregnancy. Significant associations were found with age>30 years and Hyperhomocysteinemia [p=0.017], recurrent pregnancy loss or previous adverse obstetric outcomes [p=0.067] and APLA [p=0.029]. TPO antibody levels >600 I U/ml were significantly associated with development of gestational hypertension [p=0.041] and fetal growth restriction [p=0.082]. Euthyroid women with TPO positivity were also screened periodically to counter fluctuations of the thyroid hormone levels with appropriate thyroxin supplementation. Thus, early identification along with aggressive management of thyroid dysfunction and stratification of these patients based on their TPO status with appropriate thyroxin supplementation beginning in the first trimester will aid risk modulation and also help avert complications.

Keywords: TPO antibody, subclinical hypothyroidism, anti nuclear antibody, thyroxin

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Authors: Sarra K. Ikhlef, Onita Basu

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Natural organic matter (NOM) removal efficiency using drinking water treatment biological filters can be highly influenced by backwashing conditions. Backwashing has the ability to remove the accumulated biomass and particles in order to regenerate the biological filters' removal capacity and prevent excessive headloss buildup. A lab scale system consisting of 3 biological filters was used in this study to examine the implications of different backwash strategies on biological filtration performance. The backwash procedures were evaluated based on their impacts on dissolved organic carbon (DOC) removals, biological filters’ biomass, backwash water volume usage, and particle removal. Results showed that under nutrient limited conditions, the simultaneous use of air and water under collapse pulsing conditions lead to a DOC removal of 22% which was significantly higher (p>0.05) than the 12% removal observed under water only backwash conditions. Employing a bed expansion of 20% under nutrient supplemented conditions compared to a 30% reference bed expansion while using the same amount of water volume lead to similar DOC removals. On the other hand, utilizing a higher bed expansion (40%) lead to significantly lower DOC removals (23%). Also, a backwash strategy that reduced the backwash water volume usage by about 20% resulted in similar DOC removals observed with the reference backwash. The backwash procedures investigated in this study showed no consistent impact on biological filters' biomass concentrations as measured by the phospholipids and the adenosine tri-phosphate (ATP) methods. Moreover, none of these two analyses showed a direct correlation with DOC removal. On the other hand, dissolved oxygen (DO) uptake showed a direct correlation with DOC removals. The addition of the extended terminal subfluidization wash (ETSW) demonstrated no apparent impact on DOC removals. ETSW also successfully eliminated the filter ripening sequence (FRS). As a result, the additional water usage resulting from implementing ETSW was compensated by water savings after restart. Results from this study provide insight to researchers and water treatment utilities on how to better optimize the backwashing procedure for the goal of optimizing the overall biological filtration process.

Keywords: biological filtration, backwashing, collapse pulsing, ETSW

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Authors: J. S. Bedi, J. P. S. Gill, R. S. Aulakh, Prabhjit Kaur

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The present study was undertaken to monitor organochlorine, organophosphate and synthetic pyrethroid pesticide residues in butter and ghee samples collected from six different districts of Punjab. The estimation of pesticide residues was done by multiple residue analytical technique using gas chromatography equipped with GC-ECD and GC-FTD. The confirmation of residues was done on gas chromatography mass spectrometry in both SIM and Scan mode. Results indicated the presence of HCH and pp DDE as predominant contaminant in both butter and ghee even after their ban/restriction on usage in India. Residues of HCH were detected in 25.5 and 23.2 % samples of butter and ghee, respectively, while residues of pp DDE were recorded in 29.3 and 25.0 % butter and ghee samples, respectively. More importantly, the presence of endosulfan, cypermethrin, fenvalerate, deltamethrin and chlorpyrifos was observed in few butter and ghee samples indicating the serious concerns. The spatial variation of pesticide residues occurrence indicated the cotton belt of Punjab as most affected.

Keywords: butter, ghee, pesticides residues, Punjab

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Authors: Karolina Rutkowska, Hubert Pausch, Jolanta Oprzadek, Krzysztof Flisikowski

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The placenta is one of the most important organs that plays a crucial role in the fetal growth and development. Placenta dysfunction is one of the primary cause of the intrauterine growth restriction (IUGR). Cattle have the cotyledonary placenta which consists of two anatomical parts: fetal and maternal. In the case of cattle during the first months of pregnancy, it is very easy to separate maternal caruncle from fetal cotyledon tissue, easier in fact than removing an ordinary glove from one's hand. Which in fact make easier to conduct tissue-specific molecular studies. Typically, animal models for the study of IUGR are created using surgical methods and malnutrition of the pregnant mother or in the case of mice by genetic modifications. However, proposed cattle model with MIMT1Del/WT deletion is unique because it was created without any surgical methods what significantly distinguish it from the other animal models. The primary objective of the study was to identify differential expression of selected miRNAs in the placenta from normal and intrauterine growth restricted fetuses. There was examined the expression of miRNA in the fetal and maternal part of the placenta from 24 fetuses (12 samples from the fetal part of the placenta and 12 samples from maternal part of the placenta). In the study, there was done miRNAs sequencing in the placenta of MIMT1Del/WT fetuses and MIMT1WT/WT fetuses. Then, there were selected miRNAs that are involved in fetal growth and development. Analysis of miRNAs expression was conducted on ABI7500 machine. miRNAs expression was analyzed by reverse-transcription polymerase chain reaction (RT-PCR). As the reference gene was used SNORD47. The results were expressed as 2ΔΔCt: ΔΔCt = (Ctij − CtSNORD47j) − (Cti1 − CtSNORD471). Where Ctij and CtSNORD47j are the Ct values for gene i and for SNORD47 in a sample (named j); Cti1 and CtSNORD471 are the Ct values in sample 1. Differences between groups were evaluated with analysis of variance by using One-Way ANOVA. Bonferroni’s tests were used for interpretation of the data. All normalised miRNA expression values are expressed on a value of natural logarithm. The data were expressed as least squares mean with standard errors. Significance was declared when P < 0.05. The study shows that miRNAs expression depends on the part of the placenta where they origin (fetal or maternal) and on the genotype of the animal. miRNAs offer a particularly new approach to study IUGR. Corresponding tissue samples were collected according to the standard veterinary protocols according to the European Union Normative for Care and Use of Experimental Animals. All animal experiments were approved by the Animal Ethics Committee of the State Provincial Office of Southern Finland (ESAVI-2010-08583/YM-23).

Keywords: placenta, intrauterine growth restriction, miRNA, cattle

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Authors: Kavita Khoiwal, Vatsla Dadhwal, K. Aparna Sharma, Dipika Deka, Plabani Sarkar

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Introduction: About 2.4 million (1.93 - 3.04 million) people are living with HIV/AIDS in India. Of all HIV infections, 39% (9,30,000) are among women. 5.4% of infections are from mother to child transmission (MTCT), 25,000 infected children are born every year. Besides the risk of mother to child transmission of HIV, these women are at risk of the higher adverse pregnancy outcome. The objectives of the study were to compare the obstetric and neonatal outcome in women who are HIV positive with low-risk HIV negative women and effect of antiretroviral drugs on preterm birth and IUGR. Materials and Methods: This is a retrospective case record analysis of 212 HIV-positive women delivering between 2002 to 2015, in a tertiary health care centre which was compared with 238 HIV-negative controls. Women who underwent medical termination of pregnancy and abortion were excluded from the study. Obstetric outcome analyzed were pregnancy induced hypertension, HIV positive intrauterine growth restriction, preterm birth, anemia, gestational diabetes and intrahepatic cholestasis of pregnancy. Neonatal outcome analysed were birth weight, apgar score, NICU admission and perinatal transmission.HIV-positiveOut of 212 women, 204 received antiretroviral therapy (ART) to prevent MTCT, 27 women received single dose nevirapine (sdNVP) or sdNVP tailed with 7 days of zidovudine and lamivudine (ZDV + 3TC), 15 received ZDV, 82 women received duovir and 80 women received triple drug therapy depending upon the time period of presentation. Results: Mean age of 212 HIV positive women was 25.72+3.6 years, 101 women (47.6 %) were primigravida. HIV positive status was diagnosed during pregnancy in 200 women while 12 women were diagnosed prior to conception. Among 212 HIV positive women, 20 (9.4 %) women had preterm delivery (< 37 weeks), 194 women (91.5 %) delivered by cesarean section and 18 women (8.5 %) delivered vaginally. 178 neonates (83.9 %) received exclusive top feeding and 34 neonates (16.03 %) received exclusive breast feeding. When compared to low risk HIV negative women (n=238), HIV positive women were more likely to deliver preterm (OR 1.27), have anemia (OR 1.39) and intrauterine growth restriction (OR 2.07). Incidence of pregnancy induced hypertension, diabetes mellitus and ICP was not increased. Mean birth weight was significantly lower in HIV positive women (2593.60+499 gm) when compared to HIV negative women (2919+459 gm). Complete follow up is available for 148 neonates till date, rest are under evaluation. Out of these 7 neonates found to have HIV positive status. Risk of preterm birth (P value = 0.039) and IUGR (P value = 0.739) was higher in HIV positive women who did not receive any ART during pregnancy than women who received ART. Conclusion: HIV positive pregnant women are at increased risk of adverse pregnancy outcome. Multidisciplinary team approach and use of highly active antiretroviral therapy can optimize the maternal and perinatal outcome.

Keywords: antiretroviral therapy, HIV infection, IUGR, preterm birth

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Authors: Remy Kulagowski, Tobias Sturm, Audrey Leopold, Aurelie Metay, Josephine Peigne, Alexis Thoumazeau, Alain Brauman, Bruno Fogliani, Florent Tivet

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A major challenge for agriculture is to enhance productivity while limiting the impact on the environment. Conservation agriculture (CA) is one strategy whereby both sustainability and productivity can be achieved by preserving and improving the soil quality. Soils provide and regulate a large number of ecosystem services (ES) such as agricultural productivity and climate change adaptation and mitigation. The aim of this study is to assess the impacts of contrasted CA crop management on soil functions for maize (Zea mays L.) cultivation in an eight years field experiment (2010-2018). The study included two CA practices: direct seeding in dead mulch (DM) and living mulch (LM), and conventional plough-based tillage (CT) practices on a fluvisol in New Caledonia (French Archipelago in the South Pacific). In 2018, soil quality of the cropping systems were evaluated with the Biofunctool® set of indicators, that consists in twelve integrative, in-field, and low-tech indicators assessing the biological, physical and chemical properties of soils. Main soil functions were evaluated including (i) carbon transformation, (ii) structure maintenance, and (iii) nutrient cycling in the ten first soil centimeters. The results showed significant higher score for soil structure maintenance (e.g., aggregate stability, water infiltration) and carbon transformation function (e.g., soil respiration, labile carbon) under CA in DM and LM when compared with CT. Score of carbon transformation index was higher in DM compared with LM. However, no significant effect of cropping systems was observed on nutrient cycling (i.e., nitrogen and phosphorus). In conclusion, the aggregated synthetic scores of soil multi-functions evaluated with Biofunctool® demonstrate that CA cropping systems lead to a better soil functioning. Further analysis of the results with agronomic performance of the soil-crop systems would allow to better understand the links between soil functioning and production ES of CA.

Keywords: conservation agriculture, cropping systems, ecosystem services, soil functions

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Authors: Saeed Qaisrani

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The study was conducted to observe carbon and nutrient loss by burning of rice residues on rice-wheat cropping system The rice crop was harvested to conduct the experiment in a randomized complete block design (RCBD) with factors and 4 replications with a net plot size of 10 m x 20 m. Rice stubbles were managed by two methods i.e. Incorporation & burning of rice residues. Soil samples were taken to a depth of 30 cm before sowing & after harvesting of wheat. Wheat was sown after harvesting of rice by three practices i.e. Conventional tillage, Minimum tillage and Zero tillage to observe best tillage practices. Laboratory and field experiments were conducted on wheat to assess best tillage practice and residues management method with estimation of carbon losses. Data on the following parameters; establishment count, plant height, spike length, number of grains per spike, biological yield, fat content, carbohydrate content, protein content, and harvest index were recorded to check wheat quality & ensuring food security in the region. Soil physico-chemical analysis i.e. pH, electrical conductivity, organic matter, nitrogen, phosphorus, potassium, and carbon were done in soil fertility laboratory. Substantial results were found on growth, yield and related parameters of wheat crop. The collected data were examined statistically with economic analysis to estimate the cost-benefit ratio of using different tillage techniques and residue management practices. Obtained results depicted that Zero tillage method have positive impacts on growth, yield and quality of wheat, Moreover, it is cost effective methodology. Similarly, Incorporation is suitable and beneficial method for soil due to more nutrients provision and reduce the need of fertilizers. Burning of rice stubbles has negative impact including air pollution, nutrient loss, microbes died and carbon loss. Recommended the zero tillage technology to reduce carbon losses along with food security in Pakistan.

Keywords: agricultural agronomy, food security, carbon sequestration, rice-wheat cropping system

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Authors: Tim Bolle, Franziska Kreimendahl, Thomas Gries, Stefan Jockenhoevel

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The therapeutic treatment of extensive, deep wounds is limited. Autologous split-skin grafts are used as a so-called ‘gold standard’. Most common deficits are the defects at the donor site, the risk of scarring as well as the limited availability and quality of the autologous grafts. The aim of this project is a tissue engineered dermal-epidermal skin replacement to overcome the limitations of the gold standard. A key requirement for the development of such a three-dimensional implant is the formation of a functional capillary-like network inside the implant to ensure a sufficient nutrient and gas supply. Tailored three-dimensional warp knitted spacer fabrics are used to reinforce the mechanically week fibrin gel-based scaffold and further to create a directed in vitro pre-vascularization along the parallel-oriented pile yarns within a co-culture. In this study various three-dimensional warp knitted spacer fabrics were developed in a factorial design to analyze the influence of the machine parameters such as the stitch density and the pattern of the fabric on the scaffold performance and further to determine suitable parameters for a successful fibrin gel-incorporation and a physiological performance of the scaffold. The fabrics were manufactured on a Karl Mayer double-bar raschel machine DR 16 EEC/EAC. A fine machine gauge of E30 was used to ensure a high pile yarn density for sufficient nutrient, gas and waste exchange. In order to ensure a high mechanical stability of the graft, the fabrics were made of biocompatible PVDF yarns. Key parameters such as the pore size, porosity and stress/strain behavior were investigated under standardized, controlled climate conditions. The influence of the input parameters on the mechanical and morphological properties as well as the ability of fibrin gel incorporation into the spacer fabric was analyzed. Subsequently, the pile yarns of the spacer fabrics were colonized with Human Umbilical Vein Endothelial Cells (HUVEC) to analyze the ability of the fabric to further function as a guiding structure for a directed vascularization. The cells were stained with DAPI and investigated using fluorescence microscopy. The analysis revealed that the stitch density and the binding pattern have a strong influence on both the mechanical and morphological properties of the fabric. As expected, the incorporation of the fibrin gel was significantly improved with higher pore sizes and porosities, whereas the mechanical strength decreases. Furthermore, the colonization trials revealed a high cell distribution and density on the pile yarns of the spacer fabrics. For a tailored reinforcing structure, the minimum porosity and pore size needs to be evaluated which still ensures a complete incorporation of the reinforcing structure into the fibrin gel matrix. That will enable a mechanically stable dermal graft with a dense vascular network for a sufficient nutrient and oxygen supply of the cells. The results are promising for subsequent research in the field of reinforcing mechanically weak biological scaffolds and develop functional three-dimensional scaffolds with an oriented pre-vascularization.

Keywords: fibrin-gel, skin replacement, spacer fabric, pre-vascularization

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Authors: Sagar Hedaoo, Fazle Baki, Ahmed Azab

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In supply chain management, network design for scalable manufacturing facilities is an emerging field of research. Facility location allocation assigns facilities to customers to optimize the overall cost of the supply chain. To further optimize the costs, capacities of these facilities can be changed in accordance with customer demands. A mathematical model is formulated to fully express the problem at hand and to solve small-to-mid range instances. A dedicated constraint has been developed to restrict emissions in line with the Kyoto protocol. This problem is NP-Hard; hence, a simulated annealing metaheuristic has been developed to solve larger instances. A case study on the USA-Canada cross border crossing is used.

Keywords: emission, mixed integer linear programming, metaheuristic, simulated annealing

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Authors: Fana Woldetsadik

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Ethiopia has a huge livestock population, including sheep, that has been contributing a considerable portion to the economy of the country and still promising to rally around the economic advancement of the country. However, feed shortage is a limiting factor in the production and productivity of sheep among Ethiopian smallholder farmers. Therefore, the aim of this study was to prove the role of the locally available brewery by-products called dried Atella as a supplement in feed intake, digestibility, live weight gain, carcass yield, and economic benefit in comparison with commercially purchased supplements known as niger seed cake (NSC). This on-station feeding experiment was conducted on the Zenzelma Campus of Bahir Dar University animal farm. The experimental design used for this research was a completely randomized design (CRD) with five replications. The crude protein (CP) content of dried Atella, wheat bran (WB), natural pasture hay (NPH) and NSC were about 25.07%, 16.57%, 4.48% and 38.04%, respectively, while the neutral detergent fibre (NDF),acid detergent fibre (ADF) and acid detergent lignin (ADL) content of dried Atella, WB, NPH and NSC were around 31.75%, 8.31%, 8.14%; 42.05%, 22.64%, 4.04%; 74.21%, 50.81%, 8.66%; 42.31%, 26.95% and 6.9%, respectively. The result depicted that a higher(P < 0.001) feed intake, nutrient intake, and digestibility for lambs supplemented with Atella than those supplemented with NSC. Furthermore, daily body weight gain and carcass characteristics were better (P < 0.05) for the sheep supplemented with dried Atella than NSC. On the other hand, in terms of profitability, although there was no substantial difference (P > 0.05) between T2 (animals fed NPH,NSC and WB) and T3 (animals fed NPH, Atella and WB), slightly better benefit was recorded in T3 groups. However, loss of money was recorded in T1 (animals fed NPH and WB). Hence, from the biological performance of lambs, it was concluded that Atella could be a potential supplementary feed for sheep fattening among smallholder farmers than NSC despite no profitability difference. Nevertheless, further investigation is recommended to examine the consequence of supplementation of NPH with NSC and NPH with Atella on fatty acid profile analysis, the physicochemical composition of meat, and meat composition.

Keywords: Attela, Bahir Dar university, Carcass yield, digestibility, natural pasture hay, Niger seed cake, smallholder farmers, weight gain, Ethiopia

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Authors: Farideh Doostan, Sahar Mohseni Taklloo, Mohammad Nosrati

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Chronic diseases are non-communicable and largely preventable by lifestyle changes including healthy diet consumption. They are the most common cause of death in the world and projected to increase by 15% globally between 2010 and 2020.The hazardous effects of behavioral and dietary risk factors on chronic disease have been established in prospective cohort studies and randomized trials. Because of some changes occur in college students’ lifestyle, assessment of dietary risk factors is important in these populations. Objective: This research was the first study that conducted to evaluate dietary intakes of Kermanian college students with WHO/FAO nutritional objectives. Material and Methods: In this descriptive cross-sectional study, 229 healthy college students of health faculty in Kerman University of Medical Sciences that do not intake any medical drugs were recruited using multistage sampling in 2013.Usual dietary intake was collected using a valid Food Frequency Questionnaire (FFQ) and diet quality was calculated based on WHO nutrient goals. To analysis of data between two groups, independent sample t. test and man whitney were applied. Results: Two hundred and twenty-nine college students; 151 females (65.9%) and 78 males (34.1%), the mean age of 21.9 years were studied. The mean of the Body Mass Index (Kg/m2) and Waist Circumference (cm) in males were 22.34 ±3.52 and 80.76±11.16 and in females were 21.19±2.62 and 73.67±7.65 respectively. Mean of daily cholesterol intake in males was significantly more than females (305±101 VS 268±98; P=0.008) and more than WHO/FAO recommendation (less than 300 mg/day). The mean of daily sodium intake in men and women were 10.4±1 and 10.9±5.3 respectively. These amounts were more than WHO/FAO recommendation (less than 2g/day). In addition, women were consumed fruit and vegetables more than men (839±336 VS 638±281; p ‹ 0.001) and these amounts were more than WHO/FAO recommendation (more than 400g/day) in both groups. Other intake indices were in the range of WHO/FAO recommendations, So that Percent of calories intake from total fat, saturated fatty acids, polyunsaturated fatty acids and added sugar were in compliance with WHO/FAO recommendations. Conclusion: Cholesterol intake in men and sodium intake in all participants were more than WHO/FAO recommendation. These dietary components are the most important causes of cardiovascular disease (one of the main causes of death in our population). These results indicated that proper nutritional education and interventions are needed in this population.

Keywords: college students, food intake, WHO /FAO nutrient intake goals, Kerman

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Authors: Yaw-Ling Lin

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A de Bruijn graph of order k is a graph whose vertices representing all length-k sequences with edges joining pairs of vertices whose sequences have maximum possible overlap (length k−1). Every Hamiltonian cycle of this graph defines a distinct, minimum length de Bruijn sequence containing all k-mers exactly once. A Kautz sequence is the minimal generating sequence so as the sequence of minimal length that produces all possible length-k sequences with the restriction that every two consecutive alphabets in the sequences must be different. A collection of de Bruijn/Kautz sequences are orthogonal if any two sequences are of maximally differ in sequence composition; that is, the maximum length of their common substring is k. In this paper, we discuss how such a collection of (maximal) orthogonal de Bruijn/Kautz sequences can be made and use the algorithm to build up a web application service for the synthesized DNA and other related biomolecular sequences.

Keywords: biomolecular sequence synthesis, de Bruijn sequences, Eulerian cycle, Hamiltonian cycle, Kautz sequences, orthogonal sequences

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Authors: A. H. Dikko, D. N Tsado, M. S. T. Rita, D. S. Umar

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This study was conducted on eighteen (18) New Zealand and chinchilla breeds of rabbits were used. The rabbits were allotted to 3 treatments with each treatment having six (6) animals with two (2) replicates. T1 were castrated, which both testes was removed (Bilateral); T2 were castrated, which only one testes was removed (unilateral) and T3 were not castrated (control). In nutrient digestibility, T1 and T2 (p>0.05) has a higher rate than T3. There was no significant (p<0.05) difference in live weight and dressing weight among the treatment groups. There is a significant (p > 0.05) difference in visceral organs in the treatment groups.

Keywords: New Zealand, chinchilla, castration, bilateral, unilateral

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Authors: Averalda van Graan, Joelaine Chetty, Malory Links, Agness Mwangwela, Sitilitha Masangwi, Dalitso Chimwala, Shiban Ghosh, Elizabeth Marino-Costello

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Food composition data is becoming increasingly important as dealing with food insecurity and malnutrition in its persistent form of under-nutrition is now coupled with increasing over-nutrition and its related ailments in the developing world, of which Malawi is not spared. In the absence of a food composition database (FCDB) inherent to our dietary patterns, efforts were made to develop a country-specific FCDB for nutrition practice, research, and programming. The main objective was to develop a multi-user, country-specific food composition database, and table from existing published and unpublished scientific literature. A multi-phased approach guided by the project framework was employed. Phase 1 comprised a scoping mission to assess the nutrition landscape for compilation activities. Phase 2 involved training of a compiler and data collection from various sources, primarily; institutional libraries, online databases, and food industry nutrient data. Phase 3 subsumed evaluation and compilation of data using FAO and IN FOODS standards and guidelines. Phase 4 concluded the process with quality assurance. 316 Malawian food items categorized into eight food groups for 42 components were captured. The majority were from the baby food group (27%), followed by a staple (22%) and animal (22%) food group. Fats and oils consisted the least number of food items (2%), followed by fruits (6%). Proximate values are well represented; however, the percent missing data is huge for some components, including Se 68%, I 75%, Vitamin A 42%, and lipid profile; saturated fat 53%, mono-saturated fat 59%, poly-saturated fat 59% and cholesterol 56%. A multi-phased approach following the project framework led to the development of the first Malawian FCDB and table. The table reflects inherent Malawian dietary patterns and nutritional concerns. The FCDB can be used by various professionals in nutrition and health. Rising over-nutrition, NCD, and changing diets challenge us for nutrient profiles of processed foods and complete lipid profiles.

Keywords: analytical data, dietary pattern, food composition data, multi-phased approach

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Authors: Uche A. Ibekwe

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Pension funds are pooled assets which are established to provide income for retirees. The funds are usually regulated to check excessive risk taking by fund managers. In Nigeria, the current defined contribution (DC) pension scheme appears to contain some overly stringent restrictions which might be hampering its successful implementation. Notable among these restrictions is the 25 percent maximum limit on investment in ordinary shares of quoted companies. This paper examines the extent to which these restrictions affect pension replacement rates at retirement. The study made use of both simulated and historical asset return distributions using mean-variance, regression analysis and ruin-risk analyses, the study found that the current equity investment restriction policy in Nigeria reduces replacement rates at retirement.

Keywords: equity investment, replacement rates, restrictions, ruin-risk

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Authors: Zhiqing Zhang, Xiangong Zhou, Zihan Zhou

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Ground fissures can be seen in some cities all over the world. As a special urban geological disaster, ground fissures in Xi'an have caused great harm to infrastructure. Chang'an Road Interchange in Xi'an City is a bridge across ground fissures. The damage to Chang'an Road interchange is the most serious and typical. To study the influence of ground fissures on the bridge, we established a bridge monitoring system. The main monitoring items include elevation monitoring, structural displacement monitoring, etc. The monitoring results show that the typical failure is mainly reflected in the bridge deck damage caused by horizontal tension and vertical dislocation. For the construction of urban interchange spanning ground fissures, the interchange should be divided reasonably, a simple support structure with less restriction should be adopted, and the monitoring of supports should be strengthened to prevent the occurrence of beam falling.

Keywords: bridge monitoring, ground fissures, typical disease, structural displacement

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Authors: Joselito Medina-Marin, Alexandr Karelin, Ana Tarasenko, Juan Carlos Seck-Tuoh-Mora, Norberto Hernandez-Romero, Eva Selene Hernandez-Gress

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A production line designer faces with several challenges in manufacturing system design. One of them is the assignment of buffer slots in between every machine of the production line in order to maximize the throughput of the whole line, which is known as the Buffer Allocation Problem (BAP). The BAP is a combinatorial problem that depends on the number of machines and the total number of slots to be distributed on the production line. In this paper, we are proposing a Petri Net (PN) Model to obtain the throughput in unreliable production lines, based on PN mathematical tools and the decomposition method. The results obtained by this methodology are similar to those presented in previous works, and the number of machines is not a hard restriction.

Keywords: buffer allocation problem, Petri Nets, throughput, production lines

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Authors: Li Cheng, Zhang Yilei, Cohen Yehuda

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Two motility mechanisms were introduced into iDynoMiCs software, which adopts an individual-based modeling method. Based on the new capabilities, along with the pressure motility developed before, influence of bacterial motility on biofilm formation was studied. Simulation results were evaluated both qualitatively through 3D structure inspections and quantitatively by parameter characterizations. It was showed that twitching motility increased the biofilm surface irregularity probably due to movement of cells towards higher nutrient concentration location whereas free motility, on the other hand, could make biofilms flatter and smoother relatively. Pressure motility showed no significant influence in this study.

Keywords: iDynoMics, biofilm structure, bacterial motility, motility mechanisms

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Authors: Yakup Ulusu, Faruk Ozel, Numan Eczacioglu, Abdurrahman Ozen, Sabriye Acikgoz

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GFP discovered in the mid-1970s, has been used as a marker after replicated genetic study by scientists. In biotechnology, cell, molecular biology, the GFP gene is frequently used as a reporter of expression. In modified forms, it has been used to make biosensors. Many animals have been created that express GFP as an evidence that a gene can be expressed throughout a given organism. Proteins labeled with GFP identified locations are determined. And so, cell connections can be monitored, gene expression can be reported, protein-protein interactions can be observed and signals that create events can be detected. Additionally, monitoring GFP is noninvasive; it can be detected by under UV-light because of simply generating fluorescence. Moreover, GFP is a relatively small and inert molecule, that does not seem to treat any biological processes of interest. The synthesis of GFP has some steps like, to construct the plasmid system, transformation in E. coli, production and purification of protein. GFP carrying plasmid vector pBAD–GFPuv was digested using two different restriction endonuclease enzymes (NheI and Eco RI) and DNA fragment of GFP was gel purified before cloning. The GFP-encoding DNA fragment was ligated into pET28a plasmid using NheI and Eco RI restriction sites. The final plasmid was named pETGFP and DNA sequencing of this plasmid indicated that the hexa histidine-tagged GFP was correctly inserted. Histidine-tagged GFP was expressed in an Escherichia coli BL21 DE3 (pLysE) strain. The strain was transformed with pETGFP plasmid and grown on LuiraBertoni (LB) plates with kanamycin and chloramphenicol selection. E. coli cells were grown up to an optical density (OD 600) of 0.8 and induced by the addition of a final concentration of 1mM isopropyl-thiogalactopyranoside (IPTG) and then grown for additional 4 h. The amino-terminal hexa-histidine-tag facilitated purification of the GFP by using a His Bind affinity chromatography resin (Novagen). Purity of GFP protein was analyzed by a 12 % sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The concentration of protein was determined by UV absorption at 280 nm (Varian Cary 50 Scan UV/VIS spectrophotometer). Synthesis of GFP-Polymer composite nanofibers was produced by using GFP solution (10mg/mL) and polymer precursor Polyvinylpyrrolidone, (PVP, Mw=1300000) as starting materials and template, respectively. For the fabrication of nanofibers with the different fiber diameter; a sol–gel solution comprising of 0.40, 0.60 and 0.80 g PVP (depending upon the desired fiber diameter) and 100 mg GFP in 10 mL water: ethanol (3:2) mixtures were prepared and then the solution was covered on collecting plate via electro spinning at 10 kV with a feed-rate of 0.25 mL h-1 using Spellman electro spinning system. Results show that GFP-based nano-fiber can be used plenty of biomedical applications such as bio-imaging, bio-mechanic, bio-material and tissue engineering.

Keywords: biomaterial, GFP, nano-fibers, protein expression

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Authors: Shahram Nanekarania, Majid Goodarzia

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Calpastatin and callipyge have been known as one of the candidate genes in meat quality and quantity. Calpastatin gene has been located to chromosome 5 of sheep and callipyge gene has been localized in the telomeric region on ovine chromosome 18. The objective of this study was identification of calpastatin and callipyge genes polymorphism and analysis of genotype structure in population of Lori sheep kept in Iran. Blood samples were taken from 120 Lori sheep breed and genomic DNA was extracted by salting out method. Polymorphism was identified using the PCR-RFLP technique. The PCR products were digested with MspI and FaqI restriction enzymes for calpastatin gene and callipyge gene, respectively. In this population, three patterns were observed and AA, AB, BB genotype have been identified with the 0.32, 0.63, 0.05 frequencies for calpastatin gene. The results obtained for the callipyge gene revealed that only the wild-type allele A was observed, indicating that only genotype AA was present in the population under consideration.

Keywords: polymorphism, calpastatin, callipyge, PCR-RFLP, Lori sheep

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

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

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

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Authors: M. Saleem, M. Shahnawaz, A. W. Gandahi, S. M. Bhatti

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The micronutrients boron and zinc deficiencies are extensive in the areas of rice-wheat cropping system. Optimum levels of these nutrients in soil are necessary for healthy crop growth. Since rice and wheat are major staple food of worlds’ populace, the higher yields and nutrition status of these crops has direct effect on the health of human being and economy of the country. A field study was conducted to observe the direct and residual effect of two selected micronutrients boron (B) and zinc (Zn)) on rice and wheat crop growth and its grain nutrient status. Each plot received either B or Zn at the rates of 0, 1, 2, 3 and 4 kg B ha⁻¹, and 5, 10, 15 and 20 kg Zn ha⁻¹, combined B and Zn application at 1 kg B and 5 kg Zn ha⁻¹, 2 kg B and 10 kg Zn ha⁻¹. Colemanite ore were used as source of B and zinc sulfate for Zn. The second season wheat crop was planted in the same plots after the interval period of 30 days and during this time gap soil was fallow. Boron and Zn application significantly enhanced the plant height, number of tillers, Grains panicle⁻¹ seed index fewer empty grains panicle⁻¹ and yield of rice crop at all defined levels as compared to control. The highest yield (10.00 tons/ha) was recorded at 2 Kg B, 10 Kg Zn ha⁻¹ rates. Boron and Zn concentration in grain and straw significantly increased. The application of B also improved the nutrition status of rice as B, protein and total carbohydrates content of grain augmented. The analysis of soil samples collected after harvest of rice crop showed that the B and Zn content in post-harvest soil samples was high in colemanite and zinc sulfate applied plots. The residual B and Zn were also effectual for the second season wheat crop, as the growth parameters plant height, number of tillers, earhead length, weight 1000 grains, B and Zn content of grain significantly improved. The highest wheat grain yield (4.23 tons/ha) was recorded at the residual rates of 2 kg B and 10 kg Zn ha⁻¹ than the other treatments. This study showed that one application of B and Zn can increase crop yields for at least two consecutive seasons and the mineral colemanite can confidently be used as source of B for rice crop because very small quantities of these nutrients are consumed by first season crop and remaining amount was present in soil which were used by second season wheat crop for healthy growth. Consequently, there is no need to apply these micronutrients to the following crop when it is applied on the previous one.

Keywords: residual boron, zinc, rice, wheat

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Authors: Kira Rysakova, Vitaly Novikov

Abstract:

An improved method of obtaining enzymatic collagen hydrolysate from the tissues of marine hydrobionts is proposed, which allows to obtain hydrolysate without pre-isolation of pure collagen. The method can be used to isolate enzymatic collagen hydrolysate from the waste of industrial processing of Red King crab and non-traditional objects - marine holothurias. Comparative analysis of collagen hydrolysates has shown the possibility of their use in a number of nutrient media, but this requires additional optimization of their composition and biological tests on wide sets of test strains of microorganisms.

Keywords: collagen hydrolysate, marine hydrobionts, red king crab, marine holothurias, enzymes, exclusive HPLC

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