Autonomous bioluminescence imaging of single mammalian cells with the bacterial bioluminescence system.

Bioluminescence-based imaging of living cells has become an important tool in biological and medical research. However, many bioluminescence imaging applications are limited by the requirement of an externally provided luciferin substrate and the low bioluminescence signal which restricts the sensitivity and spatiotemporal resolution. The bacterial bioluminescence system is fully genetically encodable and hence produces autonomous bioluminescence without an external luciferin, but its brightness in cell types other than bacteria has, so far, not been sufficient for imaging single cells. We coexpressed codon-optimized forms of the bacterial luxCDABE and frp genes from multiple plasmids in different mammalian cell lines. Our approach produces high luminescence levels that are comparable to firefly luciferase, thus enabling autonomous bioluminescence microscopy of mammalian cells

DNA-PAINT MINFLUX nanoscopy

MINimal fluorescence photon FLUXes (MINFLUX) nanoscopy, providing photon-efficient fluorophore localizations, has brought about three-dimensional resolution at nanometer scales. However, by using an intrinsic on–off switching process for single fluorophore separation, initial MINFLUX implementations have been limited to two color channels. Here we show that MINFLUX can be effectively combined with sequentially multiplexed DNA-based labeling (DNA-PAINT), expanding MINFLUX nanoscopy to multiple molecular targets. Our method is exemplified with three-color recordings of mitochondria in human cells

Community Survey Report: Lekhnath 2, Rithepani

Community diagnosis is defined as the pattern of disease in a community described in terms of the important factors which influence the pattern -King Maurice ed (1982). It is based on collection and interpretation of the relevant data such as age, sex, population distribution by social groups, vital statistics and incidence and prevalence of important diseases of the area.  As per the curriculum of Tribhuvan University, B.Sc Nursing 1st year, we were provided an opportunity to fulfill the practicum of community health nursing in the city of seven lakes of Rithepani-2, Lekhnath, Kaski from July 17th to Aug 11th 2016. Total population of Rithepani ward no 2 was 4529, male population was 2100 and female population was 2429 respectively. Each of the five students was assigned with ten different families for community diagnosis in order to promote the health of the individual, family and community.  The main objective of community diagnosis was to find out the health related aspects and to provide preventive, promotive curative and rehabilitative services to the individual and community as a whole. All the findings were presented among the key members of the community. On survey total population of 50 houses was found 239, among them 126 were male population and 113 were female population. Sanitation of the community was found appropriate. Moreover immunization services among under five children was adequate. Likewise, antenatal, intranatal and postnatal care was found appropriate and effective in all the houses. This signifies that people are utilizing the all health services in an appropriate manner. Journal of Gandaki Medical College Vol. 10, No. 1, 2017, Page: 63-6

Genome sequence of Burkholderia pseudomallei NCTC 13392

Here, we describe the draft genome sequence of Burkholderia pseudomallei NCTC 13392. This isolate has been distributed as K96243, but distinct genomic differences have been identified. The genomic sequence of this isolate will provide the genomic context for previously conducted functional studies

Bright ligand-activatable fluorescent protein for high-quality multicolor live-cell super-resolution microscopy

We introduce UnaG as a green-to-dark photoswitching fluorescent protein capable of high-quality super-resolution imaging with photon numbers equivalent to the brightest photoswitchable red protein. UnaG only fluoresces upon binding of a fluorogenic metabolite, bilirubin, enabling UV-free reversible photoswitching with easily controllable kinetics and low background under Epi illumination. The on- and off-switching rates are controlled by the concentration of the ligand and the excitation light intensity, respectively, where the dissolved oxygen also promotes the off-switching. The photo-oxidation reaction mechanism of bilirubin in UnaG suggests that the lack of ligand-protein covalent bond allows the oxidized ligand to detach from the protein, emptying the binding cavity for rebinding to a fresh ligand molecule. We demonstrate super-resolution single-molecule localization imaging of various subcellular structures genetically encoded with UnaG, which enables facile labeling and simultaneous multicolor imaging of live cells. UnaG has the promise of becoming a default protein for high-performance super-resolution imaging. Photoconvertible proteins occupy two color channels thereby limiting multicolour localisation microscopy applications. Here the authors present UnaG, a new green-to-dark photoswitching fluorescent protein for super-resolution imaging, whose activation is based on a noncovalent binding with bilirubin

Gliadin Nanoparticles Induce Immune Tolerance to Gliadin in Mouse Models of Celiac Disease

Peer reviewe

The Current State of Performance Appraisal Research and Practice: Concerns, Directions, and Implications

On the surface, it is not readily apparent how some performance appraisal research issues inform performance appraisal practice. Because performance appraisal is an applied topic, it is useful to periodically consider the current state of performance research and its relation to performance appraisal practice. This review examines the performance appraisal literature published in both academic and practitioner outlets between 1985 and 1990, briefly discusses the current state of performance appraisal practice, highlights the juxtaposition of research and practice, and suggests directions for further research

2012 Market Report on U.S. Wind Technologies in Distributed Applications

At the end of 2012, U.S. wind turbines in distributed applications reached a 10-year cumulative installed capacity of more than 812 MW from more than 69,000 units across all 50 states. In 2012 alone, nearly 3,800 wind turbines totaling 175 MW of distributed wind capacity were documented in 40 states and in the U.S. Virgin Islands, with 138 MW using utility-scale turbines (i.e., greater than 1 MW in size), 19 MW using mid-size turbines (i.e., 101 kW to 1 MW in size), and 18.4 MW using small turbines (i.e., up to 100 kW in size). Distributed wind is defined in terms of technology application based on a wind project’s location relative to end-use and power-distribution infrastructure, rather than on technology size or project size. Distributed wind systems are either connected on the customer side of the meter (to meet the onsite load) or directly to distribution or micro grids (to support grid operations or offset large loads nearby). Estimated capacity-weighted average costs for 2012 U.S. distributed wind installations was $2,540/kW for utility-scale wind turbines,$2,810/kW for mid-sized wind turbines, and $6,960/kW for newly manufactured (domestic and imported) small wind turbines. An emerging trend observed in 2012 was an increased use of refurbished turbines. The estimated capacity-weighted average cost of refurbished small wind turbines installed in 2012 was$4,080/kW. As a result of multiple projects using utility-scale turbines, Iowa deployed the most new overall distributed wind capacity, 37 MW, in 2012. Nevada deployed the most small wind capacity in 2012, with nearly 8 MW of small wind turbines installed in distributed applications. In the case of mid-size turbines, Ohio led all states in 2012 with 4.9 MW installed in distributed applications. State and federal policies and incentives continued to play a substantial role in the development of distributed wind projects. In 2012, U.S. Treasury Section 1603 payments and grants and loans from the U.S. Department of Agriculture’s Rural Energy for America Program were the main sources of federal funding for distributed wind projects. State and local funding varied across the country, from rebates to loans, tax credits, and other incentives. Reducing utility bills and hedging against potentially rising electricity rates remain drivers of distributed wind installations. In 2012, other drivers included taking advantage of the expiring U.S. Treasury Section 1603 program and a prosperous year for farmers. While 2012 saw a large addition of distributed wind capacity, considerable barriers and challenges remain, such as a weak domestic economy, inconsistent state incentives, and very competitive solar photovoltaic and natural gas prices. The industry remains committed to improving the distributed wind marketplace by advancing the third-party certification process and introducing alternative financing models, such as third-party power purchase agreements and lease-to-own agreements more typical in the solar photovoltaic market. Continued growth is expected in 2013