Mapping the elastic response of epithelial apical cell membranes suspended across porous array

As the elastic response of cell membranes to mechanical stimuli plays a key role in various cellular processes, novel biophysical strategies to quantify the elasticity of native membranes under physiological conditions at a nanometer scale are gaining interest. In order to investigate the elastic response of apical membranes, elasticity maps of native membrane sheets, isolated from MDCK II (Madine Darby Canine kidney strain II) epithelial cells, were recorded by local indentation with an Atomic Force Microscope (AFM). To exclude the underlying substrate effect on membrane indentation, a highly ordered gold coated porous array with a pore diameter of 1.2 μm was used to support apical membranes. Overlays of fluorescence and AFM images show that intact apical membrane sheets are attached to poly-D-lysine coated porous substrate. Force indentation measurements reveal an extremely soft elastic membrane response if it is indented at the center of the pore in comparison to a hard repulsion on the adjacent rim used to define the exact contact point. A linear dependency of force versus indentation (-dF/dh) up to 100 nm penetration depth enabled us to define an apparent membrane spring constant (kapp) as the slope of a linear fit with a stiffness value of for native apical membrane in PBS. A correlation between fluorescence intensity and kapp is also reported. Time dependent hysteresis observed with native membranes is explained by a viscoelastic solid model of a spring connected to a Kelvin-Voight solid with a time constant of 0.04 s. No hysteresis was reported with chemically fixated membranes. A combined linear and non linear elastic response is suggested to relate the experimental data of force indentation curves to the elastic modulus and the membrane thickness. Membrane bending is the dominant contributor to linear elastic indentation at low loads, whereas stretching is the dominant contributor for non linear elastic response at higher loads. The membrane elastic response was controlled either by stiffening with chemical fixatives or by softening with F-actin disrupters. Overall, the presented setup is ideally suitable to study the interactions of the apical membrane with the underlying cytoskeleton by means of force indentation elasticity maps combined with fluorescence imaging.

Data breach - services for threat detection, analysis and response

Nel mondo della sicurezza informatica, le tecnologie si evolvono per far fronte alle minacce. Non è possibile prescindere dalla prevenzione, ma occorre accettare il fatto che nessuna barriera risulterà impenetrabile e che la rilevazione, unitamente ad una pronta risposta, rappresenta una linea estremamente critica di difesa, ma l’unica veramente attuabile per poter guadagnare più tempo possibile o per limitare i danni. Introdurremo quindi un nuovo modello operativo composto da procedure capaci di affrontare le nuove sfide che il malware costantemente offre e allo stesso tempo di sollevare i comparti IT da attività onerose e sempre più complesse, ottimizzandone il processo di comunicazione e di risposta.

Response Bias Using Two-Stage Data Collection: A study of elderly participants in a program.

Researchers examining the effects of programs, in this case a state-level pharmaceutical assistance program for the elderly, sometimes must rely on multiple methods of data collection. Two-stage data collection (e.g., a telephone interview followed by a mail questionnaire) was used to obtain a full range of information. Older age groups were found to participate less frequently in the telephone interview, while certain demographic factors characterized mail questionnaire nonparticipants, all of which supports past research. Results also show that those in the poorest health are less likely to participate in the mail survey. Combining the two methods did not result in high attrition, suggesting that innovation can be successfully employed. Knowledge of the bias associated with each method will aid in targeting special groups.

Validity of using item response theory to analyze data from the Work Limitations Questionnaire

The Work Limitations Questionnaire (WLQ) is used to determine the amount of work loss and productivity which stem from certain health conditions, including rheumatoid arthritis and cancer. The questionnaire is currently scored using methodology from Classical Test Theory. Item Response Theory, on the other hand, is a theory based on analyzing item responses. This study wanted to determine the validity of using Item Response Theory (IRT), to analyze data from the WLQ. Item responses from 572 employed adults with dysthymia, major depressive disorder (MDD), double depressive disorder (both dysthymia and MDD), rheumatoid arthritis and healthy individuals were used to determine the validity of IRT (Adler et al., 2006).^ PARSCALE, which is IRT software from Scientific Software International, Inc., was used to calculate estimates of the work limitations based on item responses from the WLQ. These estimates, also known as ability estimates, were then correlated with the raw score estimates calculated from the sum of all the items responses. Concurrent validity, which claims a measurement is valid if the correlation between the new measurement and the valid measurement is greater or equal to .90, was used to determine the validity of IRT methodology for the WLQ. Ability estimates from IRT were found to be somewhat highly correlated with the raw scores from the WLQ (above .80). However, the only subscale which had a high enough correlation for IRT to be considered valid was the time management subscale (r = .90). All other subscales, mental/interpersonal, physical, and output, did not produce valid IRT ability estimates.^ An explanation for these lower than expected correlations can be explained by the outliers found in the sample. Also, acquiescent responding (AR) bias, which is caused by the tendency for people to respond the same way to every question on a questionnaire, and the multidimensionality of the questionnaire (the WLQ is composed of four dimensions and thus four different latent variables) probably had a major impact on the IRT estimates. Furthermore, it is possible that the mental/interpersonal dimension violated the monotonocity assumption of IRT causing PARSCALE to fail to run for these estimates. The monotonicity assumption needs to be checked for the mental/interpersonal dimension. Furthermore, the use of multidimensional IRT methods would most likely remove the AR bias and increase the validity of using IRT to analyze data from the WLQ.^

Bayesian estimation of multilevel item response model with missing data

This study proposed a novel statistical method that modeled the multiple outcomes and missing data process jointly using item response theory. This method follows the "intent-to-treat" principle in clinical trials and accounts for the correlation between outcomes and missing data process. This method may provide a good solution to chronic mental disorder study. ^ The simulation study demonstrated that if the true model is the proposed model with moderate or strong correlation, ignoring the within correlation may lead to overestimate of the treatment effect and result in more type I error than specified level. Even if the within correlation is small, the performance of proposed model is as good as naïve response model. Thus, the proposed model is robust for different correlation settings if the data is generated by the proposed model.^

Statistical modeling of large microarray data sets to identify stimulus-response profiles

A statistical modeling approach is proposed for use in searching large microarray data sets for genes that have a transcriptional response to a stimulus. The approach is unrestricted with respect to the timing, magnitude or duration of the response, or the overall abundance of the transcript. The statistical model makes an accommodation for systematic heterogeneity in expression levels. Corresponding data analyses provide gene-specific information, and the approach provides a means for evaluating the statistical significance of such information. To illustrate this strategy we have derived a model to depict the profile expected for a periodically transcribed gene and used it to look for budding yeast transcripts that adhere to this profile. Using objective criteria, this method identifies 81% of the known periodic transcripts and 1,088 genes, which show significant periodicity in at least one of the three data sets analyzed. However, only one-quarter of these genes show significant oscillations in at least two data sets and can be classified as periodic with high confidence. The method provides estimates of the mean activation and deactivation times, induced and basal expression levels, and statistical measures of the precision of these estimates for each periodic transcript.

Response of Catch-Per-Unit-Effort to Spatial and Environmental Factors: an Exploratory Data Analysis Using Generalized Additive Models

Understanding spatial distributions and how environmental conditions influence catch-per-unit-effort (CPUE) is important for increased fishing efficiency and sustainable fisheries management. This study investigated the relationship between CPUE, spatial factors, temperature, and depth using generalized additive models. Combinations of factors, and not one single factor, were frequently included in the best model. Parameters which best described CPUE varied by geographic region. The amount of variance, or deviance, explained by the best models ranged from a low of 29% (halibut, Charlotte region) to a high of 94% (sablefish, Charlotte region). Depth, latitude, and longitude influenced most species in several regions. On the broad geographic scale, depth was associated with CPUE for every species, except dogfish. Latitude and longitude influenced most species, except halibut (Areas 4 A/D), sablefish, and cod. Temperature was important for describing distributions of halibut in Alaska, arrowtooth flounder in British Columbia, dogfish, Alaska skate, and Aleutian skate. The species-habitat relationships revealed in this study can be used to create improved fishing and management strategies.

Analysis of head response to torso acceleration. Volume II - description of data retrieval, analysis and display software. Final report.

Transportation Systems Center, Cambridge, Mass.

CERCLIS (Comprehensive environmental response, compensation, and liability information system) site location extract data element dictionary.

PB90-215120

Foreign direct investment in the United States : an update : review and analysis of current developments : a report submitted to the Committee on Energy and Commerce, the Committee on Ways and Means, and the Committee on Foreign Affairs of the House of Representatives, to the Committee on Finance, the Committee on Commerce, Science and Transportation, and the Committee on Foreign Relations of the Senate, and to the Joint Economic Committee of the Congress in response to section 3(a) of the Foreign Direct Investment and International Financial Data Improvements Act of 1990.

"January 1995."

Effects of filter response on analysis of aircraft noise data /

Mode of access: Internet.

Informational hand book : prepared in response to numerous inquiries for statistical and other data relating directly or indirectly to the the activities of federal agencies.

Mode of access: Internet.

NYS DOH and ATSDR response to comments on: Health consultation, evaluation of environmental data collected in 2000 : Abby Street/Hickory Woods Subdivision, city of Buffalo, Erie County, New York, April 30, 2001 /

"April 8, 2004."

TCR{alpha} Genes Direct MHC Restriction in the Potent Human T Cell Response to a Class I-Bound Viral Epitope

The underlying generic properties of {alpha}β TCRs that control MHC restriction remain largely unresolved. To investigate MHC restriction, we have examined the CTL response to a viral epitope that binds promiscuously to two human leukocyte Ags (HLAs) that differ by a single amino acid at position 156. Individuals expressing either HLA-B*3501 (156Leucine) or HLA-B*3508 (156Arginine) showed a potent CTL response to the 407HPVGEADYFEY417 epitope from EBV. Interestingly, the response was characterized by highly restricted TCR β-chain usage in both HLA-B*3501+ and HLA-B*3508+ individuals; however, this conserved TRBV9+ β-chain was associated with distinct TCR {alpha}-chains depending upon the HLA-B*35 allele expressed by the virus-exposed host. Functional assays confirmed that TCR {alpha}-chain usage determined the HLA restriction of the CTLs. Structural studies revealed significant differences in the mobility of the peptide when bound to HLA-B*3501 or HLA-B*3508. In HLA-B*3501, the bulged section of the peptide was disordered, whereas in HLA-B*3508 the bulged epitope adopted an ordered conformation. Collectively, these data demonstrate not only that mobile MHC-bound peptides can be highly immunogenic but can also stimulate an extremely biased TCR repertoire. In addition, TCR {alpha}-chain usage is shown to play a critical role in controlling MHC restriction between closely related allomorphs.