Jeffrey D. Goldsmith

Severe hypoglycaemia post-gastric bypass requiring partial pancreatectomy: evidence for inappropriate insulin secretion and pancreatic islet hyperplasia.

Aims/hypothesisPostprandial hypoglycaemia following gastric bypass for obesity is considered a late manifestation of the dumping syndrome and can usually be managed with dietary modification. We investigated three patients with severe postprandial hypoglycaemia and hyperinsulinaemia unresponsive to diet, octreotide and diazoxide with the aim of elucidating the pathological mechanisms involved.MethodsGlucose, insulin, and C-peptide were measured in the fasting and postprandial state, and insulin secretion was assessed following selective intra-arterial calcium injection. Pancreas histopathology was assessed in all three patients.ResultsAll three patients had evidence of severe postprandial hyperinsulinaemia and hypoglycaemia. In one patient, reversal of gastric bypass was ineffective in reversing hypoglycaemia. All three patients ultimately required partial pancreatectomy for control of neuroglycopenia; pancreas pathology of all patients revealed diffuse islet hyperplasia and expansion of beta cell mass.Conclusions/interpretationThese findings suggest that gastric bypass-induced weight loss may unmask an underlying beta cell defect or contribute to pathological islet hyperplasia, perhaps via glucagon-like peptide 1-mediated pathways.

A Mouse Model of Clostridium difficile–Associated Disease

BACKGROUND & AIMS Infection with Clostridium difficile causes nosocomial antibiotic-associated diarrhea and colitis. Hamsters historically have been used to investigate disease pathogenesis and treatment, but are not ideal models because of the lack of hamster-specific reagents and genetically modified animals, and because they develop fulminant disease. The aim of this study was to establish a mouse model of antibiotic-induced C. difficile-associated disease (CDAD) that more closely resembles human disease. METHODS C57BL/6 mice were exposed to a mixture of antibiotics (kanamycin, gentamicin, colistin, metronidazole, and vancomycin) for 3 days. Two days later, they were given injections of clindamycin and then challenged 1 day later with different doses of C. difficile. RESULTS Mice that were exposed to antibiotics and then challenged with C. difficile developed diarrhea and lost weight. Disease severity varied from fulminant to minimal in accordance with the challenge dose. Typical histologic features of CDAD were evident. Oral vancomycin prevented CDAD in all mice, but 68% died from colitis after treatment was discontinued. All animals that survived an initial episode of CDAD showed no evidence of diarrhea or colitis after subsequent rechallenge with C. difficile. Different strains of C. difficile tested in the model showed different levels of virulence in mice. CONCLUSIONS We have developed a mouse model of CDAD that closely represents the human disease. In light of the recent substantial increases in CDAD incidence and severity, this model will be valuable in testing new treatments, examining disease pathogenesis, and elucidating mechanisms of protective immunity.

A genetic screen identifies an LKB1–MARK signalling axis controlling the Hippo–YAP pathway

The Hippo–YAP pathway is an emerging signalling cascade involved in the regulation of stem cell activity and organ size. To identify components of this pathway, we performed an RNAi-based kinome screen in human cells. Our screen identified several kinases not previously associated with Hippo signalling that control multiple cellular processes. One of the hits, LKB1, is a common tumour suppressor whose mechanism of action is only partially understood. We demonstrate that LKB1 acts through its substrates of the microtubule affinity-regulating kinase family to regulate the localization of the polarity determinant Scribble and the activity of the core Hippo kinases. Our data also indicate that YAP is functionally important for the tumour suppressive effects of LKB1. Our results identify a signalling axis that links YAP activation with LKB1 mutations, and have implications for the treatment of LKB1-mutant human malignancies. In addition, our findings provide insight into upstream signals of the Hippo–YAP signalling cascade.

Penalized Functional Regression

We develop fast fitting methods for generalized functional linear models. The functional predictor is projected onto a large number of smooth eigenvectors and the coefficient function is estimated using penalized spline regression; confidence intervals based on the mixed model framework are obtained. Our method can be applied to many functional data designs including functions measured with and without error, sparsely or densely sampled. The methods also extend to the case of multiple functional predictors or functional predictors with a natural multilevel structure. The approach can be implemented using standard mixed effects software and is computationally fast. The methodology is motivated by a study of white-matter demyelination via diffusion tensor imaging (DTI). The aim of this study is to analyze differences between various cerebral white-matter tract property measurements of multiple sclerosis (MS) patients and controls. While the statistical developments proposed here were motivated by the DTI study, the methodology is designed and presented in generality and is applicable to many other areas of scientific research. An online appendix provides R implementations of all simulations.

Fitbit®: An accurate and reliable device for wireless physical activity tracking.

Background: A smart accelerometer named the Fitbit has recently been introduced in the consumer market as a physical activity monitor that can interface wirelessly with mobile phones and a manufacturer-established website to allow consumers to track their physical activity in real-time. The purpose of this study was to examine the validity and reliability of the Fitbit for measuring energy expenditure during treadmill walking and running relative to energy expenditure assessed by indirect calorimetry. Methods: A total of 23 healthy adults (10 males, mean age: 30.6 ± 7.9 years; mean BMI: 24.7 ± 3.0 kg/m2) completed a four-phase treadmill exercise protocol (6 min/phase) under laboratory conditions. The protocol consisted of walking at slow (1.9 mph), moderate (3.0 mph), and brisk (4.0 mph) paces; and jogging (5.2 mph). Participants were fitted with three hip-based Fitbit One devices (two on right, one on left hip) and two wrist-based Fitbit Flex devices (one on right and left wrist). Energy expenditure was ...

Bone and Soft-Tissue Lesions: What Factors Affect Diagnostic Yield of Image-guided Core-Needle Biopsy?

PURPOSE To assess lesion-related and technical factors that affect diagnostic yield in image-guided core-needle biopsy (CNB) of bone and soft-tissue lesions. MATERIALS AND METHODS Institutional review board approval and verbal informed consent were obtained for a HIPAA-compliant prospective study of 151 consecutive CNBs of bone (n = 88) and soft-tissue (n = 63) lesions. Each CNB specimen was reported separately in the final pathology report. Diagnostic yield (total number of biopsies that yield a diagnosis divided by total number of biopsies) was calculated for all lesions and subgroups on the basis of lesion composition (lytic, sclerotic, soft tissue), lesion size (< or = 2, > 2 to 5, or > 5 cm), biopsy needle gauge, image guidance modality, number of specimens obtained, and specimen length (< 5, 5-10, or > 10 mm). The minimum number of specimens required to obtain a diagnosis was determined on the basis of the specimen number at which the diagnostic yield reached a plateau. Chi(2) And Wilcoxon rank-sum tests were performed in bivariate analyses to evaluate associations between each factor and diagnostic yield. Significant factors were evaluated with multivariate logistic regression. RESULTS Diagnostic yield was 77% for all lesions. Yield was 87% for lytic bone lesions and 57% for sclerotic bone lesions (P = .002). Diagnostic yield increased with larger lesions (54% for lesions < or = 2 cm, 75% for lesions > 2 to 5 cm, and 86% for lesions > 5 cm [P = .006]). There was no difference in diagnostic yield for bone versus soft-tissue lesions or according to needle gauge or image guidance modality. Diagnostic yield was 77% for bone lesions and 76% for soft-tissue lesions (P = .88). Yield was 83%, 72%, 77%, and 83% for biopsies performed with 14-, 15-, 16-, and 18-gauge needles, respectively (P = .57). Yield was 77% with computed tomographic guidance and 78% with ultrasonographic guidance (P = .99). Diagnostic yield increased with number of specimens obtained and with longer specimen length; it reached a plateau at three specimens for bone lesions and four specimens for soft-tissue lesions. CONCLUSION Diagnostic yield is higher in lytic than in sclerotic bone lesions, in larger lesions, and for longer specimens. Obtaining a minimum of three specimens in bone lesions and four specimens in soft-tissue lesions optimizes diagnostic yield.

Multispectral scanning during endoscopy guides biopsy of dysplasia in Barrett's esophagus

Esophageal cancer is increasing in frequency in the United States faster than any other cancer. Barretts esophagus, an otherwise benign complication of esophageal reflux, affects approximately three million Americans and precedes almost all cases of esophageal cancer. If detected as high-grade dysplasia (HGD), most esophageal cancers can be prevented. Standard-of-care screening for dysplasia uses visual endoscopy and a prescribed pattern of biopsy. This procedure, in which a tiny fraction of the affected tissue is selected for pathological examination, has a low probability of detection because dysplasia is highly focal and visually indistinguishable. We developed a system called endoscopic polarized scanning spectroscopy (EPSS), which performs rapid optical scanning and multispectral imaging of the entire esophageal surface and provides diagnoses in near real time. By detecting and mapping suspicious sites, guided biopsy of invisible, precancerous dysplasia becomes practicable. Here we report the development of EPSS and its application in several clinical cases, one of which merits special consideration.

Flt-1-dependent survival characterizes the epithelial-mesenchymal transition of colonic organoids

Aberrant cell survival and resistance to apoptosis are hallmarks of tumor invasion and progression to metastatic disease, but the mechanisms involved are poorly understood. The epithelial-mesenchymal transition (EMT), a process that facilitates progression to invasive cancer, provides a superb model for studying such survival mechanisms. Here, we used a unique spheroid culture system that recapitulates the structure of the colonic epithelium and undergoes an EMT in response to cytokine stimulation to study this problem. Our data reveal that the EMT results in the increased expression of both VEGF and Flt-1, a tyrosine kinase VEGF receptor, and that the survival of these cells depends on a VEGF/Flt-1 autocrine pathway. Perturbation of Flt-1 function by either a blocking antibody or adenoviral expression of soluble Flt-1, which acts in a dominant-negative fashion, caused massive apoptosis only in cells that underwent EMT. This pathway was critical for the survival of other invasive colon carcinoma cell lines, and we observed a correlative upregulation of Flt-1 expression linked to in vivo human cancer progression. A role for Flt-1 in cell survival is unprecedented and has significant implications for Flt-1 function in tumor progression, as well as in other biological processes, including angiogenesis and development.

Pregnant Patients Suspected of Having Acute Appendicitis: Effect of MR Imaging on Negative Laparotomy Rate and Appendiceal Perforation Rate

PURPOSE To investigate the effect of magnetic resonance (MR) imaging on the negative laparotomy rate (NLR) and the perforation rate (PR) in pregnant patients suspected of having acute appendicitis (AA) and to assess the need for computed tomography (CT) in this setting. MATERIALS AND METHODS The data of 148 consecutive pregnant patients (mean age, 29 years; age range, 15-42 years; mean gestational age, 20 weeks; gestational age range, 4-37 weeks) who were clinically suspected of having AA and examined with MR imaging between March 2002 and August 2007 were retrospectively analyzed in an institutional review board-approved HIPAA-compliant protocol. One hundred forty patients underwent ultrasonography (US) before MR imaging. The clinical and laboratory data and the findings of the initial US and MR image interpretations were recorded and analyzed at Student t and Fisher exact testing. The NLR and PR were calculated. RESULTS Fourteen (10%) patients had AA, and perforation occurred in three (21%) of them. US results were positive for AA in five (36%) patients with proved AA. MR results were positive in all 14 patients with AA. MR results were negative in 125 of the 134 patients without AA; there were nine false-positive cases (two positive, seven inconclusive). Among the patients without AA, the normal appendix could be visualized on US images in less than 2% (two of 126) of cases and on MR images in 87% (116 of 134) of cases (P < .0001). Twenty-seven (18%) patients underwent surgical exploration, and eight of them had negative laparotomy results, yielding an NLR of 30% and a PR of 21% (three of 14 patients). Only four (3%) patients underwent CT. CONCLUSION For pregnant patients clinically suspected of having AA, use of MR imaging yields favorable combinations of NLR and PR compared with previously reported values. The radiation exposure associated with CT examination can be avoided in most cases.

Principles of Analytic Validation of Immunohistochemical Assays Guideline From the College of American Pathologists Pathology and Laboratory Quality Center

CONTEXT Laboratories must validate all assays before they can be used to test patient specimens, but currently there are no evidence-based guidelines regarding validation of immunohistochemical assays. OBJECTIVE To develop recommendations for initial analytic validation and revalidation of immunohistochemical assays. DESIGN The College of American Pathologists Pathology and Laboratory Quality Center convened a panel of pathologists and histotechnologists with expertise in immunohistochemistry to develop validation recommendations. A systematic evidence review was conducted to address key questions. Electronic searches identified 1463 publications, of which 126 met inclusion criteria and were extracted. Individual publications were graded for quality, and the key question findings for strength of evidence. Recommendations were derived from strength of evidence, open comment feedback, and expert panel consensus. RESULTS Fourteen guideline statements were established to help pathology laboratories comply with validation and revalidation requirements for immunohistochemical assays. CONCLUSIONS Laboratories must document successful analytic validation of all immunohistochemical tests before applying to patient specimens. The parameters for cases included in validation sets, including number, expression levels, fixative and processing methods, should take into account intended use and should be sufficient to ensure that the test accurately measures the analyte of interest in specimens tested in that laboratory. Recommendations are also provided for confirming assay performance when there are changes in test methods, reagents, or equipment.