A computationally designed fluorescent biosensor for D-serine

Periplasmic solute-binding proteins (SBPs) have evolved to balance the demands of ligand affinity, thermostability and conformational change to carry out diverse functions in small molecule transport, sensing and chemotaxis. Although the ligand-induced conformational changes that occur in SBPs make them useful components in biosensors, their complexity can be difficult to emulate and they are challenging targets for protein engineering and design. Here we have engineered a fluorescent biosensor with specificity for the signalling molecule D-serine (D-SerFS) from a D-alanine-specific SBP. Through a combination of binding site and remote mutations, the affinity, specificity and thermostability were optimized to allow the detection of changes in D-serine levels using two-photon excitation fluorescence microscopy in situ and in vivo. This work illustrates the multidimensional constraints that are imposed by the trade-offs between structural dynamics, ligand affinity and thermostability, and how these must be balanced to achieve desirable activities in the engineering of complex, dynamic proteins.

Financial and Production Constraints faced by Women Entrepreneurs

Women, who try to enter the field of industry in either managerial or entrepreneurial role, are generally exposed to various constraints. Though the entrepreneurial process is the same for men and women, there are however, in practice, women have to confront many additional problems of varying dimensions and magnitudes, which prevent them from realizing their full potential as entrepreneurs. Women entrepreneurs experience multidimensional constraints. The governments at the centre and the states have established a number of specialized institutions to cater to the needs of women entrepreneurs but they still face many problems. At this juncture, the constraints faced by women entrepreneurs were studied as one of the objective. This objective was to ascertain the financial, production and labour constraints faced by women in their enterprise. To achieve the purpose of the study, 40 registered women entrepreneurs in Micro Small and Medium Enterprise (MSME) were identified in Coimbatore District of Tamil Nadu. Series of questions were asked through well structured interview schedule. Evidence in the paper shows that the most serious financial constraint faced by women was, that the entire loan was not given at a time (65.00%). In production and labour, most serious constraints faced by women entrepreneurs were labour management in various essential activities (92.5%). Methodology and design not clarified here. No recommendations The ex post - facto type of research was employed. Proportionate Random sample method” was adopted to select the respondents from the four Entrepreneurial Hubs. Women should attend training programs, seminars, workshop and conferences Related to develop professional competencies in managerial, leadership, marketing, financial, production process, profit planning, maintaining books of accounts and other skills.

Constraints, Incentives and Pockets of Local Order on the Streets of Pelotas, Brazil

This article investigates the life experiences of 19 young people on the streets of Pelotas, southern Brazil. Manifold methods were used revealing several highly interconnected aspects in their realities. This study, which was conducted for a period of seven years (2009–2016), illustrates how the studied street youths form their environments physically, socially and mentally in order to reach their goals and create the multiple means of survival in challenging street environment. It is important to understand heterogeneous, interrelated physical, sociocultural and mental elements to study subjectively experienced realities. Personally experienced multidimensional constraints and incentives and the individual’s own goals shape these realities.