Suppression of HCV-specific T cells without differential hierarchy demonstrated ex vivo in persistent HCV infection

Hepatitis C virus (HCV) has a high propensity for persistence. To better define the immunologic determinants of HCV clearance and persistence, we examined the circulating HCV-specific T-cell frequency, repertoire, and cytokine phenotype ex vivo in 24 HCV seropositive subjects (12 chronic, 12 recovered), using 361 overlapping peptides in 36 antigenic pools that span the entire HCV core, NS3-NS5. Consistent with T-cell-mediated control of HCV, the overall HCV-specific type-1 T-cell response was significantly greater in average frequency (0.24% vs. 0.04% circulating lymphocytes, P =.001) and scope (14/36 vs. 4/36 pools, P =.002) among the recovered than the chronic subjects, and the T-cell response correlated inversely with HCV titer among the chronic subjects (R = -0.51, P =.049). Although highly antigenic regions were identified throughout the HCV genome, there was no apparent difference in the overall HCV-specific T-cell repertoire or type-1/type-2 cytokine profile relative to outcome. Notably, HCV persistence was associated with a reversible CD4-mediated suppression of HCV-specific CD8 T cells and with higher frequency of CD4(+)CD25(+) regulatory T cells (7.3% chronic vs. 2.5% recovered, P =.002) that could directly suppress HCV-specific type-1 CD8 T cells ex vivo. In conclusion, we found that HCV persistence is associated with a global quantitative and functional suppression of HCV-specific T cells but not differential antigenic hierarchy or cytokine phenotype relative to HCV clearance. The high frequency of CD4(+)CD25(+) regulatory T cells and their suppression of HCV-specific CD8 T cells ex vivo suggests a novel role for regulatory T cells in HCV persistence.     

An endoscopic training model to improve accuracy of colonic polyp size measurement

Purpose

Most studies of colonic polyps rely on visual estimation when regarding polyp size; however, the reliability of a visual estimate is questionable. Our study aims to develop a training model to improve the accuracy of size estimation of colonic polyps in vivo.

Methods

Colon polyps were recorded on 160 video clips during colonoscopy. The size of each polyp was estimated by visual inspection and subsequently measured with a flexible linear measuring probe. The study included a pretest, an intervention, and a posttest. The pretest included 160 video clips, which comprised the visual-estimation portion of the study. The intervention was an educational model consisting of 30 video clips which included a visual-estimation section and a linear-measuring-probe section, designed to help the endoscopists to compare their visual estimate of size with the measured size of the polyps. The posttest included the 160 video clips used in the pretest, presented in random order. Intraobserver agreement and diagnostic accuracy were compared before and after the training session.

Results

Eight beginners and four experienced colonoscopists were enrolled. The overall kappa (κ) values of intraobserver agreement for pretest and posttest were 0.74 and 0.85 for beginner group as well as 0.83 and 0.88 for experienced group, respectively. The overall diagnostic accuracy improved from 0.52 to 0.78 for beginner group and 0.71 to 0.87 for experienced group (P?<?0.05) after education with the training model.

Conclusions

This training model could help endoscopists improve the accuracy of measurement of polyps on colonoscopy in a short period. The durability of learning effect needs further investigation.     

A comparison of intracardiac and transesophageal echocardiography to detect left atrial appendage thrombus in a swine model

Purpose

Transesophageal echocardiography (TEE) is the gold standard in the evaluation for left atrial appendage (LAA) thrombus in patients with atrial fibrillation (AF) and is often performed prior to AF ablation. We routinely use intracardiac echocardiography (ICE) to assist in AF ablation; however, standard right atrial views do not provide adequate visualization of the LAA. As the incidence of thrombus in this population is relatively low, TEE incurs additional risk, cost, and patient discomfort. Novel views of the LAA with ICE may obviate the need for TEE in this population. We tested the hypothesis that due to their proximity, imaging the LAA from the pulmonary artery (PA) would provide equivalent sensitivity and specificity to TEE in detecting LAA thrombus in a swine model.

Methods

Five domestic swine were utilized. Baseline images of the LAA with TEE were obtained. An 8Fr ICE catheter was placed in the left main PA, and imaging of the LAA was repeated. After transseptal puncture, an admixture of 2 cm³ blood and 1,000 IU of thrombin was injected into the LAA, and imaging of the LAA was repeated. Two blinded, independent reviewers experienced in ICE assessed the images and adjudicated both the presence of thrombus and the subjective image quality.

Results

The presence or absence of thrombus was correctly identified in all cases by both reviewers. Both reviewers rated the subjective quality of ICE images superior to TEE.

Conclusions

ICE is equivalent to TEE in imaging LAA thrombus in a porcine model. Whether ICE can provide similar diagnostic accuracy and safety for detecting LAA thrombus in humans remains unproven.     

Hip fracture types in men and women change differently with age

Background

Hip fractures are expensive and a frequent cause of morbidity and mortality in the elderly. In most studies hip fractures have been viewed as a unitary fracture but recently the two main types of fracture (intertrochanteric and subcapital) have been viewed as two fractures with a different etiology and requiring a different approach to prevention. The relative proportion of intertrochanteric fractures increases with age in women. In previous studies no particular pattern in men has been noted. In this study, we explored changes in the relative proportion of the two fracture types with age in the two genders.

Methods

Patients of 50 years and older, with a diagnosis of hip fracture, discharged from two local acute care hospitals over a 5 year period (n = 2150) were analyzed as a function of age and gender to explore the relative proportions of intertrochanteric and subcapital fractures, and the change in relative proportion in the two genders with age.

Results

Overall, for the genders combined, the proportion of intertrochanteric fractures increases with age (p = .007). In women this increase is significant (p < .001), but in men the opposite pattern is observed, with the proportion of intertrochanteric fractures falling significantly with age (p = .025).

Conclusions

The pattern of hip fractures is different in men and women with aging. It is likely that the pattern difference reflects differences in type and rate of bone loss in the genders, but it is conjectured that the changing rate and pattern of falling with increasing age may also be important. The two main hip fracture types should be considered distinct and different and be studied separately in studies of cause and prevention.     

Lipid membranes in poxvirus replication

Poxviruses replicate in the cytoplasm, where they acquire multiple lipoprotein membranes. Although a proposal that the initial membrane arises de novo has not been substantiated, there is no accepted explanation for its formation from cellular membranes. A subsequent membrane-wrapping step involving modified trans-Golgi or endosomal cisternae results in a particle with three membranes. These wrapped virions traverse the cytoplasm on microtubules; the outermost membrane is lost during exocytosis, the middle one is lost just prior to cell entry, and the remaining membrane fuses with the cell to allow the virus core to enter the cytoplasm and initiate a new infection.     

Brain–computer interfaces increase whole-brain signal to noise

Brain–computer interfaces (BCIs) can convert mental states into signals to drive real-world devices, but it is not known if a given covert task is the same when performed with and without BCI-based control. Using a BCI likely involves additional cognitive processes, such as multitasking, attention, and conflict monitoring. In addition, it is challenging to measure the quality of covert task performance. We used whole-brain classifier-based real-time functional MRI to address these issues, because the method provides both classifier-based maps to examine the neural requirements of BCI and classification accuracy to quantify the quality of task performance. Subjects performed a covert counting task at fast and slow rates to control a visual interface. Compared with the same task when viewing but not controlling the interface, we observed that being in control of a BCI improved task classification of fast and slow counting states. Additional BCI control increased subjects’ whole-brain signal-to-noise ratio compared with the absence of control. The neural pattern for control consisted of a positive network comprised of dorsal parietal and frontal regions and the anterior insula of the right hemisphere as well as an expansive negative network of regions. These findings suggest that real-time functional MRI can serve as a platform for exploring information processing and frontoparietal and insula network-based regulation of whole-brain task signal-to-noise ratio.     

Detection of cardiovascular autonomic neuropathy using exercise testing in patients with type 2 diabetes mellitus

AimsThis study investigated autonomic nervous system function in subjects with diabetes during exercise and recovery.MethodsEighteen type 2 diabetics (age 55 ± 2 years) and twenty healthy controls (age 51 ± 1 years) underwent two 16-min bicycle submaximal ECG stress tests followed by 45 min of recovery. During session #2, atropine (0.04 mg/kg) was administered at peak exercise, and the final two minutes of exercise and entire recovery occurred under parasympathetic blockade. Plasma catecholamines were measured throughout. Parasympathetic effect was defined as the difference between a measured parameter at baseline and after parasympathetic blockade.ResultsThe parasympathetic effect on the RR interval was blunted (P = .004) in diabetic subjects during recovery. Parasympathetic effect on QT–RR slope during early recovery was diminished in the diabetes group (diabetes 0.13 ± 0.02, control 0.21 ± 0.02, P = .03). Subjects with diabetes had a lower heart rate recovery at 1 min (diabetes 18.5 ± 1.9 bpm, control 27.6 ± 1.5 bpm, P < .001).ConclusionsIn subjects with well-controlled type 2 diabetes, even with minimal evidence of CAN using current methodology, altered cardiac autonomic balance is present and can be detected through an exercise-based assessment for CAN. The early post-exercise recovery period in diabetes was characterized by enhanced sympathoexcitation, diminished parasympathetic reactivation and delay in heart rate recovery.     

White matter tract signatures of the progressive aphasias

The primary progressive aphasias (PPA) are a heterogeneous group of language-led neurodegenerative diseases resulting from large-scale brain network degeneration. White matter (WM) pathways bind networks together, and might therefore hold information about PPA pathogenesis. Here we used diffusion tensor imaging and tract-based spatial statistics to compare WM tract changes between PPA syndromes and with respect to Alzheimer's disease and healthy controls in 33 patients with PPA (13 nonfluent/agrammatic PPA); 10 logopenic variant PPA; and 10 semantic variant PPA. Nonfluent/agrammatic PPA was associated with predominantly left-sided and anterior tract alterations including uncinate fasciculus (UF) and subcortical projections; semantic variant PPA with bilateral alterations in inferior longitudinal fasciculus and UF; and logopenic variant PPA with bilateral but predominantly left-sided alterations in inferior longitudinal fasciculus, UF, superior longitudinal fasciculus, and subcortical projections. Tract alterations were more extensive than gray matter alterations, and the extent of alteration across tracts and PPA syndromes varied between diffusivity metrics. These WM signatures of PPA syndromes illustrate the selective vulnerability of brain language networks in these diseases and might have some pathologic specificity.     

Biomechanical effects of patellar positioning on intraoperative knee joint gap measurement in total knee arthroplasty

Background

Balancing both the lateral/medial and extension/flexion joint gaps is a prerequisite for soft tissue balance in total knee arthroplasty. The purpose of this study was to quantify the effects of patellar positioning and quadriceps load during total knee arthroplasty on knee joint gap measurements.

Methods

Eight fresh-frozen cadaveric knees ranging in age from 65 to 85 years old were used. Using a medial parapatellar approach, posterior cruciate ligament sacrificing total knee arthroplasty was performed. The specimens were mounted on a custom knee testing system that allowed the femur to be locked in position for knee extension or flexion. Patellar positions of eversion, reduction, and following repair of the arthrotomy were examined. The influence of quadriceps muscle load was investigated by varying the quadriceps load from 0 to 125 N. The lateral and medial joint gaps, represented by the distance from the implanted femoral component surface to the cut tibia surface, were measured with 100 N tibial distraction force using a 3D digitizer in both extension (0°) and flexion (90°).

Findings

Both the medial and lateral joint gaps with patella eversion were significantly smaller than those with patellar reduction and arthrotomy repair (extension: all quadriceps loads, P < 0.0002; flexion: quadriceps loads less than 75 N, P < 0.0002). In patella eversion, quadriceps loading decreased the lateral joint gap more than the medial joint gap in both extension and flexion; however, the effect was greater in knee flexion with significant differences seen at all quadriceps loads, whereas in extension significant differences were only seen for quadriceps loading of 75 N and greater. Patella eversion also caused a lateral-posterior shift and external rotation of the tibia compared to the other conditions (P < 0.005). With patella reduction and repair of the arthrotomy lower quadriceps loading decreased the extension gap significantly more than the flexion gap (P < 0.01). Following repair of the arthrotomy higher quadriceps loading significantly decreased the flexion gap more than the extension gap (P < 0.04).

Interpretation

The patellar positioning and quadriceps muscle loading in total knee arthroplasty have a strong influence on intraoperative joint gap measurements.     

Restoration of Smad4 in BxPC3 Pancreatic Cancer Cells Attenuates Proliferation without Altering Angiogenesis

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive human malignancy in which the transforming growth factor beta (TGF-β) signal transducer, Smad4, is commonly mutated or deleted. BxPC3 human pancreatic cancer cells exhibit a homozygous deletion of the Smad4 gene, yet are growth inhibited by TGF-β1. In the present study, we sought to determine whether reintroduction of Smad4 into BxPC3 cells alters their behavior in vitro and in vivo. Sham transfected and Smad4 expressing BxPC3 cells exhibited similar responses to TGF-β1 with respect to p21 upregulation, hypophosphorylation of the RB protein, Smad2 phosphorylation, and Smad2/3 nuclear translocation. TGF-β1 did not alter p27 expression, and silencing of p21 with an appropriate siRNA markedly attenuated TGF-β1-mediated growth inhibition. Nonetheless, the presence of Smad4 was associated in vitro with a more prolonged doubling time, enhanced sensitivity to the growth inhibitory actions of exogenous TGF-β1, and a more flattened cellular morphology. In vivo, Smad4 expression resulted in delayed tumor growth and decreased cellular proliferation, without effects on either apoptosis or angiogenesis. These findings indicate that, in spite of the absence of Smad4, growth inhibition in BxPC3 cells by TGF-β1 is dependent on p21 upregulation and maintenance of RB in a hypophosphorylated, active state. Moreover, the presence of a functional Smad4 attenuates the capacity of BxPC3 cells to proliferate in vivo. However, this effect is transient, indicating that Smad4 growth inhibitory actions are circumvented in the later stages of pancreatic tumorigenicity.