TOPAS: network-based structural alignment of RNA sequences.
For many RNA families, the secondary structure is known to be better conserved among the member RNAs compared to the primary sequence. For this reason, it is important to consider the underlying folding structures when aligning RNA sequences, especially for those with relatively low sequence identity. Given a set of RNAs with unknown structures, simultaneous RNA alignment and folding algorithms aim to accurately align the RNAs by jointly predicting their consensus secondary structure and the optimal sequence alignment. Despite the improved accuracy of the resulting alignment, the computational complexity of simultaneous alignment and folding for a pair of RNAs is O(N6), which is too costly to be used for large-scale analysis. In order to address this shortcoming, in this work, we propose a novel network-based scheme for pairwise structural alignment of RNAs. The proposed algorithm, TOPAS, builds on the concept of topological networks that provide structural maps of the RNAs to be aligned. For each RNA sequence, TOPAS first constructs a topological network based on the predicted folding structure, which consists of sequential edges and structural edges weighted by the base-pairing probabilities. The obtained networks can then be efficiently aligned by using probabilistic network alignment techniques, thereby yielding the structural alignment of the RNAs. The computational complexity of our proposed method is significantly lower than that of the Sankoff-style dynamic programming approach, while yielding favorable alignment results. Furthermore, another important advantage of the proposed algorithm is its capability of handling RNAs with pseudoknots while predicting the RNA structural alignment. We demonstrate that TOPAS generally outperforms previous RNA structural alignment methods on RNA benchmarks in terms of both speed and accuracy. Source code of TOPAS and the benchmark data used in this paper are available at https://github.com/bjyoontamu/TOPAS.


The stoichiometry and location of troponin I- and troponin C-like proteins in the myofibril of the bay scallop, Aequipecten irradians.
Localization and quantification studies were carried out on bay-scallop (Aequipecten irradians) striated-muscle troponin C- and troponin I-like proteins. Indirect immunofluorescence microscopy of scallop myofibrils stained with either rabbit anti-(scallop troponin I) or anti-(scallop troponin C) antibodies shows staining of all I-bands observed. The results of quantification studies using sodium dodecyl sulfate poly-acrylamide-gel electrophoresis of untreated scallop myofibrils, washed scallop myofibrils, and isolated scallop thin filaments indicate an actin/tropomyosin/troponin-C molar rationn of 7:1:1. The molar ratio for troponin I could not be determined in untreated myofibrils because of interfering bands; in washed myofibrils a value of 0.6 mol of troponin I/mol of tropomyosin was found. Purified scallop troponin C binds Ca2+ and interacts with scallop troponin I to relieve troponin I-induced inhibition of actomyosin ATPase. Although scallop troponin C is an acidic protein, it appears to be less acidic than troponin C from higher organisms. A calmodulin-like protein has been isolated from scallop striated muscle that activates bovine brain phosphodiesterase to the same extent as does brain calmodulin. Its amino acid composition and its electrophoretic mobility on alkaline 6 M-urea/polyacrylamide gels differs from that of scallop troponin C, and it appears not to be associated with thin filaments.


The basal bodies of Chlamydomonas reinhardtii. Formation from probasal bodies, isolation, and partial characterization.
The assembly and composition of basal bodies was investigated in the single-celled, biflagellate green alga, Chlamydomonas reinhardtii, using the cell wall-less strain, cw15. In the presence of EDTA, both flagellar axonemes remained attached to their basal bodies while the entire basal body-axoneme complex was separated from the cell body, without cell lysis, by treatment with polyethylene glycol-400. The axonemes were then removed from the basal bodies in the absence of EDTA, leaving intact basal body pairs, free from particulate contamination from other regions of the cell. The isolated organelles produced several bands on sodium dodecyl sulfate-urea polyacrylamide gels, including two tubilin bands which co-electrophoresed with flagellar tubulin. The formation of probasal bodies was observed by electron microscopy of whole mount preparations. Synchronous cells were lysed, centrifuged onto carbon-coated grids, and either negatively stained or shadowed with platinum. The two probasal bodies of each cell appeared shortly after mitosis as thin "annuli," not visible in thin sections, each consisting of nine rudimentary triplet microtubules. Each annulus remained attached to one of the mature basal bodies by several filaments about 60 in diameter, and persisted throughout interphase until just before the next cell division. It then elongated into a mature organelle. The results revive the possibility of the nucleated assembly of basal bodies.


Use of hospital services by patients with chronic conditions in sub-Saharan Africa: a systematic review and meta-analysis.
To estimate the prevalence of individual chronic conditions and multimorbidity among adults admitted to hospital in countries in sub-Saharan Africa. We systematically searched MEDLINE®, Embase®, Global Index Medicus, Global Health and SciELO for publications reporting on patient cohorts recruited between 1 January 2010 and 12 May 2023. We included articles reporting prevalence of pre-specified chronic diseases within unselected acute care services (emergency departments or medical inpatient settings). No language restrictions were applied. We generated prevalence estimates using random-effects meta-analysis alongside 95% confidence intervals, 95% prediction intervals and I2 statistics for heterogeneity. To explore associations with age, sex, country-level income status, geographical region and risk of bias, we conducted pre-specified meta-regression, sub-group and sensitivity analyses. Of 6976 identified studies, 61 met the inclusion criteria, comprising data from 20 countries and 376 676 people. None directly reported multimorbidity, but instead reported prevalence for individual conditions. Among medical admissions, the highest prevalence was human immunodeficiency virus infection (36.4%; 95% CI: 31.3-41.8); hypertension (24.4%; 95% CI: 16.7-34.2); diabetes (11.9%; 95% CI: 9.9-14.3); heart failure (8.2%; 95% CI: 5.6-11.9); chronic kidney disease (7.7%; 95% CI: 3.9-14.7); and stroke (6.8%; 95% CI: 4.7-9.6). Among patients seeking hospital care in sub-Saharan Africa, multimorbidity remains poorly described despite high burdens of individual chronic diseases. Prospective public health studies of multimorbidity burden are needed to generate integrated and context-specific health system interventions that act to maximize patient survival and well-being.


Occupational exposures of firefighting and urinary tract cancer risk among men in the Norwegian Fire Departments Cohort.
Increased risks of bladder cancer and mesothelioma were the strongest evidence for the recent reclassification of firefighting as carcinogenic (Group 1) by the International Agency for Research on Cancer. Our study aim was to develop indicators for specific firefighting exposures and examine associations with urinary tract cancer (UTC), including bladder cancer. We developed indicators for exposure from employment at a fire department or in firefighting jobs, to fire and smoke, and to diesel exhaust for men in the Norwegian Fire Departments Cohort (n=4250). Incident UTC cases were obtained from the Cancer Registry of Norway (1960-2021). Poisson regression was used to estimate incidence rate ratios (IRR) with cumulative exposures grouped into tertiles (reference: lowest exposed tertile) with 0-year, 10-year and 15-year lagging of exposures. During 125 090 person-years of follow-up, there were 76 cases of UTC. IRRs were mostly non-significantly increased in the middle tertile and at or below 1 in the highest tertile for total duration of employment, number of fires attended and fire exposure score with and without lags. In the middle tertile for diesel exhaust exposure, UTC risk was elevated over twofold with 10-year (IRR 2.27, 95% CI 1.22 to 4.20) and 15- year (2.21, 1.18 to 4.16) lags, and near 1 in the highest tertile. Findings for bladder cancer were similar to those for UTC. Dose-response associations between the exposure indicators and UTC were not observed. Future studies using the indicators with more cases are needed.


Psychosocial adjustment of burn survivors.
The purpose of this study was to assess the magnitude and predictors of psychosocial adjustment in burn victims. It was postulated that once individuals sustain a burn, their long-term psychosocial adjustment is a function of their present coping responses, social resources, burn severity and time since burn. It was expected that these variables could also be used to identify individuals at risk for psychosocial maladjustment. A historical cohort analytical survey of 340 randomly selected adults and the mothers of 145 children who had sustained either major or minor burns during the past 12 years were administered the Coping Scale, Participation in Social and Recreational Activities Index, Social Support and the Psychosocial Adjustment to Illness Scale. The children's mothers also completed the Family Environment Scale and the Child Behaviour Checklist. In summary, the variance in psychosocial adjustment among adults was related to unemployment, loss of occupational status, avoidance coping, and little involvement in recreational activities. Together, these variables explained 40 per cent of the variance in psychosocial adjustment. Severity of the burn and time since the burn were not related to psychosocial adjustment. The prevalence of psychosocial maladjustment among the adults was 10 per cent and 15.7 per cent among children. Psychosocial adjustment among children was not related to the severity of the burn. The less adjusted children could be distinguished from adjusted children on the basis of their mothers' adjustment and methods of coping. The findings tended to refute the commonly held view that post-burn adjustment is associated with burn severity and suggests psychosocial adjustment is a function of both coping responses and social resources.


Sphingomonas formosensis sp. nov., a polycyclic aromatic hydrocarbon-degrading bacterium isolated from agricultural soil.
In the present study, a yellow-pigmented, Gram-negative, short rod-shaped novel bacterium that was capable of degrading a wide range of polycyclic aromatic hydrocarbons (naphthalene, phenanthrene and pyrene) was isolated from agricultural soil located in Yunlin County, Taiwan. Comparative 16S rRNA gene sequence analysis positioned the novel strain in the genus Sphingomonas as an independent lineage adjacent to a subclade containing Sphingomonas fennica K101(T), Sphingomonas histidinilytica UM2(T), Sphingomonas wittichii RW1(T) and Sphingomonas haloaromaticamans A175(T). 16S rRNA gene sequence analysis of strain CC-Nfb-2(T) showed highest sequence similarity to S. fennica K101(T) (96.2%), S. histidinilytica UM2(T) (96.1%), S. wittichii RW1(T) (95.9%), S. haloaromaticamans A175(T) (95.7%), and Sphingobium ummariense RL-3(T) (94.7%); lower sequence similarities were observed with strains of all other Sphingomonas species. The strain contained phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, sphingoglycolipid and diphosphatidylglycerol. The predominant fatty acids were summed feature 8 (C(18:1)ω7c and/or C(18:1)ω6c) C(16:0) and 11-methyl C(18:1)ω7c; C(14:0) 2-OH was the major 2-hydroxy fatty acid. Previously, these lipids have been found to be characteristic of members of the genus Sphingomonas. The serine palmitoyl transferase gene (spt) was also detected and sphingolipid synthesis was confirmed. The predominant isoprenoid quinone system was ubiquinone (Q-10) and the isolate contained sym-homospermidine as the major polyamine. The DNA G+C content of the isolate was 62.8±0.8 mol%. On the basis of chemotaxonomic, phenotypic and phylogenetic data, strain CC-Nfb-2(T) represents a novel species within the genus Sphingomonas, for which the name Sphingomonas formosensis sp. nov. is proposed; the type strain is CC-Nfb-2(T) (=BCRC 80272(T)=DSM 24164(T)).


Endovascular repair of traumatic mesocaval fistula.
A patient presented with a posttraumatic fistula between the superior mesenteric artery and inferior vena cava with associated pseudoaneurysm after multiple thoracoabdominal gunshot wounds. Endovascular management with placement of two overlapping stent-grafts resulted in complete resolution of the lesions with documented patency at 15 months and no known complications. This minimally invasive therapy spared the patient the morbidity of conventional open surgical repair.


Colonic stent placement as a bridge to surgery in patients with left-sided malignant large bowel obstruction. An observational study.
Acute left-sided malignant colonic obstruction is common in elderly patients, in which emergency surgery is related with high morbidity and mortality rates, and often necessitates a two-step resection. Although the use of self-expanding metallic stents (SEMS) in elderly patients has not been adequately described yet, there are almost two international important trials which are still in progress, the stenting technique is established to be, by the international literature, an useful treatment with low morbidity and mortality. It's also a bridge to surgery, since the insertion of a SEMS can decompress the obstruction, making bowel and patient preparation possible and facilitating singlestage surgical resection. Palliative stenting can improve quality of life when compared to surgery in patients with metastasis or high co-morbidity. The aim of this study is to analyze mortality, avoidance of stoma, short- and long-term survival in patient with malignant left-sided large bowel obstruction who underwent to stent placement in our Emergency Surgery Unit, which is operative since November 2010 in our city Hospital in Ferrara. Between November 2010 and December 2012 a total of 15 patients with acute left-sided malignant large bowel obstruction suitable for colonic stent application were admitted to Emergency Surgery Unit. Among these patients, 9 underwent to self-expanding metallic stent placement (group A), the other (group B) 6 patient underwent to emergency surgery. In this observational not-randomized study we analyzed the efficacy and safety of SEMS placement for patients either as a bridge to surgery or as a palliation, beside the short term and long term outcomes, versus those patients operated straight. Self-expanding metallic stents were successfully implanted in 9 of the 15 patients with acute left-sided malignant large bowel obstruction. No acute procedure-related complication was observed. All the patients in group A kept the stent in place for an average of 7,7 days, then everyone underwent to surgery. A large bowel resection with one-time recanalization was performed in 8 of the 9 patients. None Hartmann resection was necessary. Only one underwent again to surgery because of a dehiscence, a stoma was necessary. Between the other 6 patients in group B who underwent directly to surgery, In one case was necessary an Hartmann resection, another one incurred in dehiscence of the anastomosis that required reoperation with stoma creation. Placement of SEMS seems to be an useful alternative to emergent surgery in the management of acute left-sided bowel obstruction, both as a bridge to surgery and as a palliative procedure. SEMS can provide an effective and safe therapeutic option compared to emergency surgery, most of all in elderly patients, with a lower mortality rate, a significantly higher rate of primary anastomosis and the avoidance of stoma. However, to fully determine their role for these indications, more data and more high level evidence is required.


Identification of novel roles of the cytochrome p450 system in early embryogenesis: effects on vasculogenesis and retinoic Acid homeostasis.
The cytochrome P450-dependent monooxygenase system catalyzes the metabolism of xenobiotics and endogenous compounds, including hormones and retinoic acid. In order to establish the role of these enzymes in embryogenesis, we have inactivated the system through the deletion of the gene for the electron donor to all microsomal P450 proteins, cytochrome P450 reductase (Cpr). Mouse embryos homozygous for this deletion died in early to middle gestation (approximately 9.5 days postcoitum [dpc]) and exhibited a number of novel phenotypes, including the severe inhibition of vasculogenesis and hematopoiesis. In addition, defects in the brain, limbs, and cell types where CPR was shown to be expressed were observed. Some of the observed abnormalities have been associated with perturbations in retinoic acid homeostasis in later embryogenesis. Consistent with this possibility, embryos at 9.5 dpc had significantly elevated levels of retinoic acid and reduced levels of retinol. Further, some of the observed phenotypes could be either reversed or exacerbated by decreasing or increasing maternal retinoic acid exposure, respectively. Detailed analysis demonstrated a close relationship between the observed phenotype and the expression of genes controlling vasculogenesis. These data demonstrate that the cytochrome P450 system plays a key role in early embryonic development; this process appears to be, at least in part, controlled by regional concentrations of retinoic acid and has profound effects on blood vessel formation.


Cytoskeletal cross-linking and bundling in motor-independent contraction.
Eukaryotic and prokaryotic cells use cytoskeletal proteins to regulate and modify cell shape. During cytokinesis or eukaryotic cell crawling, contractile forces are generated inside the cell to constrict the division site or to haul the rear of the cell forward, respectively. In many cases, these forces have been attributed to the activity of molecular motors, such as myosin II, which, by pulling on actin filaments, can produce contraction of the actin cytoskeleton. However, prokaryotic division is driven by the tubulin-like protein FtsZ and does not seem to require additional molecular motors to constrict the division site. Likewise, Dictyostelium discoideum and Saccharomyces cerevisiae can perform cytokinesis under motor-free conditions. In addition, many crawling cells can translocate when myosin is inhibited or absent. In this review, we point out another force-generation mechanism that can play a significant role in driving these processes in eukaryotes and prokaryotes. This mechanism is mediated by cross-linking and bundling proteins that form effective interactions between cytoskeletal filaments. Some recent studies in this area are reviewed and the physical underpinnings of this force-generation mechanism are explained.


Medical selection: Auschwitz antecedents and effluent.
Medical selection in Auschwitz represents the penultimate application of the traditional paradigm of medicine; the physician as gatekeeper, and decision maker. The historical evolution of that role is considered in the context of public health, medical police, quarantine and immigration. In Nazi Germany the physician was assigned responsibility for selection on behalf of the state. The ethical implications of medical selection are considered in the context of medicine today in an age of sophisticated biotechnology, constrained resources, and an aging population; an age in which the medical profession has yet to establish a fundamental system of values.


Role of let-7b in the regulation of N-acetylgalactosaminyltransferase 2 in IgA nephropathy.
IgA nephropathy (IgAN) is characterized by aberrant O-glycosylation in the hinge region of IgA1. The early step in O-glycan formation is the attachment of N-acetylgalactosamine (GalNAc) to the serine/threonine of the hinge region; the process is catalysed by UDP-N-acetyl-α-d-galactosamine:polypeptide N-acetylgalactosaminyltransferase 2 (GALNT2). In our previous work, the microarray analysis on peripheral blood mononuclear cells (PBMCs) identified an upregulated miRNA called let-7b. To study the molecular mechanisms in which let-7b was involved, we performed a bioinformatic analysis to predict their target genes. To validate biologically let-7b targets, we performed transient transfection experiments ex vivo using PBMCs from an independent group of IgAN patients and healthy blood donors (HBDs). Bioinformatic analysis revealed that GALNT2 is the potential target of let-7b. We found this miRNA significantly upregulated in PBMCs of IgAN patients compared with HBDs. Then, we demonstrated in ex-vivo experiments that let-7b decreased GALNT2 levels in PBMCs of IgAN patients, whereas the loss of let-7b function in PBMCs of HBDs led to an increase of GALNT2 mRNA and its protein level. Finally, we found that upregulation of let-7b occurred also in B-lymphocytes from IgAN patients. Our results give novel additional information on the abnormal O-glycosylation process of IgA1 in IgAN patients. This study provides evidence for another important miRNA-based regulatory mechanism of the O-glycosylation process in which the deregulated expression of let-7b is associated with altered expression of GALNT2. This finding could be taken into consideration for new therapeutic approaches in IgAN because other serum glycosylated proteins do not display abnormal glycosylation.


Magnetic field sensor based on a dual-frequency optoelectronic oscillator using cascaded magnetostrictive alloy-fiber Bragg grating-Fabry Perot and fiber Bragg grating-Fabry Perot filters.
A magnetic field sensor using a dual-frequency optoelectronic oscillator (OEO) incorporating cascaded magnetostrictive alloy-fiber Bragg grating-Fabry Perot (MA-FBG-FP) and FBG-FP filters is proposed and demonstrated. In the OEO resonant cavity, two microwave signals are generated, whose oscillation frequencies are determined by the FBG-FP filter and MA-FBG-FP filter filters with two ultra-narrow notches and two laser sources. Due to the characteristics of MA and FBG, the two generated microwave signals show different magnetic field and temperature sensitivities. By monitoring the variations of two oscillating frequencies and the beat signal using a digital signal processor, the simultaneous measurement for the magnetic field and temperature can be realized. The proposed sensor has the advantages of high-speed and high-resolution measurement, which make it very attractive for practical magnetic field sensing applications. The sensitivities of the proposed OEO sensor for magnetic field and temperature are experimentally measured to be as high as -38.4MHz/Oe and -1.23 or -2.45 GHz/°C corresponding to the MA-FBG-FP filter and FBG-FP filter, respectively.


Phosphorylation of Ku dictates DNA double-strand break (DSB) repair pathway choice in S phase.
Multiple DNA double-strand break (DSB) repair pathways are active in S phase of the cell cycle; however, DSBs are primarily repaired by homologous recombination (HR) in this cell cycle phase. As the non-homologous end-joining (NHEJ) factor, Ku70/80 (Ku), is quickly recruited to DSBs in S phase, we hypothesized that an orchestrated mechanism modulates pathway choice between HR and NHEJ via displacement of the Ku heterodimer from DSBs to allow HR. Here, we provide evidence that phosphorylation at a cluster of sites in the junction of the pillar and bridge regions of Ku70 mediates the dissociation of Ku from DSBs. Mimicking phosphorylation at these sites reduces Ku's affinity for DSB ends, suggesting that phosphorylation of Ku70 induces a conformational change responsible for the dissociation of the Ku heterodimer from DNA ends. Ablating phosphorylation of Ku70 leads to the sustained retention of Ku at DSBs, resulting in a significant decrease in DNA end resection and HR, specifically in S phase. This decrease in HR is specific as these phosphorylation sites are not required for NHEJ. Our results demonstrate that the phosphorylation-mediated dissociation of Ku70/80 from DSBs frees DNA ends, allowing the initiation of HR in S phase and providing a mechanism of DSB repair pathway choice in mammalian cells.


Effect of Stromal Cells in Tumor Microenvironment on Metastasis Initiation.
The cellular environment where tumor cells reside is called the tumor microenvironment (TME), which consists of borders, blood vessels, lymph vessels, extracellular matrix (ECM), stromal cells, immune/inflammatory cells, secreted proteins, RNAs and small organelles. By dynamically interacting with tumor cells, stromal cells participate in all stages of tumor initiation, progression, metastasis, recurrence and drug response, and consequently, affect the fate of patients. During the processes of tumor evolution and metastasis initiation, stromal cells in TME also experience some changes and play roles in both the suppression and promotion of metastasis, while the overall function of stromal cells is beneficial for cancer cell survival and movement. In this review, we examine the effects of stromal cells in TME on metastasis initiation, including angiogenesis, epithelial-mesenchymal transition (EMT) and invasion. We also highlight functions of proteins, RNAs and small organelles secreted by stromal cells in their influences on multiple stages of tumor metastasis.


Pharmacoepidemiology: A time for a new multidisciplinary approach to precision medicine.
The advent of the genomic age has created a rapid increase in complexity for the development and selection of drug treatments. A key component of precision medicine is the use of genetic information to improve therapeutic effectiveness of drugs and prevent potential adverse drug reactions. Pharmacoepidemiology, as a field, uses observational methods to evaluate the safety and effectiveness of drug treatments in populations. Pharmacoepidemiology by virtue of its focus, tradition, and research orientation can provide appropriate study designs and analysis methods for precision medicine. The objective of this manuscript is to demonstrate how pharmacoepidemiology can impact and shape precision medicine and serve as a reference for pharmacoepidemiologists interested in contributing to the science of precision medicine. This paper depicts the state of the science with respect to the need for pharmacoepidemiology and pharmacoepidemiological methods, tools and approaches for precision medicine; the need for and how pharmacoepidemiologists use their skills to engage with the precision medicine community; and recommendations for moving the science of precision medicine pharmacoepidemiology forward. We propose a new integrated multidisciplinary approach dedicated to the emerging science of precision medicine pharmacoepidemiology.


Coconut oil as a protective carrier of dietary vitamin A fed to ruminants.
Safflower and coconut oil were compared as protective carriers of dietary vitamin A supplemented to abomasally cannulated sheep. Vitamin A, 240,000 I.U., was predissolved in 11.7 g of safflower or coconut oil and bolused directly into the rumen of mature wethers along with 4 g of chromic oxide. The vitamin A was pre-dissolved in 0, 11.7, 23.4 or 35.0 g of coconut oil in experiment 2. Determination of carrier oil protectiveness of solubilized dietary vitamin A was based on recoveries of vitamin A supplement and chromic oxide in abomasal digesta 24 h post dosage. Vitamin A recoveries were significantly higher (P < 0.05) when dissolved in coconut oil (55.6%) compared to safflower oil (35.5%). Recoveries of vitamin A in abomasal digesta increased linearly (P < 0.01) with the amount of carrier coconut oil in experiment 2. Results of these experiments support the potential use of coconut oil as a protective carrier of ruminal labile vitamin A.


Molecular basis for the nerve dependence of limb regeneration in an adult vertebrate.
The limb blastemal cells of an adult salamander regenerate the structures distal to the level of amputation, and the surface protein Prod 1 is a critical determinant of their proximodistal identity. The anterior gradient protein family member nAG is a secreted ligand for Prod 1 and a growth factor for cultured newt blastemal cells. nAG is sequentially expressed after amputation in the regenerating nerve and the wound epidermis-the key tissues of the stem cell niche-and its expression in both locations is abrogated by denervation. The local expression of nAG after electroporation is sufficient to rescue a denervated blastema and regenerate the distal structures. Our analysis brings together the positional identity of the blastema and the classical nerve dependence of limb regeneration.


Automatic skull segmentation from MR images for realistic volume conductor models of the head: Assessment of the state-of-the-art.
Anatomically realistic volume conductor models of the human head are important for accurate forward modeling of the electric field during transcranial brain stimulation (TBS), electro- (EEG) and magnetoencephalography (MEG). In particular, the skull compartment exerts a strong influence on the field distribution due to its low conductivity, suggesting the need to represent its geometry accurately. However, automatic skull reconstruction from structural magnetic resonance (MR) images is difficult, as compact bone has a very low signal in magnetic resonance imaging (MRI). Here, we evaluate three methods for skull segmentation, namely FSL BET2, the unified segmentation routine of SPM12 with extended spatial tissue priors, and the skullfinder tool of BrainSuite. To our knowledge, this study is the first to rigorously assess the accuracy of these state-of-the-art tools by comparison with CT-based skull segmentations on a group of ten subjects. We demonstrate several key factors that improve the segmentation quality, including the use of multi-contrast MRI data, the optimization of the MR sequences and the adaptation of the parameters of the segmentation methods. We conclude that FSL and SPM12 achieve better skull segmentations than BrainSuite. The former methods obtain reasonable results for the upper part of the skull when a combination of T1- and T2-weighted images is used as input. The SPM12-based results can be improved slightly further by means of simple morphological operations to fix local defects. In contrast to FSL BET2, the SPM12-based segmentation with extended spatial tissue priors and the BrainSuite-based segmentation provide coarse reconstructions of the vertebrae, enabling the construction of volume conductor models that include the neck. We exemplarily demonstrate that the extended models enable a more accurate estimation of the electric field distribution during transcranial direct current stimulation (tDCS) for montages that involve extraencephalic electrodes. The methods provided by FSL and SPM12 are integrated into pipelines for the automatic generation of realistic head models based on tetrahedral meshes, which are distributed as part of the open-source software package SimNIBS for field calculations for transcranial brain stimulation.


Implementation outcomes of a multiinstitutional web-based ethical, legal, and social implications genetics curriculum for primary care residents in three specialties.
Medical genetics lends itself to disseminated teaching methods because of mismatches between numbers of physicians having patients with genetic disorders and availability of genetic specialists. During 3 years, we implemented an interactive, web-based curriculum on ethical, legal, and social implications in medical genetics for primary care residents in three specialties at three institutions. Residents took five (of 10) cases and three (of five) tutorials that varied by specialty. We assessed changes in self-efficacy (primary outcome), knowledge, application, and viewpoints. Overall enrollment was 69% (279/403). One institution did not complete implementation and was dropped from pre-post comparisons. We developed a six-factor ethical, legal, and social implications self-efficacy scale (Cronbach α = 0.95). Baseline self-efficacy was moderate (71/115; range: 23-115) and increased 15% after participation. Pre-post knowledge scores were high and unchanged. Residents reported that this curriculum covered ethical, legal, and social implications/genetics better than their usual curricula. Most (68-91%) identified advantages, especially in providing flexibility and stimulating self-directed learning. After participation, residents reported creating learning goals (66%) and acting on those goals (62%). Ethical, legal, and social implications genetics curricular participation led to modest self-efficacy gains. Residents reported that the curriculum covered unique content areas, had advantages over traditional curriculum, and that they applied ethical, legal, and social implications content clinically. We share lessons from developing and implementing this complex web-based curriculum across multiple institutions.


Continuously-stirred anaerobic digester to convert organic wastes into biogas: system setup and basic operation.
Anaerobic digestion (AD) is a bioprocess that is commonly used to convert complex organic wastes into a useful biogas with methane as the energy carrier. Increasingly, AD is being used in industrial, agricultural, and municipal waste(water) treatment applications. The use of AD technology allows plant operators to reduce waste disposal costs and offset energy utility expenses. In addition to treating organic wastes, energy crops are being converted into the energy carrier methane. As the application of AD technology broadens for the treatment of new substrates and co-substrate mixtures, so does the demand for a reliable testing methodology at the pilot- and laboratory-scale. Anaerobic digestion systems have a variety of configurations, including the continuously stirred tank reactor (CSTR), plug flow (PF), and anaerobic sequencing batch reactor (ASBR) configurations. The CSTR is frequently used in research due to its simplicity in design and operation, but also for its advantages in experimentation. Compared to other configurations, the CSTR provides greater uniformity of system parameters, such as temperature, mixing, chemical concentration, and substrate concentration. Ultimately, when designing a full-scale reactor, the optimum reactor configuration will depend on the character of a given substrate among many other nontechnical considerations. However, all configurations share fundamental design features and operating parameters that render the CSTR appropriate for most preliminary assessments. If researchers and engineers use an influent stream with relatively high concentrations of solids, then lab-scale bioreactor configurations cannot be fed continuously due to plugging problems of lab-scale pumps with solids or settling of solids in tubing. For that scenario with continuous mixing requirements, lab-scale bioreactors are fed periodically and we refer to such configurations as continuously stirred anaerobic digesters (CSADs). This article presents a general methodology for constructing, inoculating, operating, and monitoring a CSAD system for the purpose of testing the suitability of a given organic substrate for long-term anaerobic digestion. The construction section of this article will cover building the lab-scale reactor system. The inoculation section will explain how to create an anaerobic environment suitable for seeding with an active methanogenic inoculum. The operating section will cover operation, maintenance, and troubleshooting. The monitoring section will introduce testing protocols using standard analyses. The use of these measures is necessary for reliable experimental assessments of substrate suitability for AD. This protocol should provide greater protection against a common mistake made in AD studies, which is to conclude that reactor failure was caused by the substrate in use, when really it was improper user operation.


High WT1 expression after induction therapy predicts high risk of relapse and death in pediatric acute myeloid leukemia.
To determine whether minimal residual disease (MRD) measured by Wilms' tumor gene 1 (WT1) expression is a prognostic marker in pediatric acute myeloid leukemia (AML), we quantified WT1 transcript by real-time quantitative-polymerase chain reaction in 92 AML at diagnosis and during follow-up. Patients (median age, 6 years; cytogenetics, favorable 27%, intermediate 59%, poor 13%) were treated between 1995 and 2002 and enrolled in Leucémie aiguë Myéloblastique Enfant (LAME) 89/91, LAME 99 pilot study and Acute Promyelocytic Leukemia French collaborative protocols. With a median follow-up of 26 months, event-free survival was 56% with a standard deviation (SD) of 5% and overall survival of 62.5% with an SD of 6%. WT1 copy number was normalized by TATA box binding protein gene transcripts and expressed as WT1/TBP x 1,000 ratio. Median WT1 ratio in normal patient controls was 12 (range, 0 to 57). A level over two SD than normal bone marrow controls (ie, WT1 ratio > 50), was considered as significant overexpression. At diagnosis, WT1 overexpression was detected in 78% of patients (72 of 92 patients; median copy ratio, 2231). The WT1 values were significantly higher (P = .01) in favorable cytogenetics and lower (P < .0001) in M5-FAB subtype, 11q23 rearrangements (P < .001), and infants (P = .003) and demonstrate a strong correlation with fusion transcript AML1-ETO, PML-RARalpha expression. After induction treatment, WT1 ratio was analyzed in 46 of 72 patients and found above 50 in nine of 36 patients and five of 25 patients at D35-50 and 3 to 5 months, respectively. WT1 ratio > 50 after induction is an independent prognostic risk factor of relapse (P = .002) and death (P = .02). WT1 quantification is an informative molecular marker for MRD in pediatric AML and is now performed as prospective analysis in ELAM02 protocol.


Exterior carpi radialis longus tendinoplasty for thumb basal joint arthritis.
Thumb carpometacarpal arthritis has been successfully treated with a combination of trapezium excision, ligament reconstruction, and tendon interposition (most commonly with the flexor carpi radialis [FCR] tendon). We describe a technique using the extensor carpi radialis longus (ECRL) tendon and show, through dissection of 36 cadaver hands, the close relationship between this tendon and the intermetacarpal ligament. Of 16 patients (19 hands) managed with this technique, 95% were satisfied at a mean follow-up of 42 months. We conclude that ligament reconstruction and ECRL tendon interposition constitute a viable treatment option for carpometacarpal joint arthritis, especially when the FCR tendon is unavailable or its use is undesirable.


The origin and loss of periodic patterning in the turtle shell.
The origin of the turtle shell over 200 million years ago greatly modified the amniote body plan, and the morphological plasticity of the shell has promoted the adaptive radiation of turtles. The shell, comprising a dorsal carapace and a ventral plastron, is a layered structure formed by basal endochondral axial skeletal elements (ribs, vertebrae) and plates of bone, which are overlain by keratinous ectodermal scutes. Studies of turtle development have mostly focused on the bones of the shell; however, the genetic regulation of the epidermal scutes has not been investigated. Here, we show that scutes develop from an array of patterned placodes and that these placodes are absent from a soft-shelled turtle in which scutes were lost secondarily. Experimentally inhibiting Shh, Bmp or Fgf signaling results in the disruption of the placodal pattern. Finally, a computational model is used to show how two coupled reaction-diffusion systems reproduce both natural and abnormal variation in turtle scutes. Taken together, these placodal signaling centers are likely to represent developmental modules that are responsible for the evolution of scutes in turtles, and the regulation of these centers has allowed for the diversification of the turtle shell.


Therapeutic concentrations of mitotane (o,p'-DDD) inhibit thyrotroph cell viability and TSH expression and secretion in a mouse cell line model.
Mitotane therapy is associated with many side effects, including thyroid function perturbations mimicking central hypothyroidism, possibly due to laboratory test interference or pituitary direct effects of mitotane. We investigated whether increasing concentrations of mitotane in the therapeutic range might interfere with thyroid hormone assays and evaluated the effects of mitotane on a mouse TSH-producing pituitary cell line. TSH, free T(4), and free T(3) levels do not significantly change in sera from hypo-, hyper-, or euthyroid patients after addition of mitotane at concentrations in the therapeutic window. In the mouse TalphaT1 cell line, mitotane inhibits both TSH expression and secretion, blocks TSH response to TRH, and reduces cell viability, inducing apoptosis at concentrations in the therapeutic window. TRH is not capable of rescuing TalphaT1 cells from the inhibitory effects of mitotane on TSH expression and secretion, which appear after short time treatment and persist over time. Our results demonstrate that mitotane does not interfere with thyroid hormone laboratory tests but directly reduces both secretory activity and cell viability on pituitary TSH-secreting mouse cells. These data represent a possible explanation of the biochemical picture consistent with central hypothyroidism in patients undergoing mitotane therapy and open new perspectives on the direct pituitary effects of this drug.


Mapping bone cell distributions to assess ontogenetic origin of primate midfacial form.
Midfacial reduction in primates has been explained as a byproduct of other growth patterns, especially the convergent orbits. This is at once an evolutionary and developmental explanation for relatively short snouts in most modern primates. Here, we use histological sections of perinatal nonhuman primates (tamarin, tarsier, loris) to investigate how orbital morphology emerges during ontogeny in selected primates compared to another euarchontan (Tupaia glis). We annotated serial histological sections for location of osteoclasts or osteoblasts, and used these to create three-dimensional "modeling maps" showing perinatal growth patterns of the facial skeleton. In addition, in one specimen we transferred annotations from histological sections to CT slices, to create a rotatable 3D volume that shows orbital modeling. Our findings suggest that growth in the competing orbital and neurocranial functional matrices differs among species, influencing modeling patterns. Distinctions among species are observed in the frontal bone, at a shared interface between the endocranial fossa and the orbit. The medial orbital wall is extensively resorptive in primates, whereas the medial orbit is generally depositional in Tupaia. As hypothesized, the orbital soft tissues encroach on available interorbital space. However, eye size cannot, by itself, explain the extent of reduction of the olfactory recess. In Loris, the posterior portion of medial orbit differed from the other primates. It showed evidence of outward drift where the olfactory bulb increased in cross-sectional area. We suggest the olfactory bulbs are significant to orbit position in strepsirrhines, influencing an expanded interorbital breadth at early stages of development.


Early chick embryos in vitro.
In 1955, Denis New described a technique for the in vitro culture of early avian embryos that has formed the basis for nearly all of the experimental embryological studies performed on these species since that day. Many modifications to this technique have also been described in these four decades for specific experimental purposes. Here, we review the effects of some parameters that appear to be important for different aspects of the growth of embryos in this type of culture, and conduct a small experimental comparison between different modifications of the technique as described by various authors. We conclude that the original technique still compares favorably with its alternatives.


Profile of Hospital Admissions due to Self-Inflicted Harm in Los Angeles County from 2001 to 2010.
We aimed to describe the demographic profile of self-inflicted harm (SIH) in Los Angeles County between 2001 and 2010 and to investigate trends over this 10-year period. We used the California Hospital Discharge Data to investigate all cases of hospital admission due to SIH, including suicide attempts and if they had a concurrent psychiatric diagnosis based on ICD-9 codes. African-Americans (AA) had the highest 10-year SIH admission rates. SIH admission rates remained steady throughout the 10-year study period. Median age of SIH was significantly lower in Latinos. Episodic mood disorders were the most common psychiatric comorbidity. The use of solid/liquid poisoning was the most common SIH method among all racial/ethnic groups. We found major disparities in SIH admissions across racial/ethnic subgroups. The importance of programs to identify, prevent, and treat SIH in these groups is discussed.


Inorganic polyphosphate as an energy source in tumorigenesis.
Cancer cells have high demands for energy to maintain their exceedingly proliferative growth. However, the mechanism of energy expenditure in cancer is not well understood. We hypothesize that cancer cells might utilize energy-rich inorganic polyphosphate (polyP), as energetic reserve. PolyP is comprised of orthophosphates linked by phosphoanhydride bonds, as in ATP. Here, we show that polyP is highly abundant in several types of cancer cells, including brain tumor-initiating cells (BTICs), i.e., stem-like cells derived from a mouse brain tumor model that we have previously described. The polymer is avidly consumed during starvation of the BTICs. Depletion of ATP by inhibiting glycolysis and mitochondrial ATP-synthase (OXPHOS) further decreases the levels of polyP and alters morphology of the cells. Moreover, enzymatic hydrolysis of the polymer impairs the viability of cancer cells and significantly deprives ATP stores. These results suggest that polyP might be utilized as a source of phosphate energy in cancer. While the role of polyP as an energy source is established for bacteria, this finding is the first demonstration that polyP may play a similar role in the metabolism of cancer cells.


[Occupational cancer in Italy].
The paper is a discussion of occupational cancer in Italy. The introductory section provides the necessary context of Italian industrialization and occupational health regulation. This is followed by a review of Italian epidemiological studies of occupational cancer risks, considered in terms of relative measures of risk and Attributable Risk to carcinogenic agents or exposure circumstances. A section attempts to establish the number of workers who have been exposed to carcinogens in Italy and the intensity of their exposures. Several cohort and case-control studies have addressed the issue of occupational risks, mostly among male workers. The results of these studies suggest that the growing incidence of and mortality by mesothelioma is explained by the widespread and intense exposure to asbestos in some Italian industrial settings. A high Attributable Risk of lung tumors among male populations in industrial areas of northern Italy is explained by occupational exposures. However, insufficient data are available for the clear definition of the extent and intensity of occupational exposure to carcinogenic substances. In Italy, we need to prioritize and maximize resources in occupational cancer epidemiology and to revitalize the role of national institutions. Recent legislation has established new regulations on the handling of carcinogenic substances in industrial settings, a new list of occupational diseases, and a national registry of mesothelioma linked to asbestos exposure. These legislative changes are expected to have positive effects.


High Prevalence of Diverse Insertion Sequences within the rRNA Operons of Mycoplasma bovis.
Insertion sequences (ISs) are widespread in the genome of Mycoplasma bovis strain PG45, but no ISs were identified within its two tandemly positioned rRNA operons (rrn1 and rrn2). However, characterization of the rrn locus in 70 M. bovis isolates revealed the presence of ISs related to the ISMbov1 (IS30 family) and ISMbov4 (IS4 family) isomers in 35 isolates. ISs were inserted into intergenic region 1 (IGR-1) or IGR-3, which are the putative promoter regions of rrn1 and rrn2, respectively, and into IGR-5, located downstream of the rrl2 gene. Seven different configurations (A to G) of the rrn locus with respect to ISs were identified, including those in five annotated genomes. The transcriptional start site for the single rrn operon in M. bovis strain 88127 was mapped within IGR-1, 60 bp upstream of the rrs gene. Notably, only 1 nucleotide separated the direct repeat (DR) for ISMbov1 and the promoter -35 element in configuration D, while in configuration F, the -35 motif was a part of the ISMbov1 DR. Relative quantitative real-time (qRT) PCR analysis and growth rate comparisons detected a significant increase (P < 0.05) in the expression of the rrs genes and in the number of viable cells during log phase growth (8, 12, and 16 h) in the strains with configuration F in comparison to strains with one or two rrn operons that did not have ISs. A high prevalence of IS elements within or close to the M. bovis rrn operon-promoter region may reflect their important role in regulation of both ribosome synthesis and function. Data presented in this study show a high prevalence of diverse ISs within the M. bovis rrn locus resulting in intraspecies variability and diversity. Such abundance of IS elements near or within the rrn locus may offer a selective advantage to M. bovis Moreover, the fact that expression of the rrs genes as well as the number of viable cells increased in the group of strains with IS element insertion within a putative promoter -35 sequence (configuration F) in comparison to that in strains with one or two rrn operons that do not have ISs may serve as a basis for understanding the possible role of M. bovis IS elements in fundamental biological processes such as regulation of ribosome synthesis and function.


The novel cyst nematode effector protein 19C07 interacts with the Arabidopsis auxin influx transporter LAX3 to control feeding site development.
Plant-parasitic cyst nematodes penetrate plant roots and transform cells near the vasculature into specialized feeding sites called syncytia. Syncytia form by incorporating neighboring cells into a single fused cell by cell wall dissolution. This process is initiated via injection of esophageal gland cell effector proteins from the nematode stylet into the host cell. Once inside the cell, these proteins may interact with host proteins that regulate the phytohormone auxin, as cellular concentrations of auxin increase in developing syncytia. Soybean cyst nematode (Heterodera glycines) Hg19C07 is a novel effector protein expressed specifically in the dorsal gland cell during nematode parasitism. Here, we describe its ortholog in the beet cyst nematode (Heterodera schachtii), Hs19C07. We demonstrate that Hs19C07 interacts with the Arabidopsis (Arabidopsis thaliana) auxin influx transporter LAX3. LAX3 is expressed in cells overlying lateral root primordia, providing auxin signaling that triggers the expression of cell wall-modifying enzymes, allowing lateral roots to emerge. We found that LAX3 and polygalacturonase, a LAX3-induced cell wall-modifying enzyme, are expressed in the developing syncytium and in cells to be incorporated into the syncytium. We observed no decrease in H. schachtii infectivity in aux1 and lax3 single mutants. However, a decrease was observed in both the aux1lax3 double mutant and the aux1lax1lax2lax3 quadruple mutant. In addition, ectopic expression of 19C07 was found to speed up lateral root emergence. We propose that Hs19C07 most likely increases LAX3-mediated auxin influx and may provide a mechanism for cyst nematodes to modulate auxin flow into root cells, stimulating cell wall hydrolysis for syncytium development.


Mycobacterium seoulense sp. nov., a slowly growing scotochromogenic species.
A previously undescribed, slowly growing, scotochromogenic mycobacterium was isolated from a patient with symptomatic pulmonary infection during hsp65 sequence-based identification of Korean clinical isolates. Phenetic characteristics of this strain were generally similar to those of Mycobacterium nebraskense and Mycobacterium scrofulaceum. However, some phenetic characteristics differentiated it from these two species. Its 16S rRNA gene sequences were unique and phylogenetic analysis based on 16S rRNA gene sequences placed the organism in the slowly growing Mycobacterium group close to M. nebraskense and M. scrofulaceum. Its unique mycolic acid profiles and the results of phylogenetic analysis based on two independent alternative chronometer molecules, hsp65 and rpoB, confirmed the taxonomic status of this strain as representing a novel species. These data support the conclusion that this strain represents a novel mycobacterial species, for which the name Mycobacterium seoulense sp. nov. is proposed. The type strain is strain 03-19(T) (=DSM 44998(T)=KCTC 19146(T)).


A simulation tool to study high-frequency chest compression energy transfer mechanisms and waveforms for pulmonary disease applications.
High-frequency chest compression (HFCC) can be used as a therapeutic intervention to assist in the transport and clearance of mucus and enhance water secretion for cystic fibrosis patients. An HFCC pump-vest and half chest-lung simulation, with 23 lung generations, has been developed using inertance, compliance, viscous friction relationships, and Newton's second law. The simulation has proven to be useful in studying the effects of parameter variations and nonlinear effects on HFCC system performance and pulmonary system response. The simulation also reveals HFCC waveform structure and intensity changes in various segments of the pulmonary system. The HFCC system simulation results agree with measurements, indicating that the HFCC energy transport mechanism involves a mechanically induced pulsation or vibration waveform with average velocities in the lung that are dependent upon small air displacements over large areas associated with the vest-chest interface. In combination with information from lung physiology, autopsies and a variety of other lung modeling efforts, the results of the simulation can reveal a number of therapeutic implications.


Advanced glycation end products are associated with pulse pressure in type 1 diabetes: the EURODIAB Prospective Complications Study.
We investigated the associations of pulse pressure (a measure of arterial stiffness) with the early glycation products hemoglobin A1c (HbA1c) and Amadori albumin and the advanced glycation end products pentosidine, Nepsilon-(carboxymethyl)lysine and Nepsilon-(carboxyethyl)lysine in a large group of type 1 diabetic individuals of the EURODIAB Prospective Complications Study. We did a cross-sectional nested case-control study from the EURODIAB Prospective Complications Study of 543 (278 men) European individuals with type 1 diabetes diagnosed at <36 years of age. We used linear regression analyses to investigate the association of pulse pressure with glycation products. Pulse pressure was significantly associated with plasma levels of Nepsilon-(carboxymethyl)lysine and Nepsilon-(carboxyethyl)lysine but not with HbA1c, Amadori albumin, and urinary levels of pentosidine. Regression coefficients adjusted for age, sex, mean arterial pressure, and duration of diabetes were 0.09 mm Hg (P=0.003) per 1 microM/M lysine Nepsilon-(carboxymethyl)lysine; 0.24 mm Hg (P=0.001) and -0.03 mm Hg (P=0.62) per 1 microM/M lysine Nepsilon-(carboxyethyl)lysine (in individuals with and without complications, respectively; P interaction=0.002); and 0.50 mm Hg (P=0.16) per 1% HbA1c; 0.07 mm Hg (P=0.12) per 1 U/mL Amadori albumin; and 0.77 mm Hg (P=0.48) per 1 nmol/mmol creatinine pentosidine. In young type 1 diabetic individuals, arterial stiffness is strongly associated with the advanced glycation end products Nepsilon-(carboxymethyl)lysine and Nepsilon-(carboxyethyl)lysine. These findings suggest that the formation of advanced glycation end products is an important pathway in the development of arterial stiffness in young type 1 diabetic individuals.


Characterization of encapsulated and noncapsulated Haemophilus influenzae and determination of phylogenetic relationships by multilocus sequence typing.
A multilocus sequence typing (MLST) scheme has been developed for the unambiguous characterization of encapsulated and noncapsulated Haemophilus influenzae isolates. The sequences of internal fragments of seven housekeeping genes were determined for 131 isolates, comprising a diverse set of 104 serotype a, b, c, d, e, and f isolates and 27 noncapsulated isolates. Many of the encapsulated isolates had previously been characterized by multilocus enzyme electrophoresis (MLEE), and the validity of the MLST scheme was established by the very similar clustering of isolates obtained by these methods. Isolates of serotypes c, d, e, and f formed monophyletic groups on a dendrogram constructed from the differences in the allelic profiles of the isolates, whereas there were highly divergent lineages of both serotype a and b isolates. Noncapsulated isolates were distinct from encapsulated isolates and, with one exception, were within two highly divergent clusters. The relationships between the major lineages of encapsulated H. influenzae inferred from MLEE data could not be discerned on a dendrogram constructed from differences in the allelic profiles, but were apparent on a tree reconstructed from the concatenated nucleotide sequences. Recombination has not therefore completely eliminated phylogenetic signal, and in support of this, for encapsulated isolates, there was significant congruence between many of the trees reconstructed from the sequences of the seven individual loci. Congruence was less apparent for noncapsulated isolates, suggesting that the impact of recombination is greater among noncapsulated than encapsulated isolates. The H. influenzae MLST scheme is available at www.mlst.net, it allows any isolate to be compared with those in the MLST database, and (for encapsulated isolates) it assigns isolates to their phylogenetic lineage, via the Internet.


Puerarin Attenuates Complete Freund's Adjuvant-Induced Trigeminal Neuralgia and Inflammation in a Mouse Model via Sirt1-Mediated TGF-β1/Smad3 Inhibition.
Puerarin, an active compound of radix puerariae, is a major compound used in Chinese herbal medicines and it has been well known for its pharmacological effects, including antioxidant, anti‑inflammatory, neuroprotective and cardioprotective properties. The aim of the present study was to determine the role of puerarin (Pue) in complete Freund's adjuvant (CFA)-induced trigeminal neuralgia (TN) and the effects of this compound on Sirt1 activity and on the progression of CFA-induced TN. Mice were injected with CFA on the unilateral face to induce TN. A cell model of inflammation-associated TN was established by interleukin-1β (IL-1β; 10 ng/mL) and tumor necrosis factor-α (TNF-α; 50 ng/mL) stimulation of neurons. Reverse transcription-quantitative PCR and Western blot analyses were performed to analyze mRNA and protein expression levels in trigeminal ganglion and nerve cells. Terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) staining was used to determine nerve cell apoptosis following IL-1β/TNF-α or Pue treatment. Pue is a conceivable Sirtuin1 (Sirt1) activator used for the prevention of trigeminal nerve injury that attenuates CFA-induced TN and inflammatory cytokine-evoked overactivation of neuronal inflammation and apoptosis. Treatment of mice with inflammatory cytokines induced upregulation of cleaved caspase-3 protein expression, which was neutralized by Pue supplementation. Both in vivo and in vitro experiments led to the conclusion that Pue modulated Sirt1 activation and repressed transforming growth factor-β1 (TGF-β1) protein expression and drosophila mothers against decapentaplegic homolog3 (Smad3) phosphorylation in order to exert neuroprotection. The findings suggested that Pue functioned as a potential Sirt1 activator to improve neuroinflammation-induced TN and neuronal apoptosis via the suppression of TGF-β1/Smad3 activity. The pharmacological activity of Pue provides a new perspective for the effective prevention and treatment of TN.


Energy intake patterns in obese women with binge eating disorder.
For binge eating disorder (BED) to be accepted as a distinct diagnostic category in the Diagnostic and Statistical Manual for Mental Disorders, Fifth Edition, it must be demonstrated that the criteria identify a diagnostic entity that is distinct from bulimia nervosa and obesity. This study examined the difference in total energy intake per day, patterns of energy intake throughout the day, and nutrient content of foods consumed in obese individuals who met the criteria for BED (on binge and non-binge days) and those who did not. Twenty women, 12 who met Diagnostic and Statistical Manual for Mental Disorders, Fourth Edition, criteria for BED and 8 matched obese controls, participated in the study. All participants underwent six random 24-hour dietary recall telephone interviews conducted by experienced interviewers using the Nutrition Data Software System. The BED group ingested significantly more kilocalories on days when they had binge eating episodes than the obese control group on average. The BED group ate significantly more in the evening on binge days than their control group counterparts. There is some indication in the data that those with BED may be restricting caloric intake. Finally, data indicated that the BED group ate significantly more protein, carbohydrate, and fat on binge days than on non-binge days. However, the proportion of kilocalories from each nutrient shifted on BED binge days compared with non-binge days to favor consumption of fat over carbohydrates. More research needs to be done to determine if these findings are reproducible. Then, the neurobiological underpinnings of these differences in nutrient intake patterns and nutrient selection can be studied to help to determine the biological basis of the disorder.


Aided and unaided speech perception by older hearing impaired listeners.
The most common complaint of older hearing impaired (OHI) listeners is difficulty understanding speech in the presence of noise. However, tests of consonant-identification and sentence reception threshold (SeRT) provide different perspectives on the magnitude of impairment. Here we quantified speech perception difficulties in 24 OHI listeners in unaided and aided conditions by analyzing (1) consonant-identification thresholds and consonant confusions for 20 onset and 20 coda consonants in consonant-vowel-consonant (CVC) syllables presented at consonant-specific signal-to-noise (SNR) levels, and (2) SeRTs obtained with the Quick Speech in Noise Test (QSIN) and the Hearing in Noise Test (HINT). Compared to older normal hearing (ONH) listeners, nearly all unaided OHI listeners showed abnormal consonant-identification thresholds, abnormal consonant confusions, and reduced psychometric function slopes. Average elevations in consonant-identification thresholds exceeded 35 dB, correlated strongly with impairments in mid-frequency hearing, and were greater for hard-to-identify consonants. Advanced digital hearing aids (HAs) improved average consonant-identification thresholds by more than 17 dB, with significant HA benefit seen in 83% of OHI listeners. HAs partially normalized consonant-identification thresholds, reduced abnormal consonant confusions, and increased the slope of psychometric functions. Unaided OHI listeners showed much smaller elevations in SeRTs (mean 6.9 dB) than in consonant-identification thresholds and SeRTs in unaided listening conditions correlated strongly (r = 0.91) with identification thresholds of easily identified consonants. HAs produced minimal SeRT benefit (2.0 dB), with only 38% of OHI listeners showing significant improvement. HA benefit on SeRTs was accurately predicted (r = 0.86) by HA benefit on easily identified consonants. Consonant-identification tests can accurately predict sentence processing deficits and HA benefit in OHI listeners.


Vitamin A - application technology.
The availability of synthetic vitamin A and its esters in unlimited quantities, has enabled populations around the world, consuming inadequate amounts of this vital micro-nutrient and hence subject to potential loss of sight or other manifestations of vitamin A deficiency, to have hope for a better future life. A technology exists for the preparation of synthetic vitamin A in various application forms. Many commonly-consumed foods may be used as carriers or vehicles of vitamin A to assure deficient populations of a sufficient intake of this antixerophthalmic and anti-infective vitamin.


FASB (Financial Accounting Standards Board) proposes changes in not-for-profit reporting.
The Financial Accounting Standards Board (FASB) will soon issue two statements that will have a major impact on not-for-profit financial reporting. The first, entitled Financial Statements of Not-For-Profit Organizations, deals with the form and content of not-for-profit financial statements. The second, entitled Accounting for Contributions Received and Contributions Made, establishes standards of financial accounting and reporting for contributions.


Differential Responsiveness of Monocyte and Macrophage Subsets to Interferon.
Peripheral blood mononuclear cells (PBMCs) in systemic lupus erythematosus (SLE) patients exhibit a gene expression program (interferon [IFN] signature) that is attributed to overproduction of type I IFNs by plasmacytoid dendritic cells. Type I IFNs have been thought to play a role in the pathogenesis of SLE. This study was undertaken to examine an unexpected influence of monocyte/macrophages on the IFN signature. Proinflammatory (classic) and antiinflammatory (nonclassic) monocyte/macrophages were sorted from mice and analyzed by RNA sequencing and quantitative polymerase chain reaction (qPCR). Type I IFN-α/β/ω receptor (IFNAR-1) expression was determined by qPCR and flow cytometry. Macrophages were stimulated in vitro with IFNα, and pSTAT1was measured. Transcriptional profiling of peritoneal macrophages from mice with pristane-induced SLE unexpectedly indicated a strong IFN signature in classic, but not nonclassic, monocyte/macrophages exposed to the same type I IFN concentrations. Ifnar1 messenger RNA and IFNAR surface staining were higher in classic monocyte/macrophages versus nonclassic monocyte/macrophages (P < 0.0001 and P < 0.05, respectively, by Student's t-test). Nonclassic monocyte/macrophages were also relatively insensitive to IFNα-driven STAT1 phosphorylation. Humans exhibited a similar pattern: higher IFNAR expression (P < 0.0001 by Student's t-test) and IFNα-stimulated gene expression (P < 0.01 by paired Wilcoxon's rank sum test) in classic monocyte/macrophages and lower levels in nonclassic monocyte/macrophages. This study revealed that the relative abundance of different monocyte/macrophage subsets helps determine the magnitude of the IFN signature. Responsiveness to IFNα signaling reflects differences in IFNAR expression in classic (high IFNAR) compared to nonclassic (low IFNAR) monocyte/macrophages. Thus, the IFN signature depends on both type I IFN production and the responsiveness of monocyte/macrophages to IFNAR signaling.


Treatment of Guillain-Barré syndrome and CIDP.
Guillain-Barré syndrome (GBS) and chronic inflammatory demyelinating poly-(radiculo)neuropathy (CIDP) are immune-mediated disorders with a variable duration of progression and a range in severity of weakness. Infections can trigger GBS and exacerbate CIDP. Anti-ganglioside antibodies are important, but there is debate on the role of genetic factors in the pathogenesis of these disorders. Randomized controlled trials (RCT) have shown that intravenous immunoglobulin (IVIg) and plasma exchange (PE) are effective in both GBS and CIDP. Most CIDP patients also improve after steroid therapy. Despite current treatment options, many patients have residual deficits or need to be treated for a long period of time. Therefore, new treatment trials are highly indicated. This review focuses on the current and possible new treatment options that could be guided by recent results from laboratory experiments.


Association Between Delivery System Structure and Intensity of End-of-Life Cancer Care.
To determine whether the type of delivery system is associated with intensity of care at the end of life for Medicare beneficiaries with cancer. We used SEER registry data linked with Medicare claims to evaluate intensity of end-of-life care for patients who died of one of ten common cancers diagnosed from 2009 through 2014. Patients were categorized as receiving the majority of their care in an integrated delivery system, designated cancer center, health system that was both integrated and a certified cancer center, or health system that was neither. We evaluated adherence to seven nationally endorsed end-of-life quality measures using generalized linear models across four delivery system types. Among 100,549 beneficiaries who died of cancer during the study interval, we identified only modest differences in intensity of end-of-life care across delivery system structures. Health systems with no cancer center or integrated affiliation demonstrated higher proportions of patients with multiple hospitalizations in the last 30 days of life (11.3%), death in an acute care setting (25.9%), and lack of hospice use in the last year of life (31.6%; all P < .001). Patients enrolled in hospice had lower intensity care across multiple end-of-life quality measures. Intensity of care at the end of life for patients with cancer was higher at delivery systems with no integration or cancer focus. Maximal supportive care delivered through hospice may be one avenue to reduce high-intensity care at the end of life and may impact quality of care for patients dying from cancer.


Second hepatectomy for recurrent hepatocellular carcinoma achieved sustained virological response to interferon therapy for hepatitis C.
Interferon (IFN) improves the prognosis of HCV-related hepatocellular carcinoma (HCC) in patients. However, the effects of IFN therapy for second hepatectomy (Hx) for recurrent HCC have not been established. Subjects included 96 patients who underwent a second Hx for recurrence of HCV-related HCC. Forty-four patients received IFN therapy past or postoperatively of the first Hx. Twenty of those patients attained a sustained viral response (SVR). The other 24 were non-responders (NR) and 52 patients who had not received IFN therapy (non-IFN) were classified as the NR/non-IFN group. Overall survival (SVR group vs. NR/non-IFN group: 5-yr, 91.7 vs. 51.0%; p = 0.012) and disease-free survival (SVR group vs. NR/non-IFN group: 3-yr, 64.7 vs. 25.9%; p = 0.006) rates were significantly different in both groups. By multivariate analysis, NR/non-IFN therapy, was the independent risk factor for overall survival (p = 0.025) and disease-free survival (p = 0.006) after second Hx. SVR achieved past or postoperatively of the first Hx of HCV-related HCC significantly inhibits recurrence and consequently improves patient survival after second Hx for recurrent HCC. Patients with SVR to IFN therapy would be good candidates for second Hx for recurrent HCC.


Impairment of LTD and cerebellar learning by Purkinje cell-specific ablation of cGMP-dependent protein kinase I.
The molecular basis for cerebellar plasticity and motor learning remains controversial. Cerebellar Purkinje cells (PCs) contain a high concentration of cGMP-dependent protein kinase type I (cGKI). To investigate the function of cGKI in long-term depression (LTD) and cerebellar learning, we have generated conditional knockout mice lacking cGKI selectively in PCs. These cGKI mutants had a normal cerebellar morphology and intact synaptic calcium signaling, but strongly reduced LTD. Interestingly, no defects in general behavior and motor performance could be detected in the LTD-deficient mice, but the mutants exhibited an impaired adaptation of the vestibulo-ocular reflex (VOR). These results indicate that cGKI in PCs is dispensable for general motor coordination, but that it is required for cerebellar LTD and specific forms of motor learning, namely the adaptation of the VOR.


Theophylline therapy for near-fatal Cheyne-Stokes respiration. A case report.
Cheyne-Stokes respiration is characterized by periodic breathing that alternates with hypopnea or apnea. To describe the effect of theophylline on near-fatal Cheyne-Stokes respiration. Case report. Tertiary referral center. A 48-year-old diabetic woman with a history of three cardiorespiratory arrests, a normal coronary arteriogram, normal left ventricular function, and severe Cheyne-Stokes respiration. Oxygen saturation, intra-arterial blood pressure, central venous pressure, chest wall movement, electrocardiography, electromyography, electroencephalography, electro-oculography, minute ventilation, arterial blood gases, and serum theophylline levels. After intravenous administration of 1.2 mg of theophylline at 0.6 mg/kg per hour (serum level, 5.6 microg/mL), both Cheyne-Stokes respiration and oxygen desaturation were markedly attenuated. After infusion of 2.4 mg of theophylline (serum level, 11.6 microg/mL), Cheyne-Stokes respiration resolved completely. No change was seen with placebo. Cheyne-Stokes respiration did not recur during outpatient treatment with oral theophylline. Theophylline may be a rapid and effective therapy for life-threatening Cheyne-Stokes respiration.


Bone Marrow Stromal Cells inhibited the growth and metastasis of human U87 cells through delivering exosomal miR-506.
Glioma is one of the malignancy brain tumors, which deeply threaten the health of patients. Although the traditional therapies for glioma have improved, the outcome is still far from satisfactory. Bone Marrow Stromal Cells (BMSC)-based therapy provided novel insight in the treatment for glioma. However, the detailed molecular mechanism is still not clear. The aim of present study is to discover the novel factor in BMSC-based therapy for glioma. The cell proliferation and apoptosis were identified by using CCK-8 and flow cytometry. The invasion of glioma cells was examined by using Transwell assay and wound-healing assay respectively. qRT-PCR was used to examine the expression of miR-506. Western blot was used to examine the protein levels of CD63, TSG101, NUR77 and CXCR4. Our data suggested that BMSC-derived exosome inhibited the proliferation and contributed to apoptosis of human U87 cells after culturing with miR-506 mimic. Overexpression of miR-506 in BMSC-derived exosome inhibited the invasion of human glioma U87 cells, while these effects were deeply suppressed in the presence GW4869. Our present study demonstrated that BMSC inhibited the growth and metastasis of human glioma U87 cells through delivering exosomal miR-506, and provided the evidences to develop the BMSC-based therapy for glioma.


Regional differences in alcohol use among U. S. military personnel.
The purpose of this study was to examine differences in heavy alcohol use and alcohol-related negative effects among U.S. military personnel stationed in different world regions and to examine factors that may account for regional differences. Data were drawn from the 1998 Department of Defense Survey of Health Related Behaviors among Military Personnel, which had a large representative sample of active-duty U.S. forces. Heavy alcohol use and related negative effects (severe consequences, productivity loss and alcohol overuse) were examined in four regions (Asia, Europe, Hawaii and continental United States; N = 17,154; 86% men) using multivariate logistic regression models that controlled for demographic factors. U.S. military personnel stationed in Asia were significantly more likely to be heavy alcohol users than personnel stationed in the other regions. Productivity loss was also significantly greater in Asia, whereas severe consequences and alcohol overuse were not. Possible explanations for the findings include local regional culture, availability of alcohol, freedom from restraints, response to stress, military culture and selection effects. Heavy alcohol use of military personnel varies by region of assignment and may be attributable to a variety of factors. Future studies need to examine regional alcohol use in greater detail and to include measures specific to potential explanatory domains to permit a more complete understanding of underlying causal mechanisms of heavy drinking by U.S. forces in Asia. Current findings suggest that alcohol use prevention and early intervention programs should be tailored to take account of regional differences.


Inactivation of TEM-1 beta-lactamase by 6-acetylmethylenepenicillanic acid.
6-Acetylmethylenepenicillanic acid is a new kinetically irreversible inhibitor of various beta-lactamases. Interaction between 6-acetylmethylenepenicillanate and purified TEM-1 beta-lactamase during the inactivation process was investigated. 6-Acetylmethylenepenicillanate inhibited the enzyme in a second-order fashion with a rate constant of 0.61 microM-1 . S-1. The apparent inactivation constant decreased in the presence of increasing concentrations of the substrate benzylpenicillin. Native enzyme (pI 5.4) was converted into two inactive forms with pI 5.25 and 5.15, the latter form being transient and readily converted into the more stable form with pI 5.15. Even a 50-fold excess of inhibitor over enzyme did not produce any other inactivated species of the enzyme. All the results obtained suggest that 6-acetylmethylenepenicillanate is a potent irreversible and active-site-directed inhibitor of TEM-1 beta-lactamase.


Alpha, Dimension-Free, and Model-Based Internal Consistency Reliability.
As pointed out by Sijtsma (in press), coefficient alpha is inappropriate as a single summary of the internal consistency of a composite score. Better estimators of internal consistency are available. In addition to those mentioned by Sijtsma, an old dimension-free coefficient and structural equation model based coefficients are proposed to improve the routine reporting of psychometric internal consistency. The various ways to measure internal consistency are also shown to be appropriate to binary and polytomous items.


The E3 ligase AtCHIP positively regulates Clp proteolytic subunit homeostasis.
The caseinolytic peptidase (Clp) core proteins are essential for plant growth and development, especially for chloroplast function. Antisense or overexpression of ClpP4, which is one of the Clp core subunits, causes chlorotic phenotypes in Arabidopsis. An E3 ligase gene, AtCHIP, has previously been found to ubiquitylate ClpP4 in vitro. ClpP4 antisense and overexpressing plants that also overexpressed AtCHIP were constructed to explore the effect of AtCHIP on ClpP4. Overexpression of AtCHIP was found to rescue the chlorotic phenotypes of both ClpP4 antisense and overexpressing plants. The unbalanced levels of Clp core proteins in ClpP4 antisense and overexpressing plants with overexpression of AtCHIP were similar to wild-type levels, suggesting that AtCHIP regulates Clp core proteins. The results also show that AtCHIP can interact with ClpP3 and ClpP5 in yeast and ubiquitylate ClpP3 and ClpP5 in vitro. This suggests that AtCHIP is directly related to ClpP3 and ClpP5. Given these results, the inference is that through selective degradation of Clp subunits, AtCHIP could positively regulate homeostasis of Clp proteolytic subunits and maximize the production of functional chloroplasts. Similar results were obtained from transgenic tobacco plants, suggesting that regulation of the Clp protease by AtCHIP is conserved.


Housing First is associated with reduced use of emergency medical services.
Chronically homeless adults with severe alcohol problems are disproportionately burdened with health-care problems and are high utilizers of emergency medical services (EMS). Single-site Housing First (HF), which provides immediate, permanent, low-barrier, nonabstinence-based, supportive housing, has been associated with reduced publicly funded service utilization. The aims of the current study were to determine whether time spent in single-site HF predicted decreases in EMS contacts 2 years subsequent to single-site HF move-in, and to describe medical conditions and injuries associated with EMS contacts in a sample of chronically homeless individuals with severe alcohol problems. Participants were 91 chronically homeless adults with severe alcohol problems who were enrolled in a single-site HF program between December 2005 and March 2007 in Seattle, Washington. We obtained administrative data on exposure to HF and EMS utilization for the 2 years prior to and the 2 years subsequent to participants' move-in date. EMS utilization variables included patient type (i.e., primary presenting problem), trauma/injury mechanism (i.e., EMS classification of the cause of the trauma or injury), level of care (i.e., basic life support, advanced life support), and transport destination. After controlling for baseline EMS contacts, participants evinced 3% fewer EMS contacts for each additional month of single-site HF exposure. From the baseline to follow-up period, the mean number of EMS contacts declined from 15.85 (SD = 22.96) to 9.54 (SD = 15.08), representing a 54% reduction in the number of EMS contacts. Most calls were responded to by EMTs providing basic life support, and the majority resulted in transport to a local level I trauma center. The most common presenting difficulties were medical illness and trauma. Substance use and psychiatric difficulties were infrequently documented as the primary problem. Our findings support recent assertions that housing is health care and indicate that the amount of time spent in single-site HF is associated with significant reductions in EMS utilization for at least 2 years subsequent to move-in. These findings also underscore the high levels of medical illness and trauma exposure among chronically homeless adults with severe alcohol problems.


FAP associated thyroid carcinoma in mother and her daughter.
A 21 year-old Japanese female was referred to the hospital for evaluation of a mass in her neck. Cytologic samples obtained through fine needle aspiration (FNA) material from the mass showed pseudopapillary or sheet-like clusters. The cells had oval nuclei and columnar cytoplasms. Nuclear groove and intranuclear inclusions were scarcely seen. The cells had a cribriform or solid pattern, unlike cells in papillary thyroid carcinoma. This distinct cytological appearance is thought to belong to FAP (familial adenomatous polyposis)-associated thyroid cancer. The patient was diagnosed with FAP associated papillary thyroid carcinoma. FAP was confirmed by colonoscopy. Her mother, 48 years old, also detected an anterior neck swelling at the time of her daughter's admission. At the age of ten, the mother had undergone total colectomy because of FAP. FNA of the mass demonstrated the same cytological appearance as from her daughter's tumor. Total thyroidectomy was performed. Gross and microscopic appearances of the tumor was similar in mother and daughter. We believe it is possible to detect FAP through FNA cytology of thyroid tumors if pathologists are aware of the unique cytohistological features of thyroid tumor cells in FAP.


Behavioral characterization of quinolinate-induced lesions of the medial striatum: relevance for Huntington's disease.
Huntington's disease (HD) in an inherited neurodegenerative disorder in which the striatum undergoes marked atrophic changes. Patients with HD typically have impaired cognitive function, including deficient visuospatial skills, lack of cognitive flexibility and poor recall of memories. The relationship between these cognitive abnormalities and the striatal degeneration of HD is incompletely understood. In order to explore this issue, we studied the behavior of rats with histologically confirmed bilateral quinolinate (QUIN)-induced lesions of the medial striatum. In a series of Morris Water Maze (MWM) experiments and Delayed Alternation (DA) tests, QUIN-lesioned animals exhibited: (i) impaired acquisition of visuospatial skills; (ii) impaired "transfer of learning"; (iii) preseverative behavior; (iv) deficient retrieval or retention of stored memories. The lesioned rats were unimpaired with swimming to a visible platform, while moving spontaneously in behavior boxes, and when performing various specialized tests of motor function. These results indicate that QUIN-induced lesions of the medial striatum can produce impairments of visuospatial skills, cognitive flexibility, and recall. These are the categories of cognitive function that are disturbed in HD. This suggests that the striatal degeneration of HD could be a sufficient explanation for the cognitive abnormalities associated with the illness.


CRISPR/Cas9 Technology in Translational Biomedicine.
The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) - RNA-guided Cas9 endonuclease system has provided a fast and efficient method for precise genome editing in diverse mammalian species, including humans. The CRISPR/Cas9 technology allows generation of modifications into site-specific locations of the selected genes in one major step by carrying deletions, insertions or DNA donor-directed precise sequence modifications. Cas9 forms a nucleoprotein complex with a sequence-specific guide RNA to create double-stranded breaks in complementary DNA target. Further, double-stranded break repair machinery leads to the intended gene modifications. The CRISPR/Cas9 system is widely used technique for genome modification, editing and other biotechnology applications, such as functional annotation, a system for visualization of specific genomic loci and transcriptional control of genes. CRISPR/Cas9-mediated manipulation of the laboratory animal genomes has contributed to the understanding of gene functions and has become a popular approach for modeling human disorders. Furthermore, the growing application of CRISPR-Cas9 system to human genes emerges as an extremely powerful technology for the molecular characterization and treatment of human disease. In this review we present the essential principles of CRISPR/Cas9 technology and the recent advances in its use in translational biomedicine.


Cost effectiveness of the B type natriuretic peptide, electrocardiography, and portable echocardiography for the assessment of patients from the community with suspected heart failure.
To analyze the cost efficiency of guidelines proposed by the European Society of Cardiology for investigation of patients in the community with suspected heart failure (HF). The guidelines recommend electrocardiography (ECG) and/or measurement of N-terminal pro B type natriuretic peptide (NTproBNP) prior to referral for echocardiography. Portable echocardiography is a new but validated technique for the evaluation of HF. Accordingly, 137 suspected HF patients (mean age 71+/-13 years) from the community underwent ECG and NTproBNP estimation prior to portable echocardiography. Cost effective analysis for ECG, NTproBNP, portable echocardiography and a combination of these; to define valvular heart disease, right ventricular dysfunction and left ventricular systolic and diastolic dysfunction were compared. The cost of abnormal NTproBNP followed by portable echocardiography, abnormal ECG followed by portable echocardiography and portable echocardiography alone for the detection per case of left ventricular systolic dysfunction were 313 euro, 310 euro, and 296 euro respectively and that for detection per case of any of the aforementioned cardiac abnormalities were 198 euro, 223 euro, and 170 euro respectively. Portable echocardiography alone for the assessment of suspected HF patients resulted in a cost reduction of up to 1083 euro for the detection per case of cardiac abnormality. While a strategy where initial NTproBNP estimation is cost effective in detecting any causes of heart failure, portable echocardiography remains the most costeffective strategy to assess patients from the community with suspected heart failure.


Contributions of Farm Animals to Immunology.
By their very nature, great advances in immunology are usually underpinned by experiments carried out in animal models and inbred lines of mice. Also, their corresponding knock-out or knock-in derivatives have been the most commonly used animal systems in immunological studies. With much credit to their usefulness, laboratory mice will never provide all the answers to fully understand immunological processes. Large animal models offer unique biological and experimental advantages that have been and continue to be of great value to the understanding of biological and immunological processes. From the identification of B cells to the realization that γδ T cells can function as professional antigen presenting cells, farm animals have contributed significantly to a better understanding of immunity.


Self-assembly and induced circular dichroism in dendritic supramolecules with cholesteric pendant groups.
We report on the solid-state structural features of self-assembled chiral supramolecules based on ionic complexation of chiral cholesteric pendant groups with achiral dendritic macromolecules and show that their optical activity exhibits a systematic change in the ultraviolet/visible light (UV-vis) absorption and enhancement in the circular dichroism (CD) signal, indicating the occurrence of supramolecular chirality, also referred to as induced circular dichroism (ICD). We construct a homologous series of complexes by varying systematically from 1 to 3 the generation of dendritic units contained in dendrons, dendrimers, and dendronized polymers. The structural properties of the complexes are investigated by means of small-angle X-ray scattering (SAXS) and transmission electron microscopy (TEM). Depending on the class of dendritic molecule and the generation, lamellar, columnar hexagonal, oblique columnar, and rectangular columnar phases can be found, with a direct correlation among the degrees of freedom of the dendritic macromolecules used and the level of order achieved in the self-assembled solid-state structures. The enhancement of the optical signals of these mesoscopic structures appears to be correlated with their order in the solid state. Complexes with the longest lattice correlation lengths also show the most enhanced CD signals. These results show the unique versatility of dendritic macromolecules as supramolecular templates capable of organizing low molecular weight chiral pendant units into a variety of solid-state structures with amplified optical properties.


In vitro inhibition of vancomycin-susceptible and vancomycin-resistant Enterococcus faecium and E. faecalis in the presence of citrus essential oils.
Glycopeptide-resistant Enterococcus (GRE) is an important healthcare-acquired infection (HCAI) which costs the healthcare service many millions of pounds worldwide. In this study, lemon (Citrus limon), sweet orange (Citrus sinensis) or bergamot (Citrus bergamia) essential oils (EO) and their vapours, alone and in combination, are tested for their antimicrobial activity against vancomycin-resistant and vancomycin-sensitive strains of E. faecium and E. faecalis. A blend of 1:1 (v/v) orange and bergamot EO was the most effective of the oils and/or blends tested with a minimum inhibitory concentration (MIC), at 25 degrees C and pH 5.5, of 0.25-0.5% (v/v) and a minimum inhibitory dose (MID) of 50 mg/L, at 50 degrees C at pH 7.5, when viable counts reduced by 5.5-10 log10 colony forming units (cfu)/mL, suggesting that this blend of citrus oils is effective under a range of conditions for inhibiting the growth and survival of E. faecalis, E. faecium and VRE.


4-Phenylbutyric acid prevent cytotoxicity induced by thapsigargin in rat cardiac fibroblast.
Cardiac fibroblast (CF) survival is important for the maintenance of the extracellular matrix homeostasis in the heart; providing a functional support to cardiomyocytes necessary for the correct myocardial function. Endoplasmic reticulum (ER) stress causes cellular dysfunction and cell death by apoptosis; and thapsigargin is a well-known ER stress inducer. On the other hand, the chemical chaperone, 4-phenylbutyric acid (4-PBA) had showed to prevent ER stress; however, in cardiac fibroblast both the ER stress induced by thapsigargin and prevention by 4-PBA, have not been studied in detail. Neonate rat CF were treated with thapsigargin in presence or absence of 4-PBA, and cell viability was evaluated by trypan blue exclusion and apoptosis by flow cytometry; whereas CHOP, BIP, PDI, ATF4 and procollagen protein levels were assessed by western blot. In CF, thapsigargin triggered the unfolded protein response detected by early increases in ATF4, CHOP, PDI and BIP protein levels as well as, the accumulation of intracellular procollagen. Thapsigargin also stimulated CF death in a time and concentration-dependent manner. ER stress, CF death and apoptosis induced by thapsigargin were prevented by 4-PBA. Conclusion our data suggest that 4-PBA prevent ER stress, intracellular procollagen accumulation, CF death and apoptosis induced by thapsigargin.


Hyaluronan and CD44: modulators of chondrocyte metabolism.
Stability of cell-matrix interactions promotes tissue homeostasis, and matrix receptors provide for the assembly and retention of matrix as well as the linkage to the signal transduction pathways activated in response to changes within the extracellular matrix. Therefore, changes in cell-matrix interactions may influence cell responses to cytokines and cell survival and may facilitate tissue remodeling. Matrix components sequester or present soluble morphogens to their signaling receptors. Another layer of complexity would be the establishment of new or uncoupling of cell-matrix interactions, leading to altered cellular response to morphogens. The hyaluronan receptor CD44 serves as the critical link for the retention of hyaluronan-proteoglycan aggregates to the chondrocyte cell surface. As we continue to define functional chondrocyte CD44, future studies will need to include analysis of the variant CD44 isoform expression, phosphorylation, cytoskeletal interactions, occupancy, and turnover. Disruption of chondrocyte CD44:hyaluronan interaction will induce a cascade of events resulting in the activation of both catabolic as well as anabolic gene products. Fragments of hyaluronan produced in free radical processes have the potential to augment the production of nitric oxide in a CD44-dependent mechanism. Data also support an emerging paradigm that CD44-mediated signaling affects both chondrocyte survival pathways as well as apoptotic pathways.


Sex-related differences in prognosis after myocardial infarction: changes from 1978 to 2007.
Women with myocardial infarction (MI) have shown a 28-day survival disadvantage compared with men. However, results were less consistent when considering long-term mortality in 28-day survivors. The aim was to estimate the trends for sex-related differences in the three endpoints considered for this study: (1) 28-day mortality or severe ventricular dysfunction (acute pulmonary oedema or cardiogenic shock) during the hospital stay, (2) 28-day mortality and (3) two-year cardiovascular mortality or non-fatal MI in 28-day survivors after a first MI. A cohort of 3,982 consecutive patients with first Q-wave MI admitted to a university tertiary reference hospital between 1978 and 2007 was followed for 2 years. Short-term prognosis improved in women over the studied period; similar rates were observed in both sexes in the 2000s. After adjusting for age, co-morbidities and anterior location of MI, female sex had an odds ratio=1.71 (95% confidence interval [CI] 1.34-2.17) of short-term severe MI or death over the studied period. Overall, sex differences in long-term prognosis remained similar over the studied period (hazard ratio=1.40; 95% CI 1.02-1.91). In conclusion, short-term prognosis improved over the past 30 years for first Q-wave MI patients, becoming similar for both men and women in the most recent decade. Long-term prognosis did not improve in either men or women, indicating that secondary prevention should be reinforced to achieve consistent reductions in the number of cardiovascular events.


Stimulated flow of pancreatic and biliary secretions after intestinal exposure to cholera toxin.
1. Duodeno-jejunal intestinal loops in rats were perfused with cholera toxin and the pancreatic and biliary secretory responses studied. 2. Intraluminal cholera toxin induced a significant flow of pancreatic and biliary fluid and sustained protein (enzyme) secretion in pancreatic juice, in addition to the expected stimulation of mucosal secretion. 3. Intravenously injected cholera toxin failed to elicit a secretory flow from the pancreas and biliary tract. 4. The time-course of the enhanced secretory flow from pancreas and biliary tract after intraluminal cholera toxin corresponds closely to the secretion of intestinal fluid. 5. The results of these studies provide evidence of sustained stimulation of pancreatic and biliary secretions after mucosal exposure to cholera toxin. These observations support the suggestion that secretion from the pancreas and liver may significantly contribute to the total intestinal secretion in cholera.