A New In Vitro Model to Study Cellular Responses after Thermomechanical Damage in Monolayer Cultures

Although electrosurgical instruments are widely used in surgery to cut tissue layers or to achieve hemostasis by coagulation (electrocautery), only little information is available concerning the inflammatory or immune response towards the debris generated. Given the elevated local temperatures required for successful electrocautery, the remaining debris is likely to contain a plethora of compounds entirely novel to the intracorporal setting. A very common in vitro method to study cell migration after mechanical damage is the scratch assay, however, there is no established model for thermomechanical damage to characterise cellular reactions. In this study, we established a new in vitro model to investigate exposure to high temperature in a carefully controlled cell culture system. Heatable thermostat-controlled aluminium stamps were developed to induce local damage in primary human umbilical vein endothelial cells (HUVEC). The thermomechanical damage invoked is reproducibly locally confined, therefore allowing studies, under the same experimental conditions, of cells affected to various degrees as well as of unaffected cells. We show that the unaffected cells surrounding the thermomechanical damage zone are able to migrate into the damaged area, resulting in a complete closure of the ‘wound’ within 48 h. Initial studies have shown that there are significant morphological and biological differences in endothelial cells after thermomechanical damage compared to the mechanical damage inflicted by using the unheated stamp as a control. Accordingly, after thermomechanical damage, cell death as well as cell protection programs were activated. Mononuclear cells adhered in the area adjacent to thermomechanical damage, but not to the zone of mechanical damage. Therefore, our model can help to understand the differences in wound healing during the early phase of regeneration after thermomechanical vs. mechanical damage. Furthermore, this model lends itself to study the response of other cells, thus broadening the range of thermal injuries that can be analysed.     

A Simple and Accurate Model to Predict Responses to Multi-electrode Stimulation in the Retina

Implantable electrode arrays are widely used in therapeutic stimulation of the nervous system (e.g. cochlear, retinal, and cortical implants). Currently, most neural prostheses use serial stimulation (i.e. one electrode at a time) despite this severely limiting the repertoire of stimuli that can be applied. Methods to reliably predict the outcome of multi-electrode stimulation have not been available. Here, we demonstrate that a linear-nonlinear model accurately predicts neural responses to arbitrary patterns of stimulation using in vitro recordings from single retinal ganglion cells (RGCs) stimulated with a subretinal multi-electrode array. In the model, the stimulus is projected onto a low-dimensional subspace and then undergoes a nonlinear transformation to produce an estimate of spiking probability. The low-dimensional subspace is estimated using principal components analysis, which gives the neuron’s electrical receptive field (ERF), i.e. the electrodes to which the neuron is most sensitive. Our model suggests that stimulation proportional to the ERF yields a higher efficacy given a fixed amount of power when compared to equal amplitude stimulation on up to three electrodes. We find that the model captures the responses of all the cells recorded in the study, suggesting that it will generalize to most cell types in the retina. The model is computationally efficient to evaluate and, therefore, appropriate for future real-time applications including stimulation strategies that make use of recorded neural activity to improve the stimulation strategy.     

A Kinetic Model for Adaptation and the Light Responses of Phycomyces

A kinetic model is described consisting of two sequential first order processes connected by two parallel reaction pathways, one of which is light-catalyzed. A change in light flux changes the rate constant of the light-dependent process, whereupon the levels of two chemical intermediaries readjust. The model's output duplicates all the main features of the cell's light-growth and dark-growth responses except their latent periods. An asymmetric modification of the model reproduces the two types of phototropic inversion discovered by Reichardt and Varjú and by Dennison. Simple exponential equations describe these responses of the model, as well as the theoretical course of its light and dark adaptation. It is concluded that adaptation in Phycomyces consist in the photocatalytic adjustment of the level of a metabolic reservoir.     

Monitoring HIV Testing in the United States: Consequences of Methodology Changes to National Surveys

Objective

In 2011, the National Health Interview Survey (NHIS), an in-person household interview, revised the human immunodeficiency virus (HIV) section of the survey and the Behavioral Risk Factor Surveillance System (BRFSS), a telephone-based survey, added cellphone numbers to its sampling frame. We sought to determine how these changes might affect assessment of HIV testing trends.

Methods

We used linear regression with pairwise contrasts with 2003-2013 data from NHIS and BRFSS to compare percentages of persons aged 18-64 years who reported HIV testing in landline versus cellphone-only households before and after 2011, when NHIS revised its in-person questionnaire and BRFSS added cellphone numbers to its telephone-based sample.

Results

In NHIS, the percentage of persons in cellphone-only households increased 13-fold from 2003 to 2013. The percentage ever tested for HIV was 6%–10% higher among persons in cellphone-only than landline households. The percentage ever tested for HIV increased significantly from 40.2% in 2003 to 45.0% in 2010, but was significantly lower in 2011 (40.6%) and 2012 (39.7%). In BRFSS, the percentage ever tested decreased significantly from 45.9% in 2003 to 40.2% in 2010, but increased to 42.9% in 2011 and 43.5% in 2013.

Conclusions

HIV testing estimates were lower after NHIS questionnaire changes but higher after BRFSS methodology changes. Data before and after 2011 are not comparable, complicating assessment of trends.     

A Model of the Leaf Extension Rate of Tall Fescue in Response to Nitrogen and Temperature

The leaf extension rate (LER) of tall fescue (Festuca arundinaceaSchreb.) was studied in the field under various nitrogen andtemperature regimes. The LER was closely related to temperaturewhen N was not limiting plant growth. Two distinct relationshipsbetween the LER and the temperature were obtained, one for vegetativegrowth and one for the reproductive period. These relationships,described by a Gompertz function, were exponential at temperaturesbelow 8 °C and linear at temperatures above 8 °C. Theymade possible the calculation of an optimal LER correspondingto non-limiting N conditions for plant growth. The strong influence of the temperature on the LER was stillobserved under N limiting conditions. The N status of the swardswas described by the ratio between the actual N content (N_actual)and the optimal N content (N_optimal). The N_optimal was definedas the N content experienced at a non-limiting level of N nutritionbut without N luxury consumption. The N_optimal, expressed asa function of dry matter yield, declined during growth. Theeffect of the N status of the swards on the LER was analysedby calculating the ratio between the actual LER and the optimalLER, and relating it to the ratio between N_actual and N_optimal.It was shown that these two ratios were highly correlated. Leaf extension, Festuca arundinacea, nitrogen, temperature     

A Model for a Correlated Random Walk Based on the Ordered Extension of Pseudopodia

Cell migration in the absence of external cues is well described by a correlated random walk. Most single cells move by extending protrusions called pseudopodia. To deduce how cells walk, we have analyzed the formation of pseudopodia by Dictyostelium cells. We have observed that the formation of pseudopodia is highly ordered with two types of pseudopodia: First, de novo formation of pseudopodia at random positions on the cell body, and therefore in random directions. Second, pseudopod splitting near the tip of the current pseudopod in alternating right/left directions, leading to a persistent zig-zag trajectory. Here we analyzed the probability frequency distributions of the angles between pseudopodia and used this information to design a stochastic model for cell movement. Monte Carlo simulations show that the critical elements are the ratio of persistent splitting pseudopodia relative to random de novo pseudopodia, the Left/Right alternation, the angle between pseudopodia and the variance of this angle. Experiments confirm predictions of the model, showing reduced persistence in mutants that are defective in pseudopod splitting and in mutants with an irregular cell surface.     

A New Model for the Origin of Bipedality

Scholars have long thought that bipedality evolved gradually in response to the opening of the savanna. Recently, both parts of this concept have come into question. A variety of benefits of bipedality have been posited as responsible, but a trait can not evolve unless a useful mutation appears. Perhaps we need to stop wondering about selective pressures and consider what kind of mutation might be involved in forming a bipedal pelvis. Work on the evolution of development has shown that there are segmental control genes, alterations in which have large effects. These include the hox genes, of which there are four sets in humans, referred to as the HOX A, B, C, and D sequences. Changes in their activation in embryogenesis alter the identity of vertebrae and limb structure. An alteration in the control region of certain of the distal HOX D genes may well be responsible for the sudden appearance of bipedality by moving the boundary between the lumbar and sacral vertebrae, and so moving the position of the pelvis and lower limb origin. Pongids usually have three lumbar vertebrae; early hominids, 6. Pongids also have 48 chromosomes while we have 46. HOX D is located on our 2nd chromosome, the one that is a fusion of two pongid chromosomes. If that fusion altered the onset of perhaps HOX D 10, so that it switched on a couple of segments later, then the sacrum would form further down the vertebral column and might be shorter. In this paper I look at the chromosomal location of HOX D and examine the likelihood that the fusion of two panid chromosomes could have given rise to alterations in its control resulting in the abrupt appearance of bipedality and accompanying changes in the limbs and in the chela in which the HOX sequences are reused.     

Extension of the Functional Range of Respiratory and Gas Exchange Responses in Repetitive Hypoxia

External respiration and gas exchange were studied in healthy volunteers during a session of intermittent normobaric hypoxia (INH) consisting of three cycles of breathing alternately a hypoxic mixture (10.7% O₂) for 5 min and normal air for 5 min. The ventilatory response increased in the successive cycles of hypoxia and gradually decreased during the normoxic intervals. These changes were accompanied by an increase in carbon dioxide in lung air, which was not eliminated by the increased pulmonary ventilation during the hypoxic intervals. However, the mean oxygen consumption did not change during the INH session because the ventilatory reactivity and breathing depth, as well as the efficiency of oxygen utilization, increased from cycle to cycle.__________Translated from Fiziologiya Cheloveka, Vol. 31, No. 3, 2005, pp. 100–107.Original Russian Text Copyright © 2005 by Krivoshchekov, G. Divert, V. Divert.     

Functional and Structural Responses of a Degradative Microbial Community to Substrates with Varying Degrees of Complexity in Chemical Structure

Abstract The objective of the present study was to determine whether cultivation of a degradative community on substrates with varying degrees of chlorination and complexity in chemical structure, as well as cultivation in batch and flow cell culture, would alter the community's functional capability. The community was isolated from oil-contaminated soil and maintained in the laboratory on 2,4,6-trichlorobenzoic acid for 5 months before its ability to grow on 15 different chemicals as sole carbon source was evaluated in batch and flow cell systems. While the community could grow and develop biofilms in flow cells on all the substrates, only 11 of the 15 substrates could support growth in batch culture. Although biofilm development was less extensive on chemicals such as pentachlorophenol (2.09% average area covered by biofilm; average biofilm depth = 3 μm) than on 2,4,6-trichlorobenzoic acid (50.84% area covered; biofilm depth = 6.4 μm), no correlation was observed between the degree of chlorination, or number of rings, and the number of planktonic cells or biofilm biomass. In contrast, physicochemical characteristics such as the octanol/water partition coefficient had a significant effect on the development of biofilm biomass. In the case of planktonic communities, the degree of chlorination and ring number also had no effect on the BIOLOG carbon utilization profiles of the resulting communities. Although the sessile communities generally clustered separately from their planktonic counterparts, principal component analysis of carbon utilization profiles of the sessile communities showed different grouping between growth on chlorinated and nonchlorinated substrates. Analysis of the degradative community maintained on 2,4,6-trichlorobenzoic acid over an extended period further showed that adaptation to a new chemical environment is a rather slow process, since the substrate utilization profiles did not stabilize even after 12 months. These results demonstrate the flexibility in metabolic ability and community structure found in microbial communities. Received: 30 November 1998; Accepted: 19 May 1999     

Development of a Model to Compute the Extension of Life Supporting Zones for Earth-Like Exoplanets

A radiative convective model to calculate the width and the location of the life supporting zone (LSZ) for different, alternative solvents (i.e. other than water) is presented. This model can be applied to the atmospheres of the terrestrial planets in the solar system as well as (hypothetical, Earth-like) terrestrial exoplanets. Cloud droplet formation and growth are investigated using a cloud parcel model. Clouds can be incorporated into the radiative transfer calculations. Test runs for Earth, Mars and Titan show a good agreement of model results with observations.     

A New Testing Approach for Comparing the Overall Homogeneity of Survival Curves

The assessment of overall homogeneity of time‐to‐event curves is a key element in survival analysis. The currently commonly used methods, e.g., log‐rank and Wilcoxon tests, may have a significant loss of statistical testing power under certain circumstances. In this paper a new statistical testing approach is developed to compare the overall homogeneity of survival curves. The proposed new method has greater power than the commonly used tests to detect overall differences between crossing survival curves. The small‐sample performance of the new test is investigated under a variety of situations by means of Monte Carlo simulations. Furthermore, the applicability of the proposed testing approach is illustrated by a real data example from a kidney dialysis trial. (© 2004 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Ordinal-To-Interval Scale Conversion Tables and National Items for the New Zealand Version of the WHOQOL-BREF

The World Health Organisation Quality of Life (WHOQOL) questionnaires are widely used around the world and can claim strong cross-cultural validity due to their development in collaboration with international field centres. To enhance conceptual equivalence of quality of life across cultures, optional national items are often developed for use alongside the core instrument. The present study outlines the development of national items for the New Zealand WHOQOL-BREF. Focus groups with members of the community as well as health experts discussed what constitutes quality of life in their opinion. Based on themes extracted of aspects not contained in the existing WHOQOL instrument, 46 candidate items were generated and subsequently rated for their importance by a random sample of 585 individuals from the general population. Applying importance criteria reduced these items to 24, which were then sent to another large random sample (n = 808) to be rated alongside the existing WHOQOL-BREF. A final set of five items met the criteria for national items. Confirmatory factor analysis identified four national items as belonging to the psychological domain of quality of life, and one item to the social domain. Rasch analysis validated these results and generated ordinal-to-interval conversion algorithms to allow use of parametric statistics for domain scores with and without national items.     

An In Vitro Model for Measuring Immune Responses to Malaria in the Context of HIV Co-infection

Malaria and HIV co-infection is a growing health priority. However, most research on malaria or HIV currently focuses on each infection individually. Although understanding the disease dynamics for each of these pathogens independently is vital, it is also important that the interactions between these pathogens are investigated and understood. We have developed a versatile in vitro model of HIV-malaria co-infection to study host immune responses to malaria in the context of HIV infection. Our model allows the study of secreted factors in cellular supernatants, cell surface and intracellular protein markers, as well as RNA expression levels. The experimental design and methods used limit variability and promote data reliability and reproducibility. All pathogens used in this model are natural human pathogens (Plasmodium falciparum and HIV-1), and all infected cells are naturally infected and used fresh. We use human erythrocytes parasitized with P. falciparum and maintained in continuous in vitro culture. We obtain freshly isolated peripheral blood mononuclear cells from chronically HIV-infected volunteers. Every condition used has an appropriate control (P. falciparum parasitized vs. normal erythrocytes), and every HIV-infected donor has an HIV uninfected control, from which cells are harvested on the same day. This model provides a realistic environment to study the interactions between malaria parasites and human immune cells in the context of HIV infection.     

Numerical and Demographic Responses of Pumas to Changes in Prey Abundance: Testing Current Predictions

ABSTRACT Information on factors affecting population size of pumas (Puma concolor) can be important because their principal prey over most of the western United States are valued big game species (e.g., mule deer [Odocoileus hemionus], elk [Cervus elaphus], and bighorn sheep [Ovis canadensis]). Based on the hypothesis that puma numbers are limited by their food supply, puma populations should track changes in prey abundance by growing exponentially with increases in prey and by declining with a lag response when prey decreases. Additional predictions proposed by researchers are that body mass of pumas, female productivity, kitten survival, and adult survival should decrease after a prey decline. We used a 15-year database from a hunted population of pumas in southern Idaho and northwestern Utah to test these predictions. During the 15-year time span of the database, a major decline in mule deer abundance occurred. Estimates of puma numbers and demographic characteristics came from intensive capture and radiocollaring efforts. We calculated kitten and adult survival with MICROMORT software. We found that adult puma numbers increased exponentially at r = 0.07 during a period of increasing mule deer numbers. Four years after the mule deer abundance declined, puma numbers decreased at a rate of r = −0.06. Body mass of female pumas was lower after the decline in puma numbers (42.6 ± SE = 1.2 kg, n = 40 vs. 40.1 ± 0.64 kg, n = 34, t = 5.06, P = 0.045). Kitten survival was less after the decline in deer abundance (0.573 ± 0.016, n = 30 vs. 0.856 ± 0.015, n = 25, Z = 2.40, P < 0.01). Survival of resident females was significantly less after the decline in puma numbers (0.783 ± 0.03 vs. 0.929 ± 0.019, U = 55.0, P = 0.009). Female productivity did not differ before or after the decline in deer abundance. Our results supported the majority of the predictions concerning the impact of changing deer abundance, which supported the hypothesis that the abundance of mule deer limited our population of pumas.