Multiple-criteria decision analysis techniques for anaerobic digester technology assessment.

Effective decision-making requires the evaluation of several criteria rather than a single, preferred criterion. The best decision options (alternatives) are recommended to decision-makers when a multi-criteria decision problem is addressed. This study develops a multi-criteria selection method for the assessment of small-scale anaerobic digester technology by combining two existing methods. The Simple Multi-Attribute Rating Technique (SMART) and the Analytical Hierarchy Process (AHP) approaches of multiple-criteria decision analysis were used as a decision support tool, and the preferred anaerobic digester technology was selected from a list of eleven potential small-scale digester technologies used in low to middle-income countries. These techniques were applied under two scenarios for a case study in the South African smallholder farmers. Scenario 1 involves a subsistence smallholder farming context, while scenario 2 involves a commercially oriented smallholder farming context. The overall results revealed that the DIY Biobag and Puxin digester design models achieved 82.1 and 73.7 % preference in comparison to other digester technologies for scenarios 1 and 2, respectively. The Biobag digester technology achieved the highest ranking, which is consistent with the significant cost advantage and technical characteristics of the technology. However, for those households with sufficient access to funds for the initial expenditure, the method identifies the Puxin digester as the most appropriate alternative, excluding cases where underground construction is not possible. The AGAMA BiogasPro digester was ranked in the second position in both scenarios. A sensitivity analysis was conducted to determine the effect of changes in the assessment criteria weights and found the selected alternatives stable and robust. Finally, it can be concluded that the developed technology selection model contributed a knowledge-based framework to be used in various situations by different decision-makers, thereby providing a method applicable to particular local conditions to identify the most suitable technology choices.

Quantifying community-wide antibiotic usage via urban water fingerprinting: Focus on contrasting resource settings in South Africa.

There has been a significant increase in antimicrobial agents (AAs) usage, globally - however the relative consumption is unevenly distributed between nations. Inappropriate use of antibiotics can harbour inherent antimicrobial resistance (AMR); therefore, it is important to understand and monitor community-wide prescribing and consumption behaviours throughout different communities around the world. Wastewater-Based Epidemiology (WBE) is a novel tool enabling low cost and large scale studies focussed on AA usage patterns. The back-calculation of community antimicrobial intake was performed from quantities measured in municipal wastewater and informal settlement discharge in the city of Stellenbosch, utilising WBE. Seventeen antimicrobials, and their human metabolites, were evaluated, in concordance with prescription records corresponding to the catchment region. The proportional excretion, biological/chemical stability, and method recovery of each analyte were all crucial factors in the efficacy of the calculation. Mass per day measurements were normalised to the catchment area via population estimates. Municipal wastewater treatment plant population estimates were used to normalise the wastewater samples and prescription data (mg/day/1000 inhabitants). Population estimates for the informal settlements were less accurate due to a lack of reliable sources that were relevant to the sampling time period. Both mass loads and normalised loads suggested higher than average usage throughout the settlements, relative to municipal wastewater. This was seen most prominently in emtricitabine and lamivudine; but also, sulfamethoxazole, trimethoprim, sulfadiazine, clindamycin, ciprofloxacin, ofloxacin, and doxycycline. Urban water fingerprinting (UWF) data triangulation with prescription datasets showed good correlations for several antimicrobial agents (AAs) (e.g., clindamycin, clarithromycin, ofloxacin, and doxycycline). It also revealed discrepancies in usage for some compounds (e.g., tetracycline and sulfapyridine). This might be linked with a lack of pharma compliance in prescription datasets; erroneous association of prescription boundaries with the sewerage catchment; and/or uncertainties within the sewerage catchment (e.g., population estimations). The UWF tool provided a comprehensive overview of multiclass AAs usage, both prescription and over-the counter. For example, tetracycline was not reported in available prescription statistics, but was detected at an average of 18.4 mg/day/1000inh; and no antiviral prescriptions were obtained, but emtricitabine and lamivudine were quantified at 2415.4 and 144.4 mg/day/1000inh, respectively. A lack of clarity regarding prescriptions and a lack of inclusion of several critical (often over-the-counter) medications in public health databases makes WBE a useful and comprehensive epidemiology tool for tracking pharma usage within a catchment.

Probabilistic techno-economic assessment of anaerobic digestion predicts economic benefits to smallholder farmers with quantifiable certainty.

Anaerobic digestion (AD) technology holds numerous potential benefits for farmers, however, challenges persist in terms of implementation costs and sustainability in developing countries. This paper presents a probabilistic techno-economic assessment tool for AD. A clear distinction is made between direct financial feasibility and wider (socio) economic feasibility. The tool identifies the technical- and economic factors influencing the returns of a particular AD process as well as the sensitivity of model predictions to variations in the value of the identified factors using a Monte Carlo approach. The tool is applied to assess the feasibility of a smallholder farm-based AD installation under a variety of substrates and operating conditions as an illustrative case study, where on-going flows of costs and benefits were considered over a 15-year period and discounted at a rate of 8%. The results of the case study revealed that the installation of a 10 m3 smallholder farm-based anaerobic digester are likely to be financially and economically viable with a financial benefit-cost ratio of 1.30-1.38 and an economic benefit-cost ratio ranged from 5.49 to 6.01. Risk assessment results confirmed the strong economic feasibility of a smallholder farm-based AD implementation: under the most conservative cost estimates, there is a 73% probability of achieving a financial benefit-cost ratio > 1, while there is a 96.6% probability of achieving an economic benefit-cost ratio > 1. The case study demonstrated the utility of probabilistic techno-economic assessments for informed decision making, a tool which can be readily generalized to other settings.

Cyanobacterial Growth in Minimally Amended Anaerobic Digestion Effluent and Flue-Gas.

Anaerobic digestion (AD) is an important industrial process, particularly in a biorefinery approach. The liquid effluent and carbon dioxide in the off-gas, can be used to produce high-value products through the cultivation of cyanobacteria. Growth on AD effluent is often limited due to substrate limitation or inhibitory compounds. This study demonstrates the successful cultivation of Synechococcus on minimally amended AD effluent, supplemented with MgSO4 and diluted with seawater. An 8 L airlift reactor illustrated growth in a pilot scale setup. Higher biomass yields were observed for cyanobacteria grown in diluted AD effluent compared to minimal medium, with 60% total nitrogen removal in the effluent. It was demonstrated that controlling the pH, increasing dissolved salt concentrations and adding MgSO4 to the effluent allowed for the successful cultivation of the cyanobacterium, circumventing the addition of clean water for effluent dilution. This could ultimately increase the feasibility of anaerobic digestion-microalgae integrated biorefineries.

Frequency sensitive mechanism in low-intensity ultrasound enhanced bioeffects.

This study presents two novel theoretical models to elucidate frequency sensitive nuclear mechanisms in low-intensity ultrasound enhanced bioeffects. In contrast to the typical 1.5 MHz pulsed ultrasound regime, our group previously experimentally confirmed that ultrasound stimulation of anchored chondrocytes at resonant frequency maximized gene expression of load inducible genes which are regulatory markers for cellular response to external stimuli. However, ERK phosphorylation displayed no frequency dependency, suggesting that the biochemical mechanisms involved in enhanced gene expression is downstream of ERK phosphorylation. To elucidate such underlying mechanisms, this study presents a theoretical model of an anchored cell, representing an in vitro chondrocyte, in an ultrasound field. The model results showed that the mechanical energy storage is maximized at the chondrocyte's resonant frequency and the energy density in the nucleus is almost twice as high as in the cytoplasm. Next, a mechanochemical model was developed to link the mechanical stimulation of ultrasound and the increased mechanical energy density in the nucleus to the downstream targets of the ERK pathway. This study showed for the first time that ultrasound stimulation induces frequency dependent gene expression as a result of altered rates of transcription factors binding to chromatin.

Theoretical evaluation of the acoustic field in an ultrasonic bioreactor.

Ultrasound-assisted bioreactors that provide mechanical conditioning to cells have broad applicability in tissue engineering, but biological experiments with ultrasound are very sensitive to environmental conditions. A mathematical model was developed to complement experimental measurements, as well as to describe ultrasonic fields existing in regions where measurements are impossible, specifically, within microporous tissue engineering scaffolds. The model uniquely combines Biot theory to predict the ultrasonic field in the scaffold with an electromechanical transducer model to couple the mechanical stimulation experienced by cells to the external electrical input. In the specific example examined here, cells immobilized on scaffolds are subjected to different forms of ultrasonic stimulation due to the formation of standing wave fields and vertical high-pressure bands. The model confirms the sensitivity of the supplied acoustic power to the liquid level in sonobioreactors and identifies the input electrical impedance as a method of detecting resonance effects.

Mechanotransduction of ultrasound is frequency dependent below the cavitation threshold.

This study provides evidence that low-intensity ultrasound directly affects nuclear processes, and the magnitude of the effect varies with frequency. In particular, we show that the transcriptional induction of first load-inducible genes, which is independent of new protein synthesis, is frequency dependent. Bovine chondrocytes were exposed to low-intensity (below the cavitational threshold) ultrasound at 2, 5 and 8 MHz. Ultrasound elevated the expression of early response genes c-Fos, c-Jun and c-Myc, maximized at 5 MHz. The phosphorylated ERK inhibitor PD98059 abrogated any increase in c-series gene expression, suggesting that signaling occurs via the MAPPK/ERK pathway. However, phosphorylated ERK levels did not change with ultrasound frequency, indicating that processes downstream of ERK phosphorylation (such as nuclear transport and chromatin reorganization) respond to ultrasound with frequency dependence. A quantitative, biphasic mathematical model based on Biot theory predicted that cytoplasmic and nuclear stress is maximized at 5.2 ± 0.8 MHz for a chondrocyte, confirming experimental measurements.

Integrin-mediated mechanotransduction pathway of low-intensity continuous ultrasound in human chondrocytes.

Chondrocytes are mechanosensitive cells that require mechanical stimulation for proper growth and function in in vitro culture systems. Ultrasound (US) has emerged as a technique to deliver mechanical stress; however, the intracellular signaling components of the mechanotransduction pathways that transmit the extracellular mechanical stimulus to gene regulatory mechanisms are not fully defined. We evaluated a possible integrin/mitogen-activated protein kinase (MAPK) mechanotransduction pathway using Western blotting with antibodies targeting specific phosphorylation sites on intracellular signaling proteins. US stimulation of chondrocytes induced phosphorylation of focal adhesion kinase (FAK), Src, p130 Crk-associated substrate (p130Cas), CrkII and extracellular-regulated kinase (Erk). Furthermore, pre-incubation with inhibitors of integrin receptors, Src and MAPK/Erk kinase (MEK) reduced US-induced Erk phosphorylation levels, indicating integrins and Src are upstream of Erk in an US-mediated mechanotransduction pathway. These findings suggest US signals through integrin receptors to the MAPK/Erk pathway via a mechanotransduction pathway involving FAK, Src, p130Cas and CrkII.

Forced wave motion with internal and boundary damping.

A d'Alembert-based solution of forced wave motion with internal and boundary damping is presented with the specific intention of investigating the transient response. The dynamic boundary condition is a convenient method to model the absorption and reflection effects of an interface without considering coupled PDE's. Problems with boundary condition of the form [Formula: see text] are not self-adjoint which greatly complicates solution by spectral analysis. However, exact solutions are found with d'Alembert's method. Solutions are also derived for a time-harmonically forced problem with internal damping and are used to investigate the effect of ultrasound in a bioreactor, particularly the amount of energy delivered to cultured cells. The concise form of the solution simplifies the analysis of acoustic field problems.

Experimental Validation of a Fundamental Model for PCR Efficiency.

Recently a theoretical analysis of PCR efficiency has been published by Booth et al., (2010). The PCR yield is the product of three efficiencies: (i) the annealing efficiency is the fraction of templates that form binary complexes with primers during annealing, (ii)the polymerase binding efficiency is the fraction of binary complexes that bind to polymerase to form ternary complexes and (iii)the elongation efficiency is the fraction of ternary complexes that extend fully. Yield is controlled by the smallest of the three efficiencies and control could shift from one type of efficiency to another over the course of a PCR experiment. Experiments have been designed that are specifically controlled by each one of the efficiencies and the results are consistent with the mathematical model. The experimental data has also been used to quantify six key parameters of the theoretical model. An important application of the fully characterized model is to calculate initial template concentration from real-time PCR data. Given the PCR protocol, the midpoint cycle number (where the template concentration is half that of the final concentration) can be theoretically determined and graphed for a variety of initial DNA concentrations. Real-time results can be used to calculate the midpoint cycle number and consequently the initial DNA concentration, using this graph. The application becomes particularly simple if a conservative PCR protocol is followed where only the annealing efficiency is controlling.

Efficiency of the Polymerase Chain Reaction.

The polymerase chain reaction (PCR) has found wide application in biochemistry and molecular biology such as gene expression studies, mutation detection, forensic analysis and pathogen detection. Increasingly quantitative real time PCR is used to assess copy numbers from overall yield. In this study the yield is analyzed as a function of several processes: (1) thermal damage of the template and polymerase occurs during the denaturing step, (2) competition exists between primers and templates to either anneal or form dsDNA, (3) polymerase binding to annealed products (primer/ssDNA) to form ternary complexes and (4) extension of ternary complexes. Explicit expressions are provided for the efficiency of each process, therefore reaction conditions can be directly linked to the overall yield. Examples are provided where different processes play the yield-limiting role. The analysis will give researchers a unique understanding of the factors that control the reaction and will aid in the interpretation of experimental results.