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Covering: 1970 through June of 2023Verticillins are epipolythiodioxopiperazine (ETP) alkaloids, many of which possess potent, nanomolar-level cytotoxicity against a variety of cancer cell lines. Over the last decade, their in vivo activity and mode of action have been explored in detail. Notably, recent studies have indicated that these compounds may be selective inhibitors of histone methyltransferases (HMTases) that alter the epigenome and modify targets that play a crucial role in apoptosis, altering immune cell recognition, and generating reactive oxygen species. Verticillin A (1) was the first of 27 analogues reported from fungal cultures since 1970. Subsequent genome sequencing identified the biosynthetic gene cluster responsible for producing verticillins, allowing a putative pathway to be proposed. Further, molecular sequencing played a pivotal role in clarifying the taxonomic characterization of verticillin-producing fungi, suggesting that most producing strains belong to the genus Clonostachys ( i.e. , Bionectria ), Bionectriaceae. Recent studies have explored the total synthesis of these molecules and the generation of analogues via both semisynthetic and precursor-directed biosynthetic approaches. In addition, nanoparticles have been used to deliver these molecules, which, like many natural products, possess challenging solubility profiles. This review summarizes over 50 years of chemical and biological research on this class of fungal metabolites and offers insights and suggestions on future opportunities to push these compounds into pre-clinical and clinical development.

Objective: The contribution of somatic variants to epilepsy has recently been demonstrated, particularly in the etiology of malformations of cortical development. The aim of this study was to determine the diagnostic yield of somatic variants in genes that have been previously associated with a somatic or germline epilepsy model, ascertained from resected brain tissue from patients with multidrug-resistant focal epilepsy., Methods: Forty-two patients were recruited across three categories: (1) malformations of cortical development, (2) mesial temporal lobe epilepsy with hippocampal sclerosis, and (3) nonlesional focal epilepsy. Participants were subdivided based on histopathology of the resected brain. Paired blood- and brain-derived DNA samples were sequenced using high-coverage targeted next generation sequencing to high depth (585× and 1360×, respectively). Variants were identified using Genome Analysis ToolKit (GATK4) MuTect-2 and confirmed using high-coverage Amplicon-EZ sequencing., Results: Sequence data on 41 patients passed quality control. Four somatic variants were validated following amplicon sequencing: within CBL, ALG13, MTOR, and FLNA. The diagnostic yield across 41 patients was 10%, 9% in mesial temporal lobe epilepsy with hippocampal sclerosis and 20% in malformations of cortical development., Significance: This study provides novel insights into the etiology of mesial temporal lobe epilepsy with hippocampal sclerosis, highlighting a potential pathogenic role of somatic variants in CBL and ALG13. We also report candidate diagnostic somatic variants in FLNA in focal cortical dysplasia, while providing further insight into the importance of MTOR and related genes in focal cortical dysplasia. This work demonstrates the potential molecular diagnostic value of variants in both germline and somatic epilepsy genes., (© 2024 The Authors. Epilepsia published by Wiley Periodicals LLC on behalf of International League Against Epilepsy.)

Thoracic endovascular aortic repair (TEVAR) is a minimally invasive treatment for thoracic aortic conditions including aneurysms and is associated with a number of postoperative stent graft related complications. Computational simulations of TEVAR have the potential to predict surgical outcomes and complications preoperatively. When using simulations for stent graft design and prediction of complications in a population, it is difficult to generalize patient-specific TEVAR computational models due to patient variability. This study proposes a novel modeling framework for creating realistic population-based computational models of TEVAR focused on aneurysms that allow for developing various clinically relevant geometric configurations and scenarios that are not easily attainable with limited patient data. The framework includes a methodology for developing population-based thoracic aortic geometries and defining age-dependent aortic tissue material models, as well as detailed steps and boundary conditions for finite element modeling of stent graft deployment during TEVAR. The simulation framework is illustrated for predicting the formation of a bird-beak configuration, a wedge-shaped gap at the proximal end of the deployed stent graft in TEVAR that leads to incomplete seal. A baseline TEVAR simulation model was developed along with three simulations in which the value of aortic curvature, aortic arch angle, or aortic tissue properties varied from the baseline model. Analyzing the length and angle of the bird-beak configuration in each case shows that the bird-beak size is sensitive to different values of the aortic geometry highlighting the importance of using realistic parameter values., (© 2022 John Wiley & Sons Ltd.)

Introduction: Flame retardants (FRs) are common bodily and environmental pollutants, creating concern about their potential toxicity. We and others have found that the commercial mixture FireMaster® 550 (FM 550) or its individual brominated (BFR) and organophosphate ester (OPFR) components are potential developmental neurotoxicants. Using Wistar rats, we previously reported that developmental exposure to FM 550 or its component classes produced sex- and compound-specific effects on adult socioemotional behaviors. The underlying mechanisms driving the behavioral phenotypes are unknown., Methods: To further mechanistic understanding, here we conducted transcriptomics in parallel with a novel lipidomics approach using cortical tissues from newborn siblings of the rats in the published behavioral study. Inclusion of lipid composition is significant because it is rarely examined in developmental neurotoxicity studies. Pups were gestationally exposed via oral dosing to the dam to FM 550 or the BFR or OPFR components at environmentally relevant doses., Results: The neonatal cortex was highly sexually dimorphic in lipid and transcriptome composition, and males were more significantly impacted by FR exposure. Multiple adverse modes of action for the BFRs and OPFRs on neurodevelopment were identified, with the OPFRs being more disruptive than the BFRs via multiple mechanisms including dysregulation of mitochondrial function and disruption of cholinergic and glutamatergic systems. Disrupted mitochondrial function by environmental factors has been linked to a higher risk of autism spectrum disorders and neurodegenerative disorders. Impacted lipid classes included ceramides, sphingomyelins, and triacylglycerides. Robust ceramide upregulation in the OPFR females could suggest a heightened risk of brain metabolic disease., Conclusions: This study reveals multiple mechanisms by which the components of a common FR mixture are developmentally neurotoxic and that the OPFRs may be the compounds of greatest concern., (© 2022 S. Karger AG, Basel.)

Landfill leachate and municipal wastewater are major sources of chemical pollutants that contaminate our drinking water sources. Evaluating the dissolved organic chemical composition in wastewater treatment plants is therefore essential to understand how the discharge impacts the environment, wildlife, and human health. In this study, we utilized a nontargeted analysis method coupling liquid chromatography and tandem mass spectrometry (LC-MS/MS) to analyze chemical features at different points along two landfill leachate treatment plants (LLTPs) and two municipal wastewater treatment plants (WWTPs) in the Southeastern United States. Significant feature differences were observed for the WWTPs where activated sludge clarification was employed versus the LLTPs utilizing reverse osmosis. Specifically, even though both LLTPs had the largest number of features in their influent water, their effluent following reverse osmosis yielded a lower number of features than the WWTPs. Additionally, the clarification processes of each WWTP exhibited different efficiencies as chemical disinfection removed more features than UV disinfection. Feature identification was then made using the LC, MS, and MS/MS information. Analysis of the identified molecules showed that lipids were the most effectively removed from all plants, while alkaloid and organic nitrogen compounds were the most recalcitrant., Competing Interests: The authors declare no competing financial interest.

Background and Objectives: KCNC2 encodes Kv3.2, a member of the Shaw-related (Kv3) voltage-gated potassium channel subfamily, which is important for sustained high-frequency firing and optimized energy efficiency of action potentials in the brain. The objective of this study was to analyze the clinical phenotype, genetic background, and biophysical function of disease-associated Kv3.2 variants., Methods: Individuals with KCNC2 variants detected by exome sequencing were selected for clinical, further genetic, and functional analysis. Cases were referred through clinical and research collaborations. Selected de novo variants were examined electrophysiologically in Xenopus laevis oocytes., Results: We identified novel KCNC2 variants in 18 patients with various forms of epilepsy, including genetic generalized epilepsy (GGE), developmental and epileptic encephalopathy (DEE) including early-onset absence epilepsy, focal epilepsy, and myoclonic-atonic epilepsy. Of the 18 variants, 10 were de novo and 8 were classified as modifying variants. Eight drug-responsive patients became seizure-free using valproic acid as monotherapy or in combination, including severe DEE cases. Functional analysis of 4 variants demonstrated gain of function in 3 severely affected DEE cases and loss of function in 1 case with a milder phenotype (GGE) as the underlying pathomechanisms., Discussion: These findings implicate KCNC2 as a novel causative gene for epilepsy and emphasize the critical role of K V 3.2 in the regulation of brain excitability., (Copyright © 2022 The Author(s). Published by Wolters Kluwer Health, Inc. on behalf of the American Academy of Neurology.)

Fontan circulations are surgical strategies to treat infants born with single ventricle physiology. Clinical and mathematical definitions of Fontan failure are lacking, and understanding is needed of parameters indicative of declining physiologies. Our objective is to develop lumped parameter models of two-ventricle and single-ventricle circulations. These models, their mathematical formulations and a proof of existence of periodic solutions are presented. Sensitivity analyses are performed to identify key parameters. Systemic venous and systolic left ventricular compliances and systemic capillary and pulmonary venous resistances are identified as key parameters. Our models serve as a framework to study the differences between two-ventricle and single-ventricle physiologies and healthy and failing Fontan circulations., (© The Author(s) 2021. Published by Oxford University Press on behalf of the Institute of Mathematics and its Applications. All rights reserved.)

Objective: Psychoses affecting people with epilepsy increase disease burden and diminish quality of life. We characterized postictal psychosis, which comprises about one quarter of epilepsy-related psychoses, and has unknown causation., Methods: We conducted a case-control cohort study including patients diagnosed with postictal psychosis, confirmed by psychiatric assessment, with available data regarding epilepsy, treatment, psychiatric history, psychosis profile, and outcomes. After screening 3,288 epilepsy patients, we identified 83 with psychosis; 49 had postictal psychosis. Controls were 98 adults, matched by age and epilepsy type, with no history of psychosis. Logistic regression was used to investigate clinical factors associated with postictal psychosis; univariate associations with a p value < 0.20 were used to build a multivariate model. Polygenic risk scores for schizophrenia were calculated., Results: Cases were more likely to have seizure clustering (odds ratio [OR] = 7.59, p < 0.001), seizures with a recollected aura (OR = 2.49, p = 0.013), and a family history of psychiatric disease (OR = 5.17, p = 0.022). Cases showed predominance of right temporal epileptiform discharges (OR = 4.87, p = 0.007). There was no difference in epilepsy duration, neuroimaging findings, or antiseizure treatment between cases and controls. Polygenic risk scores for schizophrenia in an extended cohort of postictal psychosis cases (n = 58) were significantly higher than in 1,366 epilepsy controls (R 2  = 3%, p = 6 × 10 -3 ), but not significantly different from 945 independent patients with schizophrenia (R 2  = 0.1%, p = 0.775)., Interpretation: Postictal psychosis occurs under particular circumstances in people with epilepsy with a heightened genetic predisposition to schizophrenia, illustrating how disease biology (seizures) and trait susceptibility (schizophrenia) may interact to produce particular outcomes (postictal psychosis) in a common disease. ANN NEUROL 2021;90:464-476., (© 2021 The Authors. Annals of Neurology published by Wiley Periodicals LLC on behalf of American Neurological Association.)

The verticillins, a class of epipolythiodioxopiperazine alkaloids (ETPs) first described 50 years ago with the discovery of verticillin A ( 1 ), have gained attention due to their potent activity against cancer cells, noted both in vitro and in vivo. In this study, the complex scaffold afforded through optimized fermentation was used as a feedstock for semisynthetic efforts designed to explore the reactivity of the C11 and C11' hydroxy substituents. Functionality introduced at these positions would be expected to impact not only the potency but also the pharmacokinetic properties of the resulting compound. With this in mind, verticillin H ( 2 ) was used as a starting material to generate nine semisynthetic analogues ( 4 - 12 ) containing a variety of ester, carbonate, carbamate, and sulfonate moieties. Likewise, verticillin A succinate ( 13 ) was synthesized from 1 to demonstrate the successful application of this strategy to other ETPs. The synthesized compounds and their corresponding starting materials (i.e., 1 and 2 ) were screened for activity against a panel of melanoma, breast, and ovarian cancer cell lines: MDA-MB-435, MDA-MB-231, and OVCAR3. All analogues retained IC 50 values in the nanomolar range, comparable to, and in some cases more potent than, the parent compounds., Competing Interests: The authors declare the following competing financial interest(s): Nicholas Oberlies is a member of the Scientific Advisory Board of Mycosynthetix, Inc., (© 2021 American Chemical Society.)

Cells sense and respond to the heterogeneous mechanical properties of their tissue microenvironment, with implications for the development of many diseases, including cancer, fibrosis, and aortic valve disease. Characterization of tissue mechanical heterogeneity on cellular length scales of tens of micrometers is thus important for understanding disease mechanobiology. In this study, we developed a low-cost bench-top microindentation system to readily map focal microscale soft tissue mechanical properties. The device was validated by comparison with atomic force microscopy nanoindentation of polyacrylamide gels. To demonstrate its utility, the device was used to measure the focal microscale elastic moduli of normal and diseased porcine aortic valve leaflet tissue. Consistent with previous studies, the fibrosa layer of intact leaflets was found to be 1.91-fold stiffer than the ventricularis layer, with both layers exhibiting significant heterogeneity in focal elastic moduli. For the first time, the microscale compressive moduli of focal proteoglycan-rich lesions in the fibrosa of early diseased porcine aortic valve leaflets were measured and found to be 2.44-fold softer than those of normal tissue. These data provide new insights into the tissue micromechanical environment in valvular disease and demonstrate the utility of the microindentation device for facile measurement of the focal mechanical properties of soft tissues., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Fontan associated liver disease is a common complication in patients with Fontan circulation, who were born with a single functioning heart ventricle. The hepatic venous pressure gradient (HVPG) is used to assess liver health and is a surrogate measure of the pressure gradient across the entire liver (portal pressure gradient (PPG)). However, it is thought to be inaccurate in Fontan patients. The main objectives of this study were (1) to apply an existing detailed lumped parameter model (LPM) of the liver to Fontan patients using patient-specific clinical data and (2) to determine whether HVPG is a suitable measurement of PPGs in these patients. An existing LPM of the liver blood circulation was applied and tuned to simulate patient-specific liver hemodynamics. Geometries were collected from seven adult Fontan patients and used to evaluate model parameters. The model was solved and tuned using waveform measurements of flows, inlet and outlet pressures. The predicted ratio of portal to hepatic venous pressures is comparable to in vivo measurements. The results confirmed that HVPG is not suitable for Fontan patients, as it would underestimate the portal pressures gradient by a factor of 3 to 4. Our patient-specific liver model provides an estimate of the pressure drop across the liver, which differs from the clinically used metric HVPG. This work represents a first step toward models suitable to assess liver health in Fontan patients and improve its long-term management., (Copyright © 2021 by ASME.)

Background: Although morphologic abnormalities in the liver are commonly encountered post Fontan palliation, the relationships between hepatic morphology, vascular flows, and clinical status remain incompletely understood. We therefore aimed to explore flow characteristics in hepatic and intestinal vessels and to examine cardiovascular associations with liver disease., Methods: This was a retrospective study of adults post Fontan palliation undergoing clinically indicated cardiovascular magnetic resonance imaging (MRI). Patients were included if MRI flow quantification was available for cardiac, hepatic and intestinal vessels; patients were excluded if phase-contrast flow imaging was insufficient for analysis., Results: Thirty patients were studied (median age at MRI 28.5 years [range 19-47]). Eighteen subjects (60%) were classified as having morphologic liver disease according to validated criteria based on available MRI imaging. Abdominal and cardiovascular flows were quantified. Patients with morphologic liver disease had a 41% reduction in superior mesenteric artery (211 ± 124 versus 358 ± 181 mL/min/m 2 , p = .004), a 36% reduction in hepatic vein (496 ± 247 versus 778 ± 220 mL/min/m 2 , p = .01), a 31% reduction in portal vein (399 ± 133 versus 580 ± 159 mL/min/m 2 , p = .004), and an 18% reduction in Fontan pathway flows (1358 ± 429 versus 1651 ± 270 mL/min/m 2 , p = .04) compared with the remaining population. Adverse cardiovascular events were not associated with morphologic liver disease., Conclusion: Morphologic liver disease appears to be associated with flow alterations within the heart, liver and intestine post Fontan palliation. These novel observations suggest that a potential relationship exists between morphologic disease and vascular flows thereby providing further insights into the pathophysiology of liver disease in this high-risk population., Competing Interests: Declaration of Competing Interest None., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Purpose: To develop a mechanically realistic aortoiliac model to evaluate anatomic variables associated with stent-graft rotation and to assess common deployment techniques that may contribute to rotation. Materials and Methods: Idealized aortoiliac geometries were constructed either through direct 3-dimensional (3D) printing (rigid) or through casting with polyvinyl alcohol using 3D-printed molds (flexible). Flexible model bending rigidity was controlled by altering wall thickness. Three flexible patient-specific models were also created based on the preoperative computed tomography angiograms. Zenith infrarenal and fenestrated devices were used in this study. The models were pressurized to 100 mm Hg with normal saline. Deployments were performed under fluoroscopy at 37°C. Rotation was calculated by tracking the change in position of gold markers affixed to the devices. Results: In the rigid idealized models, stent-graft rotation increased with increasing torsion; torsion levels of 1.6, 2.6, and 3.6 mm -1 had mean rotations of 5.2°±0.03°, 11.2°±4.8°, and 27.6°±13.0°, respectively (p<0.001). In the flexible models, the highest rotation (58°±3.0°) was observed in models with high torsion and high rigidity (7.5 mm -1 net torsion and 254 N·m 2 flexural rigidity). No rotation was observed in the absence of torsion. Applying torque to the device during insertion significantly increased stent-graft rotation by an average of 28° across all levels of torsion (p<0.01). Multiple device insertions prior to deployment did not change the observed device rotation. The patient-specific models accurately predicted the degree of rotation seen intraoperatively to within 5°. Conclusion: Insertion technique plays an important role in the degree of stent-graft rotation during deployment. Our model suggests that in vivo correction of device orientation can increase the observed rotation and supports the concept of fully removing the device, adjusting the orientation, and subsequently reinserting. Additionally, increasing iliac artery torsion in the presence of increased vessel rigidity results in stent-graft rotation.

Objectives: Adult patients who have undergone the Fontan procedure are highly vulnerable to gradual, progressive circulatory failure, and options to reverse this situation are few. A cavopulmonary assist device could decongest the venous and lymphatic systems, overcome elevated pulmonary vascular resistance, increase cardiac output, and support some of these patients to heart transplant. This study characterizes the performance and challenges of a novel multilumen cannula coupled to an external blood pump proposed as a potential Fontan cavopulmonary assist strategy., Methods: Computational fluid dynamic simulations were conducted for 3 extracardiac Fontan geometries consisting of 1 idealized model and 2 patient-specific models. A range of physiologic flow rates and pump assist levels were simulated to calculate the pressure gain provided by the multilumen cannula. Hemolysis index was estimated for the idealized model with Lagrangian particle tracking and 2 variations of the power-law. Wall shear stresses were also examined., Results: Pressure gains up to 4 and 9 mm Hg were achieved for the idealized and patient-specific models, respectively. Pressure gains increased with both higher cardiac output and larger pump intake through the external pump. Flow-weighted hemolysis show hemoglobin damage levels to be several times lower than the 2% threshold at the highest pump intake flow cases. Wall shear stress predictions depict elevated areas in the pulmonary vessels and regions of the cannula device., Conclusions: The cannula tested in this study shows promise as a percutaneous option to bridge support in some patients with a failing extracardiac Fontan. Limitations identified will be addressed in future design iterations and in ongoing experimental tests., (Copyright © 2019 The American Association for Thoracic Surgery. Published by Elsevier Inc. All rights reserved.)

Objective: During fenestrated endovascular aneurysm repair (FEVAR), accurate alignment of the fenestration and the target artery is necessary to prevent complications. This study's objective is to determine the incidence of clinical outcomes following fenestration misalignment during FEVAR., Methods: A single-center, retrospective chart review was performed for all elective FEVARs between January 2008 and April 2015. Data were gathered from patient records and intraoperative imaging. Native vessel angles were calculated using the vessel centerlines. Intraoperative stent graft orientation was determined by changing the angle of the image intensifier as the fenestration was profiled for cannulation. Vertical fenestration misalignment was defined as 4 mm or greater and is the distance from the center of the fenestration markers to the center of the target vessel ostium at the time of cannulation. Horizontal stent graft misalignment was defined as a difference between the native vessel angle and the intraoperative fenestration angle of 15° or more. Early and late clinical outcomes were analyzed with respect to the presence of either vertical or horizontal misalignment of the renal artery fenestrations., Results: The study cohort includes 65 patients who underwent FEVAR during this study period. A horizontal misalignment of 15° or more occurred in 40% of patients (n = 26) and 30° or more in 9.2% of patients (n = 6). A vertical misalignment of 4 mm or greater occurred in 32.3% of patients (n = 21). The incidence of severe postoperative complications, defined as any in-hospital end-organ ischemia and/or death, was significantly greater for patients with stent graft misalignment-31% (n = 11) vs 3% (n = 1) in the aligned group. There was a trend toward higher rates of target vessel cannulation failure in patients with stent graft misalignment 3% (n = 99 fenestrations) vs 0% (n = 76 fenestrations). The combined incidence of any intraoperative target vessel complication was significantly higher in patients with misalignment, 8.1% (4, 15) vs 1.3% (0, 8). Long-term survival was significantly lower in patients with stent graft misalignment, which was primarily driven by high intraoperative and in-hospital mortality rates (P < .05)., Conclusions: Intraoperative stent graft misalignment is associated with higher rates of procedural target vessel complications and severe postoperative adverse events. Patients with stent graft misalignment should be considered at high risk for early postoperative complications. These results highlight an important need for improved FEVAR planning., (Copyright © 2019 Society for Vascular Surgery. Published by Elsevier Inc. All rights reserved.)

Introduction: During endovascular aneurysm repair, the iliac artery typically serves as a conduit for device delivery. The degree of tortuosity and calcification in the iliac artery ultimately determines whether the device can successfully traverse the vessel. These 2 parameters can be assessed using qualitative approaches or calculated using quantitative methods based on the Society for Vascular Surgery (SVS) reporting standards. The objective of this study was to determine whether qualitative methods are sufficient to accurately assess iliac artery tortuosity and calcification by calculating interobserver variability and comparing them to the SVS Reporting Standards., Methods: Three vascular surgeons reviewed preoperative computed tomography scans for 50 patients who underwent fenestrated endovascular aneurysm repair and qualitatively assessed left and right iliac artery tortuosity and calcification. Iliac artery geometries were segmented from these image sets. Tortuosity index and calcification length ratio were calculated and categorized based on the SVS Reporting Standards., Results: Interobserver variability was calculated for the qualitative assessments using interclass correlation coefficients. For tortuosity index, among the 3 observers, good agreement was found for the left iliac artery and fair agreement was found for the right. For calcification length ratio, excellent agreement was found for both iliac arteries. When compared to the quantitative assessment, the qualitative assessments underpredicted tortuosity in 2.3% of cases, matched the quantitative values in 16.7% of cases, and overpredicted tortuosity in 81.0% of cases. The qualitative assessments underpredicted calcification in 46.3% of cases, matched the quantitative values in 49.3% of cases, and overpredicted calcification in 4.3% of cases., Conclusion: Qualitative assessment of iliac artery tortuosity showed fair-to-good interobserver agreement and poor agreement to SVS Reporting Standards. Qualitative assessment of iliac artery calcification showed excellent interobserver agreement and fair agreement to SVS Reporting Standards. These trends should be considered when qualitative reporting methodologies are used.

Fenestrated endovascular aneurysm repair (FEVAR) is a minimally invasive method of abdominal aortic aneurysm (AAA) repair utilized in patients with complex vessel anatomies. Stent grafts (SG) used in this process contain fenestrations within the device that need to be aligned with the visceral arteries upon successful SG deployment. Proper alignment is crucial to maintain blood flow to these arteries and avoid surgical complications. During fenestrated SG deployment, rotation of the SG can occur during the unsheathing process. This leads to misalignment of the vessels, and the fenestrations and is associated with poor clinical outcomes. The aim of this study was to develop a computational model of the FEVAR process to predict SG rotation. Six patient-specific cases are presented and compared with surgical case data. Realistic material properties, frictional effects, deployment methods, and boundary conditions are included in the model. A mean simulation error of 2 deg (range 1-4 deg) was observed. This model was then used to conduct a parameter study of frictional properties to see if rotation could be minimized. This study showed that increasing or decreasing the coefficients of friction (COF) between the sheath and the vessel walls would decrease the amount of rotation observed. Our model accurately predicts the amount of SG rotation observed during FEVAR and can be used as a preoperative planning tool within the surgical workflow.

Objective: Advanced endovascular aneurysm repair (EVAR) with fenestrated and branched stent grafts is increasingly being used to repair complex aortic aneurysms; however, these devices can rotate unpredictably during deployment, leading to device misalignment. The objectives of this study were to quantify the short-term clinical outcomes in patients with intraoperative stent graft rotation and to identify quantitative anatomic markers of the arterial geometry that can predict stent graft rotation preoperatively., Methods: A prospective study evaluating all patients undergoing advanced EVAR was conducted at two university-affiliated hospitals between November 2015 and December 2016. Stent graft rotation (defined as ≥10 degrees) was measured on intraoperative fluoroscopic video of the deployment sequence. Standard preoperative computed tomography angiography imaging was used to calculate the geometric properties of the arterial anatomy. Any in-hospital and 30-day complications were prospectively documented, and a composite outcome of any end-organ ischemia or death was used as the primary end point., Results: Thirty-nine patients undergoing advanced EVAR were enrolled in the study with a mean age of 75 years (interquartile range [IQR], 71-80 years) and a mean aneurysm diameter of 64 mm (IQR, 59-65 mm). The incidence of stent graft rotation was 37% (n = 14), with a mean rotation of 25 degrees (IQR, 21-28 degrees). A nominal logistic regression model identified iliac artery torsion, volume of iliac artery calcification, and stent graft length as the primary predictive factors. The total net torsion and the total volume of calcific plaque were higher in patients with stent graft rotation, 8.9 ± 0.8 mm -1 vs 4.1 ± 0.5 mm -1 (P < .0001) and 1054 ± 144 mm 3 vs 525 ± 83 mm 3 (P < .01), respectively. The length of the implanted stent grafts was also higher in patients with intraoperative rotation, 172 ± 9 mm vs 156 ± 8 mm (P < .01). The composite outcome of any end-organ ischemia or death was also substantially higher in patients with stent graft rotation (36% vs 0%; P = .004). In addition, patients with stent graft rotation had significantly higher combined rates of type Ib and type III endoleaks (43% vs 8%; P = .03)., Conclusions: Patients with intraoperative stent graft rotation have a significantly higher rate of severe postoperative complications, and this is strongly associated with higher levels of iliac artery torsion, calcification, and stent graft length. These findings suggest that preoperative quantitative analysis of iliac artery torsion and calcification may improve risk stratification of patients before advanced EVAR., (Copyright © 2017 Society for Vascular Surgery. Published by Elsevier Inc. All rights reserved.)

Native and engineered tissue development are regulated by the integrative effects of multiple microenvironmental stimuli. Microfabricated bioreactor array platforms can efficiently dissect cue-response networks, and have recently integrated critical 2D and 3D mechanical stimulation for greater physiological relevance. However, a limitation of these approaches is that assessment of tissue functional properties is typically limited to end-point analyses. Here we report a new deformable membrane platform with integrated strain sensors that enables mechanical stretching or compression of 3D cell-hydrogel arrays and simultaneous measurement of hydrogel construct stiffness in situ. We tested the ability of the integrated strain sensors to measure the evolution of the stiffness of cell-hydrogel constructs for two cases. First, we demonstrated in situ stiffness monitoring of degradable poly (ethylene glycol)-norbornene (PEG-NB) hydrogels embedded with mesenchymal stromal cells (MSCs) and cultured with or without cyclic tensile stimulation for up to 15 days. Whereas statically-cultured hydrogels degraded and softened throughout the culture period, mechanically-stimulated gels initially softened and then recovered their stiffness corresponding to extensive cell network and collagen production. Second, we demonstrated in situ measurement of compressive stiffening of MSC-seeded PEG-NB gels cultured statically under osteogenic conditions, corresponding to increased mineralization and cellularization. This measurement technique can be generalized to other relevant bioreactor and organ-on-a-chip platforms to facilitate online, non-invasive, and high-throughput functional analysis, and to provide insights into the dynamics of engineered tissue development that are otherwise not available., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Background: Endovascular aneurysm repair is a minimally-invasive method for the treatment of abdominal aortic aneurysms. For aneurysms that involve the visceral arteries, a custom-made stent graft with fenestrations for the branch arteries is required. The purpose of the current study is to evaluate the structural impact of misaligned fenestrations with respect to luminal patency and proximal aortic neck apposition in an in vitro model., Methods: A custom apparatus was used to evaluate seven Anaconda and three Zenith fenestrated stent grafts. All stent grafts were evaluated at 10° increments of stent/fenestration misalignment up to 80°. Images were captured at each interval and the luminal cross-sectional area and wall apposition were measured., Results: The Anaconda stent graft, which has an unsupported main body, demonstrated a linear reduction in luminal patency at increasing angles of misalignment (P < 0.0001). Stent/fenestration misalignments of 20° and 80° resulted in decreases in mean luminal patency of 14% and 54% respectively. The Zenith stent graft demonstrated a similar decrease in luminal patency, starting at misalignments of ≥40° (P < 0.0001). However, with stent/fenestration misalignments of ≥30°, apposition between the Zenith stent graft and the simulated aortic neck was compromised suggesting the creation of a type Ia endoleak., Conclusions: Both the Anaconda and Zenith devices behave adversely at extreme angles of misalignment with luminal narrowing in the Anaconda device and loss of wall apposition in the Zenith device; however, both stent grafts appear to be equivalent at low angles of misalignment., (Copyright © 2016 Royal College of Surgeons of Edinburgh (Scottish charity number SC005317) and Royal College of Surgeons in Ireland. Published by Elsevier Ltd. All rights reserved.)

Introduction: A complication of fenestrated endovascular aneurysm repair is the potential for stent graft rotation during deployment causing fenestration misalignment and branch artery occlusion. The objective of this study is to demonstrate that this rotation is caused by a buildup of rotational energy as the device is delivered through the iliac arteries and to quantify iliac artery geometric properties associated with device rotation., Methods: A retrospective clinical study was undertaken in which iliac artery geometric properties were assessed from preoperative imaging for 42 cases divided into 2 groups: 27 in the nonrotation group and 15 in the rotation group. Preoperative computed tomography scans were segmented, and the iliac artery centerlines were determined. Iliac artery tortuosity, curvature, torsion, and diameter were calculated from the centerline and the segmented vessel geometry., Results: The total iliac artery net torsion was found to be higher in the rotation group compared to the nonrotation group (23.5 ± 14.7 vs 14.6 ± 12.8 mm -1 ; P = .05). No statistically significant differences were found for the mean values of tortuosity, curvature, torsion, or diameter between the 2 groups., Conclusion: Stent graft rotation occurred in 36% of the cases considered in this study. Cases with high iliac artery total net torsion were found to be more likely to have stent graft rotation upon deployment. This retrospective study provides a framework for prospectively studying the influence of iliac artery geometric properties on fenestrated stent graft rotation.

Objective: In situ fenestration of endovascular stent grafts has been used as a method for branch vessel revascularization in urgent and emergent settings. The objective of this manuscript was to review the clinical and experimental evidence related to this technique., Methods: PubMed, MEDLINE, and Embase databases were searched for papers published until December 2015 describing in situ fenestration of aortic stent grafts. Benchtop, animal, and human studies were included., Results: The literature review identified 118 articles, of which 28 studies were selected for inclusion. These included 16 clinical papers (2 case series and 14 case reports) reporting in situ fenestration of 46 aortic branch vessels in 44 patients. There were 42 retrograde and 4 antegrade instances of in situ fenestration. The most frequent target vessel for in situ fenestration was the left subclavian artery (72%), and the most frequent indication for stent graft implantation was a degenerative aortic aneurysm (43%). Technical success was reported in 44 of 46 attempted fenestrations (96%). The combined rate of perioperative mortality, stroke, and paralysis was 7%. In situ fenestration was predominantly performed with the Talent (Medtronic, Santa Rosa, Calif) stent graft (54%), followed by the Zenith (Cook Medical, Bloomington, Ind) stent graft (37%) and the TAG (W. L. Gore & Associates, Newark, Del) stent graft (9%). In vitro benchtop evaluations of in situ fenestration showed minimal change in fenestration size after 1 year of pulsatile fatigue testing. The use of energy-based fenestration techniques (radiofrequency or laser) has been associated with less fabric fraying than in needle-based techniques. The larger caliber initial fenestration created by these devices also avoids the need for cutting balloons, which have also been linked with increased fabric tears and fraying of the fibers surrounding the fenestration. In addition, the Zenith stent graft was shown in benchtop testing to be the strongest in postfenestration mechanical testing, but it was also the most resistant to balloon dilation., Conclusions: In the short to moderate term, in situ fenestration appears to be a reasonable and effective method to extend the proximal landing zone for revascularization of the left subclavian artery. However, longer follow-up is needed to fully assess the long-term durability of this procedure. Based on studies of material properties, an energy-based fenestration technique (radiofrequency or laser) is recommended, along with the avoidance of cutting balloons for dilation of the fenestration., (Copyright © 2016 Society for Vascular Surgery. Published by Elsevier Inc. All rights reserved.)

Endovascular aneurysm repair (EVAR) is a minimally invasive method for the treatment of abdominal aortic aneurysms; however, the implementation of this technique is often limited by the aortic pathology, especially in the urgent or emergent setting. An 82-year-old male with a 7.3 cm symptomatic juxtarenal aneurysm presented at our centre for assessment. He was diagnosed as a high-risk candidate for open repair and therefore, not suitable for a conventional EVAR. Fortunately, a custom two-vessel fenestrated stent graft, which was originally constructed for another patient, was available. This device was implanted with no complications and all branches remain unobstructed; clear of aneurysms at 1 year. We present the use of 'in-stock' fenestrated grafts as a potential option to be considered in the urgent or emergent repair of abdominal aortic aneurysms., (2016 BMJ Publishing Group Ltd.)

Objective: Experimentally measured pullout forces for stent grafts (SGs) are used in clinical discussions and as reference values in bench studies and computer simulations. Previous values of these forces are available from studies in which the SG was pulled out in the straight caudal direction. However, clinical and numerical studies have suggested that displacement forces acting on SGs are directed more anteriorly. The objective of this study was to measure pullout forces as a function of angulation and to test the hypothesis that pullout forces decrease with increasing angulation., Methods: Six different SGs (Bolton Treovance, Cook Zenith Flex, Cook Zenith LP, Medtronic Endurant, Medtronic Talent, and Vascutek Anaconda) were deployed in fresh bovine aortas, then pulled out by an electronic motor at 1 mm/s, while tension force was measured continuously with a digital load cell. The SG off-axis angulation was changed from 0 to 90 degrees in increments of 10 degrees. The test system was submerged in a custom-built temperature-controlled saline bath at 37°C. At least three tests were performed for each device at each angle (with the exception of the Cook Zenith Flex, which experienced plastic deformation of its barbs after a single test per device). Each aortic specimen was used only once and then discarded. Hand-sutured graft anastomoses were also tested at 0 degrees to provide a reference value., Results: A total of 374 pullout tests were performed for the SGs and anastomoses. Sixty-four tests were excluded because of failure of the aorta or apparatus before device pullout. The remaining 310 tests showed pullout forces that demonstrated a decrease in the average pullout force for all six devices from 0 to 90 degrees (Bolton Treovance from 39.3 N to 23.9 N; Cook Zenith Flex from 59.8 N to 48.9 N; Cook Zenith LP from 50.3 N to 41.8 N; Medtronic Endurant from 29.9 N to 25.8 N; Medtronic Talent from 6.0 N to 5.5 N; and Vascutek Anaconda from 37.0 N to 30.3 N). For reference, the mean pullout force for the hand-sutured anastomoses was 63 N., Conclusions: This study reports for the first time the change in pullout force with angulation, showing a general pullout force decrease with increasing angle. With a larger number of samples than in previous studies, our results provide updated benchmark data that can be used for clinical discussions, computational and experimental studies, and future device design., (Copyright © 2016 Society for Vascular Surgery. Published by Elsevier Inc. All rights reserved.)

The myocardium of the left ventricle (LV) of the heart comprises layers of muscle fibers whose orientation varies through the heart wall. Because of these fibers, accurate modeling of the myocardium stress-strain behavior requires models that are nonlinear, anisotropic, and time-varying. This article describes the development and testing of a material model of the canine LV myocardium, which will be used in ongoing simulations of the mechanics of the LV with fluid-structure interaction. The model assumes that myocardium deformation has two extreme states: one during which the muscle fibers are fully relaxed, and another during which the muscle fibers are fully contracted. During the second state, the "total" stresses are assumed to be the sum of "passive" stresses, which represent the fully relaxed muscle fibers, and "active" stresses, which are additional stresses due to the contraction of the muscle fibers. The canine LV myocardium is modeled as a transversely isotropic material for which material properties vary in the fiber and cross-fiber directions. The material behavior is considered to be hyperelastic and is modeled by a strain-energy density function in a manner that is an adaptation of an approach based on measurements of the stress-strain behavior of rabbit LV myocardia. A numerical method has been developed to calculate suitable parameter values for the passive material model using previous passive canine LV myocardium stress measurements and taking into account existing physical and numerical constraints. In the absence of published measurements of total canine LV myocardium stresses, a method has been developed to estimate these stresses from available passive and total rabbit LV myocardium stresses and then to calculate active material parameter values. Material parameter values were calculated for passive and active canine LV myocardium. Passive stresses calculated using the model compare well to previous stress measurements while active stresses calculated using the model compare well with those approximated from rabbit measurements. The adapted material model of the canine LV myocardium is deemed to be suitable for use in simulations of the operation of both idealized and realistic canine hearts. The estimated model parameter values can be easily revised to more appropriate ones if measurements of active canine LV myocardium stresses become available. The extension of this material model to a fully orthotropic one is also possible but determination of its parameters would require stress-stretch measurements in the fiber and both cross-fiber directions.

This article describes two ongoing numerical studies of fluid-structure interaction in the cardiovascular system: an idealized pulsatile ventricular assist device (VAD), consisting of two fluid chambers separated by a flexible diaphragm; and blood flow and heart wall motion during passive filling of a canine heart. Simulations have been performed for the VAD and compared with the results of a previous study and to our own preliminary experimental results. Detailed measurements of the flow field in the VAD model and additional simulations are in progress. Preliminary simulations using both an idealized model of the natural heart as well as a realistic model have identified the limitations of the current numerical methods in dealing with large three-dimensional deformations. Ongoing research aims at extending the range of simulations to include large deformations and to incorporate an anisotropic material model for the heart wall to account for the muscle fibers.

Background: Microabrasion using a paste made of acid and pumice is a technique that has been used to remove white, yellow and brown stains from enamel. The authors evaluated the technique by studying the effectiveness of a proprietary microabrasion product., Methods: One author used microabrasion to remove white, yellow and brown stains from within the outermost layer of the tooth enamel of 32 subjects. Standardized slides of the teeth were taken before and one week after treatment. Four prosthodontists evaluated the paired images, using a standardized questionnaire and visual analog scales ranging from 1 (no improvement in appearance or stain not removed at all) to 7 (exceptional improvement in appearance or stain totally removed). The evaluators were calibrated and blinded., Results: The evaluators always identified a difference between the pretreatment slides and posttreatment slides; they found no difference between the control slides. In all cases but one (97 percent), the treated teeth had improved in appearance with more uniformity in color. Analysis of variance revealed no differences between evaluator ratings (P = .146). The intraclass correlation coefficient for ratings of individual cases by different evaluators was 0.72, representing a "good" level of correlation of the ratings for improvement of appearance and for stain removal. Mean (+/- standard deviation) ratings were 5.38 (+/- 1.26) for improvement of appearance and 5.06 (+/- 1.26) for stain removal., Conclusions: This study showed that enamel microabrasion could remove stains from within the outermost layer of tooth enamel, thereby improving the appearance of the teeth., Clinical Implications: This study supports recommendations that enamel microabrasion is an effective, atraumatic method of improving the appearance of teeth with stains in the outermost layer of enamel.

Purpose: This study compared the quantity of preweighed, packaged die stone use to bulk die stone use by dental students., Materials and Methods: Starting in September 1994, all the bulk die stone was removed from the dental clinic, and only preweighed packages of die stone (Silky-Rock, Whip Mix) were available to students. At the end of August 1995 and August 1997, the amount of preweighed die stone used by the students was determined and compared with the amount of bulk die stone (DieKeen, Modern Materials) used during the period of September 1993 to August 1994., Results: Between September 1994 and August 1997, the students used 42% less per annum of the preweighed, packaged die stone than the previous annual use of bulk die stone. The laboratories were much cleaner when the preweighed packages were used, and fewer cleaning staff were required. The environment was also cleaner for the laboratory staff, who used to fill the bulk containers with gypsum products., Conclusions: Students used 42% less die stone per annum when using the preweighed packages.

We determined the prevalence of other sexually transmitted diseases (STDs) in 56 women infected with the human immunodeficiency virus (HIV) and compared the usefulness of this prevalence to that of standard risk assessment alone in identifying HIV-infected women. Of the 56 women studied, 54 were infected with HIV. These women received instruction as to the modes of transmission and were then placed in standard risk groups based on their own assessments. A history of previous or current STD was obtained and followed by appropriate testing. We found that combining standard risk assessment with a current or previous STD enabled us to identify a greater percentage of women who are infected with HIV.

Epstein-Barr virus (EBV) infection of the EBV-negative Burkitt lymphoma (BL) cell line BL2 has been studied in vitro, to assess the efficiency of stable infection and the state of the viral genome in these cells. Expression of the EBV nuclear antigen complex (EBNA 1-6) was detected by 24 h post-infection, but expression of the latent membrane protein (LMP) and of the B cell activation antigen CD23 were delayed until 8 days post-infection, and were then only detectable in a subpopulation of EBNA-positive cells. Separation of cells into CD23-positive and -negative fractions 8 days after infection showed that the CD23-positive cells expressed EBNA 1-6 and LMP and contained viral episomes, whereas the CD23-negative cells were EBNA 1-6 positive, LMP negative and had very low levels of episomal viral genome. Cloning of the CD23-positive and -negative populations showed that EBV infection in the majority of clones was lost, with only 29 out of the 164 investigated still EBNA positive after 6 months of culture. Stable infection correlated with expression of EBNA 1-6, LMP, CD23 and the presence of viral episomes. One clone stably expressed EBNA 1-6 and LMP in the absence of detectable linear or episomal viral genome; this clone was assumed to have a viral genome integrated into the cellular chromosomes. Thus integration of the viral genome following in vitro infection of BL2 cells is a rare event.

The effect of high pressure on the response to glycine or kainate of voltage-clamped Xenopus oocytes micro-injected with messenger-RNA derived from either rat spinal cord or whole brain, respectively, has been investigated. Current responses were measured at 1 bar (= 10(5) Pa), 50 bar, 100 bar and 150 bar, with PO2 fixed at 1 bar and the balance helium. Glycine elicited a depolarizing current response which was antagonized by nanomolar concentrations of strychnine. The responses reversibly desensitized, with a decay constant of 0.01 s-1, when glycine concentrations greater than 250 microM were used. The decay constant was insensitive to both glycine concentration and pressure. Resensitization was complete within 4 min. Kainate elicited a depolarizing current which was non-desensitizing. The response was slightly sensitive to glutamate diethyl ester (50 microM), which increased the EC50 by 25%. The action of glycine was highly pressure sensitive. The dose-response curves established at 50 bar, 100 bar and 150 bar were shifted progressively to the right, with no effect on the maximal current. The EC50 increased from 216 microM to 296 microM at 50 bar, to 345 microM at 100 bar, and to 425 microM at 150 bar. The action of kainate was unaffected by pressure. No shift in the dose-response curves was established, nor was there any effect on the maximum current. The EC50 was 113 microM at 1 bar, and 111 microM at both 50 bar and 100 bar.(ABSTRACT TRUNCATED AT 250 WORDS)

Epstein-Barr virus (EBV) infection of B lymphocytes in vitro gives rise to immortalized lymphoblastoid cell lines. Previous reports have shown that chronic lymphocytic leukaemia (CLL) cells, although infectable by EBV, are resistant to immortalization (1-4), although a small number of CLL cell lines have been reported (5-7). In the present study we have analysed early events occurring after EBV infection in 16 CLL samples. Out of 16 samples, 15 could be infected by the virus and expressed the full EB viral nuclear antigen (EBNA) complex but only one out of 16 expressed the latent membrane protein (LMP). The five CLLs in which we could investigate the presence of viral episomes showed circularized EBV by 16 hours after infection. The sequence of EBNA expression and genome circularization mirrored that seen in normal B cells, although genome amplification was not detected. The only CLL sample which expressed LMP after EBV infection was induced to proliferate for 2-3 weeks, but no cell line was established. Immortalized cell lines were obtained from three out of 16 samples tested, but all were polyclonal for light chain expression and had arisen from the CD5-negative, normal B-cell population. Thus the inability of EBV to induce proliferation of most CLL cells correlated with the absence of LMP expression which is invariably expressed during immortalization of normal B cells. This novel type of restricted gene expression could be compatible with evasion of host immune responses and consequent long-term survival of the cell in vivo.

This study was performed to determine the prevalence, serotypes and antibiotic susceptibility patterns of penicillin-resistant Streptococcus pneumoniae in children younger than 3 years of age in day-care centers in Houston, TX. Nasopharyngeal cultures were obtained on two occasions, in March and May, 1989, from 140 children in 4 day-care centers. All penicillin-resistant S. pneumoniae organisms isolated in this study had minimum inhibitory concentrations to penicillin of between 0.1 and 0.5 microgram/ml and were thus intermediately resistant. No highly resistant S. pneumoniae (minimum inhibitory concentration > or = 1.0 microgram/ml) was isolated in this study. Nasal carriage of S. pneumoniae occurred in 39% of children; carriage of intermediately resistant S. pneumoniae occurred in 4% of children. Of the 39% of children who carried S. pneumoniae, 11% carried intermediately resistant strains. In one day-care center with a prior history of intermediately resistant S. pneumoniae (Center 1), the prevalence of intermediate penicillin resistance was significantly (P = 0.047) higher than in the other three centers. Among children surveyed twice 15% of Center 1 children carried an intermediately penicillin-resistant strain at least once, whereas in the other centers 3% of children carried an intermediately resistant strain at least once. Sixty-two percent of intermediately penicillin-resistant strains were resistant to multiple antibiotics and all were serotype 14. Intermediately penicillin-resistant S. pneumoniae isolates were prevalent among young children in day-care centers in Houston and may persist in some day-care centers and become endemic.

The dimensions of 120 maxillary first premolar all-ceramic tooth preparation designs were measured and analyzed to determine relationships between design features and recorded differences in the strengths of restorations fabricated for each preparation design. To assess clinical feasibility, the remaining dentin thickness of each preparation design was measured. A total occlusal convergence angle of 10 degrees provided the best combination of restoration strength and remaining dentin thickness. Five degrees produced the weakest restorations, and 15 degrees and 20 degrees required greater tooth reduction and pulp proximity. Stronger restorations were noted when the shoulder finish line had a sharp axiogingival line angle, possessed minimal cervical inclination, and was located as close as possible to the same occlusocervical level on all four axial surfaces.

Six maxillary first premolar tooth preparation designs for all-ceramic restorations were tested for their effect on the strength of Dicor crowns. Ten crowns were fabricated for each preparation design, and the breaking strengths were measured. Three finish line designs (1.2-mm shoulder with sharp axiogingival line angle, 1.2-mm chamfer, and 0.8-mm chamfer) and two total occlusal convergence angles (5 degrees and 15 degrees) were tested. All preparations possessed 2.0 mm of occlusal reduction. The average breaking strengths of the 10 Dicor crowns were compared to those of 10 metal ceramic control restorations. The three groups of restorations fabricated for preparations with 5 degrees of occlusal convergence were significantly weaker than those made for preparations with 15 degrees of convergence. The metal ceramic restorations were significantly stronger (247.45 kg) than the strongest Dicor crowns (56.2 kg).

Six maxillary first premolar all-ceramic tooth preparation designs were tested for their effect on the strength of Dicor crowns. Ten crowns were fabricated for each preparation design, and their breaking strengths were measured. Three finish line designs (1.2-mm shoulder with sharp axiogingival line angle, 1.2-mm shoulder with rounded axiogingival line angle, and 0.8-mm chamfer) and two total occlusal convergence angles (10 degrees and 20 degrees) were tested. All preparations possessed 2.0 mm of occlusal reduction. The average breaking strengths of the 10 Dicor crowns were compared to that of 10 metal ceramic control restorations. The tooth preparation with a 1.2-mm shoulder finish line, sharp axiogingival line angle, and 10 degrees of total occlusal convergence produced the strongest Dicor crowns (88.6 kg). The weakest restorations were observed when a 0.8-mm chamfer finish line (66.8 kg) was used. The metal ceramic restorations were significantly stronger (247.45 kg) than the strongest Dicor crowns (88.6 kg).

We have studied the responses of chronic leukaemic B cells to infection by Epstein-Barr virus (EBV). Our results define one population of B lymphocytes, represented by prolymphocytic leukaemic (PLL) cells, which are highly susceptible to immortalization by EBV. Another B-cell type, represented by chronic lymphocytic leukaemic (CLL) cells, can be readily infected by the virus but is resistant to immortalization. Comparative studies of viral and cellular related events early after infection in these 2 cell types reveal that both express the EB viral nuclear antigen (EBNA) complex, but the immortalization-resistant CLL cells fail to express the latent membrane protein (LMP), which can be detected in PLL cells 48 hr after infection. Circularization of the linear viral genome could be detected at 7 days post infection in the PLL cells, but only in 2 out of 4 CLL cells tested. Both CLL and PLL cells show increased surface expression of CD23 and HLA-DR molecules after infection but, whereas PLL cells show an increase in size, together with RNA and DNA synthesis indicative of cell cycle progression, CLL cells appear to be arrested in the G1/S phase of the cycle. The results suggest that the outcome of infection by EBV is determined by the nature of the target cell rather than by random virus-related events. The correlation between the block in immortalization of CLL cells and their failure to express LMP suggests that expression of this protein is essential for in vitro immortalization of B cells. The failure to detect circularization in some EBV-infected CLL cells suggests that this, as well as LMP expression, may be dependent on prior activation of the B cell by EBV, an event which may vary between the different CLL samples tested.