The Leu22Pro tumor-associated variant of DNA polymerase beta is dRP lyase deficient

Approximately 30% of human tumors characterized to date express DNA polymerase beta (pol β) variant proteins. Two of the polymerase beta cancer-associated variants are sequence-specific mutators, and one of them binds to DNA but has no polymerase activity. The Leu22Pro (L22P) DNA polymerase beta variant was identified in a gastric carcinoma. Leu22 resides within the 8 kDa amino terminal domain of DNA polymerase beta, which exhibits dRP lyase activity. This domain catalyzes the removal of deoxyribose phosphate during short patch base excision repair. We show that this cancer-associated variant has very little dRP lyase activity but retains its polymerase activity. Although residue 22 has no direct contact with the DNA, we report here that the L22P variant has reduced DNA-binding affinity. The L22P variant protein is deficient in base excision repair. Molecular dynamics calculations suggest that alteration of Leu22 to Pro changes the local packing, the loop connecting helices 1 and 2 and the overall juxtaposition of the helices within the N-terminal domain. This in turn affects the shape of the binding pocket that is required for efficient dRP lyase catalysis

Estimation of Immune Cell Densities in Immune Cell Conglomerates: An Approach for High-Throughput Quantification

Determining the correct number of positive immune cells in immunohistological sections of colorectal cancer and other tumor entities is emerging as an important clinical predictor and therapy selector for an individual patient. This task is usually obstructed by cell conglomerates of various sizes. We here show that at least in colorectal cancer the inclusion of immune cell conglomerates is indispensable for estimating reliable patient cell counts. Integrating virtual microscopy and image processing principally allows the high-throughput evaluation of complete tissue slides.For such large-scale systems we demonstrate a robust quantitative image processing algorithm for the reproducible quantification of cell conglomerates on CD3 positive T cells in colorectal cancer. While isolated cells (28 to 80 microm(2)) are counted directly, the number of cells contained in a conglomerate is estimated by dividing the area of the conglomerate in thin tissues sections (< or =6 microm) by the median area covered by an isolated T cell which we determined as 58 microm(2). We applied our algorithm to large numbers of CD3 positive T cell conglomerates and compared the results to cell counts obtained manually by two independent observers. While especially for high cell counts, the manual counting showed a deviation of up to 400 cells/mm(2) (41% variation), algorithm-determined T cell numbers generally lay in between the manually observed cell numbers but with perfect reproducibility.In summary, we recommend our approach as an objective and robust strategy for quantifying immune cell densities in immunohistological sections which can be directly implemented into automated full slide image processing systems

Influence of ultrasound-assisted par-frying on crust formation and browning during the production of French fries

Frying is a key processing step during the production of French fries and important for end product quality and sensory attributes. It is governed by heat and mass transfer between the frying oil and the potato strips. Crust and color of the French fries are key quality parameters and important in consumer perception. Crust formation is a result of combined heat and mass transfer effects. Convective heat transfer from frying oil to potato strips and heat conduction within the tissue cause water evaporation. Mass transfer occurs in the form of water vapor release to the frying oil and oil absorption in the outer layers of the potato strips. Browning of French fries is related to Maillard reactions between reducing sugars and amino acids. High contents of reducing sugars are often related to an undesired dark color and bitter taste of French fries. High-intensity ultrasound transmitted to liquid media causes cavitation and microstreaming, which can influence boundary layers and cell structures and result in improved heat and mass transfer. The influence of an ultrasound treatment of potato strips during the par-frying step was investigated in order to determine the effect on the resulting changes in product quality. Improved heat transfer at the product surface due to micro-streaming in the oil and a facilitated release of vapor from the product surface was observed. A faster crust formation was found at the initial phase of frying but the crust was found to become softer at longer sonication times due to persistent mechanical ultrasound effects. French fries from ultrasound assisted par-frying had a lighter color after finish-frying in comparison to the conventionally par-fried samples due to an improved release of reducing sugars from the tissue. Ultrasound-assisted par-frying showed to be effective in modifying heat and mass transfer with an impact on crust formation and browning of French fries. Further work is required regarding the optimization of parameters and sonication times

Development of a Continuous Pulsed Electric Field (PEF) Vortex-Flow Chamber for Improved Treatment Homogeneity Based on Hydrodynamic Optimization

Pulsed electric fields (PEF) treatment is an effective process for preservation of liquid products in food and biotechnology at reduced temperatures, by causing electroporation. It may contribute to increase retention of heat-labile constituents with similar or enhanced levels of microbial inactivation, compared to thermal processes. However, especially continuous PEF treatments suffer from inhomogeneous treatment conditions. Typically, electric field intensities are highest at the inner wall of the chamber, where the flow velocity of the treated product is lowest. Therefore, inhomogeneities of the electric field within the treatment chamber and associated inhomogeneous temperature fields emerge. For this reason, a specific treatment chamber was designed to obtain more homogeneous flow properties inside the treatment chamber and to reduce local temperature peaks, therefore increasing treatment homogeneity. This was accomplished by a divided inlet into the chamber, consequently generating a swirling flow (vortex). The influence of inlet angles on treatment homogeneity was studied (final values: radial angle α = 61°; axial angle β = 98°), using computational fluid dynamics (CFD). For the final design, the vorticity, i.e., the intensity of the fluid rotation, was the lowest of the investigated values in the first treatment zone (1002.55 1/s), but could be maintained for the longest distance, therefore providing an increased mixing and most homogeneous treatment conditions. The new design was experimentally compared to a conventional co-linear setup, taking into account inactivation efficacy of Microbacterium lacticum as well as retention of heat-sensitive alkaline phosphatase (ALP). Results showed an increase in M. lacticum inactivation (maximum Δlog of 1.8 at pH 7 and 1.1 at pH 4) by the vortex configuration and more homogeneous treatment conditions, as visible by the simulated temperature fields. Therefore, the new setup can contribute to optimize PEF treatment conditions and to further extend PEF applications to currently challenging products

Amplified ambivalence: having a sibling with juvenile idiopathic arthritis

Despite increased awareness of family responses to chronic illness and disability, there is still a need to understand experiences of well siblings. We begin to address this by asking “What is it like to have a sibling with juvenile idiopathic arthritis?” (JIA).Eight families with an adolescent diagnosed with JIA participated. Four members of each family, including one healthy sibling, were interviewed and transcripts analyzed using grounded theory. Analysis suggests healthy siblings see their family as different to ‘normal’ families, forfeit time with peers, share vicariously adverse experiences of their ill sibling, and feel inadequately informed. Such experiences amplify the ambivalent nature of sibling relationships and are possibly felt most strongly during late childhood and early adolescence. Support from extended family can reduce these negative experiences and facilitate social and emotional adjustment which also occurs over time as the children mature. These findings have implications for healthcare professionals and voluntary organizations

The auditory temporal attending theory revisited

Background. The temporal attending theory predicts that tone sequences presented at a regular rhythm entrain attentional oscillations and thereby facilitate the processing of sounds presented in phase with this rhythm (Jones et al., 2002). During the past decade the theory of auditory temporal attending has become widely popular (140 ISI citations by July 2014) and has inspired both music psychology as well as neuroscience research. The aim of the current study was to replicate the findings of Jones et al. (2002).Method. The original paradigm is a pitch comparison task in which two tones - an initial standard tone and the last tone of a longer series, named the comparison tone - have to be compared. In between the two, distractor tones with variable pitch are presented at a regular sequence. A comparison tone presented in phase with the entrained rhythm is hypothesized to lead to better behavioral performance, thus higher task accuracy, compared to comparison tones presented at unexpected early or late intervals. Four different variations of the original paradigm were created and 106 participants were tested in total. The Goldsmiths Musical Sophistication Index (Gold-MSI) was included in all but the first experiment to test the influence of musicality on task performance.Results and Conclusion. Over all four experiments only 38 of the 106 participants showed the desired pattern of an inverted U-shaped profile in task accuracy, and in none of the four variations did the group average effects replicate the pattern reported by Jones et al., (2002). However, evidence for a relationship between musicality and overall behavioral performance was found. Our results question the validity of the pitch comparison task for the study of auditory temporal attending

Biofilm-like habitat at the sea-surface: A mesocosm study, Cruise No. POS537, 14.09.2019 – 04.10.2019, Malaga (Spain) – Cartagena (Spain) - BIOFILM

Biofilm-like properties can form on sea surfaces, but an understanding of the underlying processes leading to the development of these biofilms is not available. We used approaches to study the development of biofilm-like properties at the sea surface, i.e. the number, abundance and diversity of bacterial communities and phytoplankton, the accumulation of gel-like particles and dissolved tracers. During the expedition POS537 we used newly developed and free drifting mesocosms and performed incubation experiments. With these approaches we aim to investigate the role of light and UV radiation as well as the microbes themselves, which lead to the formation of biofilms. With unique microbial interactions and photochemical reactions, sea surface biofilms could be biochemical reactors with significant implications for ocean and climate research, e.g. with respect to the marine carbon cycle, diversity of organisms and oceanatmosphere interactions

Ribosome engineering reveals the importance of 5S rRNA autonomy for ribosome assembly

5S rRNA is an indispensable component of cytoplasmic ribosomes in all species. The functions of 5S rRNA and the reasons for its evolutionary preservation as an independent molecule remain unclear. Here we used ribosome engineering to investigate whether 5S rRNA autonomy is critical for ribosome function and cell survival. By linking circularly permutated 5S rRNA with 23S rRNA we generated a bacterial strain devoid of free 5S rRNA. Viability of the engineered cells demonstrates that autonomous 5S rRNA is dispensable for cell growth under standard conditions and is unlikely to have essential functions outside the ribosome. The fully assembled ribosomes carrying 23S-5S rRNA are highly active in translation. However, the engineered cells accumulate aberrant 50S subunits unable to form stable 70S ribosomes. Cryo-EM analysis revealed a malformed peptidyl transferase center in the misassembled 50S subunits. Our results argue that the autonomy of 5S rRNA is preserved due to its role in ribosome biogenesis

Liver X Receptor Agonist AZ876 Induces Beneficial Endogenous Cardiac Lipid Reprogramming and Protects Against Isoproterenol‐Induced Cardiac Damage

Background: It is known that dietary intake of polyunsaturated fatty acids may improve cardiac function. However, relatively high daily doses are required to achieve sufficient cardiac concentrations of beneficial omega-3 fatty acids. The liver X receptor (LXR) is a nuclear hormone receptor and a crucial regulator of lipid homeostasis in mammals. LXR activation has been shown to endogenously reprogram cellular lipid profiles toward increased polyunsaturated fatty acids levels. Here we studied whether LXR lipid reprogramming occurs in cardiac tissue and exerts cardioprotective actions. Methods and Results: Male 129SV mice were treated with the LXR agonist AZ876 (20 mu mol/kg per day) for 11 days. From day 6, the mice were injected with the nonselective beta-agonist isoproterenol for 4 consecutive days to induce diastolic dysfunction and subendocardial fibrosis while maintaining systolic function. Treatment with isoproterenol led to a marked impairment of global longitudinal strain and the E/e' ratio of transmitral flow to mitral annular velocity, which were both significantly improved by the LXR agonist. Histological examination showed a significant reduction in isoproterenol-induced subendocardial fibrosis by AZ876. Analysis of the cardiac lipid composition by liquid chromatography-high resolution mass spectrometry revealed a significant increase in cardiac polyunsaturated fatty acids levels and a significant reduction in saturated fatty acids by AZ876. Conclusions: The present study provides evidence that the LXR agonist AZ876 prevents subendocardial damage, improves global longitudinal strain and E/e' in a mouse model of isoproterenol-induced cardiac damage, accompanied by an upregulation of cardiac polyunsaturated fatty acids levels. Cardiac LXR activation and beneficial endogenous cardiac lipid reprogramming may provide a new therapeutic strategy in cardiac disease with diastolic dysfunction

a cross-sectional study on the association between urbanicity and the acquisition of immunity

Background Malaria incidence has declined considerably over the last decade. This is partly due to a scale-up of control measures but is also attributed to increasing urbanization. This study aimed to analyse the association between malaria and urbanization and the effect of urbanicity on the acquisition of semi-immunity. Methods In 2012, children with fever presenting to St Michael’s Hospital Pramso/Ghana were recruited. The malaria-positive-fraction (MPF) of fever cases was calculated on community-level to approximate the malaria risk. The mean age of malaria cases was calculated for each community to estimate the acquisition of semi-immunity. The level of urbanicity for the communities was calculated and associations between MPF, urbanicity and immunity were modelled using linear regression. Results Twenty-six villages were included into the study with a mean MPF of 35 %. A linear decrease of 5 % (95 % CI: 4–6 %) in MPF with every ten-point increase in urbanicity was identified. The mean age of malaria patients increased by 2.9 months (95 % CI: 1.0–4.8) with every ten-point increase in urbanicity. Discussion The results confirm an association between an increase in urbanicity and declining malaria risk and demonstrate that the acquisition of semi-immunity is heterogeneous on a micro- epidemiological scale and is associated with urbanicity