New philometrids (Nematoda,Philometridae) from head tissues of two serranid fishes (<Emphasis Type="Italic">Epinephelus morio</Emphasis> and <Emphasis Type="Italic">Mycteroperca microlepis</Emphasis>) off Florida,northern Gulf of Mexico

A new species of nematode, Philometra morii sp. nov. (Philometridae), is described from males and gravid females collected from the mouth cavity of the red grouper, Epinephelus morio (Valenciennes) (Serranidae, Perciformes), from the northen Gulf of Mexico, off Florida, USA. The new species is characterized mainly by the length of spicules (84–90 and 72–87 μm) and the length (54 μm) and structure of the gubernaculum in the male, and by the presence of three large oesophageal teeth protruding from the mouth, the number and arrangement of small cephalic papillae (8 papillae in 4 pairs of external circle and 6 single papillae of internal circle), the length (1.09–1.50 mm) and structure of the oesophagus and two large papilla-like caudal projections, and by the length of their bodies (19.34–30.07 mm). Philometra morii is the third species of this genus reported from E. morio in the Gulf of Mexico, differing from the two previously described species, in addition to morphological features, by the site of infection in this host (mouth cavity and sinuses vs oculo-orbits or gonads). From the same region (northern Gulf of Mexico, off Florida), gravid females of Philometra Costa, 1845, morphologically and biometrically similar to those of P. morii, were found in the subcutaneous tissues and sinuses of the head of another serranid fish (gag), Mycteroperca microlepis (Good et Bean). Although their conspecificity with P. morii cannot be excluded, they have been designated as Philometra sp. until conspecific males are discovered and described. Based on light and scanning electron microscopy examination (latter used only for females), both these forms are described.     

Efficacy and safety of acupuncture for the treatment of non-specific acute low back pain: a randomised controlled multicentre trial protocol [ISRCTN65814467]

Background  

Low back pain and its associated incapacitating effects constitute an important healthcare and socioeconomic problem, as well as being one of the main causes of disability among adults of working age. The prevalence of non-specific low back pain is very high among the general population, and 60–70% of adults are believed to have suffered this problem at some time. Nevertheless, few randomised clinical trials have been made of the efficacy and efficiency of acupuncture with respect to acute low back pain. The present study is intended to assess the efficacy of acupuncture for acute low back pain in terms of the improvement reported on the Roland Morris Questionnaire (RMQ) on low back pain incapacity, to estimate the specific and non-specific effects produced by the technique, and to carry out a cost-effectiveness analysis.     

Hypoxia-inducible factor 1 and VEGF upregulate CXCR4 in glioblastoma: implications for angiogenesis and glioma cell invasion

Hypoxia and hypoxia-inducible factor-1 (HIF-1) play a critical role in glioblastoma multiforme (GBMs). CXCR4 is involved in angiogenesis and is upregulated by HIF-1alpha. CXCR4 is a chemokine receptor for stromal cell-derived factor-1 (SDF-1)alpha, also known as CXCL12. We hypothesized that CXCR4 would be upregulated by hypoxia in GBMs. First, we investigated the expression of HIF-1alpha and CXCR4 in GBMs. CXCR4 was consistently found colocalized with HIF-1alpha expression in pseudopalisading glioma cells around areas of necrosis. In addition, angiogenic tumor vessels were strongly positive for CXCR4. Next, we tested the in vitro effect of hypoxia and vascular endothelial growth factor (VEGF) on the expression of CXCR4 in glioma cell lines and in human brain microvascular endothelial cells (HBMECs). Exposure to hypoxia induced significant expression of CXCR4 and HIF-1alpha in glioma cells, whereas treatment with exogenous VEGF increased CXCR4 expression in HBMECs. We also transfected U87MG glioma cells with an HIF-1alpha construct and observed that CXCR4 was upregulated in these cells even in normoxic conditions. We then used a lentivirus-mediated shRNA expression vector directed against HIF-1alpha. When exposed to hypoxia, infected cells failed to show HIF-1alpha and CXCR4 upregulation. We performed migration assays under normoxic and hypoxic conditions in the presence or absence of AMD3100, a CXCR4 inhibitor. There was a significant increase in the migration of U87MG and LN308 glioma cells in hypoxic conditions, which was inhibited in the presence of AMD3100. These studies demonstrate the critical role played by hypoxia and CXCR4 in glioma cell migration. Based on these studies, we suggest that hypoxia regulates CXCR4 in GBMs at two levels. First, through HIF-1alpha in the pseudopalisading tumor cells themselves and, secondly, by the VEGF-stimulated angiogenic response in HBMECs. We believe this knowledge may lead to a potentially important two-pronged therapy against GBM progression using chemotherapy targeting CXCR4.     

Reversible Secretion of Glycosaminoglycans and Proteoglycans by Cyclically Stretched Valvular Cells in 3D Culture

Mitral valve leaflets and chordae have been shown to contain different amounts and proportions of glycosaminoglycans (GAGs) and proteoglycans (PGs) corresponding to in vivo normal or diseased cyclic strain patterns. To understand the effect of cyclic strains on GAG/PG synthesis by valvular interstitial cells (VICs) isolated from valve leaflet and chordae separately, porcine VICs were seeded within collagen gels and alternately stretched or relaxed for 24 h periods for one week in a custom-designed tissue engineering bioreactor. We found cyclic-stretch-induced upregulation of total GAGs and of individual GAG classes secreted into the culture medium. Leaflet cells showed a delayed response to stretching compared to chordal cells, but altered the proportions of various GAG classes they secreted during the culture duration. Decorin and biglycan PGs were slightly responsive to stretch. We demonstrated that mechanical stretch and relaxation conditions reversibly regulate GAG and PG production in a novel 3D model of valve tissues. This is the first study using cyclic strains to modulate GAG/PG synthesis by valve cells and our results may have implications for the remodeling of the mitral valve as well as other tissues.     

Haploidentical Stem Cell Transplantation for Refractory/Relapsed Neuroblastoma

Pediatric patients with refractory or relapsed metastatic neuroblastoma (NBL) have a poor prognosis despite autologous stem cell transplantation (SCT). Allogeneic SCT from a haploidentical donor has a remarkable alloreactive effect in patients with leukemia; thus, we evaluated this approach in children with very high-risk NBL. We analyzed data from 2 prospective phase I/II trials. A total of 26 patients with refractory (n?=?5), metastatic relapsed (n?=?20), or locally relapsed MYCN-positive (n?=?1) NBL received a median of 17?×?10⁶/kg T/B cell-depleted CD34⁺ stem cells with 68?×?10³/kg residual T cells and 107?×?10⁶/kg natural killer cells. The conditioning regimen comprised melphalan, fludarabine, thiotepa, OKT3, and a short course of mycophenolate mofetil post-transplantation. Engraftment occurred in 96% of the patients. Event-free survival and overall survival at 5 years were 19% and 23%, respectively. No transplantation-related mortality was observed, and the single death was due to progression/subsequent relapse. The median duration of follow-up was 8.1 years. Patients in complete remission before SCT had a significantly better prognosis than those with residual tumor load (P?<?.01). All patients with progressive disease before SCT relapsed within 1 year. Grade II and grade III acute graft-versus-host disease (GVHD) occurred in 31% and 12% of the patients, respectively. Chronic limited and extensive GVHD occurred in 28% and 10%, respectively. Our data indicate that haploidentical SCT is a feasible treatment option that can induce long-term remission in some patients with NBL with tolerable side effects, and may enable the development of further post-transplantation therapeutic strategies based on the donor-derived immune system.     

Lower life satisfaction,active coping and cardiovascular disease risk factors in older African Americans: outcomes of a longitudinal church-based intervention

This study examined lower life satisfaction, active coping and cardiovascular disease risk factors (diastolic and systolic blood pressure, body mass index, and circumferences) in older African Americans over the phases of an 18-month church-based intervention, using a quasi-experimental design. Participants (n = 89) were 45 years of age and older from six churches (three treatment, three comparison) in North Florida. Lower life satisfaction had a persistent unfavorable effect on weight variables. Active coping showed a direct beneficial effect on selected weight variables. However, active coping was adversely associated with blood pressure, and did not moderate the association between lower life satisfaction and cardiovascular risk factors. The intervention had a beneficial moderating influence on the association between lower life satisfaction and weight variables and on the association between active coping and these variables. Yet, this pattern did not hold for the association between active coping and blood pressure. The relationship of lower life satisfaction and selected cardiovascular risk factors and the positive effect of active coping were established, but findings regarding blood pressure suggest further study is needed.     

A Dependable Massive Storage Service for Medical Imaging

We present the construction of Babel, a distributed storage system that meets stringent requirements on dependability, availability, and scalability. Together with Babel, we developed an application that uses our system to store medical images. Accordingly, we show the feasibility of our proposal to provide an alternative solution for massive scientific storage and describe the software architecture style that manages the DICOM images life cycle, utilizing Babel like a virtual local storage component for a picture archiving and communication system (PACS-Babel Interface). Furthermore, we describe the communication interface in the Unified Modeling Language (UML) and show how it can be extended to manage the hard work associated with data migration processes on PACS in case of updates or disaster recovery.     

Sensitive Screening Tests for Suspected Class A Carbapenemase Production in Species of Enterobacteriaceae

The detection of class A serine-carbapenemases among species of Enterobacteriaceae remains a challenging issue. Methods of identification for routine use in clinical microbiology laboratories have not been standardized to date. We developed a novel screening methodology suitable for countries with high basal levels of carbapenem resistance due to non-carbapenemase-mediated mechanisms and standardized several simple confirmatory methods that allow the recognition of bacteria producing class A carbapenemases, including KPC, Sme, IMI, NMC-A, and GES, by using boronic acid (BA) derivatives. A total of 28 genetically unrelated Enterobacteriaceae strains producing several class A carbapenemases were tested. Thirty-eight genetically unrelated negative controls were included. The isolates were tested against imipenem (IPM), meropenem (MEM), and ertapenem (ETP) by MIC and disk diffusion assays in order to select appropriate tools to screen for suspected carbapenemase production. It was possible to differentiate class A carbapenemase-producing bacteria from non-carbapenemase-producing bacteria by using solely the routine IPM susceptibility tests. The modified Hodge test was evaluated and found to be highly sensitive, although false-positive results were documented. Novel BA-based methods (a double-disk synergy test and combined-disk and MIC tests) using IPM, MEM, and ETP, in combination with 3-aminophenylboronic acid as an inhibitor, were designed as confirmatory tools. On the basis of the performance of these methods, a sensitive flow chart for suspicion and confirmation of class A carbapenemase production in species of Enterobacteriaceae was designed. By using this methodology, isolates producing KPC, GES, Sme, IMI, and NMC-A carbapenemases were successfully distinguished from those producing other classes of β-lactamases (extended-spectrum β-lactamases, AmpCs, and metallo-β-lactamases, etc). These methods will rapidly provide useful information needed for targeting antimicrobial therapy and appropriate infection control.Class A carbapenemases (KPCs, Sme, NMC-A, IMI, and some allelic variants of GES/IBC) have become more prevalent within the Enterobacteriaceae family (30, 33). Early recognition of producers of carbapenemases has become mandatory, as clinical failure associated with these enzymes has been described previously (12) and as recognition is crucial for controlling the spread of carbapenemase-producing bacteria. A uniform and standardized phenotypic tool for the detection of class A carbapenemases is still lacking. Recently, the CLSI issued recommendations for the phenotypic screening of carbapenemase producers among species of Enterobacteriaceae: MICs of ertapenem (ETP), meropenem (MEM), and imipenem (IPM) of 2, 2 to 4, and 2 to 4 μg/ml, respectively (or a zone of inhibition by ETP or MEM of ≤21 mm in diameter in the disk diffusion [DD] assay), may indicate isolates with carbapenemase production, and this phenotype should be confirmed by the Hodge method (5). In Argentina, large proportions of nosocomial Enterobacteriaceae strains meet the CLSI criteria for suspected carbapenemase production according to ETP and MEM DD assay results (about 14 and 5% of 6,700 tested strains, respectively, as reported by the WHONET-Argentina Network in 2007 (http://www.paho.org). Up to one-third of these isolates had a positive Hodge test result suggestive of carbapenemase production; still, almost all of them did not produce carbapenemases as determined by molecular methods (27). Therefore, in this work, we designed a panel composed of diverse bacterial genera with distinct carbapenem susceptibility patterns to identify potential problems in the current recommendations. In addition, we developed an optimized approach for the more accurate detection of isolates with possible carbapenemase production, suitable for countries in which strains have high baseline levels of resistance to carbapenems due to non-carbapenemase-mediated mechanisms, such as those resulting from the combination of extended-spectrum β-lactamases or AmpCs and porin mutations (2, 10, 15, 34).Boronic acids (BA) were reported in the early 1980s to be reversible inhibitors of class C enzymes (1). Unexpectedly, we discovered unusual inhibition of organisms possessing KPC by 3-aminophenylboronic acid (APB), which may be attributed solely to the presence of this carbapenemase (25). Afterwards, Doi et al. and, in a subsequent study with a larger series of strains, Tsakris et al. demonstrated the overwhelming effectiveness of APB for the detection of Klebsiella pneumoniae possessing KPC (7, 32). Given these promising results, we decided to explore the potential of APB methods as confirmatory tests for class A carbapenemase detection. To date, information on the efficiency of APB methods for KPC-possessing bacterial species different from K. pneumoniae is missing. Moreover, boronic methods for carbapenemase detection described in the literature so far have been limited to KPC enzymes and have not been evaluated for the remaining class A members. Thus, we examined the capabilities of BA-based methods to detect the entire class A carbapenemase family by using these methods to test a panel of well-characterized strains, including several bacterial hosts with KPC and also GES-, Sme-, IMI-, and NMC-A-producing bacteria.Thus, in this work we propose novel and accurate phenotypic methods, appropriate for clinical laboratories, to both screen for and confirm the presence of class A carbapenemases in species of Enterobacteriaceae.