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A complex relationship between adipose tissue and malignancy, involving an inflammatory response, has been reported. The goal of this work was to assess the prevalence of white adipose tissue (WAT) inflammation in patients with endometrial cancer (EC), and the association with circulating inflammation markers. Furthermore, the aim was to characterize the pathways activated in and the cell type composition of adipose tissue in patients with EC.Adipose tissue and blood samples were prospectively collected from 101 patients with EC at initial surgery. WAT inflammation was determined based on adipocytes surrounded by macrophages forming crown-like structures. Circulating levels of metabolic syndrome-associated and inflammatory markers were quantified. RNA-sequencing was performed on adipose samples (n = 55); differential gene expression, pathway, and cellular decomposition analyses were performed using state-of-the-art bioinformatics methods.WAT inflammation was identified in 46 (45.5%) of 101 EC patients. Dyslipidemia, hypertension, and diabetes mellitus were significantly associated with WAT inflammation (p .05). WAT inflammation was associated with greater body mass index (p .001) and higher circulating levels of leptin, high-sensitivity C-reactive protein, and interleukin-6, as well as lower levels of adiponectin and sex hormone-binding globulin (p .05). Transcriptomic analysis demonstrated increased levels of proinflammatory and pro-neoplastic-related gene expression in inflamed omental adipose tissue.WAT inflammation is associated with metabolic syndrome, obesity, and inflammatory markers, as well as increased expression of proinflammatory and proneoplastic genes.

Missense driver mutations in cancer are concentrated in a few hotspots1. Various mechanisms have been proposed to explain this skew, including biased mutational processes2, phenotypic differences3–6 and immunoediting of neoantigens7,8; however, to our knowledge, no existing model weighs the relative contribution of these features to tumour evolution. We propose a unified theoretical ‘free fitness’ framework that parsimoniously integrates multimodal genomic, epigenetic, transcriptomic and proteomic data into a biophysical model of the rate-limiting processes underlying the fitness advantage conferred on cancer cells by driver gene mutations. Focusing on TP53, the most mutated gene in cancer1, we present an inference of mutant p53 concentration and demonstrate that TP53 hotspot mutations optimally solve an evolutionary trade-off between oncogenic potential and neoantigen immunogenicity. Our model anticipates patient survival in The Cancer Genome Atlas and patients with lung cancer treated with immunotherapy as well as the age of tumour onset in germline carriers of TP53 variants. The predicted differential immunogenicity between hotspot mutations was validated experimentally in patients with cancer and in a unique large dataset of healthy individuals. Our data indicate that immune selective pressure on TP53 mutations has a smaller role in non-cancerous lesions than in tumours, suggesting that targeted immunotherapy may offer an early prophylactic opportunity for the former. Determining the relative contribution of immunogenicity and oncogenic function to the selective advantage of hotspot mutations thus has important implications for both precision immunotherapies and our understanding of tumour evolution.

SUMMARYA challenge for screening new candidate drugs to treat cancer is that efficacy in cell culture models is not always predictive of efficacy in patients. One limitation of standard cell culture is a reliance on non-physiological nutrient levels to propagate cells. Which nutrients are available can influence how cancer cells use metabolism to proliferate and impact sensitivity to some drugs, but a general assessment of how physiological nutrients affect cancer cell response to small molecule therapies is lacking. To enable screening of compounds to determine how the nutrient environment impacts drug efficacy, we developed a serum-derived culture medium that supports the proliferation of diverse cancer cell lines and is amenable to high-throughput screening. We used this system to screen several small molecule libraries and found that compounds targeting metabolic enzymes were enriched as having differential efficacy in standard compared to serum-derived medium. We exploited the differences in nutrient levels between each medium to understand why medium conditions affected the response of cells to some compounds, illustrating how this approach can be used to screen potential therapeutics and understand how their efficacy is modified by available nutrients.

Purpose: We sought to determine whether sequencing analysis of circulating cell-free DNA (cfDNA) in patients with prospectively accrued endometrial cancer captures the mutational repertoire of the primary lesion and allows for disease monitoring. Experimental Design: Peripheral blood was prospectively collected from 44 newly diagnosed patients with endometrial cancer over a 24-month period (i.e., baseline, postsurgery, every 6 months after). DNA from the primary endometrial cancers was subjected to targeted next-generation sequencing (NGS) of 468 cancer-related genes, and cfDNA to a high-depth NGS assay of 129 genes with molecular barcoding. Sequencing data were analyzed using validated bioinformatics methods. Results: cfDNA levels correlated with surgical stage in endometrial cancers, with higher levels of cfDNA being present in advanced-stage disease. Mutations in cfDNA at baseline were detected preoperatively in 8 of 36 (22%) patients with sequencing data, all of whom were diagnosed with advanced-stage disease, high tumor volume, and/or aggressive histologic type. Of the 38 somatic mutations identified in the primary tumors also present in the cfDNA assay, 35 (92%) and 38 (100%) were detected at baseline and follow-up, respectively. In 6 patients with recurrent disease, changes in circulating tumor DNA (ctDNA) fraction/variant allele fractions in cfDNA during follow-up closely mirrored disease progression and therapy response, with a lead time over clinically detected recurrence in two cases. The presence of ctDNA at baseline (P < 0.001) or postsurgery (P = 0.014) was significantly associated with reduced progression-free survival. Conclusions: cfDNA sequencing analysis in patients with endometrial cancer at diagnosis has prognostic value, and serial postsurgery cfDNA analysis enables disease and treatment response monitoring. See related commentary by Grant et al., p. 305

To compare the quality of life in patients with vulval lichen sclerosus (VLS), vulval lichen planus (VLP) and chronic vulvovaginal candidiasis (CVVC), as measured by the Vulval Quality of Life Index (VQLI).A retrospective, single-centre cohort study was conducted at a combined dermatology and gynaecology practice from March 2018 to November 2021. VQLI scores and patient data were systematically collected and recorded in an online patient database. Treatment regimens were individualised and titrated to clinical response.Over 3 years, a total of 200 women were recruited: 59 with CVVC, 79 with VLP and 62 with VLS. The median duration of follow-up for all patients was 45.43 (16.25-80.89) weeks. At baseline, the median (interquartile range [IQR]) VQLI score was 24.00 (19.00-31.00), 21.00 (12.00-26.00) and 14.00 (7.00-26.00) for CVVC, VLP and VLS, respectively. At follow-up, the median (IQR) VQLI score for CVVC, VLP and VLS was 9.00 (3.00-15.00), 9.00 (3.00-16.00) and 5.00 (2.00-10.00), respectively. All three groups showed a significant improvement in VQLI score (p 0.0001). At baseline, the highest scoring domains were 'Sexual Function' for CVVC and 'Future Health Concerns' for VLP and VLS. At follow-up, the highest scoring domains were 'Sexual Function' for CVVC and VLP, and 'Future Health Concerns' for VLS.Vulval disease has an immense impact on QOL, especially in patients with CVVC. The VQLI is useful to clinicians in identifying the unique impact of each vulval condition on a patient's QOL in order to provide better patient-focussed care.

BACKGROUND/OBJECTIVES Vulvar lichen planus is a debilitating skin condition usually complicated by delayed diagnosis due to its highly variable clinical appearance and inconsistent histopathological characteristics. This study aims to devise a clinical diagnostic tool for the disease and to correlate this with histopathology findings. METHODS The retrospective single-centre chart review was conducted for patients presenting between January 2010 and December 2019. Clinical features were compared between 243 women with clinically suspected vulvar lichen planus with available histopathology, 50 patients with biopsy-proven vulvar lichen sclerosus and 50 patients with culture-proven chronic vulvovaginal candidiasis. Features which significantly differentiated between conditions were further studied using multivariate nonlinear regression analyses to formulate a score-based diagnostic criteria. Criteria was then applied to the remaining patients with inconclusive biopsies (classified as 'normal', 'non-specific' or 'suggestive or lichenoid') to determine sensitivity and specificity. RESULTS The clinical features that significantly differentiated the conditions were the presence of erosions (P

Using stochastic dynamic simulation for railway vehicle collision still faces many challenges, such as high modelling complexity and time-consuming. To address the challenges, we introduce a novel data-driven stochastic process modelling (DSPM) approach into dynamic simulation of the railway vehicle collision. This DSPM approach consists of two steps: (i) process description, four kinds of kernels are used to describe the uncertainty inherent in collision processes; (ii) solving, stochastic variational inferences and mini-batch algorithms can then be used to accelerate computations of stochastic processes. By applying DSPM, Gaussian process regression (GPR) and finite element (FE) methods to two collision scenarios (i.e. lead car colliding with a rigid wall, and the lead car colliding with another lead car), we are able to achieve a comprehensive analysis. The comparison between the DSPM approach and the FE method revealed that the DSPM approach is capable of calculating the corresponding confidence interval, simultaneously improving the overall computational efficiency. Comparing the DSPM approach with the GPR method indicates that the DSPM approach has the ability to accurately describe the dynamic response under unknown conditions. Overall, this research demonstrates the feasibility and usability of the proposed DSPM approach for stochastic dynamics simulation of the railway vehicle collision.

Clear cell carcinoma (CCC) of the cervix (cCCC) is a rare and aggressive type of human papillomavirus (HPV)-negative cervical cancer with limited effective treatment options for recurrent or metastatic disease. Historically, CCCs of the lower genital tract were associated with in utero diethylstilbestrol exposure; however, the genetic landscape of sporadic cCCCs remains unknown. Here we sought to define the molecular underpinning of cCCCs. Using a combination of whole-exome, targeted capture, and RNA-sequencing, we identified pathogenic genetic alterations in the Hippo signaling pathway in 50% (10/20) of cCCCs, including recurrent WWTR1 S89W somatic mutations in 40% (4/10) of the cases harboring mutations in the Hippo pathway. Irrespective of the presence or absence of Hippo pathway genetic alterations, however, all primary cCCCs analyzed in this study (n = 20) harbored features of Hippo pathway deregulation at the transcriptomic and protein levels. In vitro functional analysis revealed that expression of the WWTR1 S89W mutation leads to reduced binding of TAZ to 14-3-3, promoting constitutive nuclear translocation of TAZ and Hippo pathway repression. WWTR1 S89W expression was found to lead to acquisition of oncogenic behavior, including increased proliferation, migration, and colony formation in vitro as well as increased tumorigenesis in vivo, which could be reversed by targeted inhibition of the TAZ/YAP1 complex with verteporfin. Finally, xenografts expressing WWTR1 S89W displayed a shift in tumor phenotype, becoming more infiltrative as well as less differentiated, and were found to be composed of cells with conspicuous cytoplasmic clearing as compared to controls. Our results demonstrate that Hippo pathway alterations are likely drivers of cCCCs and likely contribute to the clear cell phenotype. Therapies targeting this pathway may constitute a new class of treatment for these rare, aggressive tumors. © 2022 The Pathological Society of Great Britain and Ireland.

Myopericytomas are uncommon tumors defined by their round to spindle shaped cells often arranged in a concentric pattern of perivascular growth. They are typically well-circumscribed, nodular, slow-growing lesions that occur in the soft tissue of the extremities. Here, we present a 30-year-old female with a 2.4 cm myopericytoma occurring in the deep lobe of the parotid gland. The diagnosis was made with detailed histopathologic and immunohistochemical findings and positive identification of the specific mutation for PDGFRβ p.Asp666Lys by next generation sequencing (NGS). This is the first case report of a parotid myopericytoma with a genetic testing that shows a particular mutation that has been linked to myopericytomatosis.

High-grade serous ovarian cancer (HGSOC) is an archetypal cancer of genomic instability1–4 patterned by distinct mutational processes5,6, tumour heterogeneity7–9 and intraperitoneal spread7,8,10. Immunotherapies have had limited efficacy in HGSOC11–13, highlighting an unmet need to assess how mutational processes and the anatomical sites of tumour foci determine the immunological states of the tumour microenvironment. Here we carried out an integrative analysis of whole-genome sequencing, single-cell RNA sequencing, digital histopathology and multiplexed immunofluorescence of 160 tumour sites from 42 treatment-naive patients with HGSOC. Homologous recombination-deficient HRD-Dup (BRCA1 mutant-like) and HRD-Del (BRCA2 mutant-like) tumours harboured inflammatory signalling and ongoing immunoediting, reflected in loss of HLA diversity and tumour infiltration with highly differentiated dysfunctional CD8+ T cells. By contrast, foldback-inversion-bearing tumours exhibited elevated immunosuppressive TGFβ signalling and immune exclusion, with predominantly naive/stem-like and memory T cells. Phenotypic state associations were specific to anatomical sites, highlighting compositional, topological and functional differences between adnexal tumours and distal peritoneal foci. Our findings implicate anatomical sites and mutational processes as determinants of evolutionary phenotypic divergence and immune resistance mechanisms in HGSOC. Our study provides a multi-omic cellular phenotype data substrate from which to develop and interpret future personalized immunotherapeutic approaches and early detection research.

Growth in open-source hardware designs combined with the decreasing cost of high-quality 3D printers have supported a resurgence of in-house custom lab equipment development. Herein, we describe a low-cost (< $400), open-source CO2incubator. The system is comprised of a Raspberry Pi computer connected to a 3D printer controller board that has controls for a CO2sensor, solenoid valve, heater, and thermistors. CO2is supplied through the sublimation of dry ice stored inside a thermos to create a sustained 5% CO2supply. The unit is controlled via G-Code commands sent by the Raspberry Pi to the controller board. In addition, we built a custom software application for remote control and used the open-source Grafana dashboard for remote monitoring. Our data show that we can maintain consistent CO2and temperature levels for over three days without manual interruption. The results from our culture plates and real-time PCR indicate that our incubator performed equally well when compared to a much more expensive commercial CO2incubator. We have also demonstrated that the antibiotic susceptibility assay can be performed in this low-cost CO2incubator. Our work also indicates that the system can be connected to incubator chambers of various chamber volumes.

Missense driver mutations in cancer are concentrated in a few hotspots

Quantitative reverse transcription polymerase chain reaction (RT-qPCR) assay is the gold standard recommended to test for acute SARS-CoV-2 infection. However, it generally requires expensive equipment such as RNA isolation instruments and real-time PCR thermal cyclers. As a pandemic, COVID-19 has spread indiscriminately, and many low resource settings and developing countries do not have the means for fast and accurate COVID-19 detection to control the outbreak. Additionally, long assay times, in part caused by slow sample preparation steps, have created a large backlog when testing patient samples suspected of COVID-19. With many PCR-based molecular assays including an extraction step, this can take a significant amount of time and labor, especially if the extraction is performed manually. Using COVID-19 clinical specimens, we have collected evidence that the RT-qPCR assay can feasibly be performed directly on patient sample material in virus transport medium (VTM) without an RNA extraction step, while still producing sensitive test results. If RNA extraction steps can be omitted without significantly affecting clinical sensitivity, the turn-around time of COVID-19 tests, and the backlog we currently experience can be reduced drastically. Furthermore, our data suggest that rapid RT-PCR can be implemented for sensitive and specific molecular diagnosis of COVID-19 in locations where sophisticated laboratory instruments are not available. Our USD 300 set up achieved rapid RT-PCR using thin-walled PCR tubes and a water bath setup using sous vide immersion heaters, a Raspberry Pi computer, and a single servo motor that can process up to 96 samples at a time. Using COVID-19 positive clinical specimens, we demonstrated that RT-PCR assays can be performed in as little as 12 min using untreated samples, heat-inactivated samples, or extracted RNA templates with our low-cost water bath setup. These findings can help rapid COVID-19 testing to become more accessible and attainable across the globe.

Due to the complexity of a train crash, it is a challenging process to describe and estimate mathematically. Although different\ud mathematical models have been developed, it is still difficult to balance the complexity of models and the accuracy of estimation.\ud ,is paper proposes a nonlinear spring-mass-damper model of train frontal crash, which achieves high accuracy and maintains\ud low complexity. ,e Convolutional Long-short-term-memory Generation Adversarial Network (CLGAN) model is applied to\ud study the nonlinear parameters dynamic variation of the key components of a rail vehicle (e.g., the head car, anticlimbing energy\ud absorber, and the coupler buffer devices). Firstly, the nonlinear lumped model of train frontal crash is built, and then the physical\ud parameters are deduced in twenty different cases using D’Alembert’s principle. Secondly, the input/output relationship of the\ud CLGAN model is determined, where the inputs are the nonlinear physical parameters in twenty initial conditions, and the output\ud is the nonlinear relationship between the train crash nonlinear parameters under other initial cases. Finally, the train crash\ud dynamic characteristics are accurately estimated during the train crash processes through the training of the CLGAN model, and\ud then the crash processes under different given conditions can be described effectively. ,e estimation results exhibit good\ud agreement with finite element (FE) simulations and experimental results. Furthermore, the CLGAN model shows great potential\ud in nonlinear estimation, and CLGAN can better describe the variation of nonlinear spring damping compared with the traditional\ud model. ,e nonlinear spring-mass-damper modeling is involved in improving the speed and accuracy of the train crash estimation, as well as being able to offer guidance for structure optimization in the early design stage.

Mutations in the leptin gene (ob) result in a metabolic disorder that includes severe obesity

As the population of the United States grows increasingly diverse, health care disparities become vital to understand and mitigate. The ethical and financial implications of how groups of Americans gain access to health care have evolved into some of today's most challenging socioeconomic problems. Educators in radiology are just beginning to tackle the concepts of health care disparities, unconscious bias, and cultural competency. In July 2017, the Accreditation Council for Graduate Medical Education required that all trainees and teaching faculty of accredited training programs receive training and experience in new areas of quality improvement to include an understanding of health care disparities as part of the core competencies. To our knowledge, there is no centralized curriculum regarding health care disparities for radiology residents and fellows. Many programs, in fact, have yet to introduce the concept to their trainees, who may have difficulty recognizing that this is even a problem affecting radiology. This manuscript serves as a primer for radiology trainees on health care disparities, with the goal of defining major concepts and providing examples of how variable access to radiological care can have substantial impact on patient outcomes.

The structure-based design and optimization of a novel series of selective PERK inhibitors are described resulting in the identification of 44 as a potent, highly selective, and orally active tool compound suitable for PERK pathway biology exploration both in vitro and in vivo.

Background: An evidence-based trauma-informed resilience skills training program developed for deployed military personnel was adapted and pilot-tested with pediatric residents. We anticipated high satisfaction ratings and changes in knowledge, beliefs, and self-efficacy related to coping with stress and trauma. Methods: The intervention included 6 skill-based modules covering emotion regulation, communication with angry patients and parents, reflective narrative, inspirational goal setting, problem-solving, and developing a self-care toolbox. An optional survey was administered before and after the training. Results: After training, 76% rated resilience skills as important, 60% were satisfied, and 82% indicated the training changed how they will respond to patient-related grief and trauma. They became more likely to believe attendings are affected by patient deaths and to know what helps them cope when they disagree with the medical decision making of others, more skilled in recognizing signs of stress and trauma, and more knowledgeable about evidence-based interventions.

Meat is now becoming a ‘hot’ issue for the food industry, governments and consumers. Not without reason was an earlier era of US politics known as the triumph of ‘pork-barrel politics’. Besides there being much money in the meat trades, meat in itself has considerable symbolic power (Rogers, 2004). To reshape meat production and consumption and to bring them in line with the Earth’s capacities is a microcosm of challenges facing both the food system and the way humans live and relate with the biosphere. Because of its deep impact, meat is a test case for how and whether policy makers align the food system with sustainability goals. The evidence is strong for behaviour change, for a reorientation of production and for a refinement of supply chain management. Broadly the picture is this: first, rich societies need to eat less meat and dairy while poorer societies need to be wary about getting on to the treadmill and second, consumption is too high in developed countries and developing countries are following that lead, while all need to keep meat and dairy consumption low. As Barry Popkin and others have pointed out, there are multiple health benefits to reducing meat consumption (Popkin, 2009). And there seems to be agreement that the environmental benefits of constraining meat and dairy are high too (Sustainable Development Commission, 2009).

This article sets out the findings from research on the impact of a, UK based, chefs in schools teaching programme on food, health, nutrition and cookery. Professional chefs link with local schools, where they deliver up to three sessions to one class over a year. The research measured the impact of a standardised intervention package and changes in food preparation and consumption as well as measuring cooking confidence. The target group was 9–11 year olds in four schools. The main data collection method was a questionnaire delivered 2 weeks before the intervention and 2 weeks afterwards. There was a group of four matched control schools. Those taking part in the intervention were enthused and engaged by the sessions and the impact measures indicated an intention to change. There were gains in skills and confidence to prepare and ask for the ingredients to be purchased for use in the home. Following the session with the chef, the average reported cooking confidence score increased from 3.09 to 3.35 (by 0.26 points) in the intervention group – a statistically significant improvement. In the control group this change was not statistically significant. Children’s average reported vegetable consumption increased after the session with the chef, with the consumption score increasing from 2.24 to 2.46 points (0.22 points) again, a statistically significant increase with no significant changes in the control group. The research highlights the need to incorporate evaluation into school cooking initiatives as the findings can provide valuable information necessary to fine-tune interventions and to ensure consistency of the healthy eating messages.

To identify common alleles associated with different histotypes of epithelial ovarian cancer (EOC), we pooled data from multiple genome-wide genotyping projects totaling 25,509 EOC cases and 40,941 controls. We identified nine new susceptibility loci for different EOC histotypes: six for serous EOC histotypes (3q28, 4q32.3, 8q21.11, 10q24.33, 18q11.2 and 22q12.1), two for mucinous EOC (3q22.3 and 9q31.1) and one for endometrioid EOC (5q12.3). We then performed meta-analysis on the results for high-grade serous ovarian cancer with the results from analysis of 31,448 BRCA1 and BRCA2 mutation carriers, including 3,887 mutation carriers with EOC. This identified three additional susceptibility loci at 2q13, 8q24.1 and 12q24.31. Integrated analyses of genes and regulatory biofeatures at each locus predicted candidate susceptibility genes, including OBFC1, a new candidate susceptibility gene for low-grade and borderline serous EOC.

In a recent New York Times editorial (1/20/12, Summers), the author wished for transformation of college education in a way that would reflect the structure of society and what we now understand ab...

Recombinant human erythropoietin (EPO) has been used therapeutically for more than two decades in the treatment of anemia. Although EPO is generally well tolerated, in rare cases, patients have developed anti-EPO antibodies that can negatively impact safety and efficacy. Therefore, the detection of antibodies against EPO is a regulatory requirement during clinical development and post-approval. Although it is a rare phenomenon, antibody-mediated pure red cell aplasia (PRCA) is a serious complication than can result from antibodies that develop and neutralize EPO as well as endogenous erythropoietin. Currently, there are no universally accepted analytical methods to detect the full repertoire of binding and neutralizing anti-EPO antibodies. A number of different methods that differ in terms of antibodies detected and assay sensitivities are used by different manufacturers. There is also a lack of antibody reference reagents, and therefore no consistent basis for detecting and measuring anti-EPO antibodies. Reference reagents, with established ranges, are essential to monitor the safety and efficacy of all erythropoiesis-stimulating agents (ESAs) structurally related to human erythropoietin. This is the first report of the development and characterization of a panel of fully human antibodies against EPO suitable as reference reagents. The characteristics of antibodies within the panel were selected based on the prevalence of non-neutralizing IgG and IgM antibodies in non-PRCA patients and neutralizing IgG antibodies, including IgG1 and IgG4, in antibody-mediated PRCA subjects. The reference panel includes antibodies of high- and low-affinity with binding specificity to neutralizing and non-neutralizing erythropoietin epitopes. The subclass of human antibodies in this reference panel includes an IgG1, IgG2, and IgG4, as well as an IgM isotype. This antibody panel could help select appropriate immunogenicity assays, guide validation, and monitor assay performance. Further, this human anti-ESA antibody panel may help set the limits of each assay platform in terms of the full repertoire of the anti-ESA antibodies, and may facilitate standardization of ESA immunogenicity reporting across assay platforms.

Background Cutaneous B-cell lymphoproliferative lesions can pose diagnostic challenges. This study investigates the utlility of flow cytometry in 42 cases of suspected cutaneous B-cell lymphoma. Methods All available cases were reviewed [World Health Organization-European Organization for Research and Treatment of Cancer (WHO-EORTC) classification]. Flow cytometry, immunohistochemistry and polymerase chain reaction-immunoglobulin H (PCR-IgH) analysis of blood and/or lesional skin were performed on primary cutaneous B-cell lymphoma (pcBCL, 17 cases), secondary cutaneous BCL (scBCL, 8 cases) and atypical lymphoid hyperplasia (ALH, 17 cases). Results Flow cytometry of skin detected a B-cell clone in 3/13 cases of ALH, 8/8 cases of pcBCL and 4/4 cases of scBCL, while PCR detected a clone in 3/14 cases of ALH, 4/15 cases of pcBCL and 6/8 cases of scBCL. Of eight cases of pcBCL analyzed by both methods, all eight were positive by flow while only three were positive by PCR. All cases positive by PCR were also positive by flow. Of five cases with both flow and light chain immunohistochemistry, all five showed light chain restriction by flow, while only two were positive by immunohistochemistry. Conclusion Flow cytometry is more sensitive than PCR in detecting B-cell lymphoproliferative disorders (12/12 cases, 100% vs. 10/23 cases, 43%; p