Neuroblastoma Differentiation: The Untapped Potential of Mitochondrial Uncouplers.

While the goal of most anticancer treatments is to kill cancer cells, some therapies halt cancer progression by inducing cancer cell differentiation. For example, retinoic acid induces neuroblastoma cell differentiation in vitro and is used as maintenance therapy for children with high-risk neuroblastoma. A new study by Jiang and colleagues has revealed the mitochondrial uncoupler niclosamide ethanolamine (NEN) induces neuroblastoma cell differentiation in vitro and slows neuroblastoma tumor growth in vivo. Mitochondrial uncoupler molecules alter cell metabolism by forcing cells to "burn" more nutrients, resulting in a switch from anabolic to catabolic metabolism. NEN-induced neuroblastoma cell differentiation was associated with disruption of Warburg metabolism, epigenetic remodeling, and downregulation of key oncogenic drivers of neuroblastoma development, including MYCN. NEN is currently used as an antiparasitic worm treatment and is safe to use in children but has poor pharmacokinetic properties. However, derivatives of NEN and structurally distinct uncouplers that have improved pharmacokinetic properties are in development. Results of this study ignite the idea that mitochondrial uncouplers could be used as differentiating agents and expand the pharmacotherapy toolkit to treat cancer, including neuroblastoma. See related article by Jiang et al., p. 181.

Utilisation of anti-TNF levels in a UK tertiary IBD centre.

OBJECTIVE: To ascertain how anti-tumour necrosis factor (TNF) drug and anti-drug antibody levels testing is used in a 'real-world' setting to optimise inflammatory bowel disease (IBD) treatment. DESIGN: Retrospective cohort study of prospectively collected patient data. SETTING: Tertiary IBD centre in London, UK. PATIENTS: All patients at Guy's and St Thomas' Hospitals on anti-TNF who had levels measured between the start of testing in 2012 and October 2014. INTERVENTIONS: Anti-TNF drug and anti-drug antibody levels as part of routine monitoring. MAIN OUTCOME MEASURES: Indication for measuring levels and changes in management made as a result of the levels. RESULTS: 330 infliximab levels were carried out in 199 patients and 143 adalimumab levels were carried out in 103 patients. Levels were primarily done in those with evidence of loss of response; 37% of infliximab levels and 52% of adalimumab levels. Levels resulted in a change in management in 26% of patients in infliximab group and 25% of patients in adalimumab group; however, this was greater in those with loss of response, 62% and 61% respectively. Anti-drug antibodies were detected in 7% of patients. CONCLUSIONS: Our early experience has demonstrated that measuring anti-TNF drug and anti-drug antibody levels can be useful in the optimisation of IBD management. In an increasing number of patients, particularly those with evidence of loss of response, it allows early decisions to be made regarding changing therapy. It also offers the potential for significant cost-saving by preventing pointless dose escalation in the context of therapeutic levels or when high-level anti-drug antibodies are present.

TRIM16 acts as a tumour suppressor by inhibitory effects on cytoplasmic vimentin and nuclear E2F1 in neuroblastoma cells.

The family of tripartite-motif (TRIM) proteins are involved in diverse cellular processes, but are often characterized by critical protein-protein interactions necessary for their function. TRIM16 is induced in different cancer types, when the cancer cell is forced to proceed down a differentiation pathway. We have identified TRIM16 as a DNA-binding protein with histone acetylase activity, which is required for the retinoic acid receptor ß(2) transcriptional response in retinoid-treated cancer cells. In this study, we show that overexpressed TRIM16 reduced neuroblastoma cell growth, enhanced retinoid-induced differentiation and reduced tumourigenicity in vivo. TRIM16 was only expressed in the differentiated ganglion cell component of primary human neuroblastoma tumour tissues. TRIM16 bound directly to cytoplasmic vimentin and nuclear E2F1 in neuroblastoma cells. TRIM16 reduced cell motility and this required downregulation of vimentin. Retinoid treatment and enforced overexpression caused TRIM16 to translocate to the nucleus, and bind to and downregulate nuclear E2F1, required for cell replication. This study, for the first time, demonstrates that TRIM16 acts as a tumour suppressor, affecting neuritic differentiation, cell migration and replication through interactions with cytoplasmic vimentin and nuclear E2F1 in neuroblastoma cells.

Evaluating the impact of an intervention to reduce injuries among loggers in West Virginia, 1999-2007.

The purpose of this study was to evaluate the efficacy of a video-based safety training intervention (VBSTI) to reduce injuries among loggers over an 8-year period. WV Workers' Compensation data were used to assess the trend in injury rates, medical and indemnity costs due to logging injuries. There were 1,435 logger injury claims reported and a modest decline in the minor nature of injuries. There was no significant decline in total injury rates or in "struck by object" incidents emphasized in the VBSTI. Among the severe injuries, "head and neck" injuries showed a statistically significant decrease (p = 0.0454), but "falls" showed a statistically significant inverse change (p < 0.05). Medical and indemnity costs showed a statistical significant decrement (p < 0.05). Due to potential confounding, it is inconclusive whether the modest decreases in the claims can be attributed to the VBSTI. There still remains a need to improve the safety of workers in this high-risk employment.

ATP7A is a novel target of retinoic acid receptor beta2 in neuroblastoma cells.

Increased retinoic acid receptor beta (RARbeta(2)) gene expression is a hallmark of cancer cell responsiveness to retinoid anticancer effects. Moreover, low basal or induced RARbeta(2) expression is a common feature of many human cancers, suggesting that RARbeta(2) may act as a tumour suppressor gene in the absence of supplemented retinoid. We have previously shown that low RARbeta(2) expression is a feature of advanced neuroblastoma. Here, we demonstrate that the ABC domain of the RARbeta(2) protein alone was sufficient for the growth inhibitory effects of RARbeta(2) on neuroblastoma cells. ATP7A, the copper efflux pump, is a retinoid-responsive gene, was upregulated by ectopic overexpression of RARbeta(2). The ectopic overexpression of the RARbeta(2) ABC domain was sufficient to induce ATP7A expression, whereas, RARbeta(2) siRNA blocked the induction of ATP7A expression in retinoid-treated neuroblastoma cells. Forced downregulation of ATP7A reduced copper efflux and increased viability of retinoid-treated neuroblastoma cells. Copper supplementation enhanced cell growth and reduced retinoid-responsiveness, whereas copper chelation reduced the viability and proliferative capacity. Taken together, our data demonstrates ATP7A expression is regulated by retinoic acid receptor beta and it has effects on intracellular copper levels, revealing a link between the anticancer action of retinoids and copper metabolism.

The trapezius port: a novel approach for port access.

Placement of long-term central venous access in breast cancer patients who have undergone bilateral mastectomy presents a unique challenge. The standard anterior chest wall placement of the port may be compromised by factors such as a fresh surgical site, the possibility of postoperative radiation therapy, or the immediate reconstruction field. To avoid the potential for complications that impact the surgical field or subsequent therapy, we have developed a unique new technique for central venous port placement: the trapezius port. By placing the port in a subcutaneous pocket over the trapezius muscle, the risk of infection or cross-contamination of the surgical site is avoided.

Assessing safety awareness and knowledge and behavioral change among West Virginia loggers.

OBJECTIVE: To determine if a video used during logger training influences safety attitude, knowledge, and workplace habits. METHOD: From April 2002 to October 2003, loggers receiving training through the West Virginia Division of Forestry were given a new safety module. This consisted of a pre-training survey, viewing video, brief introduction to field safety guide, and an immediate post-training survey. Six months after training, loggers were contacted by telephone to assess workplace behavioral changes. RESULTS: 1197 loggers attended 80 training sessions and completed surveys; 21% were contacted at follow up. Pre-training surveys indicated that half said "accidents" were part of the job and had experienced a "close call" in their work. An overwhelming majority felt that safety management and periodic meetings were important. Over 75% indicated they would not take risks in order to make a profit. Several statistically significant improvements were noted in safety knowledge after viewing the video: logger's location in relation to the tree stump during fatal incidents and the pictorial identification of an overloaded truck and the safest cutting notch. At follow up, many of the loggers said they related to the real life victim stories portrayed in the video. Further, the field guide served as a quick and easy reference and taught them valuable tips on safe cutting and felling. CONCLUSIONS: Significant changes in safety knowledge and attitude among certified loggers resulted from viewing the video during training. Subsequent use of the video and field guide at the worksite encouraged positive change in self reported work habits and practices.

Co-expression of beta-adrenergic receptors and cyclooxygenase-2 in pulmonary adenocarcinoma.

Pulmonary adenocarcinoma (PAC) is the leading type of lung cancer and is highly resistant to conventional cancer therapy. A better understanding of the regulatory mechanisms which control the growth of this deadly malignancy are urgently needed to develop more effective cancer intervention strategies. Recent studies have shown that PAC frequently overexpresses cyclooxygenase-2 (COX-2). This enzyme converts arachidonic acid (AA) into several metabolites, some of which have been identified as modulators of mitogenesis and apoptosis. Accordingly, the AA cascade and COX-2 are currently widely studied as potential targets for lung cancer prevention. Recent studies by our research group have shown that cell lines derived from human PACs express beta1- and beta2-adrenergic receptors, which regulate the release of AA and DNA synthesis. Moreover, we have demonstrated that an antagonist for beta-adrenergic receptors or aspirin inhibited the development of experimentally induced PAC in a hamster model. These findings suggest that beta-adrenergic receptors may serve as upstream regulators of AA and COX-2-mediated PAC growth. However, no information is currently available on the expression of beta-adrenergic receptors and its possible correlation with the expression of COX-2 in tissue samples from human PAC, casting some doubt on the significance of these findings in vitro and in an animal model. In the current study, we have therefore analyzed tissue samples of human PACs for the expression of beta1-and beta2-adrenergic receptors as well as COX-2 by reverse transcription polymerase chain reaction (RT-PCR) or immunohistochemistry. Our data show that seven out of eight samples co-expressed COX-2 and one or both of these beta-adrenergic receptors, supporting the experimental evidence for a functional link between these neurotransmitter receptors and the AA cascade in the regulation of human PAC.

Bradykinin B2 receptors activate Na+/H+ exchange in mIMCD-3 cells via Janus kinase 2 and Ca2+/calmodulin.

We used a cultured murine cell model of the inner medullary collecting duct (mIMCD-3 cells) to examine the regulation of the ubiquitous sodium-proton exchanger, Na+/H+ exchanger isoform 1 (NHE-1), by a prototypical G protein-coupled receptor, the bradykinin B2 receptor. Bradykinin rapidly activates NHE-1 in a concentration-dependent manner as assessed by proton microphysiometry of quiescent cells and by 2'-7'-bis[2-carboxymethyl]-5(6)-carboxyfluorescein fluorescence measuring the accelerated rate of pH(i) recovery from an imposed acid load. The activation of NHE-1 is blocked by inhibitors of the bradykinin B2 receptor, phospholipase C, Ca2+/calmodulin (CaM), and Janus kinase 2 (Jak2), but not by pertussis toxin or by inhibitors of protein kinase C and phosphatidylinositol 3'-kinase. Immunoprecipitation studies showed that bradykinin stimulates the assembly of a signal transduction complex that includes CaM, Jak2, and NHE-1. CaM appears to be a direct substrate for phosphorylation by Jak2 as measured by an in vitro kinase assay. We propose that Jak2 is a new indirect regulator of NHE-1 activity, which modulates the activity of NHE-1 by increasing the tyrosine phosphorylation of CaM and most likely by increasing the binding of CaM to NHE-1.

Risk factor analysis for breast cellulitis complicating breast conservation therapy.

Women who undergo breast conservation therapy for early-stage breast cancer can develop breast cellulitis, a complication for which risk factors are undefined. A matched case-control investigation was conducted to identify risk factors for the development of breast cellulitis among patients who have undergone breast conservation therapy. Patients comprised 17 patients with cases of breast cellulitis diagnosed after partial mastectomies that had been performed from 1992 through 1997 and 34 control patients who were matched to case-patients by date of breast lumpectomy and by primary surgeon. Statistical analyses indicated the following factors were associated with breast cellulitis: drainage of a hematoma (P=.010); postoperative ecchymosis (P=.021); lymphedema (odds ratio [OR], 10. 154; 95% confidence interval [CI], 1.348-208.860); resected breast tissue volume (OR, 1.456; 95% CI, 1.035-2.168); and previous number of breast seroma aspirations (OR, 3.445; 95% CI, 1.036-19.771). This is the first matched case-control study to identify risk factors for the development of breast cellulitis after breast conservation therapy.

Complex enterocutaneous fistula: closure with rectus abdominis muscle flap.

Most enterocutaneous fistulas are caused by complications of abdominal surgery that may result from anastomotic failure, poor blood supply, or iatrogenic bowel injuries. Mortality rates are high when associated sepsis and malnutrition are uncontrolled. Fistulas that occur late and those that recur spontaneously present more difficulty and may close spontaneously in less than 30% of cases. Mortality rates in patients with complex enterocutaneous fistulas may reach 60% to 80%. When traditional conservative surgeries of fistulous tract excision, bowel mobilization, and resection with primary end-to-end anastomosis fail, a more aggressive approach is required. The rectus abdominis muscle flap has been extensively studied and used in a wide variety of abdominal, vaginal, and perineal repairs. We report successful closure of complex enterocutaneous fistulas with a rectus abdominis muscle flap in a complicated case.

Slip and fall-related injuries in relation to environmental cold and work location in above-ground coal mining operations.

BACKGROUND: The association between slip and fall-related injuries and environmental temperature was examined for mostly enclosed (inside vehicles, machinery, or buildings), outdoor (outside, not enclosed), and enclosed/outdoor jobs in the coal mining industry to see if differences existed among the three work locations that had varying exposure to cold temperatures. METHODS: Temperature data from the National Climatic Data Center and injury data from the Mine Safety and Health Administration were evaluated from 1985-1990 for seven states. Proportionate methods were used to examine the relationship between slips and falls and temperature. RESULTS: Proportionate injury ratios of slips and fall-related injuries increased as temperature declined for all three work locations. Proportion of slips and fall-related injuries that occurred while running/walking increased with declining temperature, with the ground outside as the most common source of these injuries. CONCLUSIONS: Outside movement becomes a greater hazard at freezing temperatures for workers in all locations, not just outdoor workers. Any intervention methods geared toward reducing injury incidents facilitated by cold weather must also be directed toward workers who spend time in more enclosed locations.

Hepatic angiosarcoma: long-term survival after complete surgical removal.

Angiosarcoma of the liver constitutes 2 per cent of all primary tumors of the liver. This lesion has demonstrated an intimate relationship between the environment and potential malignant transformation. The CT appearance of hepatic angiosarcoma is nonspecific, whereas arteriography provides the best imaging tool for diagnosis. Hepatic resection is rarely feasible but should be considered if the disease is limited and the remainder of the liver is relatively normal. The prognosis of patients with this malignancy is poor with a median survival of 6 months. A patient with a hepatic angiosarcoma is described. Complete surgical resection was possible and was associated with a prolonged (10-year) postoperative survival.

The Potential of F-18-FDG PET in Breast Cancer. Detection of Primary Lesions, Axillary Lymph Node Metastases, or Distant Metastases.

This retrospective study was done to evaluate the utility of 2-[F-18]fluoro-2-deoxy-D-glucose positron emission tomography (F-18-FDG PET) in identifying primary and recurrent breast cancer and lymph node metastases. One hundred whole-body PET scans of 87 patients were reviewed. PET results obtained with F-18-FDG and an ECAT/EXACT-921 or an ECAT-931 (Siemens/CTI) were based on visual interpretation, or standardized uptake values (SUVs), related to histology and also compared to computerized tomography (CT) and mammography results. The sensitivity for PET in detecting primary (N = 35 studies) and recurrent breast cancer (N = 65 studies) was 96% and 85% with a specificity of 91% and 73%. The sensitivity for lymph node metastases at the time of initial diagnosis was 100% with a specificity of 100%. Quantitative SUV information did not improve the accuracy of F-18-FDG PET in identifying primary breast cancers. The results suggest that whole-body PET is useful in detecting recurrence or metastases, may be useful in detecting lymph node metastases prior to initial axillary lymph node dissection, but is less sensitive in excluding axillary lymph nodes metastases later in the course of the disease.

A DNA probe for identification of larvae of the commercial surfclam (Spisula solidissima).

A molecular DNA probe was developed for identification of larvae of the commercial surfclam Spisula solidissima (family Mactridae), to distinguish early-stage veligers from larvae of other common bivalve species in a study of surfclam settlement and recruitment on the New Jersey continental shelf. For discrimination of S. solidissima from other common bivalve species at the study site (almost all of which belong to different families), an oligonucleotide designed from the nucleotide sequence of the 18S ribosomal RNA gene provides a useful and sensitive family-specific probe and primer. For discrimination between S. solidissima and Mulinia lateralis (both members of Mactridae), the 18S rRNA gene was examined by restriction fragment length polymorphism (RFLP) analysis. A combination of the DNA probe and RFLP analysis provides a positive identification of S. solidissima and M. lateralis larvae in coastal plankton samples.

Breast cellulitis following breast conservation therapy: a novel complication of medical progress.

Breast cellulitis is a novel complication of the recently accepted practice of breast conservation therapy. This phenomenon represents an anatomic shift from ipsilateral upper extremity cellulitis seen in past years when mastectomy with axillary lymph node dissection was performed for treatment of limited disease due to breast cancer. Thirteen episodes of breast cellulitis in nine women who underwent breast conservation therapy for stage I or II breast cancer are presented. The mean duration from the end of radiotherapy to the initial episode of cellulitis was 4.9 months. Eighty-three percent of episodes occurred in patients who had radiologically demonstrated fluid collections at the surgical lumpectomy site prior to the onset of cellulitis. Eight (61.5%) of 13 episodes occurred within 3 months of a follow-up mammogram of the treated breast. Two patients developed recurrent cellulitis within a 6-month period. Breast cellulitis may be more commonly seen in clinical practice as an increasing number of patients undergo breast-sparing procedures for treatment of limited disease due to cancer.

Ductal carcinoma in situ of the breast: correlation of pathologic and mammographic features with extent of disease.

Optimal treatment of ductal carcinoma in situ (DCIS) of the breast requires an improved understanding of its pathologic extent and propensity for local recurrence. This study was performed to analyze mammographic and pathologic features of DCIS that might predict the extent of disease within the breast and facilitate treatment selection between lumpectomy alone, lumpectomy and radiotherapy, and mastectomy. At our institution, 60 cases of DCIS were diagnosed in 59 patients from June 1985 to February 1995 and form the basis of this retrospective study. Demographic and treatment-related information was obtained from hospital and tumor registry records. Mammograms were reviewed and size estimates of the abnormalities were determined. Pathologic slides from all cases were reviewed and classified according to size group, focality, nuclear grade, necrosis, and histologic subtype. DNA ploidy status and proliferation indices were available for 28 patients. Pathologically, 43 (72%) cases were < 15 mm, 14 (23%) were 16 to 40 mm, and 3 (5%) were > 40 mm. Five (8%) of the lesions were multicentric, 28 (47%) focal, and 27 (45%) multifocal. Thirty-three (55%) patients were treated by mastectomy, 16 (27%) by lumpectomy alone, and 11 (18%) by lumpectomy and radiation therapy. Mammographic size, histologic grade, presence or absence of necrosis, histologic subtype, DNA ploidy, and proliferative index were compared with pathologic size and focality by chi 2 analysis. Mammographic size correlated significantly with pathologic size (chi 2 = 11.3; P = 0.02) but underestimated the extent of disease in 9 cases. Although focality correlated significantly with pathologic size (chi 2 = 15.8; P = 0.003), the remaining histopathologic features did not significantly correlate with pathologic size or focality. Histopathologic features, including DNA studies, do not reliably predict the pathologic extent of DCIS, but mammographic size and focality do significantly correlate with pathologic size. Nevertheless, most cases of DCIS are small focal or multifocal lesions that are amenable to breast conservation approaches; further studies are needed to determine the appropriate use of lumpectomy, radiation therapy, and mastectomy in the treatment of DCIS.

Perineal hernia repair using gracilis myocutaneous flap.

Perineal hernia formation is an infrequent but well-recognized complication of major pelvic surgery. Various methods of perineal reconstruction have been reported. This report describes one technique of perineal hernia repair using a unilateral gracilis myocutaneous flap. The gracilis myocutaneous flap provides well-vascularized tissue that is useful in many situations requiring reconstruction of the pelvis and perineum, especially when the area has been irradiated.

Expression of Epstein-Barr virus nuclear antigen-1 induces B cell neoplasia in transgenic mice.

The Epstein-Barr virus (EBV) nuclear antigen-1 (EBNA-1) is a pleiotropic protein which has been characterized extensively both biochemically and functionally. It is the only one of the identified latent protein-encoding genes to be consistently expressed in viral-associated endemic Burkitt's lymphoma cells. As such, it is the only candidate viral protein to possibly perform a maintenance function in the tumour pathology. Despite this, no oncogenic activity has been attributed to the protein in tissue culture assays. The experiments described here were initiated to explore the activity of the protein in B cells in vivo. EBNA-1 transgenic mice were generated with transgene expression directed to the B cell compartment using the mouse Ig heavy chain intron enhancer. Transgene expression was demonstrated in the lymphoid tissues of mice of two independent lines. Transgenic positive mice of both lines succumb to B cell lymphoma. The B cell tumours are monoclonal, frequently of follicular centre cell origin and remarkably similar to those induced by transgenic c-myc expression. These results demonstrate that EBNA-1 is oncogenic in vivo and suggest that the gene product may play a direct role in the pathogenesis of Burkitt's lymphoma and possibly other EBV-associated malignancies.