Maximising the Use of Scarce qPCR Master Mixes

The COVID-19 pandemic resulted in a universal, immediate, and vast demand for comprehensive molecular diagnostic testing, especially real-time quantitative (qPCR)-based methods. This rapidly triggered a global shortage of testing capacity, equipment, and reagents. Even today, supply times for chemicals from date of order to delivery are often much longer than pre-pandemic. Furthermore, many companies have ratcheted up the price for minimum volumes of reaction master mixes essential for qPCR assays, causing additional problems for academic laboratories often operating on a shoestring. We have validated two strategies that stretch reagent supplies and, whilst particularly applicable in case of scarcity, can readily be incorporated into standard qPCR protocols, with appropriate validation. The first strategy demonstrates equivalent performance of a selection of “past expiry date” and newly purchased master mixes. This approach is valid for both standard and fast qPCR protocols. The second validates the use of these master mixes at less than 1x final concentration without loss of qPCR efficiency or sensitivity.     

Characterisation of Gel-Forming Mucins Produced In Vivo and In Ex Vivo Conjunctival Explant Cultures

One key element to the health of the ocular surface encompasses the presence of gel-forming mucins in the pre-ocular tear film. Conjunctival goblet cells are specialized epithelial cells that secrete mucins necessary for tear film stability and general homeostasis. Their dysfunction can be linked to a range of ocular surface inflammation disorders and chronic injuries. To obtain new perspectives and angles to tackle mucin deficiency, the need for an accurate evaluation of their presence and corresponding mucin secretion in ex vivo conjunctival cultures has become a requisite. In vitro, goblet cells show a significant decrease in the production and secretion of gel-forming mucins, accompanied by signs of dedifferentiation or transdifferentiation. Explant cultures on laminin-treated CLP-PEG hydrogels can, however, support the production of gel-forming mucins. Together, we challenge the current paradigm to evaluate the presence of cultured goblet cells solely based on their general mucin (MUC) content through imaging analyses, showing the need for additional techniques to assess the functionality of goblet cells. In addition, we broadened the gel-forming mucin profile of in vivo goblet cells with MUC5B and MUC6, while MUC2 and MUC6 is added to the profile of cultured goblet cells.     

A meta-analytic study of social desirability distortion in computer-administered questionnaires, traditional questionnaires, and interviews.

A meta-analysis of social desirability distortion compared computer questionnaires with traditional paper-and-pencil questionnaires and face-to-face interviews in 61 studies (1967–1997; 673 effect sizes). Controlling for correlated observations, a near-zero overall effect size was obtained for computer versus paper-and-pencil questionnaires. With moderators, there was less distortion on computerized measures of social desirability responding than on the paper-and-pencil measures, especially when respondents were alone and could backtrack. There was more distortion on the computer on other scales, but distortion was small when respondents were alone, anonymous, and could backtrack. There was less distortion on computerized versions of interviews than on face-to-face interviews. Research is needed on nonlinear patterns of distortion, and on the effects of context and interface on privacy perceptions and on responses to sensitive questions. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Dielectric Properties of La3+ Doped Sr0.3–3y/2Lay Ba0.7Nb2O6 Ceramics Prepared under Different Sintering Conditions*

A study of the dielectric properties of the Lanthanum doped Sr_0.3Ba_0.7Nb₂O₆ (SBN30) ceramic according to the stoichiometric formulation Sr_0.3–3y/2La_yBa_0.7Nb₂O₆ with y = 0.01, 0.03 and 0.05, and the influence of the sintering conditions is reported. The XRD shows single phase compounds for Sr_0.285La_0.01Ba_0.7Nb₂O₆ (LSBN1) and Sr_0.225La_0.03Ba_0.7Nb₂O₆ (LSBN3) ceramics, both samples having similar microstructure, densification and dielectric properties. The density increases linearly with ln t, where t is the sintering time, and the values of the maximum ferroelectric peaks of the permittivity increase steadily with t. Using the Bruggeman model to estimate the theoretical permittivity, it is concluded that the magnitude of the experimental permittivity peaks are mainly affected by the volume fraction of porosity of the samples. In this study we also establish that pore diffusion mechanisms behave according to the Ginstling-Brownshtein equation. For the Sr_0.225La_0.05Ba_0.7Nb₂O₆ (LSBN5) sample, XRD analysis reveals the presence of isostructural compounds of the intermediate phases BaNb₂O₆ and SrNb₂O₆, and the dielectric properties start to deteriorate. This fact indicates the existence of a solubility limit of Lanthanum ions in the SBN solid solution.     

Unilateral Intrastriatal 6-Hydroxydopamine Lesion in Mice: A Closer Look into Non-Motor Phenotype and Glial Response

Parkinson’s disease (PD) is a prevalent movement disorder characterized by the progressive loss of dopaminergic neurons in substantia nigra pars compacta (SNpc). The 6-hydroxydopamine (6-OHDA) lesion is still one of the most widely used techniques for modeling Parkinson’s disease (PD) in rodents. Despite commonly used in rats, it can be challenging to reproduce a similar lesion in mice. Moreover, there is a lack of characterization of the extent of behavioral deficits and of the neuronal loss/neurotransmitter system in unilateral lesion mouse models. In this study, we present an extensive behavioral and histological characterization of a unilateral intrastriatal 6-OHDA mouse model. Our results indicate significant alterations in balance and fine motor coordination, voluntary locomotion, and in the asymmetry’s degree of forelimb use in 6-OHDA lesioned animals, accompanied by a decrease in self-care and motivational behavior, common features of depressive-like symptomatology. These results were accompanied by a decrease in tyrosine hydroxylase (TH)-labelling and dopamine levels within the nigrostriatal pathway. Additionally, we also identify a marked astrocytic reaction, as well as proliferative and reactive microglia in lesioned areas. These results confirm the use of unilateral intrastriatal 6-OHDA mice for the generation of a mild model of nigrostriatal degeneration and further evidences the recapitulation of key aspects of PD, thereby being suitable for future studies beholding new therapeutical interventions for this disease.     

Oil strategies benefits over different driving cycles using numerical simulation

95 g/kin is the allowed quantity of CO2 emission normalized to NEDC to be set in 2020.In addition,NEDC will be replaced by more severe driving cycles and will be united worldwide.To respond to those criteria,automotive industries are working on every possible field.Thermal management has been proved to be effective in reducing fuel consumption.Cold start is a primordial reason of overconsumption,as the engine highest efficiency is at its optimal temperature.At cold start,the engine's oil is at its lowest temperature and thus its higher viscosity level.A high viscosity oil generates more friction,which is one of the most important heat losses in the engine.In this paper,hot oil storage is studied.Numerical simulations on GT-suite model were done.The model consists of a 4-cylinder turbocharged Diesel engine using a storage volume of 1 liter of hot oil.Ambient temperature variation were taken into consideration as well as different driving cycles.Furthermore,different configurations of the thermal strategy (multifunction oil sump) were proposed and evaluated.Lubricant temperature and viscosity profiles are presented in the article as well as fuel consumption savings for different configurations,driving cycles and ambient temperatures.     

Accelerator boards, memory, power, coprocessor, and price in an automated audit trail and networked matching algorithm of needs

Matching the user's needs to the available supply of Accelerator Boards can be methodically time-consuming and difficult. If it is not done it may be short of satisfactory. This paper presents a method of matching user needs with the available variety of Accelerator Boards (AS). It describes a method of developing and using a Product Evaluation and Selection System (PESS). PESS ranks AS products according to how well they match user's needs.     

学习波哥大公交系统的经验

几十年来，波哥大因其无秩序的交通、空气污染、缓慢而混乱的公共交通运输而闻名于世。从2001年1月开始，波哥大的交通管理系统找到了一个解决日常交通堵塞的先进方法，具有当今最新的技术水平。作为城市交通机动性的综合战略规划的一部分，其中包括促进非机动化交通和限制汽车的使用，波哥大城市发展了一套“新世纪快速公交（TransMilenio）”系统。它是依靠许多当地机构和私营公司以合作经营的方式建立起来的。     

A prisoner's dilemma experiment on cooperation with people and human-like computers.

The authors investigated basic properties of social exchange and interaction with technology in an experiment on cooperation with a human-like computer partner or a real human partner. Talking with a computer partner may trigger social identity feelings or commitment norms. Participants played a prisoner's dilemma game with a confederate or a computer partner. Discussion, inducements to make promises, and partner cooperation varied across trials. On Trial 1, after discussion, most participants proposed cooperation. They kept their promises as much with a text-only computer as with a person, but less with a more human-like computer. Cooperation dropped sharply when any partner avoided discussion. The strong impact of discussion fits a social contract explanation of cooperation following discussion. Participants broke their promises to a computer more than to a person, however, indicating that people make heterogeneous commitments. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Computer communication as an aid to independence for older adults

Computer and communication technologies offer the potential of improving die quality of life for older people by providing them with links to information and services outside of the home. This study examined die feasibility of older people using an electronic text message system to perform routine communication tasks. In addition information was gathered to identify design parameters which facilitate the interactions of older people with such computer based systems. A specialized and simplified 'communication computer' was placed in the homes of 36 older women, aged 50-95 years. The system was provided with: a simple text-editor, basic electronic mail functions and access to news/ weather, movie reviews, and health information. Both performance data and user preference data were collected. Results indicated that the participants liked using the system, were able to use it with minimal difficulty, and that it provided a valuable means for social interaction and mental stimulation. The findings suggest that computer-based systems can be a valuable support tool for older adults if they are easy to use, and provide applications that are useful for them.