Quantitative biology of single neurons

The building blocks of complex biological systems are single cells. Fundamental insights gained from single-cell analysis promise to provide the framework for understanding normal biological systems development as well as the limits on systems/cellular ability to respond to disease. The interplay of cells to create functional systems is not well understood. Until recently, the study of single cells has concentrated primarily on morphological and physiological characterization. With the application of new highly sensitive molecular and genomic technologies, the quantitative biochemistry of single cells is now accessible.     

Triboelectric charging of fungal spores during resuspension and rebound

The triboelectric charging of fungal spores was experimentally characterized during rebound and resuspension. A fungal spore source strength tester (FSSST) was used as a primary aerosol generator for spores of three fungal species and two powders (silicon carbide and silver). The critical velocity of rebound was determined using a variable nozzle area impactor (VNAI), and the charging state of particles after resuspension and rebound was measured using the FSSST, different impactor setups, electrometers, and optical particle counters. In the impactor setups and the FSSST, five different surface materials relevant for indoor environments were used (steel, glass, polystyrene, paper, and polytetrafluoroethylene). The critical velocity of rebound was determined to be 0.57 m/s for fungal spores, which is relatively low compared to silicon carbide and previous results for micron-sized aerosol particles. Based on the rebound impactor measurements, we were able to define the crucial parameters of charge transfer for different particle–surface material pairs. A contact charge parameter, which describes the triboelectric charging during rebound, was found to have a negative correlation with the charging state of the particles after the resuspension from an impactor. This connects the triboelectric charging during rebound and resuspension to each other. Based on the contact charge parameter values, quantified triboelectric series could be formed. The results of this work show that fungal spores can be charged both positively and negatively during rebound and resuspension depending on the fungal species and surface material.

Copyright © 2016 American Association for Aerosol Research     

CAVE for collaborative patient room design: analysis with end-user opinion contrasting method

Several studies indicate that virtual reality (VR) systems are useful for end-user participation in an environmental design process. However, these systems can be costly and thus support for the decision whether to invest in a VR of some type is useful. This study presents a novel method for analysing the usefulness of a VR system for the purpose of end-user participation. We collected qualitative end-user opinion data in the real environment and then contrasted this data with the capabilities of a VR system. Additionally, to better understand the capabilities of the VR used, we examined how the end-users perceive the used virtual environment, which in this case was CAVE, an immersive VR system where projectors are directed to the walls of a room-sized cube. In this way, we analysed whether the same functions and elements identified by end-users on the actual wards could also be evaluated in the CAVE. Eleven nurses and 11 patients participated in the study by evaluating a bathroom and/or four patient rooms modelled by the CAVE and the actual hospital wards. The CAVE was convenient for evaluating most issues identified by the study participants in the actual hospital wards, i.e. aesthetics; correct location of equipment, supplies and materials; distraction by or the good companion of other patients as well as window position and size and living/workspace. However, it was not possible to evaluate with full certainty the possibilities for bracing against grab bars or other objects in the VR, and this was found to be relevant to the independent functioning of patients with limited mobility. Also, due to the relatively low luminance levels of projectors, evaluations regarding lighting were considered unreliable. Moreover, end-users were not always certain about the sizes and sufficiency of space in the CAVE. Solutions to overcome these limitations were proposed.     

Design and Collection Efficiency of a New Electrostatic Precipitator for Bioaerosol Collection

We have developed and tested a new bioaerosol sampler in which airborne microorganisms are collected by electrostatic means. In this sampler, 2 ionizers charge the incoming particles if they carry insufficient electric charge for efficient collection. The organisms are then subjected to a precipitating electric field and are collected onto 2 square agar plates positioned along the flow axis. Tests with nonbiological NaCl particles versus B. subtilis var. niger (BG) spores and vegetative cells have shown that airborne microorganisms are collected more efficiently than nonbiological particles, even when the microorganisms have first passed through an electric charge neutralizer with no additional charging applied. The difference was attributed to the natural charges contained in cell membranes or spore coats of the microorganisms. Charge-neutralized BG spores and vegetative cells were collected at 4 L/min with efficiencies close to 80%, depending on the precipitation voltage, versus 50-60% for NaCl test particles. When incoming BG spores were charged with positive ions and then collected by a precipitating voltage of + 1,300 V, about 80% of the incoming spores were collected and more than 70% of incoming spores formed colonies. These experiments with BG spores have also indicated that there were no significant particle losses inside the sampler. The collection efficiency of biological and nonbiological particles increased to 90-100% when the particles were externally charged and the precipitating voltage was increased to more than - 4,000 V. It has also been shown that the aerosolized BG spores (used as anthrax simulants for bioaerosol sensors) carry a net negative electric charge. Thus the collection efficiency depends on the polarity of the electric field applied across the agar plates. These findings indicate that the collection of airborne microorganisms is possible by electrostatic precipitation without prior electric charging if the microorganisms already carry electric charges. These are usually high immediately after their release into the air.     

Techniques for Dispersion of Microorganisms into Air

ABSTRACT

Many commercially available devices initially developed for dispersion of biologically inert particles have been adopted for aerosolization of microoganisms in laboratory settings. However, these dispersion devices are not always adequate for microbial particles, as they do not simulate natural release into air. Wet dispersion methods are appropriate for viruses and most bacteria, whereas dry methods are more suitable for most fungal and actinomycete spores. Characteristics of the resulting aerosol are dependent on the dispersing shear forces and the sensitivity and agglomeration of the tested microorganisms. Consequently, each microbial group may need a specific dispersion technique. The following devices have been developed and tested in this study: the bubbling aerosol disperser, the agar-tube disperser, and the swirling-flow disperser. Testing included the evaluation of both physical and microbiological characteristics of aerosolized microorganisms. Each of the dispersers has shown several advantages over commercially available ones. When used for the dispersion of bacteria from the liquid suspension, the bubbling aerosol disperser was found to produce considerably fewer amounts of microbial fragments and much lower levels of microbial metabolic injury than the commercially available Collison nebulizer. Fungal spores dispersed from their colonies by the agar-tube disperser were found to have a more stable aerosol concentration and a lower fraction of agglomerates than achievable by conventional powder dispersion. The swirling-flow dispersion technique was used for aerosolization of actinomycetes because the agar-tube disperser could not provide a stable concentration of these spores due to their smaller size. The tests have shown that new methods minimize the changes of properties of the microorganisms during their aerosolization in the laboratory.     

Repeatability estimates for milk coagulation traits and non-coagulation of milk in Finnish Ayrshire cows

Effects of systematic environmental factors and milk production and quality traits on milk coagulation properties (MCP), and on repeatability of those traits were estimated from 979 milk samples collected once a month over a period of 2 years from 83 Finnish Ayrshire cows. Estimation was based on a multitrait animal model and REML methodology. In addition, persistence of non-coagulation of milk in individual cows, and factors associated with it were established from a sub sample of 24 cows producing non-coagulating (NC) milk at least once. MCP were at their best during the first lactation, at the beginning and at the end of lactation, and during grazing seasons. Variation in MCP with systematic environmental factors was partly due to variation in composition and quality of milk, especially in pH and ln (somatic cell count, SCC). Coefficients of repeatability for milk coagulation time and curd firmness were 0.65 and 0.68. These estimates were of the same magnitude as those for protein content, but were higher than those for daily milk yield, fat content, pH, and SCC. Based on the repeatability estimates for the milk coagulation traits and effects of the environmental factors, cows should be sampled at least three times during a lactation to estimate reliably breeding values for the milk coagulation traits. A total of 10% of the milk samples did not coagulate in 30 min after addition of rennet. Cows that produced NC milk at least once (30% of the cows) could be classified into those that produced NC milk only a few times during a lactation and those that produced NC milk at almost every sampling. Based on logistic regression analyses, peak and mid-lactation, high milk yield, low protein and fat content and high pH increased the risk of non-coagulation of milk.     

Temperature modeling and measurement of an electrokinetic separation chip

This work presents experimental [infrared (IR) thermography] and computational (finite element model) results of temperature distributions of an electrokinetic separation chip. Thermal characteristics of both the electrolyte solution and the polymer chip (SU-8) are taken into account in modeling temperature distributions during electrokinetic flow. Multiphysics and multiscale simulation couples electrostatics, heat transfer, and fluid dynamics. The accompanying IR thermography is a non-contact method, which can measure fractional temperature differences with sub-second time resolution. Any structures or temperature marker molecules interfering with the experiment are not needed. Nominal spot size in the IR measurements is 30 μm with a field of view of several millimeters enabling both local and chip-scale temperature monitoring simultaneously. As a result, we present a computer model for electrokinetic chips, which enables simulation of fractional temperature changes during electrophoresis under real operating conditions. The accuracy of the model is within ±1°C when the deviation in electrochemical processes is taken into account. The simulation results also suggest that the temperature on the chip surface qualitatively reflects the temperature inside the microchannel with an average offset of 1–2°C.     

BET Inhibition Upregulates SIRT1 and Alleviates Inflammatory Responses

Control of histone acetylation is a part of the epigenetic mechanism that regulates gene expression and chromatin architecture. The members of the bromodomain and extra terminal domain (BET) protein family are a group of epigenetic readers that recognize histone acetylation, whereas histone deacetyl‐ ases such as sirtuin 1 (SIRT1) function as epigenetic erasers. We observed that BET inhibition by the specific inhibitor JQ1 upregulated SIRT1 expression and activated SIRT1. Moreover, we observed that BET inhibition functionally reversed the pro‐inflammatory effect of SIRT1 inhibition in a cellular lung disease model. SIRT1 activation is desirable in many age‐related, metabolic and inflammatory diseases; our results suggest that BET protein inhibition would be beneficial in treatment of those conditions. Most importantly, our findings demonstrate a novel mechanism of SIRT1 activation by inhibition of the BET proteins.     

An active single-chain antibody containing a cellulase linker domain is secreted by Escherichia coli

Single-chain antibodies consist of the variable, antigen-bindingdomains of antibodies joined to a continuous polypeptide bygenetically engineered peptide linkers. We have used the flexibleinterdomain linker region of a fungal cellulase to link togetherthe variable domains of an anti-2-phenyloxazolone IgGl and showhere that the resulting single-chain antibody is efficientlysecreted and released to the culture medium of Escherichia coli.The yield of affinity-purified single-chain antibody is 1 -2mg/1 of culture medium and its affinity and stability are comparableto those of the corresponding native IgG.