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At least 703 records · Page 39

High fructose consumption induces DNA methylation at PPARα and CPT1A promoter regions in the rat liver

DNA methylation status is affected by environmental factors, including nutrition. Fructose consumption is considered a risk factor for the conditions that make up metabolic syndrome such as dyslipidemia. However, the pathogenetic mechanism by which fructose consumption leads to metabolic syndrome is unclear. Based on observations that epigenetic modifications are closely related to induction of metabolic syndrome, we hypothesized that fructose-induced metabolic syndrome is caused by epigenetic alterations. Male SD rats were designated to receive water or 20% fructose solution for 14 weeks. mRNA levels for peroxisome proliferator-activated receptor alpha (PPARα) and carnitine palmitoyltransferase 1A (CPT1A) was analyzed using Real-time PCR. Restriction digestion and real-time PCR (qAMP) was used for the analysis of DNA methylation status. Hepatic lipid accumulation was also observed by fructose intake. Fructose feeding also significantly decreased mRNA levels for PPARα and CPT1A. qAMP analysis demonstrated the hypermethylation of promoter regions of PPARα and CTP1A genes. Fructose-mediated attenuated gene expression may be mediated by alterations of DNA methylation status, and pathogenesis of metabolic syndrome induced by fructose relates to DNA methylation status. - Highlights: • No general consensus has been reached regarding the molecular mechanisms of the pathogenesis of fructose-induced diseases. • Significant increase in hepatic total methylation level was observed after fructose-supplemented feeding. • Fructose feeding significantly decreased mRNA levels for PPARα and CPT1A. • qAMP analysis demonstrated the hypermethylation of promoter regions of PPARα and CTP1A genes. • Fructose-mediated attenuated gene expression may be mediated by alterations of DNA methylation status in rat liver.

60 APPLIED LIFE SCIENCES↗

A universal mini-vector and an annealing of PCR products (APP)-based cloning strategy for convenient molecular biological manipulations

Currently, the most widely used strategies for molecular cloning are sticky-end ligation-based cloning, TA cloning, blunt-end ligation-based cloning and ligase-independent cloning. In this study we have developed a novel mini-vector pANY1 which can simultaneously meet the requirements of all these cloning strategies. In addition, the selection of appropriate restriction digestion sites is difficult in some cases because of the presence of internal sites. In this study, an annealing of PCR products (APP)-based sticky-end cloning strategy was introduced to avoid this issue. Additionally, false positives occur during molecular cloning, which increases the workload of isolating positive clones. The plasmid pANY1 contains a ccdB cassette between multiple cloning sites, which efficiently avoids these false positives. Therefore, this mini-vector should serve as a useful tool with wide applications in biosciences, agriculture, food technologies, etc.

60 APPLIED LIFE SCIENCES↗

Introduction of a glycosylation site in the constant region decreases the aggregation of adalimumab Fab

The production of therapeutic monoclonal antibodies is costly; therefore, antigen-binding fragments (Fabs) can be used instead. However, their tendency toward aggregation can reduce the half-life in the plasma and the therapeutic effectiveness. To examine the effect of glycosylation on the properties of the Fab of a therapeutic antibody, an N-glycosylation site was introduced at position 178 of the H-chain constant region of adalimumab Fab through site-directed mutagenesis of L178 N (H:L178 N Fab), and then H:L178 N Fab was expressed in Pichia pastoris. SDS-PAGE analysis with treatment of N-glycosidase F or periodic acid–Schiff reagent showed that H:L178 N Fab contained a relatively low glycan level. Moreover, the H:L178 N mutation did not decrease the binding activity and thermal stability of Fab, and H:L178 N Fab was more resistant to protease digestion than wild-type Fab. The aggregation of Fab induced by pH-shift stress was measured by monitoring the optical density at 350 nm. Although the wild-type Fab showed a large increase in optical density with an increase of protein concentration, no such increase of turbidity during aggregation was found in H:L178 N Fab. These results demonstrated that glycosylation at position 178 of the H-chain constant region of adalimumab Fab can prevent protein aggregation, and therefore serve as a potentially effective platform for drug development.

60 APPLIED LIFE SCIENCES↗

Orally active anti-hypertensive peptides found based on enteroendocrine cell responses to a dipeptide library

Highlights: • Phe-Trp (FW) evoked the strongest enteroendocrine cell response out of 338 dipeptides. • Phe-Trp-Gly-Lys (FWGK) exhibited more potent hypotensive effects than FW in SHRs. • Vasorelaxing and antihypertensive effects of FWGK were associated with the CCK system. We previously reported that an orally administered dipeptide, Arg-Phe (RF), which causes enteroendocrine cell responses, lowered blood pressure in spontaneously hypertensive rats (SHRs). In this study, we found that Phe-Trp (FW), induced the most potent enteroendocrine cell responses out of total 338 dipeptides. An FW analogue, Phe-Trp-Gly-Lys (FWGK), which was effectively produced by tryptic digestion of bovine serum albumin, decreased blood pressure after oral administration. The minimum effective dose of FWGK (50 μg/kg) was 1/300 of that of RF (15 mg/kg). FWGK stimulated cholecystokinin (CCK) secretion in the enteroendocrine cells and exhibited vasorelaxing and antihypertensive effects via the CCK{sub 1} system.

60 APPLIED LIFE SCIENCES↗

Biological upgrading of biogas assisted with membrane supplied hydrogen gas in a three-phase upflow reactor

Biogas upgrading via CO 2 conversion to CH 4 is an emerging technology for renewable natural gas production and carbon management, but its development is limited by the low H 2 gas to liquid phase transfer. Herein, an innovative biogas upgrading system employing a three-phase design was studied for CO 2 conversion with H 2 supply via gas-permeable membrane. The system produced biogas consisted of 74.1 ± 7.1 % CH 4 and 25.9 ± 7.1 % CO 2 with intermittent injection of H 2 . When H 2 supply was continuous, the CH 4 content increased to 91.6 ± 2.2 % at a H 2 :CO 2 ratio of 4.4. Although a higher ratio of 5.5 could result in a higher CH 4 percentage of 95.2 ± 2.5 %, biogas production rate started to decrease. The removal efficiency of organic contents remained above 90 % throughout the experiment. Microbial community analysis corroborated the findings, showing that hydrogenotrophic Methanobacteriaceae was more prevalent in the biofilm (71.9 %) compared to that in anaerobic digestion (15.8 %) and effluent (14.1 %).

Agriculture↗

Zero-gap microbial electrolysis cells for efficient hydrogen production from real liquid waste streams

Zero-gap microbial electrolysis cells (MECs) have demonstrated large current and hydrogen production rates from defined substrates in synthetic media, but operation with real waste streams has yet to be proved. This study evaluated the performance and 30-days stability of zero-gap MECs operated with effluent from a single-stage anaerobic digester. The system achieved a maximum current density of 8.8 ± 0.3 A/m 2 with a hydrogen production rate of 32 ± 6 L/L-d, and during 30 days of continuous operation, sustained an average current density of 7 ± 2 A/m 2 and a hydrogen production rate of 20.8 ± 0.2 L/L-d. Carbonate precipitation was identified as a major challenge to long-term stability, and mild acid washing effectively mitigated its adverse effects. The low buffer capacity of the effluent was primarily limiting performance. Furthermore, these findings underscore the significant impact of wastewater chemistry on MEC operation and validate the feasibility of utilizing real waste streams as viable feedstocks for biohydrogen production in zero-gap configurations.

Acid wash↗

Determination of impurities in cubic boron nitride (cBN) by inductively coupled plasma mass spectrometry (ICPMS)

The phase stability of cBN is critically dependent on the content and amount of impurities present. While studies have shown qualitatively that the addition of certain impurities can either increase or decrease the cBN-hBN phase transition temperature, studies have yet to show this effect quantitatively. This is because analytical methods to characterize minor and trace impurities in cBN are currently lacking. To this end, we present flux fusion methods capable of fully digesting cBN samples for ICPMS analyses. The materials used in this study are commercially available cBN powders in three different sizes. Two types of fluxes (LiOH and NaOH·H 2 O) are utilized so that both Li and Na impurities can be determined. To ensure the accuracy of Si measurements, a column chromatography method has been developed to purify Si from flux materials in order to increase the amount of dissolved sample sent to the ICP. Detection limits have been determined using procedural blanks and are found to be below 20 μg/g for all 58 elements analyzed except for Si, for which the detection limit is 312 μg/g. Nonetheless, all cBN powders studied are found to have high Si impurity levels that are above detection limit. This study provides a much-needed analytical protocol to enable comprehensive screening of impurities in cBN.

36 MATERIALS SCIENCE↗

Distribution of chemical elements in soils and stream sediments in the area of abandoned Sb–As–Tl Allchar mine, Republic of Macedonia

The aim of this study was to investigate the distribution of some toxic elements in topsoil and subsoil, focusing on the identification of natural and anthropogenic element sources in the small region of rare As–Sb–Tl mineralization outcrop and abandoned mine Allchar known for the highest natural concentration of Tl in soil worldwide. The samples of soil and sediments after total digestion were analyzed by inductively coupled plasma–mass spectrometry (ICP–MS) and inductively coupled plasma–atomic emission spectrometry (ICP–AES). Factor analysis (FA) was used to identify and characterize element associations. Six associations of elements were determined by the method of multivariate statistics: Rb–Ta–K–Nb–Ga–Sn–Ba–Bi–Li–Be–(La–Eu)–Hf–Zr–Zn–In–Pd–Ag–Pt–Mg; Tl–As–Sb–Hg; Te–S–Ag–Pt–Al–Sc–(Gd–Lu)–Y; Fe–Cu–V–Ge–Co–In; Pd–Zr–Hf–W–Be and Ni–Mn–Co–Cr–Mg. The purpose of the assessment was to determine the nature and extent of potential contamination as well as to broadly assess possible impacts to human health and the environment. The results from the analysis of the collected samples in the vicinity of the mine revealed that As and Tl elements have the highest median values. Higher median values for Sb are obviously as a result of the past mining activities and as a result of area surface phenomena in the past. - Highlights: • Soil and river sediments were analyzed from Sb–As–Tl Allchar locality. • An increased content of certain toxic elements for environment was determined. • Highest As and Tl contents are obtained in the close vicinity of Allchar mine. • River sediments portray 160 times higher content of Sb than EU values. • The results classify Allchar as probably the highest natural Tl-deposit worldwide.

54 ENVIRONMENTAL SCIENCES↗

Association between fetal exposure to phthalate endocrine disruptor and genome-wide DNA methylation at birth

Highlights: • Humans are ubiquitously exposed to phthalates through digestion, inhalation, or dermal absorption. • DNA methylation has been hypothesized to be an important mechanism in in utero phthalate exposure. • DNA methylation changes are involved in androgen response, estrogen response, protein secretion, and spermatogenesis. • Three CpG sites (cg00935782, cg07732421, and cg00764560) are involved in genes of response to androgen. Phthalic acid esters are ubiquitous and antiandrogenic, and may cause systemic effects in humans, particularly with in utero exposure. Epigenetic modification, such as DNA methylation, has been hypothesized to be an important mechanism that mediates certain biological processes and pathogenic effects of in utero phthalate exposure.

54 ENVIRONMENTAL SCIENCES↗

Mass Spectrometry Guided In Situ Proteolysis to Obtain Crystals for X-ray Structure Determination

A strategy for increasing the efficiency of protein crystallization/structure determination with mass spectrometry has been developed. This approach combines insights from limited proteolysis/mass spectrometry and crystallization via in situ proteolysis. The procedure seeks to identify protease-resistant polypeptide chain segments from purified proteins on the time-scale of crystal formation, and subsequently crystallizing the target protein in the presence of the optimal protease at the right relative concentration. We report our experience with 10 proteins of unknown structure, two of which yielded high-resolution X-ray structures. The advantage of this approach comes from its ability to select only those structure determination candidates that are likely to benefit from application of in situ proteolysis, using conditions most likely to result in formation of a stable proteolytic digestion product suitable for crystallization.

59 BASIC BIOLOGICAL SCIENCES↗

A systematic analytical framework for multi-source municipal solid waste characterization for energy recovery

Advancing municipal solid waste (MSW) management from disposal-oriented practices toward circular, value-driven systems requires standardized methodologies capable of identifying material composition and resource recoverable potential at the point of generation. Despite extensive research, MSW characterization remains fragmented due to inconsistences in sampling methodologies, waste sorting categories, and temporal coverage across previous studies which limit cross-site comparability, reproducibility, and constrain the reliable evaluation of potential resource recovery pathways. This lack of consistency has hindered the development of a unified framework for MSW characterization and resource assessment. This study introduces a standardized, field-validated protocol for MSW sampling and composition analysis that ensures consistent, traceable data across diverse waste sources. The protocol integrates randomized spatial sampling, systematic material sorting, and controlled subsampling for multi-site and multi-season field campaigns. Validation included MSW collection from residential, grocery, restaurant, and school MSW streams across five U.S. states, including Maryland, Idaho, Virginia, Ohio, and Mississippi, to demonstrate the protocol’s ability to identify source-based composition patterns relevant to resource recovery applications. Grocery and restaurant streams were dominated by food waste and high-moisture organics, while school waste contained higher paper content and residential waste showed greater heterogeneity. Aggregation into energy-relevant fractions highlighted practical recovery pathways via anaerobic digestion or gasification, supporting data-driven planning, policy, and circular economy strategies for sustainable waste management across waste sources.

09 BIOMASS FUELS↗

Preclinical pharmacology and toxicology study of Ad-hTERT-E1a-Apoptin, a novel dual cancer-specific oncolytic adenovirus

Clinical studies have demonstrated that conditionally replicating adenovirus is safe. We constructed an oncolytic adenovirus, Ad-hTERT-E1a-Apoptin, using a cancer-specific promoter (human telomerase reverse transcriptase promoter, hTERTp) and a cancer cell-selective apoptosis-inducing gene (Apoptin). Ad-hTERT-E1a-Apoptin was proven effective both in vitro and in vivo in our previous study. In this study, the preclinical safety profiles of Ad-hTERT-E1a-Apoptin in animal models were investigated. At doses of 5.0 × 10{sup 8}, 2.5 × 10{sup 9}, and 1.25 × 10{sup 10} viral particles (VP)/kg, Ad-hTERT-E1a-Apoptin had no adverse effects on mouse behavior, muscle cooperation, sedative effect, digestive system, and nervous systems, or on beagle cardiovascular and respiratory systems at 5.0 × 10{sup 8}, 2.5 × 10{sup 9}, and 1.25 × 10{sup 10} VP/kg doses. In acute toxicity tests in mice, the maximum tolerated dose > 5 × 10{sup 10} VP/kg. There was no inflammation or ulceration at the injection sites within two weeks. In repeat-dose toxicological studies, the no observable adverse effect levels of Ad-hTERT-E1a-Apoptin in rats (1.25 × 10{sup 10} VP/kg) and beagles (2.5 × 10{sup 9} VP/kg) were 62.5- and 12.5-fold of the proposed clinical dose, respectively. The anti-virus antibody was produced in animal sera. Bone marrow examination revealed no histopathological changes. Guinea pigs sensitized by three repeated intraperitoneal injections of 1.35 × 10{sup 10} VP/mL Ad-hTERT-E1a-Apoptin each and challenged by one intravenous injection of 1.67 × 10{sup 8} VP/kg Ad-hTERT-E1a-Apoptin did not exhibit any sign of systemic anaphylaxis. Our data from different animal models suggest that Ad-hTERT-E1a-Apoptin is a safe anti-tumor therapeutic agent. - Highlights: • We use the rodents and non-rodents animal models to evaluation Ad-hTERT-E1a-Apoptin. • Ad-hTERT-E1a-Apoptin is a safe anti-tumor therapeutic agent. • Demonstrate the safety and feasibility dose of injected Ad-hTERT-E1a-Apoptin.

60 APPLIED LIFE SCIENCES↗

Determination of mercury distribution inside spent compact fluorescent lamps by atomic absorption spectrometry

Highlights: Black-Right-Pointing-Pointer New treatments for CFL are required considering the aim of Directive 202/96/CE. Black-Right-Pointing-Pointer It is shown that most of the mercury introduced into a CFL is in the phosphor powder. Black-Right-Pointing-Pointer Experimental conditions for microwave-assisted sample digestion followed by AAS measurements are described. Black-Right-Pointing-Pointer By washing the glass it is possible to reduce the concentration below legal limits. - Abstract: In this study, spent compact fluorescent lamps were characterized to determine the distribution of mercury. The procedure used in this research allowed mercury to be extracted in the vapor phase, from the phosphor powder, and the glass matrix. Mercury concentration in the three phases was determined by the method known as cold vapor atomic absorption spectrometry. Median values obtained in the study showed that a compact fluorescent lamp contained 24.52 {+-} 0.4 ppb of mercury in the vapor phase, 204.16 {+-} 8.9 ppb of mercury in the phosphor powder, and 18.74 {+-} 0.5 ppb of mercury in the glass matrix. There are differences in mercury concentration between the lamps since the year of manufacture or the hours of operation affect both mercury content and its distribution. The 85.76% of the mercury introduced into a compact fluorescent lamp becomes a component of the phosphor powder, while more than 13.66% is diffused through the glass matrix. By washing and eliminating all phosphor powder attached to the glass surface it is possible to classified the glass as a non-hazardous waste.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Review of comparative LCAs of food waste management systems - Current status and potential improvements

Highlights: Black-Right-Pointing-Pointer GHG-emissions from different treatment alternatives vary largely in 25 reviewed comparative LCAs of bio-waste management. Black-Right-Pointing-Pointer System-boundary settings often vary largely in reviewed studies. Black-Right-Pointing-Pointer Existing LCA guidelines give varying recommendations in relation to several key issues. - Abstract: Twenty-five comparative cycle assessments (LCAs) addressing food waste treatment were reviewed, including the treatment alternatives landfill, thermal treatment, compost (small and large scale) and anaerobic digestion. The global warming potential related to these treatment alternatives varies largely amongst the studies. Large differences in relation to setting of system boundaries, methodological choices and variations in used input data were seen between the studies. Also, a number of internal contradictions were identified, many times resulting in biased comparisons between alternatives. Thus, noticed differences in global warming potential are not found to be a result of actual differences in the environmental impacts from studied systems, but rather to differences in the performance of the study. A number of key issues with high impact on the overall global warming potential from different treatment alternatives for food waste were identified through the use of one-way sensitivity analyses in relation to a previously performed LCA of food waste management. Assumptions related to characteristics in treated waste, losses and emissions of carbon, nutrients and other compounds during the collection, storage and pretreatment, potential energy recovery through combustion, emissions from composting, emissions from storage and land use of bio-fertilizers and chemical fertilizers and eco-profiles of substituted goods were all identified as highly relevant for the outcomes of this type of comparisons. As the use of LCA in this area is likely to increase in coming years, it is highly relevant to establish more detailed guidelines within this field in order to increase both the general quality in assessments as well as the potentials for cross-study comparisons.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

An application of the theory of planned behaviour to study the influencing factors of participation in source separation of food waste

Highlights: ► Theory of planned behaviour (TPB) has been conducted to identify the influencing factors for participation in source separation of food waste using self administered questionnaires. ► The findings suggested several implications for the development and implementation of waste separation at home programme. ► The analysis indicates that the attitude towards waste separation is determined as the main predictors where this in turn could be a significant predictor of the repondent’s actual food waste separation behaviour. ► To date, none of similar have been reported elsewhere and this finding will be beneficial to local Authorities as indicator in designing campaigns to promote the use of waste separation programmes to reinforce the positive attitudes. - Abstract: Tremendous increases in biodegradable (food waste) generation significantly impact the local authorities, who are responsible to manage, treat and dispose of this waste. The process of separation of food waste at its generation source is identified as effective means in reducing the amount food waste sent to landfill and can be reused as feedstock to downstream treatment processes namely composting or anaerobic digestion. However, these efforts will only succeed with positive attitudes and highly participations rate by the public towards the scheme. Thus, the social survey (using questionnaires) to analyse public’s view and influencing factors towards participation in source separation of food waste in households based on the theory of planned behaviour technique (TPB) was performed in June and July 2011 among selected staff in Universiti Putra Malaysia, Serdang, Selangor. The survey demonstrates that the public has positive intention in participating provided the opportunities, facilities and knowledge on waste separation at source are adequately prepared by the respective local authorities. Furthermore, good moral values and situational factors such as storage convenience and collection times are also encouraged public’s involvement and consequently, the participations rate. The findings from this study may provide useful indicator to the waste management authorities in Malaysia in identifying mechanisms for future development and implementation of food waste source separation activities in household programmes and communication campaign which advocate the use of these programmes.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Improving hydrolysis of food waste in a leach bed reactor

Highlights: • This paper assesses leaching of food waste in a two phase digestion system. • Leaching is assessed with and without an upflow anaerobic sludge blanket (UASB). • Without the UASB, low pH reduces hydrolysis, while increased flows increase leaching. • Inclusion of the UASB increases pH to optimal levels and greatly improves leaching. • The optimal conditions are suggested as low flow with connection to the UASB. - Abstract: This paper examines the rate of degradation of food waste in a leach bed reactor (LBR) under four different operating conditions. The effects of leachate recirculation at a low and high flow rate are examined with and without connection to an upflow anaerobic sludge blanket (UASB). Two dilution rates of the effective volume of the leach bed reactors were investigated: 1 and 6 dilutions per LBR per day. The increase in dilution rate from 1 to 6 improved the destruction of volatile solids without connection to the UASB. However connection to the UASB greatly improved the destruction of volatile solids (by almost 60%) at the low recirculation rate of 1 dilution per day. The increase in volatile solids destruction with connection to the UASB was attributed to an increase in leachate pH and buffering capacity provided by recirculated effluent from the UASB to the leach beds. The destruction of volatile solids for both the low and high dilution rates was similar with connection to the UASB, giving 82% and 88% volatile solids destruction respectively. This suggests that the most efficient leaching condition is 1 dilution per day with connection to the UASB.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Life cycle costing of waste management systems: Overview, calculation principles and case studies

Highlights: • We propose a comprehensive model for cost assessment of waste management systems. • The model includes three types of LCC: Conventional, Environmental and Societal LCCs. • The applicability of the proposed model is tested with two case studies. - Abstract: This paper provides a detailed and comprehensive cost model for the economic assessment of solid waste management systems. The model was based on the principles of Life Cycle Costing (LCC) and followed a bottom-up calculation approach providing detailed cost items for all key technologies within modern waste systems. All technologies were defined per tonne of waste input, and each cost item within a technology was characterised by both a technical and an economic parameter (for example amount and cost of fuel related to waste collection), to ensure transparency, applicability and reproducibility. Cost items were classified as: (1) budget costs, (2) transfers (for example taxes, subsidies and fees) and (3) externality costs (for example damage or abatement costs related to emissions and disamenities). Technology costs were obtained as the sum of all cost items (of the same type) within a specific technology, while scenario costs were the sum of all technologies involved in a scenario. The cost model allows for the completion of three types of LCC: a Conventional LCC, for the assessment of financial costs, an Environmental LCC, for the assessment of financial costs whose results are complemented by a Life Cycle Assessment (LCA) for the same system, and a Societal LCC, for socio-economic assessments. Conventional and Environmental LCCs includes budget costs and transfers, while Societal LCCs includes budget and externality costs. Critical aspects were found in the existing literature regarding the cost assessment of waste management, namely system boundary equivalency, accounting for temporally distributed emissions and impacts, inclusions of transfers, the internalisation of environmental impacts and the coverage of shadow prices, and there was also significant confusion regarding terminology. The presented cost model was implemented in two case study scenarios assessing the costs involved in the source segregation of organic waste from 100,000 Danish households and the subsequent co-digestion of organic waste with animal manure. Overall, source segregation resulted in higher financial costs than the alternative of incinerating the organic waste with the residual waste: 1.6 M€/year, of which 0.9 M€/year was costs for extra bins and bags used by the households, 1.0 M€/year for extra collections and −0.3 M€/year saved on incineration.

09 BIOMASS FUELS↗