Enhanced thermophilic high-solids anaerobic digestion of organic fraction of municipal solid waste with spatial separation from conductive materials in a single reactor volume.

Despite benefits such as lower water and working volume requirements, thermophilic high solids anaerobic digestion (THSAD) often fails due to the rapid build-up of volatile fatty acids (VFAs) and the associated drop in pH. Use of conductive materials (CM) can promote THSAD through stimulation of direct interspecies electron transfer (DIET), while the need for their constant dosing due to poor separation from effluent impairs economic feasibility. This study used an approach of spatially separating magnetite and granular activated carbon (GAC) from the organic fraction of municipal solid waste (OFMSW) in a single reactor for THSAD. GAC and magnetite addition could both mitigate the severe inhibition of methanogenesis after VFAs build-up to ∼28-30 g/L, while negligible methane production was observed in the control group. The highest methane yield (286 mL CH4/g volatile solids (VS)) was achieved in magnetite-added reactors, while the highest maximum CH4 production rates (26.38 mL CH4/g VS/d) and lowest lag-phase (2.83 days) were obtained in GAC-added reactors. The enrichment of GAC and magnetite biofilms with various syntrophic and potentially electroactive microbial groups (Ruminiclostridium 1, Clostridia MBA03, Defluviitoga, Lentimicrobiaceae) in different relative abundances indicates the existence of specific preferences of these groups for the nature of CM. According to predicted basic metabolic functions, CM can enhance cellular processes and signals, lipid transport and metabolism, and methane metabolism, resulting in improved methane production. Rearrangement of metabolic pathways, formation of pili-like structures, enrichment of biofilms with electroactive groups and a significant improvement in THSAD performance was attributed to the enhancement of the DIET pathway. Promising results obtained in this work due to the spatial separation of the bulk OFMSW and CM can be useful for modeling larger-scale THSAD systems with better recovery of CM and cost-effectiveness.

Does carbon cloth really improve thermophilic anaerobic digestion performance on a larger scale? focusing on statistical analysis and microbial community dynamics.

Currently, the phenomenon of direct interspecies electron transfer (DIET) is of great interest in the technology of anaerobic digestion (AD) due to potential performance benefits. However, the conditions for the occurrence of DIET and its limits on improving AD under conditions close to real have not been studied enough. This research is concentrated on the effect of conductive carbon cloth (R3), in comparison with a dielectric fiberglass cloth (R2) and control (R1), on the AD performance in large (90 L) thermophilic reactors, fed with a mixture of simulated organic fraction of municipal solid waste and sewage sludge. While organic loading rate (OLR) was gradually increased from 2.4 to 8.66 kg VS/(m3 day), a statistically significant (p < 0.05) difference in biogas production was observed between R1 and both R2 and R3. However, at a maximum OLR of 12.12 kg VS/(m3 day) in R3, an increase in biogas production (p < 0.05) was observed both compared to R1 (by 8.97%) and R2 (by 4.24%). The content of volatile fatty acids in R3 as a whole was the lowest, especially at the maximum OLR. Biofilm on carbon cloth was rich in syntrophic microorganisms of the genera Tepidanaerobacter, as well as Defluviitoga, capable of DIET in mixed cultures with Methanothrix, which was the most abundant methanogen in biofilm. Suspended Bifidobacterium, Fervidobacterium and Anaerobaculum were negatively affected, while Defluviitoga, Methanothermobacter and Methanosarcina, on the contrary, were positively affected by the increase in OLR and showed, respectively, a negative and positive correlation (p < 0.05) with the main AD performance parameters.

Insights into sustainable resource and energy recovery from leachate towards emission mitigation for environmental management: A critical approach.

The exponential generation of municipal solid waste (MSW) and landfill disposal without any treatment has increased the continuous generation of landfill leachate. Improper MSW and leachate management are contributing to environmental degradation and water and soil pollution, which must be treated. Numerous works have been conducted on leachate treatments for energy and resource recovery. This review presents a comprehensive study of leachate management in which different treatment methods are discussed to analyze the suitability of processes that can be employed to treat leachate efficiently. Further, the characteristics of leachate are examined as properties of leachate may be varied depending upon the region. Still, several challenges related to leachate management and its treatments are discussed in this study. An integrated system could be a better option for treating leachate because it contains large amounts of organic and inorganic compounds. Proper leachate management would help to recover energy and value-added products (metals).

Development of a Clinical Global Impression of Change (CGI-C) and a Caregiver Global Impression of Change (CaGI-C) measure for ambulant individuals with Duchenne muscular dystrophy.

BACKGROUND: In clinical trials for rare diseases, such as Duchenne muscular dystrophy, clinical outcome assessments (COA) used to assess treatment benefit are often generic and may not be sensitive enough to detect change in specific patient populations. Thus, there is a need for disease specific COAs that track meaningful change among individuals. When developing such measures, input from clinicians, caregivers and patients is critical for assessing clinically relevant concepts and ensuring validity of the measure. METHOD: The aim of this study was to develop two Duchenne-specific global impression items for use in clinical trials. The development of the Duchenne Clinical Global Impression of Change (CGI-C) and Caregiver Global Impression of Change (CaGI-C) was informed by findings from concept elicitation (CE) interviews with clinicians, caregivers and individuals with Duchenne. Through cognitive debriefing (CD) interviews, clinicians and caregivers evaluated draft CGI-C and CaGI-C items to ensure relevance and understanding of the items and instructions. Suggestions made during the CD interviews were incorporated into the finalized CGI-C and CaGI-C measures. RESULTS: The symptoms most frequently reported by clinicians, caregivers and individuals with Duchenne were muscle weakness, fatigue, cardiac difficulties and pain. Regarding physical functioning, all three populations noted that small changes in functional ability were meaningful, particularly when independence was impacted. Caregivers and clinicians reported that changes in speed, endurance and quality of movement were important, as was improvement in the ability of individuals to keep up with their peers. A change in the ability to complete everyday activities was also significant to families. These results were used to create two global impression of change items and instruction documents for use by clinicians (CGI-C) and caregivers (CaGI-C). Overall, both items were well understood by participants. The descriptions and examples developed from the CE interviews were reported to be relevant and appropriate for illustrating different levels of meaningful change in patients with Duchenne. Modifications were made based on caregiver and clinician CD feedback . CONCLUSIONS: As part of a holistic measurement strategy, such COA can be incorporated into the clinical trial setting to assess global changes in relevant symptoms and functional impacts associated with Duchenne.

Enhanced production of acetic acid through bioprocess optimization employing response surface methodology and artificial neural network.

In this study, acetic acid bacteria (AAB) are isolated from fruit waste and cow dung on the basis of acetic acid production potential. The AAB were identified based on halo-zones produced in the Glucose-Yeast extract-Calcium carbonate (GYC media) agar plates. In the current study, maximum acetic acid yield is reported to be 4.88 g/100 ml from the bacterial strain isolated from apple waste. With the help of RSM (Response surface methodology) tool, glucose and ethanol concentration and incubation period, as independent variable showed the significant effect of glucose concentration and incubation period and their interaction on the AA yield. A hypothetical model of artificial neural network (ANN) was also used to compare the predicted value from RSM. Acetic acid production through the biological route can be the sustainable and clean approach to utilizing food waste in circular economy approach.

Invasion Genetics of the Horse-Chestnut Leaf Miner, Cameraria ohridella (Lepidoptera: Gracillariidae), in European Russia: A Case of Successful Involvement of Citizen Science in Studying an Alien Insect Pest.

Based on the intensive monitoring conducted by our team and volunteers in 2021, the secondary range of an alien horse-chestnut leaf miner, Cameraria ohridella Deschka & Dimic, 1986 (Lepidoptera: Gracillariidae), was specified in European Russia. This invasive pest was confirmed in 24 out of 58 administrative regions of Russia, which it has occupied for approximately 16 years. Analysis of the COI mtDNA gene sequenced in 201 specimens collected in 21 regions of the European part of Russia indicates the occurrence of two haplotypes (A and B), which are also present in the secondary range of C. ohridella in Eastern and Western Europe. The haplotype A dominated and was present in 87.5% of specimens from European Russia. In 2021, C. ohridella produced spectacular outbreaks in Aesculus hippocastanum in southern Russia, where it damaged more than 50% of the leaves in trees in 24 out of 30 distant localities. In the south of the country, the pest infested Acer pseudoplatanus, whereas other species of Acer of European, East Asian, and North American origin showed no signs of attacks. Taking into account that Ae. hippocastanum is present in most regions of European Russia, we expect a further range expansion of C. ohridella up to the Ural Mountains.

Biochemical hydrogen potential assay for predicting the patterns of the kinetics of semi-continuous dark fermentation.

The performance and kinetics response of thermophilic semi-continuous dark fermentation (DF) of simulated complex carbohydrate-rich waste was investigated at various hydraulic retention times (HRT) (2, 2.5, and 3 d) and compared with data obtained from biochemical hydrogen potential assay (BHP). A culture of Thermoanaerobacterium thermosaccharolyticum was used as the inoculum and dominated both in BHP and semi-continuous reactor. Both the modified Gompertz and first-order models described the DF kinetics well (R2 = 0.97-1.00). HRT of 2.5 d was found to be optimal in terms of maximum hydrogen production rate and hydrogen potential, which were 3.97 and 1.26 times higher, respectively, than in BHP. The hydrolysis constant was highest at HRT of 3 d and was closest to the value obtained in the BHP. Overall, BHP has been shown to be a useful tool for predicting H2 potential and the hydrolysis constant, while the maximum H2 production rate is greatly underestimated.

Comparative effect of conductive and dielectric materials on methanogenesis from highly concentrated volatile fatty acids.

Various conductive materials and their dielectric counterparts were used to get deeper insights into contribution of direct interspecies electron transfer (DIET) in improving methanogenesis from highly concentrated volatile fatty acids (12.5 g/L). Potential CH4 yield, maximum CH4 production rate and lag phase were significantly (up to 1.4, 3.9 and 2.0 times, respectively) improved with addition of stainless-steel mesh (SM) and carbon felt (CF) compared to both control and dielectric counterparts (p < 0.05). kapp increased by 82% for SM and 63% for CF compared to control (p < 0.05). Short thick pili-like structures up to 150 nm in width were formed only in CF and SM biofilms, however, were more abundant for SM. Ureibacillus and Limnochordia specific for SM biofilms, and Coprothermobacter and Ca. Caldatribacterium for CF biofilms, were considered electrogenic. Promotion of DIET by conductive materials is governed by many factors, including specificity of electrogenic groups to material surface.

Utilization of agricultural residues for energy and resource recovery towards a sustainable environment.

Fungal pre-treatment using Pleurotus ostreatus (PO) was carried out on individual and combinations of agro-waste wheat straw (WS), rice straw (RS), and pearl millet straw (PMS) with the addition of biochar (5%,7.5% and 10%) to reduce the pre-treatment duration. Further remaining substrate known as spent mushroom substrate (SMS) was used in anaerobic digestor (AD) for estimation enhanced biomethane yield. Equal ratios of RS + WS, WS + PMS, PMS + RS, and RS + PMS + WS and biochar addition were taken for enhancing pre-treatment, PO growth and AD process. The extent of pre-treatment was recorded with the maximum lignin removal of 40.4% for RS + PMS + WS as compared to untreated counterparts and 0.5%, 2.2%, and 3.3% times more lignin removal from individual PMS, RS, and WS respectively. Addition of biochar to the substrates reduced the total pre-treatment duration by days as compared to the non-biochar substrates. Biological efficiency (BE) used for the analysis of mushroom growth varied from 51-92%. Further, the average bio-methane yield was 187 ml/gVS for SMS of PMS + WS + RS with 10% biochar indicating an increment of 83.33% from untreated SMS of PMS + WS + RS. This, higher biomethane yield was 9.35%, 22.22% and 57.14% times higher than individual SMS of PMS, RS, and WS respectively. The current study shows that biochar not only enhances the bio-methane yield but also reduces the biological pre-treatment duration and removes the dependency on one lignocellulosic biomass for energy (bio-methane) and food (mushroom) production.

Effects of various materials used to promote the direct interspecies electron transfer on anaerobic digestion of low-concentration swine manure.

The activation of direct interspecies electron transfer (DIET) by the supplementation of conductive materials is one of the effective and available methods to enhance anaerobic digestion (AD). Microorganisms that colonize the surface of these materials form biofilms, the study of which could provide new insights into the character of the DIET process and its effect on AD. The present study focused on AD performance, microbial community, as well as morphological and topological features of biofilms on various materials used to promote DIET during AD of low-concentration swine manure. The best AD characteristics were observed in stainless steel mesh (SM)/digested cow manure (CM) and polyester felt (PF)/digested sewage sludge (SS) combinations used as material/inoculum, respectively. Thus, potential methane yields in CM-SM and SS-PF were up to 26.4% and 26.2% higher compared to the corresponding controls. Microbial analysis of biofilms revealed the dominance of putatively syntrophic bacteria of the MBA03 group of the Limnochordia class in CM inoculated reactors, and syntrophic proteolytic bacteria of the genus Coprothermobacter and acetogenic Clostridium sensu stricto 1, known for their ability to carry out DIET, in SS inoculated reactors. Biofilms on non-conductive materials contained pili-like structures, which were observed only in SS inoculated reactors. Polyester felt tended to biofoul better than carbon felt, resulting in up to 2.8, 3.2 and 1.8 higher nucleic acid, extracellular polymeric substances, and total biomass content, respectively, depending on the inoculum. These results provide new insights into the different types of DIET that can occur in low-loaded AD systems with attached growth.

Monitoring and Evaluation of Patient Engagement in Health Product Research and Development: Co-Creating a Framework for Community Advisory Boards.

PURPOSE: While patient engagement is becoming more customary in developing health products, its monitoring and evaluation to understand processes and enhance impact are challenging. This article describes a patient engagement monitoring and evaluation (PEME) framework, co-created and tailored to the context of community advisory boards (CABs) for rare diseases in Europe. It can be used to stimulate learning and evaluate impacts of engagement activities. METHODS: A participatory approach was used in which data collection and analysis were iterative. The process was based on the principles of interactive learning and action and guided by the PEME framework. Data were collected via document analysis, reflection sessions, a questionnaire, and a workshop. RESULTS: The tailored framework consists of a theory of change model with metrics explaining how CABs can reach their objectives. Of 61 identified metrics, 17 metrics for monitoring the patient engagement process and short-term outcomes were selected, and a "menu" for evaluating long-term impacts was created. Example metrics include "Industry representatives' understanding of patients' unmet needs;" "Feeling of trust between stakeholders;" and "Feeling of preparedness." "Alignment of research programs with patients' needs" was the highest-ranked metric for long-term impact. CONCLUSIONS: Findings suggest that process and short-term outcome metrics could be standardized across CABs, whereas long-term impact metrics may need to be tailored to the collaboration from a proposed menu. Accordingly, we recommend that others adapt and refine the PEME framework as appropriate. The next steps include implementing and testing the evaluation framework to stimulate learning and share impacts.

PKB and the mitochondria: AKTing on apoptosis.

Cellular homeostasis depends upon the strict regulation of responses to external stimuli, such as signalling cascades triggered by nutrients and growth factors, and upon cellular metabolism. One of the major molecules coordinating complex signalling pathways is protein kinase B (PKB), a serine/threonine kinase also known as Akt. The number of substrates known to be phosphorylated by PKB and its interacting partners, as well as our broad understanding of how PKB is implicated in responses to growth factors, metabolic pathways, proliferation, and cell death via apoptosis is constantly increasing. Activated by the insulin/growth factor-phosphatidylinositol 3-kinase (PI3K) cascade, PKB triggers events that promote cell survival and prevent apoptosis. It is also now widely accepted that mitochondria are not just suppliers of ATP, but that they participate in regulatory and signalling events, responding to multiple physiological inputs and genetic stresses, and regulate both cell proliferation and death. Thus, mitochondria are recognized as important players in apoptotic events and it is logical to predict some form of interplay with PKB. In this review, we will summarize mechanisms by which PKB mediates its anti-apoptotic activities in cells and survey recent developments in understanding mitochondrial dynamics and their role during apoptosis.

Acyl coenzyme A thioesterase Them5/Acot15 is involved in cardiolipin remodeling and fatty liver development.

Acyl coenzyme A (acyl-CoA) thioesterases hydrolyze thioester bonds in acyl-CoA metabolites. The majority of mammalian thioesterases are α/ß-hydrolases and have been studied extensively. A second class of Hotdog-fold enzymes has been less well described. Here, we present a structural and functional analysis of a new mammalian mitochondrial thioesterase, Them5. Them5 and its paralog, Them4, adopt the classical Hotdog-fold structure and form homodimers in crystals. In vitro, Them5 shows strong thioesterase activity with long-chain acyl-CoAs. Loss of Them5 specifically alters the remodeling process of the mitochondrial phospholipid cardiolipin. Them5(-/-) mice show deregulation of lipid metabolism and the development of fatty liver, exacerbated by a high-fat diet. Consequently, mitochondrial morphology is affected, and functions such as respiration and ß-oxidation are impaired. The novel mitochondrial acyl-CoA thioesterase Them5 has a critical and specific role in the cardiolipin remodeling process, connecting it to the development of fatty liver and related conditions.

Carboxy-Terminal Modulator Protein (CTMP) is a mitochondrial protein that sensitizes cells to apoptosis.

The Carboxy-Terminal Modulator Protein (CTMP) protein was identified as a PKB inhibitor that binds to its hydrophobic motif. Here, we report mitochondrial localization of endogenous and exogenous CTMP. CTMP exhibits a dual sub-mitochondrial localization as a membrane-bound pool and a free pool of mature CTMP in the inter-membrane space. CTMP is released from the mitochondria into the cytosol early upon apoptosis. CTMP overexpression is associated with an increase in mitochondrial membrane depolarization and caspase-3 and polyADP-ribose polymerase (PARP) cleavage. In contrast, CTMP knock-down results in a marked reduction in the loss of mitochondrial membrane potential as well as a decrease in caspase-3 and PARP activation. Mutant CTMP retained in the mitochondria loses its capacity to sensitize cells to apoptosis. Thus, proper maturation of CTMP is essential for its pro-apoptotic function. Finally, we demonstrate that CTMP delays PKB phosphorylation following cell death induction, suggesting that CTMP regulates apoptosis via inhibition of PKB.

The Carboxy-Terminal Modulator Protein (CTMP) regulates mitochondrial dynamics.

BACKGROUND: Mitochondria are central to the metabolism of cells and participate in many regulatory and signaling events. They are looked upon as dynamic tubular networks. We showed recently that the Carboxy-Terminal Modulator Protein (CTMP) is a mitochondrial protein that may be released into the cytosol under apoptotic conditions. METHODOLOGY/PRINCIPAL FINDINGS: Here we report an unexpected function of CTMP in mitochondrial homeostasis. In this study, both full length CTMP, and a CTMP mutant refractory to N-terminal cleavage and leading to an immature protein promote clustering of spherical mitochondria, suggesting a role for CTMP in the fission process. Indeed, cellular depletion of CTMP led to accumulation of swollen and interconnected mitochondria, without affecting the mitochondrial fusion process. Importantly, in vivo results support the relevance of these findings, as mitochondria from livers of adult CTMP knockout mice had a similar phenotype to cells depleted of CTMP. CONCLUSIONS/SIGNIFICANCE: Together, these results lead us to propose that CTMP has a major function in mitochondrial dynamics and could be involved in the regulation of mitochondrial functions.