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Chronic lymphocytic leukemia (CLL) is the most commonly encountered leukemia in the clinical laboratory. Cytoskeletal defects in CLL lymphocytes can result in the formation of up to 75% smudge cells (SCs) during blood film preparation. Failure to account for these damaged lymphocytes in the white blood cell (WBC) differential diminishes the accuracy and reproducibility of the results. Lacking clear practice standards on handling SCs in CLL, different laboratories may employ different methods to mitigate SC-induced errors. This review explores the pathophysiology of SCs, their effect on WBC differentials in CLL, and how these results can impact clinical decisions. The pros and cons of various SC corrective methods are described to assist laboratories in developing an optimized protocol to reduce errors and inconsistencies in WBC differentials. Finally, the potential utility of SC enumeration as an indicator of CLL prognosis is discussed in terms of laboratories with differing access to technology.

INTRODUCTION: The IMMray PanCan-d test combines an 8-plex biomarker signature with CA19-9 in a proprietary algorithm to detect pancreatic ductal adenocarcinoma (PDAC) in serum samples. This study aimed to validate the clinical performance of the IMMray PanCan-d test and to better understand test performance in Lewis-null (le/le) individuals who cannot express CA19-9. METHODS: Serum samples from 586 individuals were analyzed with the IMMray PanCan-d biomarker signature and CA19-9 assay, including 167 PDAC samples, 203 individuals at high risk of familial/hereditary PDAC, and 216 healthy controls. Samples were collected at 11 sites in the United States and Europe. The study was performed by Immunovia, Inc (Marlborough, MA), and sample identity was blinded throughout the study. Test results were automatically generated using validated custom software with a locked algorithm and predefined decision value cutoffs for sample classification. RESULTS: The IMMray PanCan-d test distinguished PDAC stages I and II (n = 56) vs high-risk individuals with 98% specificity and 85% sensitivity and distinguished PDAC stages I-IV vs high-risk individuals with 98% specificity and 87% sensitivity. We identified samples with a CA19-9 value of 2.5 U/mL or less as probable Lewis-null (le/le) individuals. Excluding these 55 samples from the analysis increased the IMMray PanCan-d test sensitivity to 92% for PDAC stages I-IV (n = 157) vs controls (n = 379) while maintaining specificity at 99%; test sensitivity for PDAC stages I and II increased from 85% to 89%. DISCUSSION: These results demonstrate the IMMray PanCan-d blood test can detect PDAC with high specificity (99%) and sensitivity (92%).

e16258 Background: The IMMray® PanCan-d blood test for early detection of PDAC has been commercially available in 45 U.S. states since the laboratory’s licensure in August 2021. This is the first clinically-available blood test dedicated to the early detection of PDAC, combining an 8-plex biomarker signature with CA19-9 in a proprietary algorithm. Here we report the initial results of this assay. Methods: All orders received and results reported were reviewed for the IMMray PanCan-d test between August 4, 2021 and February 11, 2022. Serum specimens were processed at the Immunovia, Inc. laboratory in Marlborough, Massachusetts. Demographic information of individuals undergoing testing is based on the completed test requisition submitted by their health care providers. All information was de-identified prior to this analysis. Results: The median age of those tested was 59 years and 46% were male. 76% of patients were referred because of a history of familial pancreatic cancer. Germline mutation prevalence is reported in the table below. 12% of patients had IPMNs or mucinous cystic neoplasms, 2% had chronic pancreatitis and 3% had new onset Diabetes Mellitus. Table. Demographics of Individuals Tested with the IMMray PanCan-d test 80.5% of IMMray PanCan-d results were negative, 1.1% were high-risk signature present, and 11.5% were borderline. These results are fully consistent with validation studies for this assay in high-risk surveillance populations (Brand RE et al Clin Transl Gastroenterol, doi: 10.14309/ctg.0000000000000468). 6.9% of individuals tested had a CA19-9 value of less than 2.5 U/ml, which we identify as presumptively Lewis null (le/le) and CA19-9 non-secretors. Given that CA19-9 contributes significantly to calculated decision values for the IMMray PanCan-d test, individuals with a CA19-9 less than 2.5 U/ml receive a result of test not performed. Conclusions: The commercial availability of a biomarker assay for the early detection of PDAC resulted in a testing population with demographics and referral criteria similar to that of PDAC high-risk surveillance programs in the U.S. IMMray PanCan-d test results were as expected based on prior validation studies.[Table: see text]

Artificial Intelligence (AI) research and regulation seek to balance the benefits of innovation against any potential harms and disruption. However, one unintended consequence of the recent surge in AI research is the potential re-orientation of AI technologies to facilitate criminal acts, which we term AI-Crime (AIC). We already know that AIC is theoretically feasible thanks to published experiments in automating fraud targeted at social media users, as well as demonstrations of AI-driven manipulation of simulated markets. However, because AIC is still a relatively young and inherently interdisciplinary area—spanning socio-legal studies to formal science—there is little certainty of what an AIC future might look like. This article offers the first systematic, interdisciplinary literature analysis of the foreseeable threats of AIC, providing law enforcement and policy-makers with a synthesis of the current problems, and a possible solution space.

Schools worldwide have implemented many different peer-led interventions with mixed results, but the evidence base on their effectiveness as mental health interventions remains limited. This study combines a scoping review and systematic review to map the variations of peer-led interventions in schools and to evaluate the quality of the existing evidence base. This scoping review and systematic review evaluated the existing literature across 11 academic databases. Studies were included if they reported a peer-led intervention that aimed to address a mental health or wellbeing issue using a peer from the same school setting. Data were extracted from published and unpublished reports and presented as a narrative synthesis. 54 studies met eligibility criteria for the scoping review, showing that peer-led interventions have been used to address a range of mental health and wellbeing issues globally. 11 studies met eligibility criteria for the systematic review with a total of 2,239 participants eligible for analysis (929 peer leaders; 1,310 peer recipients). Two studies out of seven that looked at peer leaders showed significant improvements in self-esteem and social stress, with one study showing an increase in guilt. Two studies out of five that looked at peer recipient outcomes showed significant improvements in self-confidence and in a quality of life measure, with one study showing an increase in learning stress and a decrease in overall mental health scores. The findings from these reviews show that despite widespread use of peer-led interventions, the evidence base for mental health outcomes is sparse. There appear to be better documented benefits of participation for those who are chosen and trained to be a peer leader, than for recipients. However, the small number of included studies means any conclusions about effectiveness are tentative.

Isoprene concentrations were measured at four heights below, within and above the forest canopy in Wytham Woods (UK) throughout the summer of 2018 using custom-built gas chromatographs (the iDirac). These observations were complemented with selected ancillary variables, including air temperature, photosynthetically active radiation (PAR), occasional leaf gas exchange measurements and satellite retrievals of normalized difference vegetation and water indices (NDVI and NDWI). The campaign overlapped with a long and uninterrupted heatwave accompanied by moderate drought. Peak isoprene concentrations during the heatwave-drought were up to a factor of 4 higher than those before or after. Higher temperatures during the heatwave could not account for all the observed isoprene; the enhanced abundances correlated with drought stress. Leaf-level emissions confirmed this and also included compounds associated with ecosystem stress. This work highlights that a more in-depth understanding of the effects of drought stress is required to better characterize isoprene emissions.

Background: Recent estimates suggest that one in ten young people worldwide experiences a diagnosable mental health disorder, with many more suffering subsyndromal levels of psychological distress. As young people spend much of their time in schools, the role of educational settings in the delivery of mental health provision is increasingly recognised. Advances in neurodevelopmental research have highlighted the important and complex role of peer influence on adolescent behaviours, suggesting peer-led support schemes have high potential utility. Schools worldwide have implemented peer-led interventions with mixed results, but the global evidence base on their effectiveness remains limited. This systematic review aims to examine the evidence base of the outcomes of school-based peer-led interventions on the mental health of young people aged 4–18.Methods: Eligible studies will be randomised controlled trials and observational studies that report on the mental health outcomes of a peer-led school-based intervention. Eligible participants will be aged 4–18 and will be current students of the intervention school. Individual- and group-based interventions will be included. The following 11 databases will be screened based upon their reach in healthcare and education: PsycINFO, MEDLINE, EMBASE, CINAHL, CENTRAL, BEI, Scopus, Web of Science, ERIC, SSCI and Social Care Online. There will be no restriction placed on publication period, original language or country of publication. Papers will be systematically screened for eligibility by two review authors. Data will be presented as a descriptive report. A meta-analysis will be carried out if a subset of studies allows, given the anticipated high levels of heterogeneity.Discussion: This systematic review will be the first to synthesise the global evidence on the mental health outcomes of peer-led interventions for children and adolescents in a school setting. It will analyse the available data in order to understand the role of these interventions in schools, inform future developers of peer support programmes and identify gaps in current research. This review will be of value to policy makers, health and education services, researchers and those involved in delivering peer support initiatives.

Artificial Intelligence (AI) research and regulation seek to balance the benefits of innovation against any potential harms and disruption. However, one unintended consequence of the recent surge in AI research is the potential re-orientation of AI technologies to facilitate criminal acts, which I term AI-Crime (AIC). We already know that AIC is theoretically feasible thanks to published experiments in automating fraud targeted at social media users, as well as demonstrations of AI-driven manipulation of simulated markets. However, because AIC is still a relatively young and inherently interdisciplinary area of research—spanning socio-legal studies to formal science—there is little certainty of what an AIC future might look like. This article offers the first systematic, interdisciplinary literature analysis of the foreseeable threats of AIC, providing law enforcement and policy-makers with a synthesis of the current problems.

Artificial intelligence (AI) research and regulation seek to balance the benefits of innovation against any potential harms and disruption. However, one unintended consequence of the recent surge in AI research is the potential re-orientation of AI technologies to facilitate criminal acts, term in this article AI-Crime (AIC). AIC is theoretically feasible thanks to published experiments in automating fraud targeted at social media users, as well as demonstrations of AI-driven manipulation of simulated markets. However, because AIC is still a relatively young and inherently interdisciplinary area—spanning socio-legal studies to formal science—there is little certainty of what an AIC future might look like. This article offers the first systematic, interdisciplinary literature analysis of the foreseeable threats of AIC, providing ethicists, policy-makers, and law enforcement organisations with a synthesis of the current problems, and a possible solution space.

Institutions governing multi-agent systems (MASs) are a pervasive means to guide agents towards the aims of the MAS (e.g. collecting data) with regulations on the outcomes of agents’ behaviour. Yet, wider organisations/governments often intend to guide the design of institutions governing MAS in meeting different aims (e.g. preserving the rights of agents). A pervasive means to guide the design of MAS-governing institutions (or any institution, for that matter) is to use institutions at higher tiers of governance (e.g. directives, constitutions) to regulate the regulations of institutions at lower tiers of governance (e.g. national legislation, software policies). A recent innovation has been an automated means to determine the compliance of a lower-tier institution’s regulations with a higher-tier’s. However, for a designer of a non-compliant institution there remains a dilemma: be punished for non-compliant regulations or arduously determine and rectify the underlying causes of non-compliance. In this paper we propose a way to automatically determine how to revise an institution to be compliant that also minimises the change in the regulations’ outcomes thus keeping as closely as possible to the institution designers’ original intentions.

Rationale: Interferon-γ release assays have significant advantages over tuberculin skin testing in many clinical situations. However, recent studies have called into question their reliability in serial testing of healthcare workers because of reportedly high rates of positivity and high conversion/reversion rates on retesting.Objectives: To define the performance characteristics of the T-SPOT.TB test, an interferon-γ release assay, during serial screening programs of healthcare workers at 19 U.S. hospitals.Methods: A total of 42,155 T-SPOT.TB test results from healthcare workers at 19 geographically diverse hospitals obtained for routine tuberculosis screening programs were analyzed to determine the rates of positivity, reversion, and conversion in serial testing data.Measurements and Main Results: In 19,630 evaluable serial pairs from 16,076 healthcare workers, the mean test positivity rate was 2.3% (range, 0.0–27.4%). The mean conversion rate was 0.8% (range, 0.0–2.5%), and the mean reversion rate was ...

An institution typically comprises constitutive rules, which give shape and meaning to social interactions and regulative rules, which prescribe agent behaviour in the society. Regulative rules guide social interaction, in particular when they are coupled with reward and punishment regulations that are enforced for (non-)compliance. Institution examples include legislation and contracts. Formal institutional reasoning frameworks automate ascribing social meaning to agent interaction and determining whether those actions have social meanings that comprise (non-)compliant behaviour. Yet, institutions do not just govern societies. Rather, in what is called multi-level governance, institutional designs at lower governance levels (e.g., national legislation at the national level) are governed by higher level institutions (e.g., directives, human rights charters and supranational agreements). When an institution design is found to be non-compliant, punishments can be issued by annulling the legislation or imposing fines on the responsible designers (i.e., government). In order to enforce multi-level governance, higher governance levels (e.g., courts applying human rights) must check lower level institution designs (e.g., national legislation) for compliance; in order to avoid punishment, lower governance levels (e.g., national governments) must check their institution designs are compliant with higher-level institutions before enactment. However, checking non-compliance of institution designs in multi-level governance is non-trivial. In particular, because institutions in multi-level governance operate at different levels of abstraction. Lower level institutions govern with concrete regulations whilst higher level institutions typically comprise increasingly vague and abstract regulations. To address this issue, in this paper we propose a formal framework with a novel semantics that defines compliance between concrete lower level institutions and abstract higher level institutions. The formal framework is complemented by a sound and complete computational framework that automates compliance checking, which we apply to a real-world case study.

An 82-year-old Japanese nonsmoking man presented with persistent dry cough and small left apical pneumothorax. High resolution CT scan of the chest demonstrated bilateral upper lobe pleuroparenchymal thickening and architectural distortion. Serial imaging revealed mild progression and development of small bilateral pneumothoraces, and pneumomediastinum. A surgical lung biopsy was required to confirm the diagnosis.

This paper deals with the problem of automating the contribution of resources owned by people to do work for others. This is by providing a means for owners of resources to maintain autonomy over how, when and to whom their resources are used with the specification of use policies governing resources. We give representations of requests for resource usage as a set of conditional norms and a use policy as specifying what norms should and should not be imposed on a resource i.e. a set of meta-norms. Our main contribution is a reasoner built on the Event Calculus, that detects conflicts between requests and use policies, determining whether the request can be accepted.

GABP is a heteromeric transcription factor complex which consists of the ets related protein, GABPalpha, and the Notch-related protein, GABPbeta. We isolated a human genomic DNA fragment which is highly homologous and colinear with human GABPalpha cDNA, but which lacks introns. This processed pseudogene, psihGABPalpha, is expressed as RNA in U937 human myeloid cells, but a mutation at the site that corresponds to the ATG start methionine codon prevents its translation into protein. The pseudogene was localized to chromosome 7 using a somatic cell hybrid mapping panel and it is not syntenic with authentic GABPalpha, which was localized to chromosome 21. We have identified psihGABPalpha, a novel, GABPalpha-related processed pseudogene which is expressed as a RNA transcript in human myeloid cells.

Background We hypothesized that the method of breast cancer margin assessment may be associated with different rates of positive margins and residual carcinoma. Methods A total of 178 breast cancer specimens were divided into 2 groups (A and B) based on the margin assessment method used. Rates of positive margins, re-excision, and residual carcinoma at re-excision were compared and analyzed statistically. Results At least 1 margin was positive in 64.7% in group A and in 65.2% in group B. At directed re-excision 54% in group A and 51% in group B had residual carcinoma. The lateral margin was positive in 44% in group A compared with 26% in group B ( P = .06). The posterior margin was positive in 19% in group A and in 51% in group B ( P = .001). Conclusions Two different breast cancer specimen margin assessment methods had comparable rates of positive margins and residual carcinoma at re-excision. Different patterns of specific margin positivity suggest that the method of margin assessment may alter results.

Sets of related norms (normative systems) are likely to evolve due to changing goals of an organization or changing values of a society, this may introduce incoherence, such as the simultaneous prohibition and obligation of an action or a set of deadlocked duties. This paper presents a compositional framework that may be used for detecting whether normative systems are coherent by analysing traces of actions and their legality. Unlike other mechanisms for checking normative system coherence, the framework makes it possible to re-check just those parts of the system that have changed, without re-checking the entirety.

Abstract Vascular smooth muscle (VSM) contains a bidirectional isoform of 11β-hydroxysteroid dehydrogenase (11β-HSD), the enzyme that can metabolize endogenous glucocorticoids to their respective 11-dehydro derivatives. 11βOH-progesterone (11βOH-P), a compound that can be produced in vivo, is as potent or more potent than licorice derivatives in inhibiting renal and hepatic 11β-HSD. When studied in homogenates prepared from primary cultures of rat VSM, 11βOH-P and its derivative, 11-keto-progesterone (11-keto-P), proved to be potent, directionally specific inhibitors of vascular 11β-HSD. 11βOH-P selectively inhibited the forward dehydrogenase reaction (corticosterone→11-dehydrocorticosterone), whereas 11-keto-P selectively blocked the reverse oxidoreductase reaction. To test the physiological effects, vascular rings were prepared from rat aorta. Rings were incubated in culture media containing either a submaximal concentration of corticosterone (10 nmol/L), 11-dehydrocorticosterone (100 nmol/L), 11βOH-P (1 μmol/L), 11-keto-P (1 μmol/L), or a combination of glucocorticoid and inhibitor for 24 hours. After the 24-hour incubation, rings were briefly stimulated sequentially with phenylephrine (10 nmol/L to 1 μmol/L) and angiotensin II (1 μmol/L). The immediate contractile response in rings incubated with both corticosterone and 11βOH-P was greater than in rings previously incubated with either the corticosterone or 11βOH-P alone (eg, response to 100 nmol/L phenylephrine in milligrams of force, mean±SE: corticosterone, 728±56, n=9; 11βOH-P, 325±105, n=4; both, 1132±122, n=8; corticosterone versus both, P

Paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) and concurrent radiation (paclitaxel/RT) are being actively evaluated as treatment for locally advanced lung, pancreatic, and gastric cancers. Paclitaxel is a potent chemotherapeutic agent that interferes with mitotic spindle function to synchronize cells at G 2 M, the most radiosensitive phase of the cell cycle. Because G 2 M arrest is necessary but not sufficient for paclitaxel radiosensitization, other cellular factors must interact with paclitaxel to produce radiosensitization. We have investigated whether molecular genetic alterations that disturb cell cycle regulation in tumor cells affect the action of paclitaxel as a radiosensitizer. The p53 tumor suppressor gene serves a critical role at the G 1 S transi tion of the cell cycle by blocking entry into S phase under conditions that would result in genetic instability. Mutations in the tumor suppressor gene, p53, confer resistance to radiation therapy and most chemotherapeutic agents, including cisplatin and 5-fluorouracil. In striking contrast, we have now shown that p53 mutations in non-small cell lung cancer do not confer resistance to paclitaxel/RT. The effect of p53 mutations on response to paclitaxel/RT in pancreatic and gastric cancers is currently being investigated. Knowledge of specific genetic alterations in individual tumors may permit the selection of optimal therapeutic strategy for individual patients.

Over-expression of the multifunctional zinc-finger transcription factor Yin Yang 1 (YY1) has been associated with cellular proliferation and resistance to apoptotic stimuli. In this study, we report that YY1 was uniformly highly over-expressed in a wide range of human cancer cell lines and in human colon cancer tissue samples. The examination of YY1-specific mRNA expression demonstrated at least six mRNA isoforms ubiquitously expressed in normal human adult and fetal tissues. Substantial over-expression of two specific mRNA isoforms of 7.5 and 2.9 kb size, respectively, was detected in gastrointestinal and other cancer cells in vitro, whereby mRNA stability differed significantly between various cell lines. YY1 protein expression levels were similar in different colon cancer cell lines. Using FISH analysis of several colorectal cancer cell lines, the human YY1 locus was expectedly identified on chromosome 14q32 and no evidence of gene amplification and chromosomal translocation was observed. However, varying degree of aneuploidy was noted in vitro. YY1 immunoreactivity in human colon tumor samples was found more intense in poorly differentiated tumors than in moderately and well differentiated colon cancers and lower expression levels tended to be associated with shorter survival. In conclusion, YY1 was over-expressed in colon cancer in the absence of gene amplification and chromosomal translocation. YY1 mRNA and protein stability are important regulatory mechanisms of YY1 expression in colon cancer.

Extracorporeal membrane oxygenation has demonstrated effectiveness for cardiopulmonary support in a variety of clinical situations. This article reviews the cases in which extracorporeal membrane oxygenation was used as an adjunct to pediatric cardiac transplantation. Twenty children, aged 7 days to 17 years, with cardiac failure refractory to conventional therapy received extracorporeal membrane oxygenation for 6 to 192 hours. In 4 cases it was used as a bridge to transplantation; in 10 cases it facilitated resuscitation of the cardiac allograft in the immediate postoperative period; and in 6 cases it complemented therapy for severe rejection in the late postoperative period. Twelve patients survived extracorporeal membrane oxygenation, 7 of whom lived more than 8 months. One long-term survivor was in the bridge-to-transplant group, 4 in the immediate postoperative group, and 2 in the rejection group. All survivors have normal cardiac allograft function. These data suggest that extracorporeal membrane oxygenation can be used to support profound cardiac failure in the pediatric heart transplant patient as a bridge to transplantation, in the resuscitation of the cardiac allograft, or to supplement a rejecting allograft.

Fractions of hydroxyethyl starch (HES), 100,000-1,000,000 daltons, have been shown to reduce increased permeability changes in postcapillary venules associated with ischemia-reperfusion injury in canine myocardium and rat skel etal muscle. The authors studied the effect of intravenous administration of two fractions of the biodegradable HES, Fm (100,000-300,000 daltons) and FLm (300,000-1,000,000 daltons), in the prevention of acute renal failure in a rat warm renal ischemia model. A series of renal artery cross-clamp times was used to determine the threshold for irreversible damage (thirty minutes). After thirty minutes of warm renal ischemia, 2.5 cc of 6 % Fm or FLm solution, 5% human serum albumin, or 9% normal saline was administered. The effect of the solutions was evaluated by four criteria: (1) forty-eight hours' serum BUN/creatinine, (2) forty-eight hours' urine output, (3) percent of water content in the kidney, and (4) a histopathologic grading. There were no differences among the animals in any of the four groups in renal function at forty-eight hours, percent of water content of the kidneys, urine output, and extent of medullary and cortical injury. The authors attribute the lack of protective effect by the HES macromolecules to intrinsic properties of the kidney such as the extensive endothelial cell surface area of the well-vascularized kidney and the hypersensitivity of the kidney to hypoxic injury.

We identified significantly hypermethylated genes in clear cell renal cell carcinoma.We previously identified a set of under expressed genes in renal cell carcinoma tissue through transcriptional profiling and a robust computational screen. We selected 19 of these genes for hypermethylation analysis using a rigorous search for the best candidate regions, considering CpG islands and transcription factor binding sites. The genes were analyzed for hypermethylation in the DNA of 38 matched clear cell renal cell carcinoma and normal samples using matrix assisted laser desorption ionization time-of-flight mass spectrometry. The significance of hypermethylation was assessed using 3 statistical tests. We validated the down-regulation of significantly hypermethylated genes at the RNA and protein levels in a separate set of patients using reverse transcriptase-polymerase chain reaction, immunohistochemistry and Western blots.We found 7 significantly hypermethylated regions from 6 down-regulated genes, including SFRP1, which was previously shown to be hypermethylated in renal cell carcinoma and other cancer types.To our knowledge we report for the first time that another 5 genes (SCNN1B, SYT6, DACH1, and the tumor suppressors TFAP2A and MT1G) are hypermethylated in renal cell carcinoma. Robust computational screens and the high throughput methylation assay resulted in an enriched set of novel genes that are epigenetically altered in clear cell renal cell carcinoma. Overall the detection of hypermethylation in these highly down-regulated genes suggests that assaying for their methylation using cells from urine or blood could provide the basis for a viable diagnostic test.

Normal mature quiescent human B lymphocytes, isolated as a function of buoyant density, require activation for up-regulation of IL-13R constituents. Cell activation through a combination of surface Ig and CD40 receptor ligation leads to the most substantial message production for IL-13Rα1. Functional consequences of this receptor variation, in initially quiescent cells, includes demonstrable effects on cellular proliferation in response to ligand exposure. Variations in the method of surface activation, with particular emphasis on the CD40 receptor, reveals that immobilized CD40 ligand may be sufficient, in and of itself, to up-regulate IL-13Rα1, which may bear significance for B-lymphocyte bystander proliferation. Regulation of the IL-13Rα1 protein and message also differs as a function of cellular phenotype. Although values are greater in memory than naive B cells, as they are initially isolated from extirpated tonsils, variations in the magnitude of message and protein, as a function of surface stimulation, are more substantial in the naive subset. The magnitude of variation in message production in naive cells is associated with a more vigorous proliferative response to IL-13 than seen in memory lymphocytes. The cellular response to IL-13, as a function of activation and phenotype, is the converse of that demonstrated for IL-2. Evaluation of proliferation, receptor message, ligand binding protein production, and the response to putatively synergistic cytokines reveals that IL-2 is the predominant lymphokine utilized by memory cells. This is in contradistinction to IL-13, which along with IL-4, are the predominant moieties for naive lymphocytes.

Polymerase chain reaction (PCR) amplification and product analysis for the detection of chromosomal translocations, such as the t(14;18), has traditionally been a two-step process. PCR product detection has generally entailed gel electrophoresis and/or hybridization or sequencing for confirmation of assay specificity. Using a microvolume fluorimeter integrated with a thermal cycler and a PCR-compatible double-stranded DNA (dsDNA) binding fluorescent dye (SYBR Green I), we investigated the feasibility of simultaneous thermal amplification and detection of MBR/JH translocation products by fluorescence melting curve analysis. We analyzed DNA from 30 cases of lymphoproliferative disorders comprising 19 cases of previously documented MBR/JH-positive follicle center lymphoma and 11 reactive lymphadenopathies. The samples were coded and analyzed blindly for the presence of MBR/JH translocations by fluorescence melting curve analysis. We also performed dilutional assays using the MBR/JH-positive cell line SUDHL-6. Multiplex PCR for MBR/JH and β-globin was used to simultaneously assess sample adequacy. All (100%) of the 19 cases previously determined to be MBR/JH positive by conventional PCR analysis showed a characteristic sharp decrease in fluorescence at ∼90°C by melting curve analysis after amplification. Fluorescence melting peaks obtained by plotting the negative derivative of fluorescence over temperature (− d F/ d T) versus temperature ( T ) showed melting temperatures ( T m ) at 88.85 ± 1.15°C. In addition, multiplex assays using both MBR/JH and β-globin primers yielded easily distinguishable fluorescence melting peaks at ∼90°C and 81.2°C, respectively. Dilutional assays revealed that fluorescence melting curve analysis was more sensitive than conventional PCR and agarose gel electrophoresis with ultraviolet transillumination by as much as 100-fold. Simultaneous amplification and fluorescence melting curve analysis is a simple, reliable, and sensitive method for the detection of MBR/JH translocations. The feasibility of specific PCR product detection without electrophoresis or utilization of expensive fluorescently labeled probes makes this method attractive for routine molecular diagnostics.