Elevated coding mutation rate during the reprogramming of human somatic cells into induced pluripotent stem cells

Mutations in human induced pluripotent stem cells (iPSCs) pose a risk for their clinical use due to preferential reprogramming of mutated founder cell and selection of mutations during maintenance of iPSCs in cell culture. It is unknown, however, if mutations in iPSCs are due to stress associated with oncogene expression during reprogramming. We performed whole exome sequencing of human foreskin fibroblasts and their derived iPSCs at two different passages. We found that in vitro passaging contributed 7% to the iPSC coding point mutation load, and ultradeep amplicon sequencing revealed that 19% of the mutations preexist as rare mutations in the parental fibroblasts suggesting that the remaining 74% of the mutations were acquired during cellular reprogramming. Simulation suggests that the mutation intensity during reprogramming is ninefold higher than the background mutation rate in culture. Thus the factor induced reprogramming stress contributes to a significant proportion of the mutation load of iPSCs.     

Determinants and Reference Ranges of Serum Immunoglobulins in Middle-Aged and Elderly Individuals: a Population-Based Study

Purpose

In clinical practice, currently one reference range for serum immunoglobulin (Ig) A, G, and M is applied to all adults, although various factors may influence Ig serum levels. Population-based data on determinants of IgA, IgG, and IgM and recommendations for subgroup specific reference ranges are lacking. We aimed to provide an overview of determinants of IgA, IgG, and IgM in community-dwelling middle-aged and elderly individuals and explore determinants that influence Ig reference ranges.

Methods

Within the Rotterdam Study, we performed linear regression analyses for the association of demographic, lifestyle, and cardiovascular factors with serum IgA, IgG, and IgM. We furthermore calculated Ig reference ranges (based on percentiles), both overall and within relevant subgroups.

Results

We included 8768 participants (median age 62 years). IgA and IgG increased non-linearly with higher age (P?<?.0001 for both). Women had lower IgA (beta:???0.24; 95% confidence interval [95% CI]:???0.29;???0.20) and IgG (beta:???0.33; 95% CI:???0.44;???0.23), but higher IgM levels (beta: 0.08; 95% CI: 0.04;0.13) than men. Former and particularly current smoking were associated with lower IgA and IgG (betas between???0.07 and???1.03). Higher alcohol consumption was associated with lower IgG (beta for heavy drinking:???0.70; 95% CI:???0.91;???0.48). Corticosteroid use was associated with lower IgG (beta:???1.12; 95% CI:???1.58;???0.66). Associations with cardiovascular factors were heterogeneous and differed between sexes.

Conclusion

Age, sex, smoking, alcohol consumption, corticosteroid use, and cardiovascular factors are determinants that should be considered when interpreting serum Ig levels in middle-aged and elderly individuals and may require adjusted reference ranges.

     

Prosthesis-Patient Mismatch Increases Early and Late Mortality in Low Risk Aortic Valve Replacement

PPM was highly associated with long-term all-cause mortality. Small Aortic Valve Annulus MDT (SAVA-MDT) exploring options to prevent PPM, including the implantation of newer generation prosthetic valves, aortic root enlargement and TAVI, should be established.

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Factors predicting tracer uptake in somatostatin receptor and MIBG scintigraphy of metastatic gastroenteropancreatic neuroendocrine tumors.

Radiolabeled octreotide analogs (Oct) and metaiodobenzylguanidine (MIBG) offer 2 different approaches for imaging and targeting metastatic gastroenteropancreatic neuroendocrine tumors (GEP-NET). Despite successful establishment of the revised World Health Organization (WHO) classification, which distinguishes between low- and high-grade malignant GEP-NET, there is a lack of scintigraphic studies comparing uptake behavior on the basis of this categorization. This study aims to define predisposing factors of tracer uptake for both imaging principles implementing the updated tumor criteria of the current WHO classification. METHODS: Fifty-seven consecutive patients with histologically confirmed metastatic GEP-NET evaluated with both 111In-pentetreotide and 123I/131I-MIBG scintigraphy were included in this study. Intensity of tracer uptake was graded according to the different metastatic regions. Patients were classified as overall positive when avid uptake in the clinically relevant tumor lesions was present. Correlation was tested between the proportion of positive patients and tumor origin, function, and malignancy. RESULTS: Overall, 52 patients (91.2%) were Oct positive and 28 patients (49.1%) were MIBG positive. The proportion of tracer-positive patients was significantly higher (P < 0.05) in low-grade malignant tumors for both tracers and in functioning as well as in gastroenteral NET for MIBG. Five patients were negative for both tracers. None of the Oct-negative patients proved to be MIBG positive. CONCLUSION: Oct affinity is observed with high frequency throughout the subgroups of metastatic GEP-NET, whereas corresponding MIBG uptake is overall less prevalent and more group dependent. Tumor differentiation significantly impacts both Oct and MIBG uptake, whereas functionality predisposes only for MIBG accumulation. Though clearly inferior to Oct-based radioimaging in most GEP-NET, MIBG achieves a remarkable rate of radioligand accumulation in functioning midgut enterochromaffin cell metastases (>80% of patients positive). These results may have implications for patient management and potentially for selection and performance of targeted therapy.     

Safety and efficacy of transvenous lead extraction of very old leads

OBJECTIVESTransvenous lead extraction using mechanical rotational- or laser sheaths is an established procedure. Lead dwell time has been recognized as a risk factor for extraction failure and procedure-related complications. We therefore investigated the safety and efficacy of transvenous extraction of leads with an implant duration of more than 10 years.Open in a separate windowMETHODSBetween January 2013 and March 2017, a total of 403 patients underwent lead extraction in 2 high-volume lead extraction centres. One hundred and fifty-four patients with extraction of at least 1 lead aged over 10 years were included in this analysis. Laser lead extraction was the primary extraction method, with additional use of mechanical rotational sheaths or femoral snares, if necessary. All procedural- and patient-based data were collected into a database and retrospectively analysed.RESULTSMean patient’s age was 65.8 ± 15.8 years, 68.2% were male. Three hundred and sixty-two leads had to be extracted. The mean lead dwell time of treated leads was 14.0 ± 6.1 years. Complete procedural success was achieved in 91.6% of cases, while clinical success was achieved in 96.8%. Failure of extraction occurred in 3.2%. Leads that could not be completely removed had a significantly longer lead dwell time (18.2 vs 13.2 years; P = 0.016). Additional mechanical rotational sheaths or femoral snares were used in 26 (16.9%) patients. Overall complication rate was 4.6%, including 5 (3.3%) major and 2 (1.3%) minor complications. There was no procedure-related mortality.CONCLUSIONSTransvenous lead extraction in leads aged over 10 years is safe and effective when performed in specialized centres and with use of multiple tools and techniques. Leads that could not be completely extracted had a statistically significant longer lead dwell time.     

Chronic mitral regurgitation detected on cardiac MDCT: differentiation between functional and valvular aetiologies

Objective

To determine whether cardiac computed tomography (MDCT) can differentiate between functional and valvular aetiologies of chronic mitral regurgitation (MR) compared with echocardiography (TTE).

Methods

Twenty-seven patients with functional or valvular MR diagnosed by TTE and 19 controls prospectively underwent cardiac MDCT. The morphological appearance of the mitral valve (MV) leaflets, MV geometry, MV leaflet angle, left ventricular (LV) sphericity and global/regional wall motion were analysed. The coronary arteries were evaluated for obstructive atherosclerosis.

Results

All control and MR cases were correctly identified by MDCT. Significant differences were detected between valvular and control groups for anterior leaflet length (30?±?7 mm vs. 22?±?4 mm, P?<?0.02) and thickness (3.0?±?1 mm vs. 2.2?±?1 mm, P?<?0.01). High-grade coronary stenosis was detected in all patients with functional MR compared with no controls (P?<?0.001). Significant differences in those with/without MV prolapse were detected in MV tent area (?1.0?±?0.6 mm vs. 1.3?±?0.9 mm, P?<?0.0001) and MV tent height (?0.7?±?0.3 mm vs. 0.8?±?0.8 mm, P?<?0.0001). Posterior leaflet angle was significantly greater for functional MR (37.9?±?19.1° vs. 22.9?±?14°, P?<?0.018) and less for valvular MR (0.6?±?35.5° vs. 22.9?±?14°, P?<?0.017). Sensitivity, specificity, and positive and negative predictive values of MDCT were 100%, 95%, 96% and 100%.

Conclusion

Cardiac MDCT allows the differentiation between functional and valvular causes of MR.     

Race and Health Disparities in Patient Refusal of Surgery for Early-Stage Pancreatic Cancer: An NCDB Cohort Study

Aim

To identify factors associated with refusal of surgery in patients with early-stage pancreatic cancer and estimate the impact of this decision on survival.

Methods

Using the National Cancer Data Base, 26,358 patients were identified with potentially resectable tumors (pretreatment clinical stage I: T1 or T2 N0M0). Multivariate models were employed to identify factors predicting failure to undergo surgery and assess the impact on survival.

Results

Of early-stage patients who were recommended surgery, 7.8% (N?=?992) refused surgery for resectable early-stage pancreatic cancer. On multivariable analysis, patients were more likely to refuse surgery if they were older [odds ratio (OR)?=?1.18; 95% confidence interval (CI) 1.16–1.19], female (OR?=?1.52; 95% CI 1.33–1.73), African American (vs White, OR?=?1.79; 95% CI 1.37–2.34), on Medicare/Medicaid (vs private, OR?=?2.75; 95% CI 1.54–4.92) or had higher Charlson–Deyo score (2 vs 0, OR?=?1.33; 95% CI 1.03–1.72). Patients were also significantly more likely to refuse surgery if they were seen at a center that is not an academic/research program (OR 1.9; 95% CI 1.6–2.27). Patients who were recommended surgery but refused had significantly worse survival than those with stage I who received surgery [median survival 6.8 vs 24 months, Cox hazard ratio (HR) 3.41; 95% CI 3.12–3.60].

Conclusions

The percentage of patients refusing surgery for operable early-stage pancreatic cancer has been decreasing in the last decade but remains a significant issue that affects survival. Disparities in refusal of surgery are independently associated with several variables including gender, race, and insurance. To mitigate national disparities in surgical care, future studies should focus on exploring potential reasons for refusal and developing communication interventions.

     

Assessment of Osstell? and Periotest? systems in measuring dental implant stability (in vitro study)

Objectives

The objectives of this in vitro study were to investigate the sensitivity and reliability of the Osstell™ systems (Resonance Frequency Analysis – RFA) compared to the Periotest® system in implant bone simulated conditions. Three conditions were simulated: (1) the direct fixture-bone contact and fibrous tissue fixture contact, (2) The different levels of horizontal bone loss, and (3) The hardening implant–bone interface.

Materials and methods

Forty-nine dental implant fixtures were placed in the center of acrylic cubes. In Part I seven fixtures were placed in direct contact with acrylic and another seven were placed in contact with polyvinyl siloxane impression material as soft interface. Part II: four sets of 0, 2, 4 and 6 mm horizontally exposed fixture samples were made (seven fixtures in each set). Part III: seven fixtures were placed in contact with a thin mix of autopolymerizing resin. The stability of these fixtures was measured using Osstell™ and Periotest® systems.

Results

The mean Periotest® value(PTV) and Osstell™ measurements showed a significant difference between the direct contact and soft interface (P < 0.001). These values also showed statistically significant difference between the different levels of horizontally exposed fixture groups (P < 0.001). The level of horizontal fixture exposure was strongly correlated with the PTVS (r = 0.967) and strongly negatively correlated with Implant Stability Quotient (r = −0.946). A strong correlation was found between the Osstell™ readings and the change in the stiffness of the autopolymerizing resin fixture interface group (r = 0.986).

Conclusions

Both Osstell™ and Periotest® systems proved to be sensitive in measuring dental implant stability in hard and in soft interfaces. Osstell™ also proved to be sensitive in detecting changes in the fixture interface stiffness. Osstell™ system proved to be more reliable compared to Periotest® system in measuring dental implant stability in hard and in soft interfaces.     

Treatment options for renal cell carcinoma in renal allografts: a case series from a single institution

Renal cell carcinoma (RCC) is more common in renal transplant and dialysis patients than the general population. However, RCC in transplanted kidneys is rare, and treatment has previously consisted of nephrectomy with a return to dialysis. There has been recent interest in nephron‐sparing procedures as a treatment option for RCC in allograft kidneys in an effort to retain allograft function. Four patients with RCC in allograft kidneys were treated with nephrectomy, partial nephrectomy, or radiofrequency ablation. All of the patients are without evidence of recurrence of RCC after treatment. We found nephron‐sparing procedures to be reasonable initial options in managing incidental RCCs diagnosed in functioning allografts to maintain an improved quality of life and avoid immediate dialysis compared with radical nephrectomy of a functioning allograft. However, in non‐functioning renal allografts, radical nephrectomy may allow for a higher chance of cure without the loss of transplant function. Consequently, radical nephrectomy should be utilized whenever the allograft is non‐functioning and the patient's surgical risk is not prohibitive.     

External anal sphincter contraction during cough: not a simple spinal reflex

AIMS: To assess whether the anal contraction during voluntary coughing is a simple spinal reflex-mediated activity or not. To address this question we studied the external intercostal (EIC) muscle activity and external anal sphincter (EAS) response to cough. MATERIALS AND METHODS: Electromyographic recordings were made from pre-gelled disposable surface electrodes. EAS electromyographic recordings were made from the EAS of the pelvic floor in 15 continent women all suffering from urgency and/or frequency without urge or stress urinary incontinence, and referred for urodynamic investigation. Electromyographic signal was immediately integrated (EMGi). The abdominal pressure was recorded with bladder and rectal pressure. EAS EMGi was recorded during successive voluntary cough. In three women, we have also recorded EIC EMGi activity since it is synchronous with diaphragmatic EMG activity during cough initiation. RESULTS: In all subjects, EAS EMGi activity precedes the onset of the abdominal pressure increase. The mean latency of EAS EMGi was 615 msec (+/-278). In the three subjects whose EMGi activity was recorded both on EAS and EIC, the onset of EAS EMGi activity occurred before the EIC EMGi activity (latency ranging from 40 to 780 msec) and before the increase in the abdominal pressure. CONCLUSIONS: The present study suggests that during coughing, EAS EMG activity increases before external intercostal muscle EMGi activity. The contraction of the EAS preceding the activation of muscles involved in coughing indicates that this response is not a result of a simple spinal reflex, but more likely the result of a more intricate reflex involving complex integrative centers.