A Fast and Accessible Methodology for Micro-Patterning Cells on Standard Culture Substrates Using Parafilm™ Inserts

Micropatterning techniques provide direct control over the spatial organization of cells at the sub-mm scale. Regulation of these spatial parameters is important for controlling cell fate and cell function. While micropatterning has proved a powerful technique for understanding the impact of cell organization on cell behaviour, current methods for micropatterning cells require complex, specialized equipment that is not readily accessible in most biological and bioengineering laboratories. In addition, currently available methods require significant protocol optimization to ensure reliable and reproducible patterning. The inaccessibility of current methods has severely limited the widespread use of micropatterning as a tool in both biology and tissue engineering laboratories. Here we present a simple, cheap, and fast method to micropattern mammalian cells into stripes and circular patterns using Parafilm™, a common material found in most biology and bioengineering laboratories. Our method does not require any specialized equipment and does not require significant method optimization to ensure reproducible patterning. Although our method is limited to simple patterns, these geometries are sufficient for addressing a wide range of biological problems. Specifically, we demonstrate i) that using our Parafilm™ insert method we can pattern and co-pattern ARPE-19 and MDCK epithelial cells into circular and stripe micropatterns in tissue culture polystyrene (TCPS) wells and on glass slides, ii) that we can contain cells in the desired patterns for more than one month and iii) that upon removal of the Parafilm™ insert we can release the cells from the containment pattern and allow cell migration outward from the original pattern. We also demonstrate that we can exploit this confinement release feature to conduct an epithelial cell wound healing assay. This novel micropatterning method provides a reliable and accessible tool with the flexibility to address a wide range of biological and engineering problems that require control over the spatial and temporal organization of cells

Copyright and cultural work: an exploration

This article first discusses the contemporary debate on cultural “creativity” and the economy. Second, it considers the current state of UK copyright law and how it relates to cultural work. Third, based on empirical research on British dancers and musicians, an analysis of precarious cultural work is presented. A major focus is how those who follow their art by way of “portfolio” work handle their rights in ways that diverge significantly from the current simplistic assumptions of law and cultural policy. Our conclusions underline the distance between present top-down conceptions of what drives production in the cultural field and the actual practice of dancers and musicians

Comparison of p53 and DNA content abnormalities in adenocarcinoma of the oesophagus and gastric cardia.

This study examined the association between 17p allelic loss, p53 gene mutation, p53 protein expression and DNA aneuploidy in a series of adenocarcinomas arising in the oesophagus and gastric cardia. 17p allelic loss was detected in 79% (15 of 19) of oesophageal and in 83% (29 of 35) of gastric adenocarcinomas. p53 mutations were detected in 70% (14 of 20) and 63% (26 of 41) of oesophageal and of gastric adenocarcinomas respectively. Both tumour types were associated with a predominance of base transitions at CpG dinucleotides. In five cases of oesophageal adenocarcinoma, the same mutation was detected both in tumour and in adjacent dysplastic Barrett's epithelium. Diffuse p53 protein expression was detected in 65% (13 of 20) and 59% (24 of 41) of oesophageal and of gastric tumours, respectively, and was associated with the presence of p53 missense mutation (Chi-squared, P < 0.0001). DNA aneuploidy was detected in 80% (16 of 20) of oesophageal and in 70% (28 of 40) of gastric tumours. No association was found between p53 or DNA content abnormalities and tumour stage or histological subtype. In conclusion, this study detected a similar pattern of p53 alterations in adenocarcinoma of the oesophagus and gastric cardia--molecular data consistent with the observation that these tumours demonstrate similar clinical and epidemiological features

Alteration of immune function in women collegiate soccer players and college students

The purpose of this study was to monitor the stress-induced alteration in concentrations of salivary immunoglobulin (S-IgA) and cortisol and the incidence of upper respiratory tract infections (URTI) over the course of a 9-week competitive season in college student-athletes and college students. The subjects consisted of 14 NCAA Division III collegiate female soccer athletes (19.8 ± 1.0 years, mean ± SD) and 14 female college students (22.5 ± 2.6 years). Salivary samples were collected for 9 weeks during a competitive soccer season. S-IgA and cortisol concentrations were determined by enzyme linked immunosorbent assay (ELISA). A training and performance questionnaire was given to the subjects every week, to record the subjects’ session rating of perceived exertion (RPE) for all the training, load, monotony and strain, as well as any injuries or illnesses experienced. The between groups ANOVA procedure for repeated measures showed no changes in salivary concentrations of IgA and cortisol. Chisquare analysis showed that during the 9-week training season injury and illness occurred at a higher rate among the soccer players. There was a significant difference at baseline between soccer and control SIgA levels (p ≤ 0.05). Decreased levels of SIgA and increases in the indices of training (load, strain and monotony) were associated with an increase in the incidence of illness during the 9-week competitive soccer season

Open defecation and childhood stunting in India: an ecological analysis of new data from 112 districts.

Poor sanitation remains a major public health concern linked to several important health outcomes; emerging evidence indicates a link to childhood stunting. In India over half of the population defecates in the open; the prevalence of stunting remains very high. Recently published data on levels of stunting in 112 districts of India provide an opportunity to explore the relationship between levels of open defecation and stunting within this population. We conducted an ecological regression analysis to assess the association between the prevalence of open defecation and stunting after adjustment for potential confounding factors. Data from the 2011 HUNGaMA survey was used for the outcome of interest, stunting; data from the 2011 Indian Census for the same districts was used for the exposure of interest, open defecation. After adjustment for various potential confounding factors--including socio-economic status, maternal education and calorie availability--a 10 percent increase in open defecation was associated with a 0.7 percentage point increase in both stunting and severe stunting. Differences in open defecation can statistically account for 35 to 55 percent of the average difference in stunting between districts identified as low-performing and high-performing in the HUNGaMA data. In addition, using a Monte Carlo simulation, we explored the effect on statistical power of the common practice of dichotomizing continuous height data into binary stunting indicators. Our simulation showed that dichotomization of height sacrifices statistical power, suggesting that our estimate of the association between open defecation and stunting may be a lower bound. Whilst our analysis is ecological and therefore vulnerable to residual confounding, these findings use the most recently collected large-scale data from India to add to a growing body of suggestive evidence for an effect of poor sanitation on human growth. New intervention studies, currently underway, may shed more light on this important issue

Stringy black holes and energy conditions

The energy condition inequalities for the matter stress energy comprised out of the dilaton and Maxwell fields in the dilaton-Maxwell gravity theories emerging out of string theory are examined in detail. In the simplistic 1+1 dimensional models, $R\le 0$ (where $R$ is the Ricci scalar), turns out to be the requirement for ensuring focusing of timelike geodesics. In 3+1 dimensions, we outline the requirements on matter for pure dilaton theories-these in turn constrain the functional forms of the dilaton. Furthermore, in charged dilaton gravity a curious opposite behaviour of the matter stress energy w.r.t the violation/conservation of the Weak Energy Condition is noted for the electric and magnetic black hole metrics written in the string frame of reference. We also investigate the matter that is necessary for creating certain specific non-asymptotically flat black holes. For the electric and magnetic black hole metrics, strangely, matter satisfies the weak energy condition in the string frame. Finally, the Averaged Null Energy Condition is evaluated along radial null geodesics for each of these black hole spacetimes.Comment: 23 pages, RevTex, no figures, corrected and extended,to appear in Physical Review

Flat World of Dilatonic Domain Walls

We study dilatonic domain walls specific to superstring theory. Along with the matter fields and metric the dilaton also changes its value in the wall background. We found supersymmetric (extreme) solutions which in general interpolate between isolated superstring vacua with non-equal value of the matter potential; they correspond to the static, planar domain walls with {\it flat} metric in the string (sigma model) frame. We point out similarities between the space-time of dilatonic walls and that of charged dilatonic black holes. We also comment on non-extreme solutions corresponding to expanding bubbles.Comment: 11 pgs (+2 figures available upon request), UPR-560-

Selective influences of maximum dynamic strength and bar-power output on team sports performance: a comprehensive study of four different disciplines

This study examined the selective influences of one-repetition maximum (1RM) values [assessed in the half-squat (HS)] and bar-power production [assessed in both HS and jump squat (JS) exercises] on the physical performance of male and female team sport athletes from four different sports. Three-hundred and three elite players (31 Olympians) from four different disciplines (47 male soccer players, 58 female soccer players, 28 male handball players, 58 female handball players, 49 male rugby players, and 63 male futsal players) participated in this study. The physical tests were performed over 2 consecutive days for soccer and rugby players, and in 1 day for the remaining athletes. On the first day, rugby and soccer athletes performed squat jumps (SJ), countermovement jumps (CMJ), and HS 1RM. On the second day, they executed HS and JS tests (to assess the maximum bar-power output) and the linear and change-of-direction (COD) speed tests. For the other players, the sequence of the measurements was the same; however, they did not perform the HS exercise. Athletes were separated, using a median split analysis, into two distinct groups, according to their bar-power output in both JS and HS exercises and their performance in HS 1RM. The magnitude-based inferences method was used to examine the differences between “higher” and “lower” performance groups. Overall, the bar-power outputs were better connected to improved acceleration, speed, and jump performance than the 1RM measures. From these findings, it is possible to infer that players able to produce higher bar-power outputs are likely to sprint faster and jump higher. Therefore, coaches involved in team sports are strongly encouraged to use the bar-power method to evaluate the athletic performance of their players

Beyond the Singularity of the 2-D Charged Black Hole

Two dimensional charged black holes in string theory can be obtained as exact (SL(2,R)xU(1))/U(1) quotient CFTs. The geometry of the quotient is induced from that of the group, and in particular includes regions beyond the black hole singularities. Moreover, wavefunctions in such black holes are obtained from gauge invariant vertex operators in the SL(2,R) CFT, hence their behavior beyond the singularity is determined. When the black hole is charged we find that the wavefunctions are smooth at the singularities. Unlike the uncharged case, scattering waves prepared beyond the singularity are not fully reflected; part of the wave is transmitted through the singularity. Hence, the physics outside the horizon of a charged black hole is sensitive to conditions set behind the past singularity.Comment: 19 pages, 5 figures; v2: refs added, minor typos corrected; v3: references on the infinite blue shift at the inner horizon and minor corrections adde