Suppression of DNA-damage checkpoint signaling by Rsk-mediated phosphorylation of Mre11.

Ataxia telangiectasia mutant (ATM) is an S/T-Q-directed kinase that is critical for the cellular response to double-stranded breaks (DSBs) in DNA. Following DNA damage, ATM is activated and recruited by the MRN protein complex [meiotic recombination 11 (Mre11)/DNA repair protein Rad50/Nijmegen breakage syndrome 1 proteins] to sites of DNA damage where ATM phosphorylates multiple substrates to trigger cell-cycle arrest. In cancer cells, this regulation may be faulty, and cell division may proceed even in the presence of damaged DNA. We show here that the ribosomal s6 kinase (Rsk), often elevated in cancers, can suppress DSB-induced ATM activation in both Xenopus egg extracts and human tumor cell lines. In analyzing each step in ATM activation, we have found that Rsk targets loading of MRN complex components onto DNA at DSB sites. Rsk can phosphorylate the Mre11 protein directly at S676 both in vitro and in intact cells and thereby can inhibit the binding of Mre11 to DNA with DSBs. Accordingly, mutation of S676 to Ala can reverse inhibition of the response to DSBs by Rsk. Collectively, these data point to Mre11 as an important locus of Rsk-mediated checkpoint inhibition acting upstream of ATM activation.

Association between DNA damage response and repair genes and risk of invasive serous ovarian cancer.

BACKGROUND: We analyzed the association between 53 genes related to DNA repair and p53-mediated damage response and serous ovarian cancer risk using case-control data from the North Carolina Ovarian Cancer Study (NCOCS), a population-based, case-control study. METHODS/PRINCIPAL FINDINGS: The analysis was restricted to 364 invasive serous ovarian cancer cases and 761 controls of white, non-Hispanic race. Statistical analysis was two staged: a screen using marginal Bayes factors (BFs) for 484 SNPs and a modeling stage in which we calculated multivariate adjusted posterior probabilities of association for 77 SNPs that passed the screen. These probabilities were conditional on subject age at diagnosis/interview, batch, a DNA quality metric and genotypes of other SNPs and allowed for uncertainty in the genetic parameterizations of the SNPs and number of associated SNPs. Six SNPs had Bayes factors greater than 10 in favor of an association with invasive serous ovarian cancer. These included rs5762746 (median OR(odds ratio)(per allele) = 0.66; 95% credible interval (CI) = 0.44-1.00) and rs6005835 (median OR(per allele) = 0.69; 95% CI = 0.53-0.91) in CHEK2, rs2078486 (median OR(per allele) = 1.65; 95% CI = 1.21-2.25) and rs12951053 (median OR(per allele) = 1.65; 95% CI = 1.20-2.26) in TP53, rs411697 (median OR (rare homozygote) = 0.53; 95% CI = 0.35 - 0.79) in BACH1 and rs10131 (median OR( rare homozygote) = not estimable) in LIG4. The six most highly associated SNPs are either predicted to be functionally significant or are in LD with such a variant. The variants in TP53 were confirmed to be associated in a large follow-up study. CONCLUSIONS/SIGNIFICANCE: Based on our findings, further follow-up of the DNA repair and response pathways in a larger dataset is warranted to confirm these results.

Patient expectation is the most important predictor of discharge destination after primary total joint arthroplasty.

The purpose of this study was to identify preoperative predictors of discharge destination after total joint arthroplasty. A retrospective study of three hundred and seventy-two consecutive patients who underwent primary total hip and knee arthroplasty was performed. The mean length of stay was 2.9 days and 29.0% of patients were discharged to extended care facilities. Age, caregiver support at home, and patient expectation of discharge destination were the only significant multivariable predictors regardless of the type of surgery (total knee versus total hip arthroplasty). Among those variables, patient expectation was the most important predictor (P < 0.001; OR 169.53). The study was adequately powered to analyze the variables in the multivariable logistic regression model, which had a high concordance index of 0.969.

Unipedal balance is affected by lower extremity joint arthroplasty procedure 1 year following surgery.

Lower Extremity Joint Arthroplasty (LEJA) surgery is an effective way to alleviate painful osteoarthritis. Unfortunately, these surgeries do not normalize the loading asymmetry during the single leg stance phase of gait. Therefore, we examined single leg balance in 234 TJA patients (75 hips, 65 knees, 94 ankles) approximately 12 months following surgery. Patients passed if they maintained single leg balance for 10s with their eyes open. Patients one year following total hip arthroplasty (THA-63%) and total knee arthroplasty (TKA-69%) had similar pass rates compared to a total ankle arthroplasty (TAA-9%). Patients following THA and TKA exhibit better unilateral balance in comparison with TAA patients. It may be beneficial to include a rigorous proprioception and balance training program in TAA patients to optimize functional outcomes.

Percent body fat more associated with perioperative risks after total joint arthroplasty than body mass index

Copyright © 2014 Elsevier Inc. All rights reserved.Understanding the impact of obesity on elective total joint arthroplasty (TJA) remains critical. Perioperative outcomes were reviewed in 316 patients undergoing primary TJA. Higher percent body fat (PBF) was associated with postoperative blood transfusion, increased hospital length of stay (LOS) >3 days, and discharge to an extended care facility while no significant differences existed for BMI. Additionally, PBF of 43.5 was associated with a 2.4× greater likelihood of blood transfusion, PBF of 36.5 with a 1.9× greater likelihood for LOS >3 days, and PBF of 36.0 with a 1.4× greater likelihood for discharge to an extended care facility. PBF may be a more effective measure than BMI to use in screening for perioperative risks and acute outcomes associated with obese total joint patients.

Later Life Consequences of Developmental Mitochondrial DNA Damage in C. elegans

Mitochondria are responsible for producing the vast majority of cellular ATP, and are therefore critical to organismal health [1]. They contain thir own genomes (mtDNA) which encode 13 proteins that are all subunits of the mitochondrial respiratory chain (MRC) and are essential for oxidative phosphorylation [2]. mtDNA is present in multiple copies per cell, usually between 103 and 104 , though this number is reduced during certain developmental stages [3, 4]. The health of the mitochondrial genome is also important to the health of the organism, as mutations in mtDNA lead to human diseases that collectively affect approximately 1 in 4000 people [5, 6]. mtDNA is more susceptible than nuclear DNA (nucDNA) to damage by many environmental pollutants, for reasons including the absence of Nucleotide Excision Repair (NER) in the mitochondria [7]. NER is a highly functionally conserved DNA repair pathway that removes bulky, helix distorting lesions such as those caused by ultraviolet C (UVC) radiation and also many environmental toxicants, including benzo[a]pyrene (BaP) [8]. While these lesions cannot be repaired, they are slowly removed through a process that involves mitochondrial dynamics and autophagy [9, 10]. However, when present during development in C. elegans, this damage reduces mtDNA copy number and ATP levels [11]. We hypothesize that this damage, when present during development, will result in mitochondrial dysfunction and increase the potential for adverse outcomes later in life.

To test this hypothesis, 1st larval stage (L1) C. elegans are exposed to 3 doses of 7.5J/m2 ultraviolet C radiation 24 hours apart, leading to the accumulation of mtDNA damage [9, 11]. After exposure, many mitochondrial endpoints are assessed at multiple time points later in life. mtDNA and nucDNA damage levels and genome copy numbers are measured via QPCR and real-time PCR , respectively, every 2 day for 10 days. Steady state ATP levels are measured via luciferase expressing reporter strains and traditional ATP extraction methods. Oxygen consumption is measured using a Seahorse XFe24 extra cellular flux analyzer. Gene expression changes are measured via real time PCR and targeted metabolomics via LC-MS are used to investigate changes in organic acid, amino acid and acyl-carnitine levels. Lastly, nematode developmental delay is assessed as growth, and measured via imaging and COPAS biosort.

I have found that despite being removed, UVC induced mtDNA damage during development leads to persistent deficits in energy production later in life. mtDNA copy number is permanently reduced, as are ATP levels, though oxygen consumption is increased, indicating inefficient or uncoupled respiration. Metabolomic data and mutant sensitivity indicate a role for NADPH and oxidative stress in these results, and exposed nematodes are more sensitive to the mitochondrial poison rotenone later in life. These results fit with the developmental origin of health and disease hypothesis, and show the potential for environmental exposures to have lasting effects on mitochondrial function.

Lastly, we are currently working to investigate the potential for irreparable mtDNA lesions to drive mutagenesis in mtDNA. Mutations in mtDNA lead to a wide range of diseases, yet we currently do not understand the environmental component of what causes them. In vitro evidence suggests that UVC induced thymine dimers can be mutagenic [12]. We are using duplex sequencing of C. elegans mtDNA to determine mutation rates in nematodes exposed to our serial UVC protocol. Furthermore, by including mutant strains deficient in mitochondrial fission and mitophagy, we hope to determine if deficiencies in these processes will further increase mtDNA mutation rates, as they are implicated in human diseases.

Numerical modelling of solder joint formation

Solder materials are used to provide a connection between electronic components and printed circuit boards (PCBs) using either the reflow or wave soldering process. As a board assembly passes through a reflow furnace the solder (initially in the form of solder paste) melts, reflows, then solidifies, and finally deforms between the chip and board. A number of defects may occur during this process such as flux entrapment, void formation, and cracking of the joint, chip or board. These defects are a serious concern to industry, especially with trends towards increasing component miniaturisation and smaller pitch sizes. This paper presents a modelling methodology for predicting solder joint shape, solidification, and deformation (stress) during the assembly process.

A computational modelling framework to predict macroscopic phenomena in solder joint formation

A computational model of solder joint formation and the subsequent cooling behaviour is described. Given the rapid changes in the technology of printed circuit boards, there is a requirement for comprehensive models of solder joint formation which permit detailed analysis of design and optimization options. Solder joint formation is complex, involving a range of interacting phenomena. This paper describes a model implementation (as part of a more comprehensive framework) to describe the shape formation (conditioned by surface tension), heat transfer, phase change and the development of elastoviscoplastic stress. The computational modelling framework is based upon mixed finite element and finite volume procedures, and has unstructured meshes enabling arbitrarily complex geometries to be analysed. Initial results for both through-hole and surface-mount geometries are presented.

An integrated modeling approach to solder joint formation

The attachment of electronic components to printed circuit boards using solder material is a complex process. This paper presents a novel modeling methodology, which integrates the governing physics taking place. Multiphysics modeling technology, imbedded into the simulation tool—PHYSICA is used to simulate fluid flow, heat transfer, solidification, and stress evolution in an integrated manner. Results using this code are presented, detailing the mechanical response of two solder materials as they cool, solidify and then deform. The shape that a solder joint takes upon melting is predicted using the SURFACE EVOLVER code. Details are given on how these predictions can be used in the PHYSICA code to provide a modeling route by which the shape, solidification history, and resulting stress profiles can be predicted.

Optimisation modelling for flip-chip solder joint reliability

This paper details and demonstrates integrated optimisation-reliability modelling for predicting the performance of solder joints in electronic packaging. This integrated modelling approach is used to identify efficiently and quickly the most suitable design parameters for solder joint performance during thermal cycling and is demonstrated on flip-chip components using “no-flow” underfills. To implement “optimisation in reliability” approach, the finite element simulation tool – PHYSICA, is coupled with optimisation and statistical tools. This resulting framework is capable of performing design optimisation procedures in an entirely automated and systematic manner.

Computer modelling of the reliability of flip chips with metal column bumping

Recently, research has been carried out to test a novel bumping method which omits the under bump metallurgy forming process by bonding copper columns directly onto the Al pads of the silicon dies. This bumping method could be adopted to simplify the flip chip manufacturing process, increase the productivity and achieve a higher I/O count. This paper describes an investigation of the solder joint reliability of flip-chips based on this new bumping process. Computer modelling methods are used to predict the shape of solder joints and response of flip chips to thermal cyclic loading. The accumulated plastic strain energy at the comer solder joints is used as the damage indicator. Models with a range of design parameters have been compared for their reliability. The parameters that have been investigated are the copper column height, radius and solder volume. The ranking of the relative importance of these parameters is given. For most of the results presented in the paper, the solder material has been assumed to be the lead-free 96.5Sn3.5Ag alloy but some results for 60Sn40Pb solder joints have also been presented.

Lifetime assessment of electronic components for high reliability aerospace applications

This paper details a modelling approach for assessing the in-service (field) reliability and thermal fatigue life-time of electronic package interconnects for components used in the assembly of an aerospace system. The Finite Element slice model of a Plastic Ball Grid Array (PBGA) package and suitable energy based damage models for crack length predictions are used in this study. Thermal fatigue damage induced in tin-lead solder joints are investigated by simulating the crack growth process under a set of prescribed field temperature profiles that cover the period of operational life. The overall crack length in the solder joint for all different thermal profiles and number of cycles for each profile is predicted using a superposition technique. The effect of using an underfill is also presented. A procedure for verifying the field lifetime predictions for the electronic package by using reliability assessment under Accelerated Thermal Cycle (ATC) testing is also briefly outlined.