Diversity of bacteria associated with Bursaphelenchus xylophilus and other nematodes isolated from Pinus pinaster trees with pine wilt disease.
The pinewood nematode (PWN), Bursaphelenchus xylophilus, has been thought to be the only causal agent of pine wilt disease (PWD), however, since bacteria have been suggested to play a role in PWD, it is important to know the diversity of the microbial community associated to it. This study aimed to assess the microbial community associated with B. xylophilus and with other nematodes isolated from pine trees, Pinus pinaster, with PWD from three different affected forest areas in Portugal. One hundred and twenty three bacteria strains were isolated from PWN and other nematodes collected from 14 P. pinaster. The bacteria strains were identified by comparative analysis of the 16S rRNA gene partial sequence. All except one gram-positive strain (Actinobacteria) belonged to the gram-negative Beta and Gammaproteobacteria. Most isolates belonged to the genus Pseudomonas, Burkholderia and to the family Enterobacteriaceae. Species isolated in higher percentage were Pseudomonas lutea, Yersinia intermedia and Burkholderia tuberum. The major bacterial population associated to the nematodes differed according to the forest area and none of the isolated bacterial species was found in all different forest areas. For each of the sampled areas, 60 to 100% of the isolates produced siderophores and at least 40% produced lipases. The ability to produce siderophores and lipases by most isolates enables these bacteria to have a role in plant physiological response. This research showed a high diversity of the microbial community associated with B. xylophilus and other nematodes isolated from P. pinaster with PWD.


Recent synthetic transport systems.
This critical review covers progress with synthetic transport systems, particularly ion channels and pores, between January 2006 and December 2009 in a comprehensive manner. This is the third part of a series launched in the year 2000, covering a rich collection of structural and functional motifs that should appeal to a broad audience of non-specialists, including to organic, biological, supramolecular and polymer chemists. Impressive breakthroughs have been achieved over the past four years in part because of a fruitful expansion toward new types of interactions, including metal-organic, π-π, aromatic electron donor-acceptor, anion-π or anion-macrodipole interactions as well as dynamic covalent bonds (169 references).


Major emerging problems with minor meloidogyne species.
Root-knot nematodes (Meloidogyne spp.) represent one of the most polyphagous genera of plant-parasitic nematodes. To date, close to 100 valid species are recognized. In contrast to the size of the genus, the majority of past research focused on a small number of species, i.e., the so-called 'major' species M. arenaria, M. hapla, M. incognita, and M. javanica. This review highlights recent work aimed at 'minor' root-knot nematodes: M. chitwoodi, M. fallax, M. minor, M. enterolobii (=M. mayaguensis), M. exigua, and M. paranaensis. Some of these species have been described only recently. After a brief profile of each species, identification methods and their application in Meloidogyne spp. are summarized. Intraspecific variation and its impact on plant resistance breeding are discussed and interactions between M. enterolobii and Fusarium solani are highlighted as an example of synergistic interactions with other plant pathogens. Future research on Meloidogyne spp. is not only shaped by recent breakthroughs such as completing the genome sequences of M. hapla and M. incognita, but is also influenced by changes in agriculture. Taken together, the aim of this review is to draw attention to previously neglected and newly described Meloidogyne spp. that are developing into major problems for agriculture in tropical and temperate climates.


How we manage variceal hemorrhage in cirrhotic patients. Key practical messages from the British Guidelines.
Variceal bleeding is a serious complication of portal hypertension with high morbidity and mortality. Advances in our understanding of screening and risk stratification along with evidence-based management strategies for acute variceal bleeding as well as primary and secondary prevention have improved overall outcomes in patients with portal hypertension. The guidelines recently published by the British Society of Gastroenterology (BSG) and Baveno 6 consensus have aimed to enhance the standard of care in the management of varices and their complications. This concise review focuses on the key practical messages for screening and management of varices and variceal bleeding in light of these guidelines. The review also takes into account important evidence published since the BSG guidelines and Baveno 6 consensus.


[Intrinsic waxes of saprophytic mycobacteria].
The ability of soil mycobacteria for the synthesis of true waxes may be revealed on media with hydrocarbons. Most strains produced only traces of wax, and some strains (2 out of 62) in considerable amounts (over 1% of dry biomass). The chemical composition of the wax depends on the structrue of oxidized substrate: a mixture of two to three homologues with the molecular weights of 452, 480 and 508, respectively, is formed on a medium with hexadecane, and a mixture of seven homologues with the homologous difference of 14 in the case of glucose.


Pathways for collaboratively strengthening water and sanitation systems.
Collaborative approaches are seen as a promising way to strengthen Water, Sanitation, and Hygiene (WASH) service delivery systems when challenges exceed the mandates and capabilities of any single entity. While collaborative approaches are well studied in high-income country contexts, current understanding of their application to international development contexts is limited. This paper uses fuzzy-set Qualitative Comparative Analysis to assess what conditions and pathways drove or impeded progress within eleven collaborative approaches for WASH service delivery in Eastern Africa. Evidence supported three main findings: (1) Government uptake of recommendations is necessary for progress but cannot be guaranteed solely by government participation in the collaboration, (2) different forms of problem identification are possible; problem scopes are often predefined to align with funders and partner government agendas, but flexible scopes that foster collective problem identification can reap benefits, and (3) hub convening power can be critical and convening power can be gained in different ways. Political dynamics, shifting priorities, and turnover undermine collaborative efforts, but collaborative approaches can still make progress in spite of turnover if funds are available for implementation of activities (i.e. in addition to funds for meetings and hub roles) and program implementers either facilitate collective problem identification or establish a hub with convening power. Yet even these tactics are vulnerable to instability, thus in highly unstable contexts, stakeholders and funders should be realistic from the outset about what they may be able to achieve. Building on existing theories of collaborative approaches, this work revealed that there is no single best design for collaborative approaches in WASH, rather, core elements worked together in different ways depending on the context.


Violence perpetration among urban american Indian youth: can protection offset risk?
To predict the likelihood of violence perpetration given various combinations of the most statistically salient risk and protective factors related to violence perpetration. Urban Indian Youth Health Survey, conducted from October 9, 1995, to March 30, 1998, consisting of 200 forced-choice items exploring values, cultural identity, relationships, decision-making skills, and health and well-being. Urban schools and an after-school youth development program at an urban American Indian center. Five hundred sixty-nine urban American Indian youth enrolled in grades 3 through 12. Violence perpetration dichotomized in 2 ways: (1) level of violence perpetration (ie, hitting someone 1-2 times in the past year vs picking fights, hitting repeatedly, participating in group fights, or shooting or stabbing someone in the past year) and (2) having shot and/or stabbed someone during the past year. In the final multivariate models with age as a covariate, most protective against violence perpetration were connections to school (odds ratio [OR], 0.17), positive affect (OR, 0.29), and peer prosocial behavior norms against violence (OR, 0.35). School connectedness (OR, 0.01) and positive affect (OR, 0.46) were also protective against shooting and/or stabbing someone, as was parental prosocial behavior norms against violence (OR, 0.23). The strongest risk factors for violence perpetration were substance use (OR, 2.60) and suicidal thoughts/behaviors (OR, 2.71); for shooting and/or stabbing, it was substance use (OR, 5.26). The likelihood of violence perpetration increased markedly (from 10% to 85%) as the exposure to risk factors increased and protective factors decreased. For shooting or stabbing someone, the probabilities ranged from 3% (0 risks and 3 protective factors) to 64% (1 risk and 0 protective factors). The dramatic reduction in the likelihood of violence involvement when risk was offset with protective factors in the probability profiles suggests the utility of a dual strategy of reducing risk while boosting protection.


Acute effects of tobacco smoke on alveolar macrophages cultured in gas phase.
With a culture technique in gaseous phase which permits a direct contact between cells and the atmosphere, without any liquid interposition, we studied the short-term effects of tobacco smoke on guinea pig alveolar macrophages (AM). Tobacco smoke had a cytotoxic effect on AM as evaluated by the decrease in cell ATP content, total smoke appearing more toxic than the gas phase. Toxicity of smoke gas phase is also ascertained by morphologic cell changes in electron microscopy and by a decrease in cell bactericidal activity. The large variation in AM susceptibility is not related to cell content in superoxide dismutase but seems to be in good correlation with AM glutathione content, especially when cells are cultured with N-acetylcysteine. However, the protective glutathione action is not total and seems less important than the one observed with a low concentration of NO2 (0.2 ppm/24 hours).


Donut: Augmentation Technique for Enhancing The Efficacy of Glaucoma Suspect Screening.
Glaucoma is the second most common cause of blindness. A glaucoma suspect has risk factors that increase the possibility of developing glaucoma. Evaluating a patient with suspected glaucoma is challenging. The "donut method" was developed in this study as an augmentation technique for obtaining high-quality fundus images for training ConvNeXt-Small model. Fundus images from GlauCUTU-DATA, labelled by randomizing at least 3 well-trained ophthalmologists (4 well-trained ophthalmologists in case of no majority agreement) with a unanimous agreement (3/3) and majority agreement (2/3), were used in the experiment. The experimental results from the proposed method showed the training model with the "donut method" increased the sensitivity of glaucoma suspects from 52.94% to 70.59% for the 3/3 data and increased the sensitivity of glaucoma suspects from 37.78% to 42.22% for the 2/3 data. This method enhanced the efficacy of classifying glaucoma suspects in both equalizing sensitivity and specificity sufficiently. Furthermore, three well-trained ophthalmologists agreed that the GradCAM++ heatmaps obtained from the training model using the proposed method highlighted the clinical criteria.Clinical relevance- The donut method for augmentation fundus images focuses on the optic nerve head region for enhancing efficacy of glaucoma suspect screening, and uses Grad-CAM++ to highlight the clinical criteria.


Effect of neplanocin A on differentiation, nucleic acid methylation, and c-myc mRNA expression in human promyelocytic leukemia cells.
The effect of the cyclopentenyl adenosine analog neplanocin A (NPC) on cell growth and differentiation was examined in the human promyelocytic leukemia cell line HL-60. Continuous exposure of HL-60 cells to 0.1-3.3 microM NPC resulted in a progressive reduction in cell growth which was accompanied by an increase in differentiation to cells with a myelocyte and neutrophil morphology. The latter effect was expressed as an increase in the capacity of cells to reduce nitro blue tetrazolium and reached a level of 40% of the total cell population. Preceding the phenotypic changes was the preferential inhibition of RNA and DNA methylation in comparison to inhibition of their synthesis which coincided with the formation of a metabolite of NPC with the chromatographic characteristics of S-adenosyl-L-methionine (AdoMet). In addition, c-myc mRNA expression, which is amplified in HL-60 cells, was markedly reduced following NPC treatment. These results indicate that NPC is an effective inhibitor of RNA and DNA methylation resulting from its conversion to an analog of AdoMet, and that these effects appear to be responsible for reduced c-myc RNA expression and the induction of myeloid differentiation in this cell line.


Carbon allocation in ectomycorrhizal plants at limited optimal N supply: an attempt aat unraveling conflicting theories.
With regard to mycorrhiza, conflicting theories try to explain how the balance between fungal demand for carbohydrates and the plant's needs for nutrients varies, resulting in conflicting predictions. In order to evaluate current concepts, we investigated some metabolic parameters, which are indicative for plant carbon allocation in response to mycorrhization at limited and optimal N supply. Pinus pinaster seedlings were inoculated with living or dead (control) cultures of Pisolithus tinctorius, supplied with ammonium at 4 (limiting) or 7% d−1 (non-limiting) N relative addition rate (RARN), and followed development for 29 days. Mycorrhizal colonization of roots was quantified by the determination of ergosterol. A series of enzymes (sucrose and trehalose metabolism, anaplerosis) and metabolites (soluble carbohydrate, including trehalose; fructose 2,6 bisphosphate, free amino acids) relevant in the C/N exchange between symbionts, and in the carbon allocation and sink strength within the plant were assayed for 2-day-intervals for up to 14 days, and at 5-day-intervals for the rest of the experiment. The first 10 days reflected the establishment of mycorrhizal interaction, and the carbon allocation to the root was higher in M plants independent of N supply. Following this period, carbon allocation became N-related, higher at low, and lower at high N supply. The belowground C investment of M plants was dependent on N availability, but not on N gain. Finally, increased belowground C allocation was accompanied by a shift from plant to fungal metabolism.


Host suitability affects odor association in Cotesia marginiventris: implications in generalist parasitoid host-finding.
Insect herbivores often induce plant volatile compounds that can attract natural enemies. Cotesia marginiventris (Hymenoptera: Braconidae) is a generalist parasitoid wasp of noctuid caterpillars and is highly attracted to Spodoptera exigua-induced plant volatiles. The plasticity of C. marginiventris associative learning to volatile blends of various stimuli, such as host presence, also has been shown, but little is known about how this generalist parasitoid distinguishes between host species of varying suitability. Spodoptera exigua is an excellent host that yields high parasitoid emergence, while Trichoplusia ni serves as a sub-optimal host species due to high pre-imaginal wasp mortality. We have found that S. exigua and T. ni induce different volatile blends while feeding on cotton. Here, wind tunnel flight assays were used to determine the importance of differentially induced volatiles in host-finding by C. marginiventris. We found that, while this generalist parasitoid wasp can distinguish between the two discrete volatile blends when presented concurrently, a positive oviposition experience on the preferred host species (S. exigua) is more important than host-specific volatile cues in eliciting flight behavior towards plants damaged by either host species. Furthermore, wasps with oviposition experience on both host species did not exhibit a deterioration in positive flight behavior, suggesting that oviposition in the sub-optimal host species (T. ni) does not cause aversive odor association.


Learning viewpoint-invariant face representations from visual experience in an attractor network.
In natural visual experience, different views of an object or face tend to appear in close temporal proximity as an animal manipulates the object or navigates around it, or as a face changes expression or pose. A set of simulations is presented which demonstrate how viewpoint-invariant representations of faces can be developed from visual experience by capturing the temporal relationships among the input patterns. The simulations explored the interaction of temporal smoothing of activity signals with Hebbian learning in both a feedforward layer and a second, recurrent layer of a network. The feedforward connections were trained by competitive Hebbian learning with temporal smoothing of the post-synaptic unit activities. The recurrent layer was a generalization of a Hopfield network with a low-pass temporal filter on all unit activities. The combination of basic Hebbian learning with temporal smoothing of unit activities produced an attractor network learning rule that associated temporally proximal input patterns into basins of attraction. These two mechanisms were demonstrated in a model that took grey-level images of faces as input. Following training on image sequences of faces as they changed pose, multiple views of a given face fell into the same basin of attraction, and the system acquired representations of faces that were approximately viewpoint-invariant.


In vitro activity of azole derivatives and griseofulvin against planktonic and biofilm growth of clinical isolates of dermatophytes.
As shown by recent research, most of the clinically relevant fungi, including dermatophytes, form biofilms in vitro and in vivo, which may exhibit antimicrobial tolerance that favour recurrent infections. The aim of this study was to determine the minimum inhibitory concentrations (MICs) of itraconazole (ITC), voriconazole (VCZ) and griseofulvin (GRI) against Trichophyton rubrum, Trichophyton tonsurans, Trichophyton mentagrophytes, Microsporum canis and Microsporum gypseum in planktonic and biofilm growth. For the planktonic form, susceptibility testing was performed according to the Clinical and Laboratory Standards Institute (CLSI), document M38-A2, while biofilm susceptibility was evaluated using the XTT colorimetric essay. The planktonic growth of all strains was inhibited, with MIC values ranging from 0.00195 to 0.1225 μg/mL for VRC, 0.00195 to 0.25 μg/mL for ITC and <0.0039 to 4 μg/mL for GRI, while a 50-fold increase in the MIC was required to significantly reduce the metabolic activity (P < .05) of dermatophyte biofilms. In brief, the ability of dermatophytes to form biofilms may be a contributing factor for the recalcitrance of dermatophytoses or the dissemination of the disease.


Renovascular Hypertension: Novel Insights.
Renovascular hypertension (RVH) remains among the most prevalent and important, but also potentially reversible, causes of secondary hypertension. The predominant causes of renal artery stenosis (RAS) are atherosclerotic renovascular arterial stenosis (ARAS) and renal fibromuscular dysplasia. This condition can lead to progressive renal injury, cardiovascular complications and 'flash pulmonary edema'. Duplex Doppler ultrasonography, computed tomographic angiography and magnetic resonance angiography are the most commonly used diagnostic methods. There are three therapeutic options available: medical therapy including renin-angiotensin-aldosterone system antagonists, lipid-lowering agents, and antiplatelet therapy, percutaneous angioplasty with or without stent placement and surgical revascularization. Three large trials failed to demonstrate the superiority of renal artery revascularization over pharmaceutical therapy in controlling blood pressure and preserving renal function. For this reason, today revascularization is only recommended for patients with progressive worsening of renal function, recurrent 'flash pulmonary edema' and rapid increase in antihypertensive requirement in patients with previously well-controlled hypertension. However, more properly designed trials are needed in order to identify which patient populations would probably benefit from renal revascularization.


[Organization of surgical care to the wounded in a modern armed conflict: surgical care at the different echelons of care (Report 1)].
The main statements of organization of care to the wounded in modern armed conflicts are presented for the aim of an efficient discussion ahead of publishing a new edition of "Guidelines for War Surgery", Ministry of Defense of the Russian Federation. Lessons learned from the latest armed conflicts, military reform, development of medical units and medical treatment facilities, as well as appearance of new samples of medical equipment and medical kits approved by the Russian Army resulted in the necessity of changes in surgical care to the wounded.


The role of platelet alpha-granular proteins in the regulation of thrombopoietin messenger RNA expression in human bone marrow stromal cells.
Thrombopoietin (TPO), the specific cytokine that regulates platelet production, is expressed in human bone marrow (BM), kidney, and liver. There appears to be no regulation of TPO in the kidney and liver, but TPO messenger RNA (mRNA) expression can be modulated in the stromal cells of the BM. In this study, we used primary human BM stromal cells as a model to study the regulation of TPO mRNA expression in response to various platelet alpha-granular proteins. We showed that platelet-derived growth factor (PDGF) BB and fibroblast growth factor (FGF) 2 stimulated TPO mRNA expression in both a dose-dependent and time-dependent manner. The addition of 50 ng/mL of PDGF and 20 ng/mL of FGF resulted in maximal induction of TPO mRNA expression in 4 hours. We also found that platelet factor 4 (PF4), thrombospondin (TSP), and transforming growth factor-beta (TGF-beta) are negative modulators of megakaryocytopoiesis. We observed suppression in TPO mRNA expression with 1 microg/mL of both PF4 and TSP and 50 ng/mL of TGF-beta, with maximal suppression occurring 4 hours after the addition of these proteins. Finally, the addition of whole-platelet lysate produced a dose-dependent inhibition of TPO expression. On the basis of these findings, we propose that the platelet alpha-granular proteins studied may regulate TPO gene expression in BM stromal cells by means of a feedback mechanism.


Point-of-care systems for rapid DNA quantification in oncology.
The development of new powerful applications and the improvement in fabrication techniques are promising an explosive growth in lab-on-chip use in the upcoming future. As the demand reaches significant levels, the semiconductor industry may enter in the field, bringing its capability to produce complex devices in large volumes, high quality and low cost. The lab-on-chip concept, when applied to medicine, leads to the point-of-care concept, where simple, compact and cheap instruments allow diagnostic assays to be performed quickly by untrained personnel directly at the patient's side. In this paper, some practical and economical considerations are made to support the advantages of point-of-care testing. A series of promising technologies developed by STMicroelectronics on lab-on-chips is also presented, mature enough to enter in the common medical practice. The possible use of these techniques for cancer research, diagnosis and treatment are illustrated together with the benefits offered by their implementation in point-of-care testing.


A novel serum metabolomics-based diagnostic approach for colorectal cancer.
To improve the quality of life of colorectal cancer patients, it is important to establish new screening methods for early diagnosis of colorectal cancer. We performed serum metabolome analysis using gas-chromatography/mass-spectrometry (GC/MS). First, the accuracy of our GC/MS-based serum metabolomic analytical method was evaluated by calculating the RSD% values of serum levels of various metabolites. Second, the intra-day (morning, daytime, and night) and inter-day (among 3 days) variances of serum metabolite levels were examined. Then, serum metabolite levels were compared between colorectal cancer patients (N = 60; N = 12 for each stage from 0 to 4) and age- and sex-matched healthy volunteers (N = 60) as a training set. The metabolites whose levels displayed significant changes were subjected to multiple logistic regression analysis using the stepwise variable selection method, and a colorectal cancer prediction model was established. The prediction model was composed of 2-hydroxybutyrate, aspartic acid, kynurenine, and cystamine, and its AUC, sensitivity, specificity, and accuracy were 0.9097, 85.0%, 85.0%, and 85.0%, respectively, according to the training set data. In contrast, the sensitivity, specificity, and accuracy of CEA were 35.0%, 96.7%, and 65.8%, respectively, and those of CA19-9 were 16.7%, 100%, and 58.3%, respectively. The validity of the prediction model was confirmed using colorectal cancer patients (N = 59) and healthy volunteers (N = 63) as a validation set. At the validation set, the sensitivity, specificity, and accuracy of the prediction model were 83.1%, 81.0%, and 82.0%, respectively, and these values were almost the same as those obtained with the training set. In addition, the model displayed high sensitivity for detecting stage 0-2 colorectal cancer (82.8%). Our prediction model established via GC/MS-based serum metabolomic analysis is valuable for early detection of colorectal cancer and has the potential to become a novel screening test for colorectal cancer.


Non-guideline-recommended prescribing of proton pump inhibitors: implications for the future and reducing over usage.
Drs Godman and Fadare review the paper by Mares-Garcia and colleagues in Spain regarding the extent of non-guideline recommended prescribing (NGRP) of proton pump inhibitors (PPIs) in ambulatory care. NGRP of PPIs is an increasing public health concern given the implications of overuse of PPIs on infection rates, osteoporosis and chronic kidney disease as well as increasing polypharmacy rates. Costs are less of an issue especially where there are low prices for generic PPIs; the main focus is on improving the quality of PPI prescribing. Encouragingly, Mares-Garcia and colleagues identified key factors associated with NGRP, providing direction to others.


Pain and quality of life among older people with rheumatoid arthritis and/or osteoarthritis: a literature review.
The aim of this study was to review the research literature on pain and quality of life (QoL) and the relationship between these variables among people aged 75 years and above with rheumatoid arthritis and/or osteoarthritis. A Medline and CINAHL search was carried out using MeSH terms rheumatoid arthritis, osteoarthritis, QoL and pain in various combinations. Seventeen articles were identified that met the requirements for methodological quality and inclusion criteria. No study focused only on respondents aged 75 years or over. The studies had varying representation of this age group. Pain was common in both groups and was found to increase with age and disease duration among those with rheumatoid arthritis but not among those with osteoarthritis. Increased pain could lead to depression. Pain, functional limitation and increased age were found to decrease QoL among those with rheumatoid arthritis and osteoarthritis alike. Social support was found to buffer against negative effects on QoL among those with osteoarthritis while no moderating effects were found in rheumatoid arthritis. Increased age was found to relate to pain (rheumatoid arthritis) and decrease QoL (both rheumatoid arthritis and osteoarthritis). It is, however, hard to draw any firm conclusions about older people's pain and QoL because of the lack of studies including respondents aged 75 years or over. Thus, research about pain and QoL, especially focusing on the old and the very elderly with rheumatoid arthritis/osteoarthritis, is needed. It also seems justified to say that nursing care should especially focus on older people and that these people should be assessed for their level of pain, functional limitations and QoL especially in the case of having rheumatoid arthritis and/or osteoarthritis.


Synthesis, characterization and anticancer activity in vitro evaluation of novel dicyanoaurate (I)-based complexes.
Molecular structures containing gold, such as auranofin, have been extensively studied in the diagnosis and treatment of many diseases, including cancer treatment. The pharmacological properties of the newly synthesized unique gold-ligand structures have been reported for different cancer cell lines. However, findings on bishydeten-metal salt complexes with gold are rare. In this work, the synthesis of five novel cyanide-bridged coordination compounds having the closed formulae [Ni(bishydeten)][Au(CN)2]2 (1), [Cu(bishydeten)][Au(CN)2]2 (2), [Zn(bishydeten)2Au3(CN)4][Au2(CN)3] (3), [Cd(bishydeten)0,5]2[Au(CN)2]4.2H2O (4), and [Cd(bishydeten)2][Au(CN)2]2 (5) (where bisyhdeten = N,N-bis(2-hydroxyethyl)ethylene diamine), and their characterization by elemental, infrared, ESI-MS, X-ray (for 2) and thermic measurement methods were performed. Complexes 1 and 3 are thermally more stable than the other three complexes. For these, pharmacological adequacies were also tested. The nucleic acid and protein binding affinities of the Au (I) compounds were also estimated by spectroscopic and electrophoretic techniques. Au (I) complexes were identified as strong chemotherapeutic with mild cytotoxicity, and they demonstrated a dose-dependent inhibition on the growth of cancer cells with IC50 at 0.11 to 0.47 μM. Investigation of mechanisms of action on cells revealed that Au (I) compounds managed to inhibit cell migration and led to a decrease in cytoskeletal proteins such as CK7 and CK20. However, Au (I) compounds failed to inhibit DNA topoisomerase I. Overall, and we suggest that potent antiproliferative activity, mild cytotoxicity, good solubility, and micromolar dosage of Au (I) compounds containing bisyhdeten-metal derivatives render them the potential focus of further studies as chemotherapeutic agents.


Fluoroscopically guided balloon dilation or temporary stent placement for patients with gastric conduit strictures after esophagectomy with esophagogastrostomy.
The purpose of this study was to evaluate the clinical features of patients with gastric conduit strictures after esophagectomy and to report our initial experience in the management of these strictures with fluoroscopically guided balloon dilation, temporary stent placement, or both. From January 1996 to January 2012, 12 patients with gastric conduit stricture after esophagectomy underwent fluoroscopic balloon dilation, temporary stent placement, or both. Stent placement was indicated in patients who had gastro-respiratory fistulas or recurrent symptoms within 2 weeks of balloon dilation. Technical and clinical success, complications, and length of strictures were retrospectively evaluated. The mean length of strictures in all patients was 49 mm (range, 21-76 mm). The strictures were accompanied by gastrorespiratory fistulas in four patients. Seven patients with gastric conduit strictures without fistulas underwent 23 balloon dilations. Five patients underwent stent placement owing to gastrorespiratory fistula formation (n = 4) and frequent recurrent symptoms after balloon dilation (n = 1). During a mean follow-up period of 8 months (range, 2-28 months), clinical success was achieved in 11 patients (91.6%) after a single balloon dilation (n = 1), multiple balloon dilations (n = 5), or stent placement (n = 5). Complications occurred in 3 of 10 patients (30%) after balloon dilation and in one of five patients (20%) after stent placement. Gastric conduit strictures are characterized by their substantial length and tend to be accompanied by gastrorespiratory fistulas. Despite relatively high complication and recurrence rates, this study may offer a viable treatment of gastric conduit strictures by using fluoroscopically guided balloon dilation and stent placement.


Neutron production associated with radiotherapy linear accelerators using intensity modulated radiation therapy mode.
A number of experiments were conducted on a Clinac 21EX radiotherapy accelerator using an IMRT treatment plan to determine neutron dose equivalent as a function of both patient dose delivered and machine workload. It was determined that IMRT mode is more neutron dose intensive as a function of patient dose when compared to a similar standard non-IMRT treatment. It was found that when the neutron production is normalized to workload, the measured neutron dose equivalents are similar. It is therefore recommended that neutron production be reported as a function of workload when considering IMRT treatment modes.


Extrinsic and intrinsic control by EKLF (KLF1) within a specialized erythroid niche.
The erythroblastic island provides an important nutritional and survival support niche for efficient erythropoietic differentiation. Island integrity is reliant on adhesive interactions between erythroid and macrophage cells. We show that erythroblastic islands can be formed from single progenitor cells present in differentiating embryoid bodies, and that these correspond to erythro-myeloid progenitors (EMPs) that first appear in the yolk sac of the early developing embryo. Erythroid Krüppel-like factor (EKLF; KLF1), a crucial zinc finger transcription factor, is expressed in the EMPs, and plays an extrinsic role in erythroid maturation by being expressed in the supportive macrophage of the erythroblastic island and regulating relevant genes important for island integrity within these cells. Together with its well-established intrinsic contributions to erythropoiesis, EKLF thus plays a coordinating role between two different cell types whose interaction provides the optimal environment to generate a mature red blood cell.


Panniculitis in a patient with metastatic renal cell carcinoma on a tyrosine kinase inhibitor.
A 71-year-old female was diagnosed with localized renal cell carcinoma in July 2008 with subsequent metastasis in 2012 to the right adrenal gland, lungs, and brain. Due to disease progression, she was started on pazopanib 800 mg daily in October 2012. In November 2016, the patient developed an ill-defined, red, 10 × 15 cm indurated plaque on the left lateral upper thigh with a discrete 3 cm firm tender tumor without ulceration. An incisional biopsy was performed and showed panniculitis with features resembling sclerosing lipogranuloma. Alternative causes including rheumatologic disease and trauma were ruled out. We report the first case of pazopanib-induced panniculitis. Key clinical and histopathological features include tender subcutaneous nodules, exclusion of other causes, and fatty microcysts within a densely sclerotic background on pathology. As targeted therapies are becoming increasingly common in the field of oncology, prompt identification and reporting of adverse reactions is critical for proper management.


Observations on pathology trends in 62,537 prostate biopsies obtained from urology private practices in the United States.
To evaluate pathology trends of 62,537 first-time prostate needle-core biopsies submitted by office-based urologists, processed at a single pathology laboratory. Prostate biopsy cases obtained over a 2-year period were assessed. Patient information included age, digital rectal examination (DRE) status, and prostate-specific antigen (PSA) serum levels. Biopsy pathology results included the number of tissue samples per case, Gleason score, presence of Gleason grades 4 or 5, percent of biopsy length with evidence of cancer, number of samples with cancer per biopsy, and determination of DNA ploidy status using microspectrophotometry. Adenocarcinoma, suspicious lesions, and isolated high-grade prostatic intraepithelial neoplasia (PIN) were diagnosed in 38.3%, 2.9%, and 4.1% of the biopsies, respectively. For each serum PSA and age range assessed, the positive biopsy rate and incidence of critical pathologic features increased consistently. The average percentage of biopsy length with evidence of tumor, the percentage of cases with Gleason grades 4 or 5, and the percentage of cases with an abnormal DNA ploidy all decreased significantly over the 2-year period (P <0.01). The number of tissue cores and anatomic sites (locations) being sampled per biopsy are increasing. The tumor size detected and percentage of cases with Gleason grades 4 and 5 are decreasing. There has been a slight increase in the number of biopsies performed on men younger than 60 years of age and a slight decrease in biopsies performed on men older than 70 years of age. The decline in meaningful pathologic features observed in biopsies over time may be clinically relevant to improved disease management.


Lack of potassium effect on Na-Cl cotransport in the medullary thick ascending limb.
The effect of potassium on sodium chloride uptake into rabbit renal medullary thick ascending limb of Henle's loop (mTALH) cells was studied to assess whether K participates in the Na-Cl cotransport system. Na uptake into the mTALH cells was inhibited 70% at 3 min by 1 mM furosemide. The total and furosemide-sensitive Na uptake was stimulated by Cl. Additionally, Cl uptake into the mTALH cells was stimulated by Na gradients and inhibited 42% at 3 min by 1 mM furosemide. Na uptake was studied in the presence of 0,5, or 140 mM external K gradients. Na uptake was similar in the absence and presence of K. Additionally, furosemide inhibited Na uptake as effectively in the absence or presence of K. Similar studies were conducted to study the effects of Na on 86Rb uptake. Na did not stimulate 86Rb uptake. The uptake of 86Rb was similar in the presence of 0,5, or 140 mM Na gradients. Furosemide had no significant inhibitory effect on 86Rb uptake. Barium (5 mM), an inhibitor of K conductance pathways, inhibited total 86Rb uptake by 19%. In the presence of 5 mM BaCl2, Na still did not have a stimulatory effect on 86Rb uptake. The results confirm the existence of a Na-Cl cotransport system in mTALH cells, but a direct effect of K on the NaCl cotransport system could not be demonstrated under the experimental conditions we used.


Characterization and identification of ubiquitin conjugation sites with E3 ligase recognition specificities.
In eukaryotes, ubiquitin-conjugation is an important mechanism underlying proteasome-mediated degradation of proteins, and as such, plays an essential role in the regulation of many cellular processes. In the ubiquitin-proteasome pathway, E3 ligases play important roles by recognizing a specific protein substrate and catalyzing the attachment of ubiquitin to a lysine (K) residue. As more and more experimental data on ubiquitin conjugation sites become available, it becomes possible to develop prediction models that can be scaled to big data. However, no development that focuses on the investigation of ubiquitinated substrate specificities has existed. Herein, we present an approach that exploits an iteratively statistical method to identify ubiquitin conjugation sites with substrate site specificities. In this investigation, totally 6259 experimentally validated ubiquitinated proteins were obtained from dbPTM. After having filtered out homologous fragments with 40% sequence identity, the training data set contained 2658 ubiquitination sites (positive data) and 5532 non-ubiquitinated sites (negative data). Due to the difficulty in characterizing the substrate site specificities of E3 ligases by conventional sequence logo analysis, a recursively statistical method has been applied to obtain significant conserved motifs. The profile hidden Markov model (profile HMM) was adopted to construct the predictive models learned from the identified substrate motifs. A five-fold cross validation was then used to evaluate the predictive model, achieving sensitivity, specificity, and accuracy of 73.07%, 65.46%, and 67.93%, respectively. Additionally, an independent testing set, completely blind to the training data of the predictive model, was used to demonstrate that the proposed method could provide a promising accuracy (76.13%) and outperform other ubiquitination site prediction tool. A case study demonstrated the effectiveness of the characterized substrate motifs for identifying ubiquitination sites. The proposed method presents a practical means of preliminary analysis and greatly diminishes the total number of potential targets required for further experimental confirmation. This method may help unravel their mechanisms and roles in E3 recognition and ubiquitin-mediated protein degradation.


Cytometric DNA analysis and prognostic biomarkers in breast carcinoma. Expression of P53 product in the different ploidy classes.
The aim of the study was to correlate the DNA Index (DI) and S-phase fraction (SPF) values determined by multiparametric flow cytometry in breast cancer (mainly T1 and T2 stages) with several clinico-pathologic variables and other biological parameters. For this purpose, a total of 136 breast cancers were submitted to flow cytometry and to several types of immunohistochemical analyses. Among clinico-pathologic data we considered pT, pN and grade and among immunohistochemical markers, hormonal receptors, P53, c-erbB-2 and MIB-1. We found that DNA aneuploidy was strongly correlated with high tumoral grade, absence of hormonal receptors, high proliferation, as shown by high MIB-1 (> or = 36%) and SPF values (> or = 13.3%), and P53 positivity. Grouping the tumours according to their DI values, we observed a relative significantly higher correlation of the near-triploid range carcinomas (DI 1.40-1.60) with immunohistochemical expression of P53 (p = 0.0001). Near-triploid DI was also associated with a high proliferative activity, expressed both by SPF (p = 0.0001) and MIB-1 reactivity (p = 0.0001), high tumoral grade (p = 0.0001) and presence of axillary metastases (p = 0.03). These data suggest that DNA near-triploid tumours in breast cancer may have a more aggressive behaviour in comparison with other DI classes.


Relationship between different stages of the corpus luteum and the expression of the peroxisome proliferator-activated receptor gamma protein in bovine large lutein cells.
Lutein cells produce progestins that support pregnancy. Steroidogenesis requires coordination of the anabolic and catabolic pathways of lipid metabolism. Peroxisome proliferator-activated receptors (PPAR) are transcription factors that are central in the regulation of lipid metabolism. Hence, they may play a role in regulation of the development and regression of the corpus luteum. The present study investigated the expression of PPAR-gamma, n during different stages of the corpus luteum. Lutein cells were isolated mechanically from non-pregnant and pregnant heifers on days 5, 12 and 20 of the oestrous cycle (n = 3 for each day). PPAR-gamma in single cells was analysed by flow cytometry. PPAR-gamma 1 and PPAR-gamma 2 isoforms were distinguished by immunoblotting. The cell cycle of the lutein cells was measured by the flow cytometric quantification of DNA in single cells, using propidium iodide staining after ethanol fixation and RNAse treatment, and by the detection of the proliferating cell nuclear antigen (PCNA). The response of the cells to PPAR-gamma agonist 15-deoxy-delta 12,14 prostaglandin J2 (15dPGJ2, 200 and 490 nmol l-1) with and without changing the cell cycle by the anti-apoptogenic drug aurintricarboxylic acid (ATA, 10 mumol l-1) was used as an in vitro model to study the relationship between the cell cycle and PPAR-gamma. The concentration of PPAR-gamma per cell from non-pregnant heifers was significantly higher on day 5 (3.40 +/- 0.30 fmol) compared with that on day 12 (1.34 +/- 0.18 fmol, P < 0.05) and day 20 (0.55 +/- 0.2 fmol, P < 0.05). In pregnant heifers, the concentration of PPAR-gamma was significantly (P < 0.01) higher than in non-pregnant heifers. A decrease in the PPAR-gamma 1 isoform relative to PPAR-gamma 2 was observed in cells on day 12 of the oestrous cycle compared with day 5. The cell cycle (S phase portion in cells on days 5, 12 and 20: 16 +/- 4%, 6 +/- 4% and 4 +/- 3%, respectively) and the portion of cells with PCNA correlated with the amount of PPAR-gamma in non-pregnant heifers. ATA promoted the S phase in cells of non-pregnant heifers (day 12) and the endogenous agonist of PPAR-gamma, 15dPGJ2, inhibited the response to ATA in a dose-dependent manner, indicating that PPAR-gamma plays a role in the arrest of the cell cycle in lutein cells to maintain their differentiated state.


Pediatric residents' perceptions about the impact of the COVID-19 pandemic on their training.
Introduction. Medical residency programs suffered changes that may have affected academic training during the COVID-19 pandemic. Objectives. To describe the perceptions of pediatric residents about the pandemic's impact on their education. To transculturally adapt and validate the COVID-19 Resident Education and Experience Survey into Spanish. Materials and methods. Observational, cross-sectional study. Participants were pediatric residents from hospitals across the country. The survey by Ostapenko et al. was used. A descriptive analysis was done using the SPSS software, version 21. The project was approved by the Institutional Ethics Committee. Results. The survey was completed by 127 residents. Most did more than 50 hours of health care activities before and during the pandemic. Also, 43.3% (95% CI: 35-52) spent at least 1 hour a day studying individually before the pandemic, while 63% (95% CI: 54.3-70.9) did so during the pandemic. In relation to the time spent doing academic work, 75.6% (95% CI: 67.4-82.2) reported that, before the pandemic, they spent at least 4 hours a week doing academic activities, dropping to 41.7% (95% CI: 33.5-50.4) during the pandemic. More than 60% (95% CI: 54.3-70.1) perceived that the pandemic impaired their training to become a specialist and 93.7% (95% CI: 88.1-96.8), that their stress levels increased. Conclusions. The perception was that participants spent less hours doing academic activities during the pandemic. Most surveyed participants mentioned that their stress levels increased and that the pandemic impaired their training to become a specialist.


Pneumothorax and/or Pneumomediastinum Worsens the Prognosis of COVID-19 Patients with Severe Acute Respiratory Failure: A Multicenter Retrospective Case-Control Study in the North-East of Italy.
Pneumothorax (PNX) and pneumomediastinum (PNM) are potential complications of COVID-19, but their influence on patients' outcomes remains unclear. The aim of the study was to assess incidence, risk factors, and outcomes of severe COVID-19 complicated with PNX/PNM. A retrospective multicenter case-control analysis was conducted in COVID-19 patients admitted for respiratory failure in intermediate care units of the Treviso area, Italy, from March 2020 to April 2021. Clinical characteristics and outcomes of patients with and without PNX/PNM were compared. Among 1213 patients, PNX and/or PNM incidence was 4.5%. Among these, 42% had PNX and PNM, 33.5% only PNX, and 24.5% only PNM. COVID-19 patients with PNX/PNM showed higher in-hospital (p = 0.02) and 90-days mortality (p = 0.048), and longer hospitalization length (p = 0.002) than COVID-19 patients without PNX/PNM. At PNX/PNM occurrence, one-third of subjects was not mechanically ventilated, and the respiratory support was similar to the control group. PNX/PNM occurrence was associated with longer symptom length before hospital admission (p = 0.005) and lower levels of blood lymphocytes (p = 0.017). PNX/PNM are complications of COVID-19 associated with a worse prognosis in terms of mortality and length of hospitalization. Although they are more frequent in ventilated patients, they can occur in non-ventilated, suggesting that mechanisms other than barotrauma might contribute to their presentation.


Effect of Peroxisome Proliferator-Activated Receptor-γ Coactivator-1 Alpha Variants on Spontaneous Clearance and Fibrosis Progression during Hepatitis C Virus Infection in Moroccan Patients.
Hepatitis C virus (HCV) is still one of the main causes of liver disease worldwide. Metabolic disorders, including non-alcoholic fatty liver disease (NAFLD), induced by HCV have been shown to accelerate the progression of fibrosis to cirrhosis and to increase the risk of hepatocellular carcinoma. An optimal peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PPARGC1A) activity is crucial to prevent NAFLD installation. The present study aims to investigate the associations between two common PPARGC1A polymorphisms (rs8192678 and rs12640088) and the outcomes of HCV infection in a North African context. A series of 592 consecutive Moroccan subjects, including 292 patients with chronic hepatitis C (CHC), 100 resolvers and 200 healthy controls were genotyped using a TaqMan allelic discrimination assay. PPARGC1A variations at rs8192678 and rs12640088 were not associated with spontaneous clearance of HCV infection (adjusted ORs = 0.76 and 0.79 respectively, P > 0.05, for both). Furthermore, multivariable logistic regression analysis showed that both SNPs were not associated with fibrosis progression (OR = 0.71; 95% CI 0.20-2.49; P = 0.739; OR = 1.28; 95% CI 0.25-6.54; P = 0.512, respectively). We conclude that, in the genetic context of South Mediterranean patients, rs8192678 and rs12640088 polymorphisms of PPARGC1A are neither associated with spontaneous clearance nor with disease progression in individuals infected with HCV.


Pathway Analysis of ChIP-Seq-Based NRF1 Target Genes Suggests a Logical Hypothesis of their Involvement in the Pathogenesis of Neurodegenerative Diseases.
Nuclear respiratory factor 1 (NRF1) serves as a transcription factor that activates the expression of a wide range of nuclear genes essential for mitochondrial biogenesis and function, including mitochondrial respiratory complex subunits, heme biosynthetic enzymes, and regulatory factors involved in the replication and transcription of mitochondrial DNA. Increasing evidence indicates that mitochondrial function is severely compromised in the brains of aging-related neurodegenerative diseases. To identify the comprehensive set of human NRF1 target genes potentially relevant to the pathogenesis of neurodegenerative diseases, we analyzed the NRF1 chromatin immunoprecipitation followed by deep sequencing (ChIP-Seq) dataset retrieved from the Encyclopedia of DNA Elements (ENCODE) project. Overall, we identified 2,470 highly stringent ChIP-Seq peaks on protein-coding genes in SK-N-SH human neuroblastoma cells. They were accumulated in the proximal promoter regions with an existence of the NRF1-binding consensus sequence. The set of ChIP-Seq-based NRF1 target genes included known NRF1 targets such as EIF2S1, EIF2S2, CYCS, FMR1, FXR2, E2F6, CD47, and TOMM34. By pathway analysis, the molecules located in the core pathways related to mitochondrial respiratory function were determined to be highly enriched in NRF1 target genes. Furthermore, we found that NRF1 target genes play a pivotal role in regulation of extra-mitochondrial biological processes, including RNA metabolism, splicing, cell cycle, DNA damage repair, protein translation initiation, and ubiquitin-mediated protein degradation. We identified a panel of neurodegenerative disease-related genes, such as PARK2 (Parkin), PARK6 (Pink1), PARK7 (DJ-1), and PAELR (GPR37) for Parkinson's disease, as well as PSENEN (Pen2) and MAPT (tau) for Alzheimer's disease, as previously unrecognized NRF1 targets. These results suggest a logical hypothesis that aberrant regulation of NRF1 and its targets might contribute to the pathogenesis of human neurodegenerative diseases via perturbation of diverse mitochondrial and extra-mitochondrial functions.


Kawasaki disease on PDGF expression and VSMC proliferation.
The effect of serum of patients with Kawasaki disease (KD) on expression of platelet-derived growth factor (PDGF) B chain protein in vascular endothelial cells (VEC) was studied by immunocytochemical method. Meanwhile, the effects of the endothelial cell conditioned media (ECM) on expression of PDGF receptor mRNA in vascular smooth muscle cells (VSMC) and on cell cycle of VSMC were investigated by the methods of nucleic acid hybridization and flow cytometry (FCM). The results showed that the serum of patients with KD induced the expression of PDGF-B chain protein significantly. ECM significantly promoted the expression of PDGF receptor mRNA and induced the proliferation of VSMC. These data suggest that the activation of PDGF-PDGF receptor may play a role in the pathogenesis of coronary complication of KD.


Glycogen synthase kinase 3 inhibition stimulates human cartilage destruction and exacerbates murine osteoarthritis.
To assess the role of glycogen synthase kinase 3 (GSK-3) as a regulator of cartilage destruction in human tissue and a murine model of osteoarthritis (OA). Surgical destabilization of the medial meniscus (DMM) was performed to induce experimental murine OA, and joint damage was assessed histologically. Bovine nasal and human OA cartilage samples were incubated with interleukin-1 (IL-1) plus oncostatin M (OSM) and GSK-3 inhibitor. Collagen and proteoglycan release was assessed by hydroxyproline measurement and dye binding assay, collagenase activity was assessed by bioassay, and gene expression was analyzed by real-time polymerase chain reaction. Human articular chondrocytes were isolated by enzymatic digestion and cultured prior to gene silencing and immunoblotting of cell lysates and nuclear fractions. Mice treated with GSK-3 inhibitor exhibited significantly greater cartilage damage compared with sham-operated control mice. GSK-3 inhibition in bovine cartilage dramatically accelerated IL-1 plus OSM-stimulated degradation, concomitant with a profound increase in collagenase activity. GSK-3 inhibitor induced collagen release from human OA cartilage in the presence of IL-1 plus OSM and increased proteoglycan loss. Gene expression profiling of resorbing OA cartilage revealed a marked procatabolic switch in gene expression upon GSK-3 inhibition. This was mirrored in human articular chondrocytes following GSK3 silencing, particularly with the GSK-3β isoform. GSK-3 inhibition or silencing led to enhanced IL-1 plus OSM-stimulated abundance and activity of Jun, and silencing of c-jun ameliorated GSK-3 inhibitor-mediated procatabolic gene expression. GSK-3 is an important regulator of matrix metalloproteinase (MMP)-mediated joint destruction, the inhibition of which by proinflammatory stimuli de-represses catabolic gene expression. Therapeutic strategies that maintain cartilage GSK-3 activity may therefore help curtail aberrant MMP activity during pathologic joint destruction.


Assignment and NOE analysis of 2'-hydroxyl protons in RNA: implications for stabilization of RNA A-form duplexes.
The ribose 2'-OH hydroxyl group distinguishes RNA from DNA. The 2'-OH hydroxyl protons are responsible for differences in conformation, hydration, and thermodynamic stability of RNA and DNA oligonucleotides. Additionally, the 2'-OH group plays a central role in RNA catalysis. This important group lies in the shallow groove of RNA, where it is involved in a network of hydrogen bonds with water molecules stabilizing RNA A-form duplexes. Structural and dynamical information on 2'-OH hydroxyl protons is essential to understand their respective roles. Here we report the 2'-OH hydroxyl proton assignments for a 30mer RNA, the HIV-2 transactivation region, in water using solution NMR techniques. We provide structural information on 2'-OH hydroxyl groups in the form of orientational preferences contradicting the paradigm that the 2'-OH hydroxyl typically points away from the ribose H1' proton.


Green Toxicology: a strategy for sustainable chemical and material development.
Green Toxicology refers to the application of predictive toxicology in the sustainable development and production of new less harmful materials and chemicals, subsequently reducing waste and exposure. Built upon the foundation of "Green Chemistry" and "Green Engineering", "Green Toxicology" aims to shape future manufacturing processes and safe synthesis of chemicals in terms of environmental and human health impacts. Being an integral part of Green Chemistry, the principles of Green Toxicology amplify the role of health-related aspects for the benefit of consumers and the environment, in addition to being economical for manufacturing companies. Due to the costly development and preparation of new materials and chemicals for market entry, it is no longer practical to ignore the safety and environmental status of new products during product development stages. However, this is only possible if toxicologists and chemists work together early on in the development of materials and chemicals to utilize safe design strategies and innovative in vitro and in silico tools. This paper discusses some of the most relevant aspects, advances and limitations of the emergence of Green Toxicology from the perspective of different industry and research groups. The integration of new testing methods and strategies in product development, testing and regulation stages are presented with examples of the application of in silico, omics and in vitro methods. Other tools for Green Toxicology, including the reduction of animal testing, alternative test methods, and read-across approaches are also discussed.


Computational Insights into the Mechanism of the Selective Catalytic Reduction of NO x : Fe- versus Cu-Exchanged Zeolite Catalysts.
We computationally investigate the mechanism of the reduction half-cycle of the selective catalytic reduction of nitrogen oxides with ammonia. We compare both Fe- and Cu-exchanged zeolite catalysts and aim at exploring all accessible reaction pathways. From our calculations, a comprehensive picture emerges that unifies several previous mechanistic proposals. We find that both for Fe and for Cu catalysts different reaction pathways are feasible but some of the possible reaction pathways differ in these two cases. Our computational results provide a basis for the interpretation of in situ spectroscopic investigations that can possibly distinguish the different mechanistic pathways.


Determining the accommodative response from wavefront aberrations.
The purpose of this study was to evaluate some of the methods used to calculate objective refractions from wavefront aberrations, to determine their applicability for accommodation research. A wavefront analyzer was used to measure the ocular aberrations of 13 emmetropes and 17 myopes at distance, and 4 near target vergences: 2, 3, 4, and 5 D. The accommodative response was calculated using the following techniques: least squares fitting (Zernike defocus), paraxial curvature matching (Seidel defocus), and 5 optical quality metrics (PFWc, PFSc, PFCc, NS, and VSMTF). We also evaluated a task-specific method of determining optimum focus that used a through-focus procedure to select the image that best optimized both contrast amplitude and gradient (CAG). Neither Zernike nor Seidel defocus appears to be the best method for determining the accommodative response from wavefront aberrations. When the eye has negative spherical aberration, Zernike defocus tends to underestimate, whereas Seidel defocus tends to overestimate the accommodative response. A better approach is to first determine the best image plane using a suitable optical quality metric and then calculate the accommodative error relative to this plane. Of the metrics evaluated, both NS and VSMTF were reasonable choices, with the CAG algorithm being a less preferred alternate.


Inflammation, heat shock proteins and periodontal pathogens in atherosclerosis: an immunohistologic study.
Inflammation is a significant component of atherosclerosis lesions. Bacteria, including periodontopathogens, have been demonstrated in atherosclerotic plaques and cross-reactivity of the immune response to bacterial GroEL with human heat shock protein 60 has been suggested as a link between infections and atherosclerosis. In this study, the nature of the inflammatory infiltrate and the presence of human heat shock protein 60 and GroEL were examined in 31 carotid endarterectomy specimens. Additionally, monoclonal antibodies were used to detect the presence of six bacteria, including those implicated in periodontal disease. The inflammatory cell infiltrate of the lesions was dominated by CD14(+) macrophages and CD4(+) T cells. Most cells of the infiltrate as well as the endothelium were HLA-DR(+), indicating activation; however, there was an absence of CD25 expression, demonstrating that the activated T cells were not proliferating. Few CD1a(+) and CD83(+) cells were noted. Human heat shock protein 60 expression was evident on endothelial cells and cells with the appearance of smooth muscle cells and lymphocytes. GroEL and bacteria were detected within intimal cells. Chlamydia pneumoniae, Porphyromonas gingivalis, Fusobacterium nucleatum, Tannerella forsythia, Prevotella intermedia, and Actinobacillus actinomycetemcomitans were found in 21%, 52%, 34%, 34%, 41%, and 17% of arteries, respectively. These results give evidence for a specific immune response associated with atherosclerosis. Whether bacteria initiate the observed inflammation in atherosclerotic lesions is not clear; however, the present study shows that maintenance of inflammation may be enhanced by the presence of periodontopathic bacteria.


Thixotropy and shear thinning of lubricated contacts with confined membranes.
We have modeled the nonlinear dynamics and the rheological behavior of a system under shear containing a membrane confined between two attractive walls. The presence of the membrane induces additional tangential forces on the walls that always increase the global friction. At low shear rates, the membrane exhibits chaotic dynamics with slow coarsening leading to thixotropy, i.e. to a slow decrease of the membrane-induced tangential forces on the walls. At intermediate shear rates, the membrane profile presents stationary periodic patterns. At higher shear rates, membrane dynamics are governed by a nonlinear evolution equation which is similar to the Kuramoto-Sivashinski equation, but with a sixth-order stabilizing term. The membrane experiences chaotic dynamics without coarsening. As a consequence of the nonlinear dynamics of the membrane at intermediate and large shear rates, the system exhibits shear thinning.


How Can We Improve Disease Education in People with Gout?
Gout management is currently suboptimal despite excellent available therapy. Gout patient education has been shown to enhance medication adherence and self-management, but needs improvement. We explored the literature on gout patient education including gaps in gout patient knowledge; use of written materials; in-person individual and group sessions; education via nurses, pharmacists, or multi-disciplinary groups; and use of phone, web-based, mobile health app, and text messaging educational efforts. Nurse-led interventions have shown significant improvement in reaching urate goals. Pharmacist-led programs have likewise succeeded, but to a lesser degree. A multi-disciplinary approach has shown feasibility. Needs-assessments, patient questionnaires, and psychosocial evaluations can enhance targeted education. An interactive and patient-centered approach can enhance gout educational interventions. Optimal programs will assess for and address educational needs related to knowledge gaps, health literacy, race, gender, socio-economic status, and level of social support.


Regulation of calcium dynamics and propagation velocity by tissue microstructure in engineered strands of cardiac tissue.
Disruptions to cardiac tissue microstructure are common in diseased or injured myocardium and are known substrates for arrhythmias. However, we have a relatively coarse understanding of the relationships between myocardial tissue microstructure, propagation velocity and calcium cycling, due largely to the limitations of conventional experimental tools. To address this, we used microcontact printing to engineer strands of cardiac tissue with eight different widths, quantified several structural and functional parameters and established correlation coefficients. As strand width increased, actin alignment, nuclei density, sarcomere index and cell aspect ratio decreased with unique trends. The propagation velocity of calcium waves decreased and the rise time of calcium transients increased with increasing strand width. The decay time constant of calcium transients decreased and then slightly increased with increasing strand width. Based on correlation coefficients, actin alignment was the strongest predictor of propagation velocity and calcium transient rise time. Sarcomere index and cell aspect ratio were also strongly correlated with propagation velocity. Actin alignment, sarcomere index and cell aspect ratio were all weak predictors of the calcium transient decay time constant. We also measured the expression of several genes relevant to propagation and calcium cycling and found higher expression of the genes that encode for connexin 43 (Cx43) and a subunit of L-type calcium channels in thin strands compared to isotropic tissues. Together, these results suggest that thinner strands have higher values of propagation velocity and calcium transient rise time due to a combination of favorable tissue microstructure and enhanced expression of genes for Cx43 and L-type calcium channels. These data are important for defining how microstructural features regulate intercellular and intracellular calcium handling, which is needed to understand mechanisms of propagation in physiological situations and arrhythmogenesis in pathological situations.


CXCL1/CXCR2 Paracrine Axis Contributes to Lung Metastasis in Osteosarcoma.
Osteosarcoma, the most common of all bone malignancies, has a high likelihood of lung metastasis. Up until now, the molecular mechanisms involved in osteosarcomas with lung metastases are not clearly understood. Recent observations have shown that the chemokine CXCL1 and its receptor CXCR2 assist with the homing of neutrophils into the tumor microenvironment. Here, we show that the CXCL1/CXCR2 paracrine axis is crucial for lung metastasis in osteosarcoma. In an in vivo lung metastasis model of osteosarcoma, lung blood vessels expressed CXCL1 and osteosarcoma cells expressed the CXCR2 receptor. CXCR2 expression was higher in osteosarcoma cell lines than in normal osteoblast cells. Immunohistochemistry staining of clinical osteosarcoma specimens revealed positive correlations between CXCR2 expression and pathology stage and also vascular cell adhesion molecule 1 (VCAM-1) expression. High levels of CXCL1 secreted by human pulmonary artery endothelial cells (HPAECs) promoted osteosarcoma cell mobility, which was mediated by the upregulation of VCAM-1 expression. When HPAECs-conditioned media was incubated in osteosarcoma cells, we observed that the CXCR2 receptor and FAK/PI3K/Akt/NF-κB signaling cascade were required for VCAM-1 expression. Our findings illustrate a molecular mechanism of lung metastasis in osteosarcoma and indicate that CXCL1/CXCR2 is worth targeting in treatment schemas.


Prenatal valproate exposure and adverse neurodevelopmental outcomes: Does sex matter?
Prenatal exposure to the antiepileptic drug (AED) valproic acid (VPA) is associated with an increased risk of impaired postnatal neurodevelopment, including autism spectrum disorder (ASD) and attention-deficit/hyperactivity disorder (ADHD). We aimed to evaluate the influence of sex and drug dosage on the association between prenatal VPA exposure and postnatal behavioral outcomes. The Australian Pregnancy Register of AEDs was interrogated to identify children aged 4-11 years prenatally exposed to AEDs. Parents reported on their child's behavior using the Autism Spectrum Quotient-Children's Version and the National Institute for Children's Health Quality Vanderbilt Assessment Scale for ADHD. General linear mixed-effects models were used to investigate the relationship between clinicodemographic variables and psychometric scores. A total of 121 children were studied: 54 prenatally exposed to VPA (28 males, 26 females; mean dose ± SD: 644 ± 310 mg/day) and 67 exposed to other AEDs. There was a main effect of sex showing higher ASD scores in males compared to females (p = .006). An interaction between sex and VPA exposure revealed that males had higher ASD symptoms among children exposed to AEDs other than VPA (p = .01); however, this typical sex dynamic was not evident in VPA-exposed children. There was no evidence of any dose-response relationship between VPA exposure and ASD symptoms. Males had higher ADHD scores compared to females, but there was no evidence for a link between ADHD symptoms and VPA exposure. Prenatal VPA exposure seems to negate the usual male sex-related predominance in the incidence of ASD. These initial findings deepen the concept of VPA as a "behavioral teratogen" by indicating that its effect might be influenced by sex, with females appearing particularly sensitive to the effects of VPA. No association between VPA doses and adverse postnatal behavioral outcomes was detected, possibly related to the low VPA doses used in this study.


Studying vapor-liquid transition using a generalized ensemble.
Homogeneous vapor-liquid nucleation is studied using the generalized Replica Exchange Method (gREM). The generalized ensemble allows the study of unstable states that cannot directly be studied in the canonical ensemble. Along with replica exchange, this allows for efficient sampling of the multiple states in a single simulation. Statistical Temperature Weighted Histogram Analysis Method is used for postprocessing to get a continuous free energy curve from bulk vapor to bulk liquid. gREM allows the study of planar, cylindrical, and spherical interfaces in a single simulation. The excess Gibbs free energy for the formation of a spherical liquid droplet in vapor for a Lennard-Jones system is calculated from the free energy curve and compared against the umbrella sampling results. The nucleation free energy barrier obtained from gREM is then used to calculate the nucleation rate without relying on any classification scheme for separating the vapor and liquid.


A thermoresponsive biodegradable polymer with intrinsic antioxidant properties.
Oxidative stress in tissue can contribute to chronic inflammation that impairs wound healing and the efficacy of cell-based therapies and medical devices. We describe the synthesis and characterization of a biodegradable, thermoresponsive gel with intrinsic antioxidant properties suitable for the delivery of therapeutics. Citric acid, poly(ethylene glycol) (PEG), and poly-N-isopropylacrylamide (PNIPAAm) were copolymerized by sequential polycondensation and radical polymerization to produce poly(polyethylene glycol citrate-co-N-isopropylacrylamide) (PPCN). PPCN was chemically characterized, and the thermoresponsive behavior, antioxidant properties, morphology, potential for protein and cell delivery, and tissue compatibility in vivo were evaluated. The PPCN gel has a lower critical solution temperature (LCST) of 26 °C and exhibits intrinsic antioxidant properties based on its ability to scavenge free radicals, chelate metal ions, and inhibit lipid peroxidation. PPCN displays a hierarchical architecture of micropores and nanofibers, and contrary to typical thermoresponsive polymers, such as PNIPAAm, PPCN gel maintains its volume upon formation. PPCN efficiently entrapped and slowly released the chemokine SDF-1α and supported the viability and proliferation of vascular cells. Subcutaneous injections in rats showed that PPCN gels are resorbed over time and new connective tissue formation takes place without signs of significant inflammation. Ultimately, this intrinsically antioxidant, biodegradable, thermoresponsive gel could potentially be used as an injectable biomaterial for applications where oxidative stress in tissue is a concern.


A virus-neutralising antibody is not cytotoxic in vitro.
Hybridoma cell lines are characterized by a preferential loss of the heavy chain gene. This observation has led to the theory that the immunoglobulin heavy chain possesses an intrinsic cytotoxic activity in some cell types. We have generated transgenic mice expressing the heavy and light chain genes of the virus-neutralising antibody A1 carrying constant domains of the human gamma1 and kappa isotype. Heavy chain and light chain transgenes were under trancriptional control of identical promoter regions derived from the mammary gland specific ovine beta-lactoglobulin gene. The copy number of the heavy chain transgene was consistently lower than the copy number of the light chain gene in all lines of transgenic mice. Moreover, the light chain gene was expressed in significant excess of the heavy chain gene in the lactating mammary gland in all transgenic lines. In several transgenic lines, the differences in antibody expression were greater than could be explained by the differences in transgene copy number. One potential cause of this phenotype could be a cytotoxic effect of free heavy chain protein in embryonic cells (resulting in differences in copy number) or mammary epithelial cells (resulting in differences in transgene expression). We therefore directly assessed the effect of the expression of free A1 heavy chain protein in epithelial cell lines and in murine embryonic stem cells. However, full-length A1 heavy chain mRNA and protein could be expressed transiently and stably in both epithelial and embryonic stem cells and had no detectable effect on cell viability. Taken together, these findings argue against an inherent cytotoxicity of the free A1 heavy chain protein in epithelial or embryonic cells.


Putative role of adhesion molecules in metabolic disorders.
Diabetes mellitus is associated with an increased risk of premature vascular disease. Vascular research is focusing on a potential role of adhesion molecules in diabetes mellitus, since classical risk factors including hyperlipidemia and hypertension do not completely account for the increased incidence of atherosclerosis in diabetes. After the expression of adhesion molecules on the cell surface, they are shed into plasma. Thus plasma concentrations of circulating adhesion molecules may be representative for endothelial activation, damage or turnover. Recently, evidence has been accumulating that increased plasma levels of adhesion molecules may predict cardiovascular disease, may be pathognomonic for diabetic microangiopathy or may even play a functional pathophysiologic role. The purpose of this article is to briefly summarise the role of (circulating) adhesion molecules in diabetes mellitus.


Leukocyte extracellular vesicle concentration is inversely associated with liver fibrosis severity in NAFLD.
The enhanced liver fibrosis (LFS) score and the nonalcoholic fatty liver disease (NAFLD) fibrosis score (NFS) are algorithmic-derived scores for diagnosing severe (F3/F4) liver fibrosis. In a pilot, substudy of the Wessex Evaluation of fatty Liver and Cardiovascular markers in NAFLD with OMacor thErapy (WELCOME) trial, we tested whether measurements of plasma platelet-, endothelial-, and leukocyte-derived extracellular vesicles (EVs) counts are (a) associated with, and predict, F3/F4 fibrosis and (b) able to improve risk prediction of F3/F4 fibrosis in NAFLD, building upon LFS or NFS algorithms. Twenty-six individuals with NAFLD had liver fibrosis severity determined by Kleiner scoring after liver biopsy. Plasma samples stained with CD41a, CD42b, CD31, CD105, CD14, CD16, and CD284 antibodies were analyzed using flow cytometry to measure platelet-, endothelial-, and leukocyte-derived EVs counts. The independence of associations between EVs and F3/F4 fibrosis were tested using logistic regression. Receiver operator characteristic (ROC) curves were used to evaluate F3/F4 fibrosis prediction models. LFS was more strongly associated with F3/F4 fibrosis than NFS (χ2= 15.403, P < 0.0001, and χ2= 6.300, P = 0.012, respectively). The association between LFS and F3/F4 fibrosis was further improved by addition of CD14+ EVs (χ2=20.847,P = 0.016 vs. χ2=12.803,P = 0.015, respectively) or CD16+ EVs (χ2=22.205,P = 0.009 vs. χ2=17.559,P = 0.001, respectively), and the area under the ROC for LFS (AUC = 0.915, se = 0.055, P = 0.001) was increased by the addition of CD14+ or CD16+ EVs (AUC = 0.948, se = 0.042, and P < 0.001 and AUC = 0.967, se = 0.055, P < 0.001, respectively) as predictor variables. In this small preliminary study, CD14+ and CD16+ EV counts show potential to predict liver fibrosis severity with either marker improving the ability of the LFS to identify F3/F4 fibrosis in this small preliminary cohort study.


Dynamic cellular finite-element method for modelling large-scale cell migration and proliferation under the control of mechanical and biochemical cues: a study of re-epithelialization.
Computational modelling of cells can reveal insight into the mechanisms of the important processes of tissue development. However, current cell models have limitations and are challenged to model detailed changes in cellular shapes and physical mechanics when thousands of migrating and interacting cells need to be modelled. Here we describe a novel dynamic cellular finite-element model (DyCelFEM), which accounts for changes in cellular shapes and mechanics. It also models the full range of cell motion, from movements of individual cells to collective cell migrations. The transmission of mechanical forces regulated by intercellular adhesions and their ruptures are also accounted for. Intra-cellular protein signalling networks controlling cell behaviours are embedded in individual cells. We employ DyCelFEM to examine specific effects of biochemical and mechanical cues in regulating cell migration and proliferation, and in controlling tissue patterning using a simplified re-epithelialization model of wound tissue. Our results suggest that biochemical cues are better at guiding cell migration with improved directionality and persistence, while mechanical cues are better at coordinating collective cell migration. Overall, DyCelFEM can be used to study developmental processes when a large population of migrating cells under mechanical and biochemical controls experience complex changes in cell shapes and mechanics.


4-(N-hydroxyphenyl)retinamide can selectively induce apoptosis in human epidermoid carcinoma cells but not in normal dermal fibroblasts.
The retinoid 4-(N-hydroxyphenyl)retinamide (4HPR, fenretinide) has both growth inhibitory and apoptosis-inducing effects on a number of cancer cell lines in vitro and in vivo and has been entered into a number of oncological trials. However, little is known about its mechanism(s) of action or its effects on normal cells such as fibroblasts. In this study, the effects of fenretinide on both epidermoid carcinoma cells of vulva (cell line A431) and normal human dermal fibroblasts, both as monolayers and also grown in 3D cell culture systems, have been investigated. The 3D cell culture system contained normal human fibroblasts embedded in a type I collagen gel with the carcinoma cells seeded on top of the collagen gel, which mimics the epidermoid carcinoma. Fenretinide significantly inhibited the rate of DNA synthesis of carcinoma cells, while there was little effect on fibroblasts on monolayers, at 10(-6)-10(-5) M, which are clinically attainable doses. Fenretinide at 5 x 10(-6) M induced apoptosis characterised by cell shrinkage, membrane blebbing, nuclear condensation and/or fragmentation, and cell detachment in carcinoma cells, but not fibroblasts from monolayers. Fenretinide also reduced the viability of carcinoma cells in the 3D cell culture system without affecting fibroblasts. These data show that fenretinide may preferentially induce apoptosis in epidermoid carcinoma cells.


Carcinoma erysipeloides from prostate cancer presenting as cellulitis.
We report a 72-year-old man with carcinoma erysipeloides presenting as cellulitis of the thigh. Immunohistochemical study of the skin biopsy showed positive staining to prostate-specific antigen, confirming that the tumor originated in the prostate.


Novel therapeutic approaches in polycythemia vera.
Polycythemia vera (PV) is the most common Philadelphia chromosome-negative myeloproliferative neoplasm. Whereas low-risk patients are treated with aspirin and phlebotomy, high-risk patients receive cytoreductive therapy, which most commonly consists of hydroxyurea in the United States. Concerns about the long-term safety of hydroxyurea, as well as a desire for more efficacious and targeted therapy, have led to the development of novel therapies for high-risk patients with PV. Pegylated interferon (IFN) has shown promise in phase 2 studies of PV, and preliminary data from ongoing phase 3 studies suggest noninferiority as a frontline therapy. Efficient count control, tolerability, and even molecular responses as a salvage therapy have been demonstrated. Ropeginterferon-α-2b, a monopegylated IFN with a longer half-life and less frequent dose interval compared with recombinant or pegylated IFN, is an impressive agent in development. Ruxolitinib has a proven role as second-line therapy for PV, but an ongoing trial combining ruxolitinib and IFN as salvage therapy is under way. Early-phase clinical trials have also suggested that MDM2 inhibitors such as idasanutlin and histone deacetylase inhibitors should continue in their development. If these novel agents are able to modify the natural history of PV, the treatment paradigm in newly diagnosed patients will evolve from risk-adapted or reactive treatment toward early interventions.


Decontamination of autogenous bone grafts collected during dental implant site preparation: a pilot study.
Dental implant site preparation produces bone particles that can be used as autogenous bone graft material for the reconstruction of alveolar bone defects; however, collected bone particles are contaminated with oral microorganisms that may cause augmentation failure due to complications associated with infection. The stringent aspiration protocol, preoperative oral chlorhexidine rinsing, and antibiotic prophylaxis were implemented before collecting bone particles. Nonetheless, collected bone particles were still contaminated with bacteria, and, therefore, decontamination of the collected bone particles with chlorhexidine or clindamycin was considered. The aims of this study were to quantitatively determine the degree of bacterial contamination of collected bone particles and to quantitatively evaluate the efficacy of treating collected bone particles with clindamycin or chlorhexidine solutions. Both of the agents effectively decontaminated the collected bone particles. Comparison between these antimicrobials in further studies could be useful in determining which is most effective.


Peripheral blood mononuclear cell subsets in patients with severe inherited forms of epidermolysis bullosa.
Epidermolysis bullosa (EB) is a group of inherited disorders in which slight trauma to the skin results in blister formation. Patients with severe types of EB suffer cutaneous infections that sometimes progress to septicemia and cutaneous and gastrointestinal carcinomas that are locally aggressive and frequently metastasize. Previous studies have shown deficits in natural killer (NK) cell activity as well as in lymphokine and monokine production in patients with severe forms of EB. Alterations in peripheral blood mononuclear cells, however, which may reflect on immune functions in patients with EB, have received little attention. A prospective study was designed to ascertain if differences existed between subsets of peripheral blood mononuclear cells in patients with severe forms of EB vs healthy control subjects. Thirty patients with clinical and histologic diagnoses of EB and 30 healthy volunteers were studied. Flow cytometric analysis of labeled cells was performed. Absolute numbers of CD3+, CD2+, CD4+, CD19+, NK+, CD29+, and CD45R+ cells were lower in patients with severe types of EB in comparison with controls. The T cells showed decreased numbers of interleukin 2 receptors. An increase in numbers of CD20+, CD4+ CD8+, and CD4-CD8- cells was also observed in patients with severe types of EB. Alterations in monocyte and lymphocyte subsets known to affect host immune response were observed in patients with severe forms of EB. Quantitative changes relative to controls included decreased total numbers of T cells with greater decreases in helper cells, decreased NK cells, and a diminished number of interleukin 2 receptors. Such changes have been associated previously with a lower resistance to infections and to neoplasia. The changes in subsets correlated with the severity of the cutaneous and extracutaneous disease in the patients with EB.


Ethnic differences in primary angle-closure glaucoma.
Observational studies from different countries have shown that populations of East Asian origin have a higher frequency of primary angle-closure glaucoma compared with those of European or African descent. As half of all cases of glaucoma reside in Asia, and with primary angle-closure glaucoma carrying a higher rate of visual morbidity, primary angle-closure glaucoma poses an important public health problem; however, the inconsistent use of techniques and definitions to detect and diagnose primary angle-closure glaucoma has resulted in difficulties in interpreting the accuracy and comparability of such data. Therefore it is important to review these studies in the light of a consistent classification system. There are increasing reports that support previous findings on the incidence and prevalence of primary angle-closure glaucoma in different ethnic groups. There have also been further investigations into the mechanism and natural history of primary angle-closure glaucoma in Asian populations. International investigations into primary angle-closure glaucoma have demonstrated reproducible evidence that ethnic variations do exist. Cross-sectional studies in this area have also suggested that differences in anterior chamber depth, together with its association with peripheral anterior synechiae, may be part of the underlying mechanism behind these differences. The ideas generated need to be further explored with longitudinal data of changes in anterior chamber depth and peripheral anterior synechiae in different populations. The detailed mechanisms behind the development of angle-closure and primary angle-closure glaucoma should also be investigated.


Randomized phase II study of clinical effects of ghrelin after esophagectomy with gastric tube reconstruction.
Ghrelin is a peptide hormone with pleiotropic functions including stimulation of growth hormone secretion and appetite, and its levels decrease after esophagectomy. The aim of this study was to evaluate whether exogenous ghrelin administration can meliorate the postoperative decrease of oral food intake and body weight, which are serious complications after esophagectomy. This prospective randomized, placebo-controlled, clinical trial assigned a total of 20 patients with thoracic esophageal cancer who underwent radical operation into either a ghrelin (n =10) or placebo (n =10) group. Synthetic human ghrelin (3 microg/kg) or 0.9% saline placebo was administered intravenously twice daily for 10 days from the day after the start of food intake. The primary end point was calories of food intake. Comparison of appetite and changes in weight and body composition were also made between the 2 groups. Intake of food calories was greater in ghrelin group than placebo group (mean 874 vs 605 kcal per day; P =.015). The appetite score tended to be greater in ghrelin group than placebo group (P =.094). Loss of weight was less in ghrelin group (-1% vs -3%; P =.019) and this attenuation was due largely to a decrease of lean body weight loss (0% vs -4%; P =.012). No side effects were observed in either groups. These preliminary results suggest that administration of ghrelin after esophagectomy increased oral food intake and attenuated weight loss together with maintenance of lean body weight.


A Multicenter, Randomized, Observer-blinded, Active-controlled Study Evaluating the Safety and Effectiveness of Ceftaroline Compared With Ceftriaxone Plus Vancomycin in Pediatric Patients With Complicated Community-acquired Bacterial Pneumonia.
The broad-spectrum cephalosporin ceftaroline, a metabolite of the prodrug ceftaroline fosamil, has shown in vitro activity against clinical isolates from pediatric patients. This multicenter, randomized, observer-blinded, active-controlled study (NCT01669980) assessed the safety and effectiveness of ceftaroline fosamil compared with ceftriaxone plus vancomycin in patients between 2 months and 17 years of age with complicated community-acquired bacterial pneumonia. Patients were randomized 3:1 (stratified by age cohort) to receive either ceftaroline fosamil or ceftriaxone plus vancomycin (comparator) as intravenous therapy for ≥3 days. Patients who met specific study criteria on or after Study Day 4 were permitted to switch to an oral study drug. Safety assessments were treatment-emergent adverse events, and the effectiveness of treatment was assessed by clinical and microbiologic outcomes. The median duration of intravenous treatment was 9.0 (range, 3.0-19.0) days in the ceftaroline fosamil group (N=30) and 7.5 (5.0-13.0) days in the comparator group (N=10). At least one treatment-emergent adverse event was experienced by 12/30 patients (40%) in the ceftaroline fosamil group and 8/10 (80%) in the comparator group; most treatment-emergent adverse events in both groups were mild to moderate in intensity. Clinical response rates in the modified intent-to-treat population were 52% (15/29 patients) in the ceftaroline fosamil group and 67% in the comparator group (6/9); clinical stability at Study Day 4 was 21% (6/29) and 22% (2/9), respectively. Ceftaroline fosamil was well tolerated and showed similar clinical response rates to ceftriaxone plus vancomycin in pediatric patients with complicated community-acquired bacterial pneumonia.


[A quantitative diagnosis of chronic gastritis].
To explore a definite diagnostic criterion of chronic gastritis, we undertook a quantitative study of the gastric biopsy specimens taken from 131 subjects. Seven stages were classified from normal gastric mucosa to mild, moderate and severe superficial and atrophic gastritis. The results shown the number of inflammatory cell was much greater in the mild superficial gastritis (128 +/- 84.37/HP) than in normal gastric mucosa (22 +/- 10.54HP, P less than 0.001). In these stages, the numbers of pyloric and fundic gland progressively decreased, the size of the gastric gland gradually enlarged and the percentage of intestinal metaplasia by degrees increased. According to these, we establish the quantitative diagnostic criterion of chronic gastritis.


Short hairpin RNA gene silencing of prolyl hydroxylase-2 with a minicircle vector improves neovascularization of hindlimb ischemia.
In this study, we target the hypoxia inducible factor-1 alpha (HIF-1-alpha) pathway by short hairpin RNA interference therapy targeting prolyl hydroxylase-2 (shPHD2). We use the minicircle (MC) vector technology as an alternative for conventional nonviral plasmid (PL) vectors in order to improve neovascularization after unilateral hindlimb ischemia in a murine model. Gene expression and transfection efficiency of MC and PL, both in vitro and in vivo, were assessed using bioluminescence imaging (BLI) and firefly luciferase (Luc) reporter gene. C57Bl6 mice underwent unilateral electrocoagulation of the femoral artery and gastrocnemic muscle injection with MC-shPHD2, PL-shPHD2, or phosphate-buffered saline (PBS) as control. Blood flow recovery was monitored using laser Doppler perfusion imaging, and collaterals were visualized by immunohistochemistry and angiography. MC-Luc showed a 4.6-fold higher in vitro BLI signal compared with PL-Luc. BLI signals in vivo were 4.3×10(5)±3.3×10(5) (MC-Luc) versus 0.4×10(5)±0.3×10(5) (PL-Luc) at day 28 (p=0.016). Compared with PL-shPHD2 or PBS, MC-shPHD2 significantly improved blood flow recovery, up to 50% from day 3 until day 14 after ischemia induction. MC-shPHD2 significantly increased collateral density and capillary density, as monitored by alpha-smooth muscle actin expression and CD31(+) expression, respectively. Angiography data confirmed the histological findings. Significant downregulation of PHD2 mRNA levels by MC-shPHD2 was confirmed by quantitative polymerase chain reaction. Finally, Western blot analysis confirmed significantly higher levels of HIF-1-alpha protein by MC-shPHD2, compared with PL-shPHD2 and PBS. This study provides initial evidence of a new potential therapeutic approach for peripheral artery disease. The combination of HIF-1-alpha pathway targeting by shPHD2 with the robust nonviral MC plasmid improved postischemic neovascularization, making this approach a promising potential treatment option for critical limb ischemia.


Determinants of clinical performance in a physical therapy program.
In admission decisions, allied health programs evaluate various factors that may include overall grade-point average (GPA), science GPA, GPA in the most recent 60 hours of course work, volunteer/work experience, and performance during an admission interview. Most use mathematical formulas based on these parameters, but the rationale for weighting each factor is not always clear. To determine whether pre-admission variables can differentiate between good and poor clinical performances, the authors compared pre-admission data from 118 graduates of a physical therapy (PT) baccalaureate program with their clinical performances. GPA in the PT curriculum was included to determine whether it was related to clinical performance, which was taken as the average of overall performance scores assigned to each student by his or her instructors at four clinical education sites. The students were divided into quartiles by clinical scores. One-way ANOVA was used to identify differences between quartiles for each variable (alpha level of p < 0.05). The interview score was the only pre-admission variable that differentiated between students who did and did not perform well in the clinic (p < 0.003). Those who performed better in the clinic also had higher PT GPAs (p < 0.0001). The interview format and scoring system used may easily be adapted for other programs. Recommendations are made regarding how to utilize interview scores in the admission decision.


Implication of enterohepatic re-circulation on single dose bioequivalence evaluation of two brands of clonidine hydrochloride tablets in healthy human volunteers.
A single dose, crossover bioequivalence study of two different brands of clonidine hydrochloride 25 mug tablets was conducted in 24 (+2 stand by) healthy, adult, male, Indian subjects under fasting conditions to check the implication of enterohepatic re-circulation on assessment of bioequivalence. After an overnight fasting of at least 10 h, the subjects received single oral dose of test or reference product with either of the product as per randomization schedule in each period with a washout period of 10 days. The pre-dose blood sample was collected within a period of one h before dosing. The post-dose blood samples were collected at specified time intervals up to 96 h. The plasma concentrations of clonidine were quantified by validated LCMS/MS method and pharmacokinetic parameters were computed. The 90% confidence intervals of test/reference ratios for C(max) and area under the plasma-concentration- time-curve AUC under 0-t were found to be between 0.80 and 1.25 for log-transformed data. Analysis of variance did not show significant difference to these parameters. No meaningful values of K(el) and therefore AUC under 0-infinity could be calculated for significant number of subjects due to enterohepatic re-circulation. Based on the results obtained, two different brands of clonidine 25 mug tablets have comparable rate and extent of absorption after oral administration but failed to show bioequivalence as per regulatory requirement of Food and Drugs Administration-united states.


[Hyperplastic changes and aldosterone content in the adrenal cortex in essential hypertension and primary aldosteronism].
Autopsy and operative material (adrenalectomy for hyperaldosteronism) was studied to elucidate morphology, incidence of nodules, aldosterone content in the adrenal of patients with essential hypertension (EH). It was established than nodular masses in the adrenals in the form of micro and macronodules are present in 80% of EH patients. Aldosterone content in the adrenals in both nodules and in the adjacent cortex is significantly higher than in the adrenals of patients without EH. This fact as well as increased cell nuclei size in the fascicular and glomerular zones indicate high secretory activity of the adrenals in EH. Clinicomorphological comparisons in patients after adrenalectomy because of hyperaldosteronism syndrome allow to conclude that grave forms of EH may be followed by aldosteronism syndromes with nodular hyperplasia of the adrenal cortex being the basis of the syndrome. Indications to adrenalectomy require clear criteria of differential diagnosis with primary aldosteronism.