Model for intravital microscopic evaluation of the effects of arterial occlusion-caused ischemia in bone.
An in vivo model has been developed for chronic observation of the effects of ischemia on cortical bone remodeling and perfused vascularity. Diaphragm occluders were implanted around the right common iliac artery of four rabbits and inflated to produce 10 h of ischemia to the limb. Microcirculation was monitored with intravital microscopy of injected fluorescent microspheres and FITC-Dextran 70 through a bone window, the tibial bone chamber implant (BCI). Bone resorption and apposition in the BCI were indicated with mineralization dyes. Between 2 and 12 h following release of the occluder, secondary ischemia/no-reflow and other evidence of reperfusion injury were observed. Vessel damage was suggested by abnormally high leakage of FITC-D70 from the few vessels perfused during secondary ischemia. In the weeks following occluder release perfused vasculature increased beyond pre-occlusion levels. Net bone resorption reached a maximum when vascularity passed normal levels. In order to further validate the arterial occlusion model for osteonecrosis, techniques for (1) confirming bone death and (2) detecting increased leukocyte adherence to endothelial cells were added. The dead cell stain Ethidium homodimer-1 was used to tag dead osteocytes immediately after occlusion and produced a measure designated "osteonecrosis index." To detect leukocytes adhering to vessel walls, carboxyfluorescein diacetate, succinimidyl ester was injected at occluder release. An increase in the number of adherent leukocytes was detected.


Dopamine uptake inhibitors but not dopamine releasers induce greater increases in motor behavior and extracellular dopamine in adolescent rats than in adult male rats.
Most life-long drug addiction begins during adolescence. Important structural and functional changes in brain occur during adolescence and developmental differences in forebrain dopamine systems could mediate a biologic vulnerability to drug addiction during adolescence. Studies investigating age differences in psychostimulant responses have yielded mixed results, possibly because of different mechanisms for increasing extracellular dopamine. Recent research from our laboratory suggests that adolescent dopamine systems may be most affected by selective dopamine uptake inhibitors. We investigated age-related behavioral responses to acute administration of several dopamine uptake inhibitors [methylphenidate, 1-{2-[bis-(4-fluorophenyl)methoxy]ethyl}-4-(3-phenylpropyl)piperazine (GBR12909), and nomifensine] and releasing agents [amphetamine and methylenedioxymethamphetamine (MDMA)] in adolescent and adult male rats. Methylphenidate and amphetamine effects on stimulated dopamine efflux were determined using fast-scan cyclic voltammetry in vivo. Dopamine uptake inhibitors but not dopamine releasing agents induced more locomotion and/or stereotypy in adolescent relative to adult rats. MDMA effects were greater in adults at early time points after dosing. Methylphenidate but not amphetamine induced much greater dopamine efflux in periadolescent relative to adult rats. Periadolescent male rats are particularly sensitive to psychostimulants that are DAT inhibitors but are not internalized and do not release dopamine. Immaturity of DAT and/or DAT associated signaling systems in adolescence specifically enhances behavioral and dopaminergic responses in adolescence.


Collagen-EGCG Combination Synergistically Prevents UVB-Induced Skin Photoaging in Nude Mice.
Ultraviolet (UV) radiation is a major cause of skin photoaging through generating excessive oxidative stress and inflammation. One of the strategies is to use photo-chemoprotectors, such as natural products with antioxidant and anti-inflammatory properties, to protect the skin from photo damage. The present study investigates the photoprotective potentials of topical administration of unhydrolyzed collagen, epigallocatechin gallate (EGCG), and their combination against ultraviolet B (UVB)-induced photoaging in nude mice. It is found that both the solo and combined pretreatments could recover UVB-induced depletion of antioxidative enzymes, including superoxide dismutase and glutathione peroxidase (GSH-Px), as well as an increase of lipid peroxide malondialdehyde and inflammatory tumor necrosis factor-α. Meanwhile, the UVB-stimulated skin collagen degradation is attenuated significantly with drug treatments, which is evidenced by expression analysis of matrix metalloproteinase-1 and hydroxyproline. Additionally, the mouse skin histology shows that the drug-pretreated groups possess decreased epidermis thickness and normal collagen fiber structure of the dermis layer. These results demonstrate that both EGCG and collagen can protect the skin against UVB-induced skin photoaging. Synergistically, the combination of them shows the maximum prevention to skin damage, showing its potential in the application of anti-photoaging formulation products.


Development of a bioclimatic wind rose tool for assessment of comfort wind resources in Sydney, Australia for 2013 and 2030.
This study assessed the effect of wind on human thermal comfort by preforming outdoor urban climatic comfort simulations using state-of-the-art heat-balance models of human thermo-physiology (Universal Thermal Climate Index-UTCI). A series of simulations for computing "wind cooling potential" have been performed using the UTCI index temperatures. The comfort cooling effect of wind has been estimated by modelling with wind taken into account, and under calm wind (0.05 m/s) (ΔUTCI). A novel wind rose biometeorological data visualisation tool that integrates an additional thermal comfort dimension into the conventional climatology wind rose visualisation was developed in this study. The new wind rose graphic tool identifies "predominant" wind directions, and whether or not they are "desirable" from the human thermal comfort point of view. This tool's utility lies in its identification of the optimal building orientation in its surrounding urban morphology, based on the cooling potential of wind resources when enhanced ventilation is desirable for thermal comfort.


Rapid detection of group B streptococci directly from vaginal swabs.
Duplicate vaginal swabs were obtained from patients who attended obstetric or gynecologic clinics affiliated with the Magee Womens Hospital in Pittsburgh. One swab was cultured semiquantitatively on 5% sheep blood agar to detect group B streptococci (GBS). The other swab was subjected to a rapid method (25 min) for antigen detection and micronitrous acid exposure to extract the GBS antigen, followed by latex particle agglutination. A total of 464 swabs were evaluated by direct plating. Fifty-two swabs (11.2%) were found to contain GBS. Overall, the rapid method detected 21 of 52, or 40.4%, positive specimens. The sensitivity of the rapid method for identifying the most heavily colonized samples was 85.7%. This method can be used to identify maternity patients who are heavily colonized with GBS and are at high risk of delivering septic infants.


Antibiotic losses from unprotected manure stockpiles.
Manure management is a major concern in livestock production systems. Although historically the primary concerns have been nutrients and pathogens, manure is also a source of emerging contaminants, such as antibiotics, to the environment. There is a growing concern that antibiotics in manure are reaching surface and ground waters and contributing to the development and spread of antibiotic resistance in the environment. One such pathway is through leaching and runoff from manure stockpiles. In this study, we quantified chlortetracycline, monensin, and tylosin losses in runoff from beef manure stockpiles during two separate but consecutive experiments representing different weather conditions (i.e., temperature and precipitation amount and form). Concentrations of chlortetracycline, monensin, and tylosin in runoff were positively correlated with initial concentrations of antibiotics in manure. The highest concentrations of chlortetracycline, monensin, and tylosin in runoff were 210, 3175, and 2544 microg L(-1), respectively. Relative antibiotic losses were primarily a function of water losses. In the experiment that had higher runoff water losses, antibiotic losses ranged from 1.2 to 1.8% of total extractable antibiotics in manure. In the experiment with lower runoff water losses, antibiotic losses varied from 0.2 to 0.6% of the total extractable antibiotics in manure. Manure analysis over time suggests that in situ degradation is an important mechanism for antibiotic losses. Degradation losses during manure stockpiling may exceed cumulative losses from runoff events. Storing manure in protected (i.e., covered) facilities could reduce the risk of aquatic contamination associated with manure stockpiling and other outdoor manure management practices.


Methionine oxidation inhibits assembly and promotes disassembly of apolipoprotein C-II amyloid fibrils.
Methionine residues are linked to the pathogenicity of several amyloid diseases; however, the mechanism of this relationship is largely unknown. These diseases are characterized, in vivo, by the accumulation of insoluble proteinaceous plaques, of which the major constituents are amyloid fibrils. In vitro, methionine oxidation has been shown to modulate fibril assembly in several well-characterized amyloid systems. Human apolipoprotein (apo) C-II contains two methionine residues (Met-9 and Met-60) and readily self-assembles in vitro to form homogeneous amyloid fibrils, thus providing a convenient system to examine the effect of methionine oxidation on amyloid fibril formation and stability. Upon oxidation of the methionine residues of apoC-II with hydrogen peroxide, fibril formation was inhibited. Oxidized apoC-II molecules did not inhibit native apoC-II assembly, indicating that the oxidized molecules had a reduced ability to interact with the growing fibrils. Single Met-Val substitutions were performed and showed that oxidation of Met-60 had a more significant inhibitory effect than oxidation of Met-9. In addition, Met-Gln substitutions designed to mimic the effect of oxidation on side chain hydrophilicity showed that a change in hydrophobicity at position 60 within the core region of the fibril had a potent inhibitory effect. The oxidation of preformed apoC-II fibrils caused their dissociation; however, mutants in which the Met-60 was substituted with a valine were protected from this peroxide-induced dissociation. This work highlights an important role for methionine in the formation of amyloid fibril structure and gives new insight into how oxidation affects the stability of mature fibrils.


Gonadotropin-mediated regulation of the murine VEGF expression in MA-10 Leydig cells.
Presence of vascular endothelial growth factor (VEGF) is not only limited to cells directly involved in angiogenesis but has also been demonstrated in steroidogenic cells like testicular Leydig cells. Because Leydig cells are subjected to regulation by gonadotropic hormones and produce steroid hormones, we have investigated here the effects of human chorionic gonadotropin (hCG) or steroid hormones on VEGF expression in cultured mouse tumor Leydig cells (MA-10 cells) and have then analyzed the underlying molecular mechanisms. Northern blot analysis and enzyme-linked immunosorbent assays revealed increases in VEGF mRNA and protein levels, respectively, over 3-20 hours in MA-10 cells after stimulation with hCG or 8-Br-cAMP. Although MA-10 cells lack the classical progesterone receptor, progesterone was able to stimulate VEGF expression. Promoter analyses and antibody supershift experiments suggested that the proximal region is able to constitutively bind the transcription factors Sp1 and Sp3. Mutations of 2 potential Sp1 binding sites in the proximal region showed the requirement of these motifs for stimulation of VEGF by hCG and 8-Br-cAMP. The distal cytosine-rich sequence interacts with so far-unidentified faster migrating factors. Following stimulation with hCG or 8-Br-cAMP, the binding of these proteins was increased in the complexes formed in the proximal and distal regions. VEGF expression in Leydig cells is regulated by gonadotropin via a cAMP-dependent mechanism, and the transcription factors Sp1 and Sp3 appear to be involved in the activation of the promoter. Progesterone also appears to play a role in the regulation of VEGF, acting presumably via a nonconventional receptor that remains to be characterized yet.


A writer's guide to education scholarship: Quantitative methodologies for medical education research (part 1).
Quantitative research is one of the many research methods used to help educators advance their understanding of questions in medical education. However, little research has been done on how to succeed in publishing in this area. We conducted a scoping review to identify key recommendations and reporting guidelines for quantitative educational research and scholarship. Medline, ERIC, and Google Scholar were searched for English-language articles published between 2006 and January 2016 using the search terms, "research design," "quantitative," "quantitative methods," and "medical education." A hand search was completed for additional references during the full-text review. Titles/abstracts were reviewed by two authors (BT, PC) and included if they focused on quantitative research in medical education and outlined reporting guidelines, or provided recommendations on conducting quantitative research. One hundred articles were reviewed in parallel with the first 30 used for calibration and the subsequent 70 to calculate Cohen's kappa coefficient. Two reviewers (BT, PC) conducted a full text review and extracted recommendations and reporting guidelines. A simple thematic analysis summarized the extracted recommendations. Sixty-one articles were reviewed in full, and 157 recommendations were extracted. The thematic analysis identified 86 items, 14 categories, and 3 themes. Fourteen quality evaluation tools and reporting guidelines were found. Discussion This paper provides guidance for junior researchers in the form of key quality markers and reporting guidelines. We hope that quantitative researchers in medical education will be informed by the results and that further work will be done to refine the list of recommendations.


Non-Contact, Simple Neonatal Monitoring by Photoplethysmography.
This paper presents non-contact vital sign monitoring in neonates, based on image processing, where a standard color camera captures the plethysmographic signal and the heart and breathing rates are processed and estimated online. It is important that the measurements are taken in a non-invasive manner, which is imperceptible to the patient. Currently, many methods have been proposed for non-contact measurement. However, to the best of the authors' knowledge, it has not been possible to identify methods with low computational costs and a high tolerance to artifacts. With the aim of improving contactless measurement results, the proposed method based on the computer vision technique is enhanced to overcome the mentioned drawbacks. The camera is attached to an incubator in the Neonatal Intensive Care Unit and a single area in the neonate's diaphragm is monitored. Several factors are considered in the stages of image acquisition, as well as in the plethysmographic signal formation, pre-filtering and filtering. The pre-filter step uses numerical analysis techniques to reduce the signal offset. The proposed method decouples the breath rate from the frequency of sinus arrhythmia. This separation makes it possible to analyze independently any cardiac and respiratory dysrhythmias. Nine newborns were monitored with our proposed method. A Bland-Altman analysis of the data shows a close correlation of the heart rates measured with the two approaches (correlation coefficient of 0.94 for heart rate (HR) and 0.86 for breath rate (BR)) with an uncertainty of 4.2 bpm for HR and 4.9 for BR (k = 1). The comparison of our method and another non-contact method considered as a standard independent component analysis (ICA) showed lower central processing unit (CPU) usage for our method (75% less CPU usage).


Construction of a High-Density Genetic Map and Analysis of Seed-Related Traits Using Specific Length Amplified Fragment Sequencing for Cucurbita maxima.
Seed traits are agronomically important for Cucurbita breeding, but the genes controlling seed size, seed weight and seed number have not been mapped in Cucurbita maxima (C. maxima). In this study, 100 F2 individual derived from two parental lines, "2013-12" and "9-6", were applied to construct a 3,376.87-cM genetic map containing 20 linkage groups (LGs) with an average genetic distance of 0.47 cM using a total of 8,406 specific length amplified fragment (SLAF) markers in C. maxima. Ten quantitative trait loci (QTLs) of seed width (SW), seed length (SL) and hundred-seed weight (HSW) were identified using the composite interval mapping (CIM) method. The QTLs affecting SW, SL and HSW explained a maximum of 38.6%, 28.9% and 17.2% of the phenotypic variation and were detected in LG6, LG6 and LG17, respectively. To validate these results, an additional 150 F2 individuals were used for QTL mapping of SW and SL with cleaved amplified polymorphic sequence (CAPS) markers. We found that two major QTLs, SL6-1 and SW6-1, could be detected in both SLAF-seq and CAPS markers in an overlapped region. Based on gene annotation and non-synonymous single-nucleotide polymorphisms (SNPs) in the major SWand SL-associated regions, we found that two genes encoding a VQ motif and an E3 ubiquitin-protein ligase may be candidate genes influencing SL, while an F-box and leucinerich repeat (LRR) domain-containing protein is the potential regulator for SW in C. maxima. This study provides the first high-density linkage map of C. maxima using SNPs developed by SLAF-seq technology, which is a powerful tool for associated mapping of important agronomic traits, map-based gene cloning and marker-assisted selection (MAS)-based breeding in C. maxima.


[Electrocardiographic changes in pancreatitis].
Of 400 patients with acute, chronic or chronic relapsing pancreatitis surveyed in the present study, only 54 had had ECG in their files. Among these, 80% showed ECG alterations, mostly sinus tachycardia and diffuse disturbances of ventricular repolarization. The causes of these alterations are, as yet controversial. Some explanations for these alterations are hypovolemia, sepsis and acute inflammatory state. Other important findings in the patients were bundle branch block, not encountered either before the pancreatic crisis or after its resolution, nor was dielectric effect and lesion current observed in either the acute and chronic forms. The possibility of the presence of previous cardiopathy in patients with high alcoholic intake, Chagas' disease, high blood pressure or diabetes, which are quite likely in these patients, should be recalled as important factors: marked electrolytes disorders were not frequent and did not correlate with ECG findings. The aim of this study is to highlight the importance of ECG during systematic search in the follow-up of patients with pancreatitis, in order to better understand associated cardiac disorders and to improve diagnosis, prevention and treatment.


EliXR-TIME: A Temporal Knowledge Representation for Clinical Research Eligibility Criteria.
Effective clinical text processing requires accurate extraction and representation of temporal expressions. Multiple temporal information extraction models were developed but a similar need for extracting temporal expressions in eligibility criteria (e.g., for eligibility determination) remains. We identified the temporal knowledge representation requirements of eligibility criteria by reviewing 100 temporal criteria. We developed EliXR-TIME, a frame-based representation designed to support semantic annotation for temporal expressions in eligibility criteria by reusing applicable classes from well-known clinical temporal knowledge representations. We used EliXR-TIME to analyze a training set of 50 new temporal eligibility criteria. We evaluated EliXR-TIME using an additional random sample of 20 eligibility criteria with temporal expressions that have no overlap with the training data, yielding 92.7% (76 / 82) inter-coder agreement on sentence chunking and 72% (72 / 100) agreement on semantic annotation. We conclude that this knowledge representation can facilitate semantic annotation of the temporal expressions in eligibility criteria.


The actions of serotonergic agonists and antagonists on the activity of brisk ganglion cells in the rabbit retina.
Extracellular electrophysiological recordings were obtained from rabbit retinal ganglion cells in either a superfused eyecup or an in vivo preparation. Selective antagonists or agonists of serotonin at the 5-HT2 or 5-HT1A receptors were applied, and the changes in light-evoked and spontaneous activity were studied. Both 5-HT1A agonists and 5-HT2 antagonists reduced the ON-components of the light-evoked responses of all classes of brisk ganglion cell; spontaneous activity was reduced in these cells as well. These results suggest that the indoleamine-accumulating amacrine cells of the rabbit retina serve to facilitate the output of the depolarizing rod bipolar cell and thereby increase the efficacy of transmission between this and other cells in the rabbit retina, and that this process is mediated by 5-HT2 receptors. On the basis of the similarity of the actions of the 2 classes of drug studied, we hypothesize further that 5-HT1A receptors mediate an inhibitory process that serves to terminate the indoleamine-induced facilitation. This process may be located either in the bipolar terminal or presynaptic to it in the terminal of the putative indoleaminergic cells.


[Dynamics of energy metabolism in ontogenesis of striped shield bug (Graphosoma lineatum L.) and cabbage moth (Mamestra brassicae L.)].
Dynamics of growth and oxygen consumption during ontogenesis of insects with direct (striped shield bug Graphosoma lineatum L.) and indirect (cabbage moth Mamestra brassicae L.) development have been compared. The correlation between a character of energy metabolism alteration and peculiarities of development of the insects has been shown. Cyclic decrease of oxygen consumption during molt and sharp dropping during metamorphosis have been observed in insects with indirect development. The decrease of oxygen consumption has been observed in insects with direct development only during molts. The coefficient a of allometric dependence of oxygen consumption on body weight of imago for cabbage moth was two times higher than that for striped shield bug.


Farm safety issues in Old Order Anabaptist communities: unique aspects and innovative intervention strategies.
The Amish and other Old Order Anabaptists have been inseparably linked with agriculture since coming to America. However few efforts have been identified which analyze the issues involved with Old Order Anabaptist farm injuries or present best practices for addressing these problems. As part of an effort to develop culturally appropriate and effective injury prevention strategies for use within the Old Order Anabaptist community, this article identifies important cultural issues that should be considered in understanding and attempting to reduce farm injures in this population, summarizes statistics concerning farm-related fatalities among Old Order Anabaptists, and discusses methods which have been effectively used to address farm injuries within these communities. In the statistical analysis, 92 cases were identified that were classified as being work-related or occurring in a farm work-related setting during the period 1980 to 2000. Approximately 64% of the identified cases were children 15 years and under; and approximately 77% were male. Run-over incidents were the most frequent primary cause of the fatal injuries, while animal-related behavior was the most frequent secondary cause, reflecting agricultural practices that remain highly dependent upon the use of horses and mules. The authors believe evidence suggests that certain Old Order Anabaptist choices concerning farm safety issues may be directly related to their socio-religious beliefs, and that effective intervention strategies must also be sensitive to socio-religious beliefs.


Effects of intermittent pneumatic calf compression in normal and postphlebitic legs.
The changes in common femoral vein flow produced by three different intermittent pneumatic compression devices were recorded with a Doppler velocity detector in 20 normal legs and 20 with postphlebitic syndrome. Mean and peak velocity increases were measured and expressed as a percent of resting baseline values. There was no significant difference in the peak velocity increase produced by the three devices in normals and abnormals; however, there were differences in the mean velocity increases. The devices worked as well on postphlebitic legs as on normal ones. Correct cuff application was more critical than indicated by the manufacturers, suggesting that some of the failures of intermittent pneumatic compression may have resulted from improper cuff placement. The results show that different designs of intermittent pneumatic compression equipment accelerate venous flow in the leg.


[The effect of "animal hypnosis" on the motor dominant created by the action of direct current on the cortex of the left hemisphere].
The state of "animal hypnosis" in rabbits was created by the DC current anode applied to the sensorimotor cortical area of the left hemisphere. It was impossible to elicit the "animal hypnosis" during the optimum of the dominant. The state of "animal hypnosis" could be easily elicited against the background of weak motor dominant on the next day after its creation. The "animal hypnosis" restored the "left" motor dominant after its extinction.


A Scoping Review of the Clinical Evidence for the Health Benefits of Culinary Doses of Herbs and Spices for the Prevention and Treatment of Metabolic Syndrome.
Metabolic syndrome (MetS) is a growing global health problem. Evidence suggests that diets rich in phytochemical-containing herbs and spices can contribute to reducing the risk of chronic diseases. This review assesses the scope of evidence supporting the use of herbs and spices in the diet for the prevention or treatment of MetS and its associated health conditions. A search of the PubMed, Scopus and Google Scholar databases was carried out to assess the available clinical evidence for culinary doses of commonly used herbs and spices. Trials that were measuring health factors related to metabolic disorders in healthy individuals, or the health of individuals with MetS or associated diseases, were included. Out of a total of 1738 papers identified, there were 142 relevant studies on black pepper, chilli, cardamom, cinnamon, coriander, cumin, fennel, fenugreek, garlic, ginger, nigella seed, rosemary, sage and turmeric. No relevant research was found for cloves, mint, oregano, parsley or thyme. Cinnamon, fenugreek and ginger were the herbs/spices with the most published trials on them and that showed promise for glycaemic control. Cardamom appears to have potential to reduce inflammatory markers, and cinnamon, ginger and turmeric to reduce blood lipids. Patients with type 2 diabetes were the population most likely to be included in studies, but the preventative benefits of herbs/spices in healthy populations were also investigated, particularly for chilli, ginger and cinnamon. There is evidence for the beneficial effect of culinary doses of many common herbs/spices in the prevention and treatment of MetS and associated disorders.


Purification and characterization of monomeric lysine decarboxylase from soybean (Glycine max) axes.
Lysine decarboxylase (EC 126.96.36.199) was purified 364-fold from 2-day-old soybean (Glycine max) axes. The enzyme was a monomeric protein having a molecular mass of 95,000 Da and an isoelectric point of 4.0. The K(m) for L-lysine was 1.17 mM. The optimal temperature and pH of the enzyme were 37 degrees C and 7.5, respectively. Storage of the enzyme at temperature ranging from 0 to 4 degrees C caused a 50% loss of the activity in 24 h. The enzyme was competitively inhibited by Cl- with a Ki value of 1.46 mM. However, the activity of the purified enzyme was not inhibited by F-, Br-, I-, H2PO4-, HPO4(2-), or SO4(2-). Cadaverine at 1 mM inhibited the enzyme activity by 35%.


Sprouty-related Ena/vasodilator-stimulated phosphoprotein homology 1-domain-containing protein (SPRED1), a tyrosine-protein phosphatase non-receptor type 11 (SHP2) substrate in the Ras/extracellular signal-regulated kinase (ERK) pathway.
SHP2 is a tyrosine phosphatase involved in the activation of the Ras/ERK signaling pathway downstream of a number of receptor tyrosine kinases. One of the proposed mechanisms involving SHP2 in this context is to dephosphorylate and inactivate inhibitors of the Ras/ERK pathway. Two protein families bearing a unique, common domain, Sprouty and SPRED proteins, are possible candidates because they have been reported to inhibit the Ras/ERK pathway upon FGF activation. We tested whether any of these proteins are likely substrates of SHP2. Our findings indicate that Sprouty2 binds to the C-terminal tail of SHP2, which is an unlikely substrate binding site, whereas SPRED proteins bind to the tyrosine phosphatase domain that is known to be the binding site for its substrates. Overexpressed SHP2 was able to dephosphorylate SPREDs but not Sprouty2. Finally, we found two tyrosine residues on SPRED1 that are required, when phosphorylated, to inhibit Ras/ERK activation and identified Tyr-420 as a specific dephosphorylation target of SHP2. The evidence obtained indicates that SPRED1 is a likely substrate of SHP2, whose tyrosine dephosphorylation is required to attenuate the inhibitory action of SPRED1 in the Ras/ERK pathway.


Colorectal cancer-specific cytochrome P450 2W1: intracellular localization, glycosylation, and catalytic activity.
Cytochrome P450 2W1 (CYP2W1) is expressed at high levels in colorectal cancer cells. Moreover, we have shown previously that a higher tumor expression is associated with less survival. In this study, we characterize post-translational modification, inverted endoplasmic reticulum (ER) topology, and catalytic activity of CYP2W1. The analysis of colorectal normal and cancer tissues and CYP2W1 overexpressing human embryonic kidney (HEK) 293 cells showed that a fraction of CYP2W1 is modified by N-glycosylation. Bioinformatic analysis identified Asn177 as the only possible glycosylation site of CYP2W1, which was supported by the inability of an N177A mutant to be glycosylated in HEK 293 cells. Analysis of the membrane topology indicated that unlike other cytochromes P450, CYP2W1 in HEK 293-transfected cells and in nontransfected Caco2TC7 and HepG2 cells is oriented toward the lumen of the ER, a topology making CYP2W1 available to the ER glycosylation machinery. Immunofluorescence microscopy and cell surface biotinylation experiments revealed approximately 8% of the CYP2W1 on the cell surface. Despite the reverse orientation of CYP2W1 in the ER membrane, apparently making functional interactions with NADPH-cytochrome P450 reductase impossible, CYP2W1 in HEK 293 cells was active in the metabolism of indoline substrates and was able to activate aflatoxin B1 into cytotoxic products. The study identifies for the first time a cytochrome P450 enzyme with a luminal ER orientation and still retaining catalytic activity. Together, these results suggest the possibility of using CYP2W1 as a drug target in the treatment of colon cancer using antibodies and/or specific CYP2W1 activated prodrugs.


Calcium hydroxide: study based on scientific evidences.
The characteristics of calcium hydroxide come from its dissociation into calcium and hydroxyl ions. The action of these ions on tissues and bacteria explains the biological and antimicrobial properties of this substance. Under the conditions of this retrospective of literature, based on scientific evidences, it is possible to state that: 1. Dentin is considered the best pulpal protective, and calcium hydroxide has proved, through numerous studies, its capability of inducing the formation of a mineralized bridge over pulpal tissue. 2. It is necessary, whenever possible, to provide time for calcium hydroxide paste to manifest its potential of action on the microorganisms present in endodontic infections. The maintenance of a high concentration of hydroxyl ions can change bacteria enzymatic activity and promote its inactivation. 3. The site of action of hydroxyl ions of calcium hydroxide includes the enzymes in the cytoplasmic membrane. This medication has a large scope of action, and therefore is effective on a wide range of microorganisms, regardless their metabolic capability. In microbial world, cytoplasmic membranes are similar, irrespective from microorganisms morphological, tinctorial and respiratory characteristics, which means that this medication has a similar effect on aerobic, anaerobic, Gram-positive and Gram-negative bacteria. 4. Calcium hydroxide as temporary dressing used between appointments promotes better results on the periapical healing process than the treatment in one appointment.


Nucleotide-free MalK drives the transition of the maltose transporter to the inward-facing conformation.
The complex MalFGK2 hydrolyzes ATP and alternates between inward- and outward-facing conformations during maltose transport. It has been shown that ATP promotes closure of MalK2 and opening of MalFG toward the periplasm. Yet, why the transporter rests in a conformation facing the cytosol in the absence of nucleotide and how it returns to this state after hydrolysis of ATP is unknown. The membrane domain MalFG may be naturally stable in the inward-facing conformation, or the ABC domain may catalyze the transition. We address this question by analyzing the conformation of MalFG in nanodiscs and in proteoliposomes. We find that MalFG alone exists in an intermediate state until MalK binds and converts the membrane domain to the inward-facing state. We also find that MalK, if overly-bound to MalFG, blocks the transition of the transporter, whereas suppressor mutations that weaken this association restore transport. MalK therefore exploits hydrolysis of ATP to reverse the conformation of MalFG to the inward-facing conformation, a step essential for release of maltose in the cytosol.


Graph-Based Predictive Modelling of Chronic Disease Development: Type 2 DM Case Study.
This study proposes a graph-based method for representing the dynamics of chronic diabetes as a complex process with different characteristics. The study was based on the case histories of 6864 patients with diabetes mellitus, 90% of whom suffer from type 2 diabetes. Our method allows to predict the sequence of events during the development of type 2 diabetes for each patient. Typical developmental trajectories of the disease were investigated, their clustering was carried out, the trajectory patterns were identified and studied. Based on the constructed directed graph reflecting transitions between different conditions of the patients, the clustering of diabetic statuses was carried out using the Modularity Class method; 8 clusters were selected, each of them was interpreted and studied. The method of the disease developmental trajectories creation by means of machine learning methods was described. Unlike static models of a disease course, this method considers complete past information on the patient and his or her previous events, using each event of the course of disease to predict the next event.


Safety of homeopathic injectables for subcutaneous administration: a documentation of the experience of prescribing practitioners.
The aim of this study was to chart the experiences of homeopathic injectables prescribing practitioners with regard to safety issues and the extent in which these practitioners would feel restricted in case subcutaneously administered homeopathics were banned. This was a survey among practitioners who prescribe homeopathic injectables in 12 European countries. Data were gathered from 1693 doctors experienced in the use of homeopathic injectables for subcutaneous use. The data are based on experience with an estimated 36 million patient contacts. Of the 1693 doctors 1594 (94.2%) choose subcutaneous administration because of its therapeutic effect. 96.4% of the doctors never, very rarely, or rarely, observed any adverse reactions because of the subcutaneous form of application. The reported adverse reactions were mostly harmless (local redness, hematoma, local pain). Of the doctors, 98.1% never, very rarely, or rarely observed any adverse reactions caused by the specific homeopathic medicinal product used. In addition, 99.5% of the doctors desire homeopathic injectables for subcutaneous use to stay on the market and 89% would be severely or very severely limited in their profession if homeopathic injectables were not available. The study suggests that homeopathic injectables have a very low risk profile. A very small number of severe adverse reactions (anaphylactic reaction, feverish symptoms, aversion/anxiety against injections, and asthma) have been reported with products with a concentration higher than 1:10,000.


Training Cardiac Surgeons: Safety and Requirements.
To analyze whether cardiac surgical residents can perform their first surgeries without compromising patients' safety or outcomes, by comparing their performance and results to those of senior surgeons. All documented CABGs conducted between 2002 and 2020 were included. Surgeries were divided according to the experience level of the main surgeon (defined by the number of CABG conducted by him/her) using the following thresholds: 1000; 150; 80 and 35. This resulted in 5 groups: senior surgeons (the reference group); attending surgeons; fellow surgeons; advanced residents and new residents. Primary endpoint was 30 day mortality. Secondary endpoints included a list of intra and post-operative parameters (including in-hospital complications). A multivariable analysis was conducted. 16,486 CABG were conducted by 66 different surgeons over a period of 18 years. Multivariable analysis did not find significant differences between both the primary and the secondary endpoints. Skin-to-skin time correlated significantly with experience level, as new residents needed almost 30% more time than senior surgeons (234 vs 180 minutes). With a suitable supervision by experienced surgeons, patient selection and sufficient resources (longer duration of surgery), surgical residents can perform CABGs with good results and without compromising the patient's outcome.


Addition of intravenous aminophylline to beta2-agonists in adults with acute asthma.
Aminophylline has been used extensively in acute asthma, but its role is unclear especially with respect to any additional benefit when added to beta2-agonists. To determine the magnitude of effect of the addition of intravenous aminophylline to beta2-agonists in adult patients with acute asthma treated in the emergency setting. Studies were identified from the following sources: The Cochrane Airways Group register (derived from MEDLINE, EMBASE, CINAHL standardised searches), hand searched respiratory journals and meeting abstracts. Potentially relevant articles were obtained, and their bibliographic lists were hand searched for additional articles. The search included searches of the database up to 1999. Randomised controlled trials comparing intravenous aminophylline versus placebo in adults with acute asthma and treated with beta-adrenergic agonists. Patients could be treated with or without corticosteroids or other bronchodilators. A total of 210 abstracts were identified. Two independent reviewers selected a total of 27 eligible studies for possible inclusion, in which quality assessment was performed and a third reviewer was used to adjudicate disagreements. Peak expiratory flow (PEFR) and forced expiratory volume in the first second (FEV1) data were extracted and entered in Review Manager from these studies. Information not obtained from the authors was estimated from graphs. All data were entered and double checked by two reviewers. Results are reported as weighted mean differences (WMD) or odds ratio (OR), both with 95% confidential intervals (CI). Fifteen trials were included. Overall, the quality of the studies was only moderate; concealment of allocation was assessed as clearly adequate in only seven (45%) of the trials. The doses of aminophylline and other medications and the severity of asthma varied between studies. There was no statistically significant effect of aminophylline on airflow outcomes at any time period. The aminophylline treated group had higher values of PEFR at 12 (PEFR 8 L/min or 2.3%) and 24 hours (PEFR 22 L/min or 6.4%), but these were not significant (p>0.05). Two subgroup analyses were performed by grouping studies according to mean baseline airflow limitation (n = 11 studies) and the use of any steroids (n = 9 studies). There was no relationship between baseline airflow limitation nor the use of steroids on the effect of aminophylline. Aminophylline treated patients reported more palpitations/arrhythmias (OR: 2.9; 95% CI: 1.5 to 5.7) and vomiting (OR: 4.2; 95% CI 2.4 to 7.4), but no difference was found in tremor or hospital admissions. In acute asthma, the use of intravenous aminophylline did not result in any additional bronchodilation compared to standard care with beta-agonists. The frequency of adverse effects was higher with aminophylline. No subgroups in which aminophylline might be more effective could be identified. These results should be added to consensus statements and guidelines.


The safety goals of the U.S. Nuclear Regulatory Commission.
In August 1986, after 6 years of effort, the U.S. Nuclear Regulatory Commission adopted a Policy Statement on safety goals for nuclear power reactors. The commission's qualitative goals state that individual members of the public should be provided a level of protection from the consequences of nuclear power plant operation such that they bear no significant additional risk to life and health, and societal risks to life and health from nuclear power should be comparable to or less than the risks of generating electricity by viable competing technologies and should not be a significant addition to other societal risks. The commission's safety goal Policy Statement also includes quantitative design objectives.


Mutations in short stature homeobox containing gene (SHOX) in dyschondrosteosis but not in hypochondroplasia.
Dyschondrosteosis (DCO) and hypochondroplasia (HCH) are common skeletal dysplasias characterized by disproportionate short stature. The diagnosis of these conditions might be difficult to establish especially in early childhood. Point mutations and deletions of the short stature homeobox containing gene (SHOX) are detected in DCO and idiopathic short stature with some rhizomelic body disproportion, whereas mutations in the fibroblast growth factor receptor 3 (FGFR3) gene are found in 40-70% of HCH cases. In this study, we performed mutational analysis of the coding region of the SHOX gene in five DCO and 18 HCH patients, all of whom tested negative for the known HCH-associated FGFR3 mutations. The polymorphic CA-repeat analysis, direct sequencing and Southern blotting were used for detection of deletions and point mutations. The auxological and radiological phenotype of these patients was carefully determined. Three novel mutations in DCO patients were found: (1) a deletion of one base (de1272G) (according to GenBank accession nos. Y11536, Y11535), resulting in a premature stop codon at position 75 of the amino acid sequence; (2) the transversion C485G resulting in the substitution Leu132Val; and (3) the transversion G549T causing an Arg153Leu substitution. These substitutions segregate with the DCO phenotype and affect evolutionarily conserved homeodomain residues, based on a comparison of homeobox containing proteins in 13 species. Moreover, these changes were not found in 80 unrelated, unaffected individuals. This strongly suggests that these mutations are pathogenic. The phenotype of our patients with DCO and HCH varied from mild to severe shortness and body disproportion. These results further support clinical and genetic heterogeneity of dyschondrosteosis and hypochondroplasia.


[Relationship between the 1st experience with the use of an information bank to optimize management of a territorial psychiatric service and the task of rehabilitating mental patients].
The paper concerns results of the use of an information bank for purposes of optimizing, from the standpoint of a total systems approach, the structure and activity of psychiatric services in the region and enhancing the effectivity of the rehabilitative programmes. The main principles of forming the bank of information, including computer analogues of case histories, the system of managing the bases and models of optimizing the activities of facilities and their subunits, are described. A systemic coordination of such models provided a complete continuity in the activities of hospitals, dispensaries and intermediate subunits. This circumstance allowed an implementation of successful outpatient treatment in a large group of patients with a protracted development of diseases and consequently to enhance the efficacy of the activities of mental health services in the region.


Fibrinogen gamma gene rs2066865 and risk of cancer-related venous thromboembolism.
Venous thromboembolism (VTE) is a frequent complication in patients with cancer. Homozygous carriers of the fibrinogen gamma gene (FGG) rs2066865 have a moderately increased risk of VTE, but the effect of the FGG variant in cancer is unknown. We aimed to investigate the effect of the FGG variant and active cancer on the risk of VTE. Cases with incident VTE (n=640) and a randomly selected age-weighted sub-cohort (n=3,734) were derived from a population-based cohort (the Tromsø study). Cox-regression was used to estimate hazard ratios (HR) with 95% confidence intervals (CI) for VTE according to categories of cancer and FGG In those without cancer, homozygosity at the FGG variant was associated with a 70% (HR 1.7, 95% CI: 1.2-2.3) increased risk of VTE compared to non-carriers. Cancer patients homozygous for the FGG variant had a two-fold (HR 2.0, 95% CI: 1.1-3.6) higher risk of VTE than cancer patients without the variant. Moreover, the six-months cumulative incidence of VTE among cancer patients was 6.4% (95% CI: 3.5-11.6) in homozygous carriers of FGG and 3.1% (95% CI: 2.3-4.7) in those without risk alleles. A synergistic effect was observed between rs2066865 and active cancer on the risk of VTE (synergy index: 1.81, 95% CI: 1.02-3.21, attributable proportion: 0.43, 95% CI: 0.11-0.74). In conclusion, homozygosity at the FGG variant and active cancer yielded a synergistic effect on the risk of VTE.


Detection of true pathologic stage I lung cancer in a screening program and the effect on survival.
One-hundred-sixty-nine lung cancers have occurred to date among 10,040 cigarette smoking men who participated in the New York Lung Cancer Detection Program. Almost 40% of the cases, 65, were still Stage I when their disease was diagnosed; 62 had thoracotomy and resection, and in 57, mediastinal node dissection confirmed that the mediastinum was free of metastases ("true pathologic" Stage I). Fifty-four of the 62 (87%) are still alive at this time, while only 15 of 104 (14%) of those with Stage II and III lung cancers are alive. Only two patients of the 62 in Stage I who were treated by resection died of lung cancer, both with T2 tumors. Two others are alive with metastases, one died postoperatively, and five died of other causes without evidence of lung cancer. The estimated probability of survival for true Stage I lung cancer is over 90% at five years, and close to 40% of all lung cancers can be detected in this favorable stage by present radiologic and cytologic screening techniques.


Characterization of cells transformed by the human polyomavirus JC virus.
One unique feature of the prototype JC virus (JCV) (Mad 1) genome is the occurrence of a second TATA sequence within the early promoter region. A naturally occurring oncogenic variant of JCV (Mad 4) lacks this second TATA box. Several cell lines transformed by Mad 1, Mad 4 and simian virus 40 were characterized, in part to investigate whether the second TATA sequence is functional. S1 nuclease mapping of early JCV gene transcription products revealed a major set of start sites common to both Mad 1 and Mad 4 mRNAs. In addition, a second set of early transcripts was found exclusively in Mad 1 transformants, presumably positioned by the second TATA box. The presence of these unique mRNAs in the Mad 1-transformed cells did not appear to have any bearing on the other parameters investigated, including size and quantity of early viral proteins, integration patterns of viral DNA and growth properties of the cells.


Measuring empowerment in patients with psoriasis: the Psoriasis Empowerment Enquiry in the Routine Practice (PEER) questionnaire.
While empowerment is an outcome pursued by advocacy groups and health care providers, no data are yet available about empowerment in patients with psoriasis in the literature. Our aim was to develop a tool for measuring levels of empowerment in psoriatic patients, and to assess its validity and reliability. A cross-sectional study among outpatients with psoriasis was carried out. Principal component analysis was performed to validate the questionnaire and to measure any associations with demographic and clinical data. The 12-item questionnaire (Psoriasis Empowerment Enquiry in the Routine practice, or PEER) showed acceptable consistency and reliability, and three distinct and interpretable factors have been identified: Knowledge, Experience, Skills. Patients older than 44 years and patients with psoriasis for more than one year showed statistically significantly higher levels of empowerment. Neither educational level nor the number of consultations influenced the global empowerment score (GES). A link between GES and prescribed therapy was found, suggesting that patients receiving systemic drugs have better knowledge, more insight and superior skills to manage their psoriasis. We conclude that PEER is an appropriate tool to assess empowerment among patients with psoriasis and may be used to evaluate the efficacy of educational interventions. Further studies are needed to assess its convergent validity and to improve its reliability.


Neuropharmacologic management of behavior sequelae in head injury: a case report.
A patient with traumatic head injury was confused, agitated, and belligerent. Initial treatment with sedatives (diazepam) and antipsychotics (haloperidol and thioridazine hydrochloride) did not diminish this irritability or destructive behavior. A single case experimental methodology was adopted to evaluate the behavioral response to other sequentially introduced neuroactive drugs. Methylphenidate produced increased agitation. No response was noted to propranolol. Lithium, however, was associated with significant calming of the patient, successful completion of rehabilitation, and discharge to home where the patient safely cared for himself while his wife worked.


Carbon disulfide neurotoxicity in rats: IV. Increased mRNA expression of low-affinity nerve growth factor receptor--a sensitive and early indicator of PNS damage.
Expression of the low-affinity nerve growth factor receptor (NGF-R) in the peripheral nervous system is regulated by Schwann cell-axonal contact. Steady-state mRNA levels for NGF-R are very low in the mature peripheral nervous system, but are markedly upregulated in sciatic nerve during both primary demyelination (tellurium exposure) and secondary demyelination (Wallerian degeneration). Upregulation also occurs in various subdegenerative axonopathy models where there is axonal atrophy, suggesting its usefulness as a marker for subtle perturbations in normal axon-Schwann cell interactions (Roberson et al., Mol Brain Res 1995; 28:231-238). To further test this hypothesis, we examined NGF-R mRNA expression in sciatic nerves of rats exposed to carbon disulfide (CS2), a toxicant known to cause a distal axonopathy. Adult rats were exposed to CS2 gas (50, 500, or 800 ppm, 6 hr/day, 5 days/wk) for 2-13 weeks. RNA was isolated from sciatic nerves and levels of mRNA for NGF-R determined by Northern blot analysis. NGF-R mRNA expression increased in a dose- and time-dependent manner. Message levels were already increased after 2 wks of exposure to 800 ppm CS2, and increased further with continued exposure. Morphological alterations were not apparent in the sciatic nerve, even at the highest dosage levels with the longest exposure times. Upregulation of NGF-R mRNA is thus an indicator of subtle alterations in the normal axon-Schwann cell relationship and provides a sensitive measure of CS2 neurotoxicity. Assay of this marker may also be useful as a rapid and very sensitive general screen for other compounds which are potentially toxic to the peripheral nervous system.


A web-based positive-affect intervention to reduce stress and improve well-being in women living with HIV - feasibility and acceptability of a single-arm, pilot study.
Women living with HIV (WLWH) experience high rates of depression but are underrepresented in mental health research. Positive emotions are associated with beneficial health outcomes in WLWH and should be a targeted component of psychological interventions in this population. Positive psychological interventions aim to increase positive emotions through the use of simple exercises, such as keeping a gratitude journal. We conducted a single-arm feasibility/acceptability study of a five-week, self-guided, web-based positive affect skills intervention in a sample of WLWH (N = 23) who also participate in a longitudinal observational study, the Women's Interagency HIV Study. The intervention was feasible as measured by home practice and post-intervention assessment completion, and acceptable as measured by exit interview feedback regarding recommendation of the program to friends or others living with HIV. On average, participants completed home practice for about 8 out of 9 skills. The mean response for recommendation of the program to a friend was 9.26/10 (SD = 1.63) and the mean response for recommendation of the program to others living with HIV was 9.68/10 (SD = 0.82). Participant feedback will be used to adapt and enhance the delivery of this intervention. Further studies are needed to assess efficacy and impact on psychological outcomes.


A quantitative assessment of the role of the parasite Amoebophrya in the termination of Alexandrium fundyense blooms within a small coastal embayment.
Parasitic dinoflagellates of the genus Amoebophrya infect free-living dinoflagellates, some of which can cause harmful algal blooms (HABs). High prevalence of Amoebophrya spp. has been linked to the decline of some HABs in marine systems. The objective of this study was to evaluate the impact of Amoebophrya spp. on the dynamics of dinoflagellate blooms in Salt Pond (MA, USA), particularly the harmful species Alexandrium fundyense. The abundance of Amoebophrya life stages was estimated 3-7 days per week through the full duration of an annual A. fundyense bloom using fluorescence in situ hybridization coupled with tyramide signal amplification (FISH-TSA). More than 20 potential hosts were recorded including Dinophysis spp., Protoperidinium spp. and Gonyaulax spp., but the only dinoflagellate cells infected by Amoebophrya spp. during the sampling period were A. fundyense. Maximum A. fundyense concentration co-occurred with an increase of infected hosts, followed by a massive release of Amoebophrya dinospores in the water column. On average, Amoebophrya spp. infected and killed ∼30% of the A. fundyense population per day in the end phase of the bloom. The decline of the host A. fundyense population coincided with a dramatic life-cycle transition from vegetative division to sexual fusion. This transition occurred after maximum infected host concentrations and before peak infection percentages were observed, suggesting that most A. fundyense escaped parasite infection through sexual fusion. The results of this work highlight the importance of high frequency sampling of both parasite and host populations to accurately assess the impact of parasites on natural plankton assemblages.


Double mating: its use to study heritable factors in dental caries.
When Osborne-Mendel female rats (white) were mated with both an Osborne-Mendel and an NIH Black rat male during the same breeding period, litters were born which contained both Osborne-Mendel (white) and crossbred (grey to black) offspring. The Osborne-Mendel and crossbred animals developed widely different levels of caries activity even though they were exposed to identical environmental conditions during the intrauterine, preweaning, and experimental periods. These findings are indicative of a strong heritable influence on the development of dental caries.


The combination of tetracyclines effectively ameliorates liver fibrosis via inhibition of EphB1/2.
Eph receptor tyrosine kinase EphB1/2 contributes to the development of liver fibrosis, suggesting the rationale that EphB1/2 inhibitors may be effective in liver fibrosis therapy. Since tetracycline antibiotics were recently demonstrated as EphB kinase inhibitors, in present study we investigated their therapeutic potential against liver fibrosis. Our results showed that the tetracycline combination of demeclocycline (D), chlortetracycline (C), and minocycline (M) inhibited the activation of hepatic stellate cells (HSCs) in vitro and alleviated CCl4-induced animal model of liver fibrosis in vivo. Mechanistically, DCM combination inhibited EphB1/2 phosphorylation and subsequent activation of the MAPK signaling. Moreover, we found that short-term and low-dose DCM combination treatment decreased tissue inflammation and improved liver fibrosis in mice. Thus, our study indicates that tetracyclines may be repurposed for the treatment of liver fibrosis.


Childhood cancer incidence and trends in Minnesota, 1988-1994.
Childhood cancer incidence patterns for Minnesota, obtained from the Minnesota Cancer Surveillance System, were compared with national rates as well as with historic data from eight Minnesota counties. In total, 1,140 neoplasms were diagnosed in children (ages 0 to 14) between 1988 and 1994. Leukemias were the most common diagnosis for boys (30.3%) and girls (29.6%), followed by central nervous system tumors. The average annual age-adjusted incidence rates for all cancer sites were 167.2 and 136.2 per million for boys and girls, respectively. These rates were somewhat higher than national rates. In particular, the incidence rate for astrocytoma in boys was significantly elevated. Childhood cancer incidence, particularly brain tumors, has increased in the eight-county region from 1969 to 1994. This analysis demonstrated the Minnesota's childhood cancer incidence patterns are similar to national patterns.


Gluten sensitivity: new epidemic or new myth? Every major change in our diet carries with it the possibility of unforeseen risks.
Fueled in part by recent best-selling books that warn of the evils of gluten in our diets, a significant proportion of our population is now either avoiding foods that contain gluten or eliminating gluten entirely from their diets, and these numbers continue to grow. The gluten-free trend-and the accompanying multibillion dollar industry it has created-stems from the spreading belief that eating foods containing wheat or other gluten-laden grains may not only result in weight gain and obesity but can also lead to a laundry list of ailments ranging from depression and anxiety to arthritis and autism. One popular book contends that current recommendations for a high-grain, low-fat diet underlie much of today's chronic health problems and that a low-carbohydrate, high-fat/cholesterol diet is ideal. Every major change in our diet carries with it the possibility of unforeseen risks. Concern about the impact of such popularized dietary recommendations on overall well-being-and on cardiovascular health in particular-warrants discussion in the medical community.


Probing methane in air with a midinfrared frequency comb source.
We employed a midinfrared frequency comb source for methane detection in ambient air. The transmitted spectra over a bandwidth of about 500 nm were recorded with an optical spectrum analyzer under various experimental conditions of different path lengths. The normalized absorption spectra were compared and fitted with simulations, yielding quantitative values of concentrations of methane and water vapor in the ambient air. The 3σ detection limit was ∼6.6×10-7 cm-1 in ambient air for a broad spectral range, achieved with a path length of ∼590 m. This approach provides a broad spectral range, a large dynamic range, high sensitivity, and accurate calibration. The performed analysis of the residuals shows that an excellent agreement between the measured and calculated spectral profiles was obtained.


Assessing the feasibility of using uniaxial accelerometers with an online support platform in the delivery of a community-based exercise referral scheme.
Exercise referral schemes are established within community-based health care; however, they have been criticized for failing to evidence long-term behavior change relative to usual care. As such, recent reviews have called for refinement of their delivery with a focus on embedded strategies targeting client motivation. This research letter presents findings from an initial pilot trial conducted within Wales' National Exercise Referral Scheme (NERS), examining the feasibility of using validated physical activity monitoring devices and an accompanying online platform within standard scheme delivery. 30 individuals referred to generic or cardiovascular pathways were offered the system; of these 17 agreed to participate. Common reasons for declining were clustered into lack of technology literacy or access, condition severity, or fear of costs associated with losing the device. Analysis of follow-up interviews after 4 weeks of use indicated that while participants found the monitoring devices practical and informative, only a minority (n = 4) were using the system in full. Crucially, the system element most aligned with contemporary theories of motivation (the online portal) was not used as expected. In addition, feedback from exercise referral professionals indicated that there were demands for support from clients, which might be mitigated by more effective independent system use. Recommendations for larger scale trials using similar systems include consideration of targeted patient groups, equity of access, and providing adequate technological support that is currently beyond the capacity of the NERS system.


Collapse of the ear canal during masking for bone conduction.
Although it is well recognized that collapse of the external meatus can mimic a conductive hearing loss in the test ear, attention must also be directed to the masked ear during masked bone conduction threshold testing. Failure to recognize collapse of the masked ear may result in a false air-bone gap in the test ear. Two such cases are presented and the mechanism analyzed. Suggestions as to identification and resolution of this problem are discussed.


Simultaneous thermal and optical imaging of two-phase flow in a micro-model.
In the study of non-equilibrium heat transfer in multiphase flow in porous media, parameters and constitutive relations, like heat transfer coefficients between phases, are unknown. In order to study the temperature development of a relatively hot invading immiscible non-wetting fluid and, ultimately, approximate heat transfer coefficients, a transparent micro-model is used as an artificial porous medium. In the last few decades, micro-models have become popular experimental tools for two-phase flow studies. In this work, the design of an innovative, elongated, PDMS (polydimethylsiloxane) micro-model with dimensions of 14.4 × 39 mm(2) and a constant depth of 100 microns is described. A novel setup for simultaneous thermal and optical imaging of flow through the micro-model is presented. This is the first time that a closed flow cell like a micro-model is used in simultaneous thermal and optical flow imaging. The micro-model is visualized by a novel setup that allowed us to monitor and record the distribution of fluids throughout the length of the micro-model continuously and also record the thermal signature of the fluids. Dynamic drainage and imbibition experiments were conducted in order to obtain information about the heat exchange between the phases. In this paper the setup as well as analysis and qualitative results are presented.


Crosstalk between lysine-specific demethylase 1 (LSD1) and histone deacetylases mediates antineoplastic efficacy of HDAC inhibitors in human breast cancer cells.
Our previous studies demonstrated that lysine-specific demethylase 1 (LSD1) and histone deacetylases (HDACs) closely interact in controlling growth of breast cancer cells. However, the underlying mechanisms are largely unknown. In this study, we showed that knockdown of LSD1 expression (LSD1-KD) by RNAi decreased mRNA levels of HDAC isozymes in triple-negative breast cancer (TNBC) cells. Small interfering RNA (siRNA)-mediated depletion of HDAC5 expression induced the most significant accumulation of H3K4me2, a specific substrate of LSD1. Combined treatment with LSD1 inhibitor, pargyline, and HDAC inhibitor, SAHA (Vorinostat), led to superior growth inhibition and apoptotic death in TNBC cells, but exhibited additive or antagonistic effect on growth inhibition in non-TNBC counterparts or non-tumorigenic breast cells. Additionally, LSD1-KD enhanced SAHA-induced reexpression of a subset of aberrantly silenced genes, such as NR4A1, PCDH1, RGS16, BIK, and E-cadherin whose reexpression may be tumor suppressive. Genome-wide microarray study in MDA-MB-231 cells identified a group of tumor suppressor genes whose expression was induced by SAHA and significantly enhanced by LSD1-KD. We also showed that concurrent depletion of RGS16 by siRNA reduced overall cytotoxicity of SAHA and blocked the reexpression of E-cadherin, CDKN1C and ING1 in LSD1-deficient MDA-MB-231 cells. Furthermore, cotreatment with RGS16 siRNA reversed the downregulation of nuclear factor-kappaB expression induced by combined inhibition of LSD1 and HDACs, suggesting a crucial role of RGS16 in controlling key pathways of cell death in response to combination therapy. Taken together, these results provide novel mechanistic insight into the breast cancer subtype-dependent role of LSD1 in mediating HDAC activity and therapeutic efficacy of HDAC inhibitor.


Thioridazine cures extensively drug-resistant tuberculosis (XDR-TB) and the need for global trials is now!
Thioridazine (TDZ) has been shown to have in vitro activity against multidrug-resistant (MDR) and extensively drug-resistant (XDR) strains of Mycobacterium tuberculosis, to promote the killing of intracellular MDR and XDR strains and to cure the mouse of antibiotic-susceptible and -resistant pulmonary tuberculosis (TB) infections. Recently, TDZ was used to cure 10 of 12 XDR-TB patients in Buenos Aires, Argentina. At the time of writing, it is being used for the therapy of non-antibiotic-responsive terminal XDR-TB patients in Mumbai, India, on the basis of compassionate therapy and although it is too early to determine a cure, the patients have improved appetite, weight gain, are afebrile and free of night sweats, and their radiological picture shows great improvement. Because XDR-TB is essentially a terminal disease in many areas of the world and no new effective agents have yet to yield successful clinical trials, global clinical trials for the therapy of XDR-TB are urgently required.


Maximizing wound healing with silver-impregnated porcine xenograft.
1. Grossly contaminated or chronically infected wounds require adequate debridement of dead tissue and wound coverage or closure. 2. Porcine xenograft is a temporary wound covering that prevents evaporation and thereby creates an environment for new cells to propagate instead of being sacrificed by desiccation. 3. The addition of silver ions to the porcine xenograft provides a potent antimicrobial agent that is only active at the surface where it is needed, has no systemic effects, and allows epithelialization to proceed in half the time of uncovered or unsilvered methods. 4.Silver-impregnated porcine xenograft provides a temporary wound covering that is antimicrobial, physiologic, and allows even the most massive and chronically contaminated wounds of years duration to be healed by careful, consistent dressing changes without skin grafts or flaps.


Generation of dendritic cells from peripheral blood adherent cells in medium with human serum.
Dendritic cells (DC) provide an effective pathway for presenting antigens to T cells, both self-antigens during T-cell development and foreign antigens during immunity. As such, these cells may be promising adjuvants for immunotherapy. Thus, it is important to establish simple and fast method(s) to generate sufficient numbers of human DC in medium free of calf serum so that the cells can be used for both experimental and clinical purposes. In this report, we used peripheral blood adherent cells, without laborious cell purification or depletion, as the starting population and cultured them in medium supplemented with granulocyte/macrophage colony-stimulating factor and interleukin-4. Substantial numbers of cells with the phenotypical and functional characteristics of immature DC were obtained in a 7-day culture. We then compared DC cultured in medium supplemented with either fetal calf serum or pooled human ABRh+ serum and found no difference in cell yields and in their ability to stimulate alloreactive T cells or to present soluble antigens to T cells. Irradiated cells were less efficient than non-irradiated cells in antigen presentation and stimulation of T cells. Finally, we have examined DC with or without additional tumour necrosis factor-alpha treatment and found that antigen-pulsed mature cells could as efficiently present antigen to T cells as did immature cells. This method is suitable for the generation of DC in studies of large clinical materials.


DNA-based identification of a hepatic trematode in an elk calf.
Liver fluke infection was identified as a probable cause of clinical disease in an approximately 6-mo-old elk (Cervus elaphus) in coastal Oregon. Clinical pathology and necropsy findings are described. The alcohol-fixed flukes that were submitted for identification were similar in size to Fasciola hepatica, but their shape resembled Fascioloides magna in that they lacked a distinctive anterior cone. A few structures consistent with the eggs of F. magna were observed in liver lesions, suggesting that at least some of the worms were sexually maturing. Due to difficulties in morphologic identification associated with improper fixation technique, DNA analysis was used to compare small subunit (SSU) and internal transcribed spacer 2 ribosomal RNA gene sequences of the recovered parasites with those of F. hepatica and F. magna, confirming these small, but sexually mature flukes were F. magna. This is the first publication of the SSU gene sequence for F. magna. Phylogenetic analysis showed that it is related to, but is an outlier, to the genus Fasciola. Due to the high mortality rate associated with this disease outbreak, the overall significance of trematodiasis in the herd is unclear.


Morphology of initial lesions of enamel treated with different commercial dentifrices using a pH cycling model: scanning electron microscopy observations.
The aim of this study was to evaluate and compare the 'calcium fluoride-like reaction products' formed, both on the surface and in the subsurface caries-like lesions of enamel, from different commercial dentifrices manufactured locally from developed and developing countries. The experiment was conducted by using the in vitro single-section technique under the pH-cycling system used for 10 days. The tested dentifrices include; Maxam DFP Maxam Tartar Controls, First, Tianqui Medicated (China); Vicco Vajradenti (India); Colgate MFP2. Crest Tartar Control (USA); and a non-fluoride dentifrice, Jie Yin (China) used as control. The surface and subsurface of the lesions, before and after treatment, were examined using scanning electron microscopy. Varying degrees of fine globular patterns of calcium fluoride-like material were observed on the enamel surfaces of five out of eight groups and in the subsurface of four groups. Two treatment groups showed a 'similar appearance' compared with the control and untreated groups. This study suggests and supports that, when compared with 'multinational dentifrices', some Chinese and Indian dentifrices manufactured locally failed to induce the typical morphological appearance of globules as seen with fluoride dentifrices.


Is the development of desalination compatible with sustainable development of the Arabian Gulf?
The development of desalination has been essential to the rapid economic development of the countries bordering the Arabian Gulf. The current production capacity of sea water desalination plants drawing water from Gulf is over 20 million m3 day-1, which may rise to 80 million m3 day-1 by 2050. Whilst supporting aspects of sustainable development related to water and sanitation, desalination impacts the marine environment through impingement and entrainment of organisms in intakes, and through thermal, brine and chemical discharges. This may compromise other objectives for sustainable development related to sustainable use of the oceans. Under business as usual scenarios, by 2050, the impact of individual desalination plants will combine causing a regional scale impact. Without mitigating actions to avoid the business as usual scenario, by 2050, desalination in combination with climate change, will elevate coastal water temperatures across more than 50% of the Gulf by at least 3 °C, and a volume of water equivalent to more than a third of the total volume of water between 0 and 10 m deep will pass through desalination plants each year. This will adversely impact the coastal ecosystem of the Gulf, with impacts on biodiversity, fisheries and coastal communities and may cause potential loss of species and habitats from the Gulf. Given the significant implications of these preliminary findings, and in light of the precautionary approach to management, it is recommended that mitigating options addressing behavioural, regulatory and technological change are rapidly evaluated and implemented to avoid the development of desalination in the region along a business as usual pathway, and multidisciplinary research studies should be conducted to reduce uncertainty in predictions of future impacts.


Treatment of depression in patients with epilepsy.
In this article, we review the current best evidence for the treatment of depression in patients with epilepsy. Depression is a common epilepsy comorbidity, but it is often unrecognized. The most important step in appropriately managing mood disorders in this population is making the diagnosis. Clinical vigilance and routine use of a validated screening tool can improve detection and quality of care. As is increasingly the case for the general population, persons with epilepsy are often interested in exploring alternative therapies for chronic conditions, including depression. Unfortunately, the benefit of complementary and alternative therapies for depression currently is largely unproven for persons with a seizure history, although an early study of exercise for mild depression has shown some benefit. Concerns about drug interactions, side effects, and expense may be barriers to the prescription of antidepressant medications for people requiring chronic antiepileptic drug (AED) therapy. For this reason, use of an AED with mood-stabilizing properties has appeal and may be appropriate for selected individuals with mild depressive symptoms. Undue fear of lowering seizure threshold should not preclude the prescription of an antidepressant medication, as the perceived risks are often overestimated and rarely outweigh the risk of leaving depression untreated. At present, the best evidence for efficacy and safety support the use of citalopram, sertraline, or mirtazapine as initial pharmacotherapy, whereas bupropion should be avoided. Start low, go slow, and use the lowest effective dose. Cognitive behavioral therapy is a valuable adjunct to antidepressant therapy in this population. For people with refractory partial epilepsy and refractory depression, vagus nerve stimulation has some appeal, in that it may be beneficial for both conditions, but the efficacy of vagus nerve stimulation in improving mood in patients with epilepsy remains unclear.


Statistical analysis of global connectivity and activity distributions in cellular networks.
Various molecular interaction networks have been claimed to follow power-law decay for their global connectivity distribution. It has been proposed that there may be underlying generative models that explain this heavy-tailed behavior by self-reinforcement processes such as classical or hierarchical scale-free network models. Here, we analyze a comprehensive data set of protein-protein and transcriptional regulatory interaction networks in yeast, an Escherichia coli metabolic network, and gene activity profiles for different metabolic states in both organisms. We show that in all cases the networks have a heavy-tailed distribution, but most of them present significant differences from a power-law model according to a stringent statistical test. Those few data sets that have a statistically significant fit with a power-law model follow other distributions equally well. Thus, while our analysis supports that both global connectivity interaction networks and activity distributions are heavy-tailed, they are not generally described by any specific distribution model, leaving space for further inferences on generative models. Supplementary Material is available online at www.liebertonline.com.


Mild traumatic brain injury impacts associations between limbic system microstructure and post-traumatic stress disorder symptomatology.
Post-traumatic stress disorder (PTSD) is a psychiatric disorder that afflicts many individuals, yet the neuropathological mechanisms that contribute to this disorder remain to be fully determined. Moreover, it is unclear how exposure to mild traumatic brain injury (mTBI), a condition that is often comorbid with PTSD, particularly among military personnel, affects the clinical and neurological presentation of PTSD. To address these issues, the present study explores relationships between PTSD symptom severity and the microstructure of limbic and paralimbic gray matter brain regions, as well as the impact of mTBI comorbidity on these relationships. Structural and diffusion MRI data were acquired from 102 male veterans who were diagnosed with current PTSD. Diffusion data were analyzed with free-water imaging to quantify average CSF-corrected fractional anisotropy (FA) and mean diffusivity (MD) in 18 limbic and paralimbic gray matter regions. Associations between PTSD symptom severity and regional average dMRI measures were examined with repeated measures linear mixed models. Associations were studied separately in veterans with PTSD only, and in veterans with PTSD and a history of military mTBI. Analyses revealed that in the PTSD only cohort, more severe symptoms were associated with higher FA in the right amygdala-hippocampus complex, lower FA in the right cingulate cortex, and lower MD in the left medial orbitofrontal cortex. In the PTSD and mTBI cohort, more severe PTSD symptoms were associated with higher FA bilaterally in the amygdala-hippocampus complex, with higher FA bilaterally in the nucleus accumbens, with lower FA bilaterally in the cingulate cortex, and with higher MD in the right amygdala-hippocampus complex. These findings suggest that the microstructure of limbic and paralimbic brain regions may influence PTSD symptomatology. Further, given the additional associations observed between microstructure and symptom severity in veterans with head trauma, we speculate that mTBI may exacerbate the impact of brain microstructure on PTSD symptoms, especially within regions of the brain known to be vulnerable to chronic stress. A heightened sensitivity to the microstructural environment of the brain could partially explain why individuals with PTSD and mTBI comorbidity experience more severe symptoms and poorer illness prognoses than those without a history of brain injury. The relevance of these microstructural findings to the conceptualization of PTSD as being a disorder of stress-induced neuronal connectivity loss is discussed.


HAMA: a multiplexed LC-MS/MS assay for specificity profiling of adenylate-forming enzymes.
Adenylation enzymes selecting substrates for ribosomal and nonribosomal protein and peptide biosynthesis have been popular targets of enzyme engineering. Previous standard assays for adenylation specificity have been cumbersome and failed to reflect the competition conditions inside a cell because they measure substrates one at a time. We have developed an adenylation assay based on hydroxamate quenching and LC-MS/MS detection of hydroxamate products testing dozens of competing amino acid substrates in parallel. Streamlined specificity profiling of adenylation enzymes will facilitate engineering and directed evolution of ribosomal and nonribosomal peptide synthesis.


PhenoPlasm: a database of disruption phenotypes for malaria parasite genes.
Two decades after the first Plasmodium transfection, attempts have been made to disrupt more than 3,151 genes in malaria parasites, across five Plasmodium species. While results from rodent malaria transfections have been curated and systematised, empowering large-scale analysis, phenotypic data from human malaria parasite transfections currently exists as individual reports scattered across a the literature. To facilitate systematic analysis of published experimental genetic data across Plasmodium species, we have built PhenoPlasm ( http://www.phenoplasm.org), a database of phenotypes generated by transfection experiments in all Plasmodium parasites. The site provides a simple interface linking citation-backed Plasmodium reverse-genetic phenotypes to gene IDs. The database has been populated with phenotypic data on 367 P. falciparum genes, curated from 176 individual publications, as well as existing data on rodent Plasmodium species from RMgmDB and PlasmoGEM. This is the first time that all available data on P. falciparum transfection experiments has been brought together in a single place. These data are presented using ortholog mapping to allow a researcher interested in a gene in one species to see results across other Plasmodium species. The collaborative nature of the database enables any researcher to add new phenotypes as they are discovered. As an example of database utility, we use the currently available datasets to identify RAP (RNA-binding domain abundant in Apicomplexa)-domain containing proteins as crucial to parasite survival.


Block of T lymphocyte differentiation by activation of the cAMP-dependent signal transduction pathway.
Differentiation of T lymphocytes is a complex and finely tuned process. Here we show that treatment of mouse fetal thymus organ cultures with agents activating the cAMP-dependent signalling pathway results in the block of thymocyte differentiation. This is due to severe impairment of maturation beyond the CD4-/CD8- stage. In addition, rearrangements at the TCR alpha gene locus, but not at the TCR beta locus, are completely inhibited. The cAMP effect is reversible and is restricted to TCR alpha beta+ cells. cAMP acts both by triggering apoptosis and by inducing cell-cycle block in thymocytes. Thus, activation of the cAMP pathway provides a mechanism to modulate thymic function for hormones and ligands whose receptors are coupled to adenylate cyclase.


Insulin production and resistance in cystic fibrosis: effect of age, disease activity, and genotype.
To assess the major determinants of glucose tolerance between age, genotype, and clinical status in cystic fibrosis (CF) patients, and study if defects of insulin secretion and insulin sensitivity were associated with the onset of CF-related diabetes (CFRD). One hundred and nineteen patients, in stable clinical condition were studied. They were subdivided into 3 groups based on age, and 2 groups based on Schwachman-Kulczycki clinical score. All patients were genotyped, and subsequently divided into 3 groups. Ninety-four healthy normal-weight controls, comparable for sex and age were also studied. All subjects had baseline blood samples taken for glucose and insulin, C-peptide, and glycated hemoglobin. Homeostasis model assessment of insulin resistance (HOMA-IR), fasting glucose/insulin ratio (FGIR) were calculated as indices of IR and insulinogenic index as a marker of pancreatic β-cell function. All patients underwent an oral glucose tolerance test, and 57 underwent an IVGTT for the calculation of first-phase (FPIR) and acute insulin responses (AIR). The F508del homozygous patients had an increased chance of developing impaired glucose tolerance (IGT) and significantly lower FPIR, decreased HOMA-IR, and insulinogenic index. Heterozygote F508del patients had an increased chance of having normal glucose tolerance. HOMA-IR, FGIR, and insulinogenic index did not change with age or clinical score. HOMAIR correlated with FPIR. FPIR correlated positively with insulinogenic index. AIR correlated negatively with FGIR, and positively with C-reactive protein. In multiple linear regression analyses, glucose tolerance was related to the agegroup, and to the HOMA-IR and insulinogenic indexes. IGT and CFRD were related mainly to genotype, although, as expected, the prevalence increased with age. The data suggested a possible combined contribution of insulin deficiency, β-cell function, and reduced insulin sensitivity to the onset of CFRD; however, further studies are warranted to better elucidate this aspect.


Transforaminal approach in lumbar disc herniations: transforaminal microdiscectomy (TFMD) technique.
The endoscopic foraminal approach in foraminal, lateral and far-lateral disc hernias, is a contemporary minimal invasive approach. This study was performed to show that the approach is possible with using the microscope without an endoscope, and even intervention on the discs within the spinal canal is possible by having access through the foramen. Forty-two cases with disc hernias in the medial of the pedicle were included in this study. Surgeries were performed with the transforaminal approach and microsurgically. Extraforaminal disc hernias were not included in the study. Access was made through the Kambin's triangle. The foramen was enlarged and the spinal canal was entered. The procedure took 65 minutes on average, and the mean bleeding amount was about 100 ml. Patients were mobilized within the same day postoperatively. No permenant complications were seen. Follow-up periods ranged between 5 and 84 months and the mean followup period was 30.2 months. Transforaminal microdiscectomy is a method that can be performed in any clinic with standard neurosurgical instruments. It does not require additional equipment or a high amount of investment. Application of transforaminal inter-body fusion is possible when required.


Effective Magnetic MOFs Adsorbent for the Removal of Bisphenol A, Tetracycline, Congo Red and Methylene Blue Pollutions.
A magnetic metal-organic frameworks adsorbent (Fe3O4@MIL-53(Al)) was prepared by a typical solvothermal method for the removal of bisphenol A (BPA), tetracycline (TC), congo red (CR), and methylene blue (MB). The prepared Fe3O4@MIL-53(Al) composite adsorbent was well characterized by scanning electron microscope (SEM), transmission electron microscope (TEM), X-ray diffraction (XRD), and fourier transform infrared spectrometer (FTIR). The influence of adsorbent quantity, adsorption time, pH and ionic strength on the adsorption of the mentioned pollutants were also studied by a UV/Vis spectrophotometer. The adsorption capacities were found to be 160.9 mg/g for BPA, 47.8 mg/g for TC, 234.4 mg/g for CR, 70.8 mg/g for MB, respectively, which is superior to the other reported adsorbents. The adsorption of BPA, TC, and CR were well-fitted by the Langmuir adsorption isotherm model, while MB followed the Freundlich model, while the adsorption kinetics data of all pollutants followed the pseudo-second-order kinetic models. The thermodynamic values, including the enthalpy change (ΔH°), the Gibbs free energy change (ΔG°), and entropy change (ΔS°), showed that the adsorption processes were spontaneous and exothermic entropy-reduction process for BPA, but spontaneous and endothermic entropy-increasing processes for the others. The Fe3O4@MIL-53(Al) was also found to be easily separated after external magnetic field, can be a potential candidate for future water treatment.


[Calculation of lavage volume in transurethral prostate resection (TURP)].
Evaluation of the relationship between ethanol concentration in expired air and the amount of absorbed irrigation fluid in patients who undergo a transurethral resection of the prostate. 76 patients were evaluated in a prospective study. The irrigation fluid contained 1.92 w/w% of ethanol. The ethanol concentration in breath was measured at least every 10 minutes with a Alcotest 7410 (Dräger) or a Biomed 3010 (Biotest). Documented values: breath ethanol concentration, mean arterial pressure, heart rate, haemoglobin concentration, serum sodium concentration, the amount of irrigation fluid, the time of operation, special events, therapeutic interventions. A modification of the Widmark formula was used for calculating the amount of absorbed irrigation fluid from the breath ethanol concentration. A rise of the ethanol level above 0.2/1000 was observed in 10 patients. No clinically significant difference in heart rate and blood pressure during surgery between the 24 patients with no alcohol in breath and the 10 patients with more than 0.2/1000 was observed. Prolonged duration of resection and greater amount of used irrigation fluid correlated with higher ethanol concentration in breath. During the study period no patient developed the clinical features of a transurethral resection syndrome. If 2 w/w% of ethanol is added to the irrigation fluid, breath ethanol concentration under 0.2/1000 may be considered harmless, while values above 0.2/1000 are predictors of massive absorption of irrigation fluid. In our patients, the addition of ethanol to the irrigation fluid has been a suitable and easy way to monitor the absorption of irrigation fluid, if the ethanol concentration in the breath is measured every ten minutes. If 2 w/w% of ethanol have been added to the irrigation fluid, the absorbed volume (E) can be estimated using the formula: E = 37.5 x body weight (kg) x ethanol concentration (/1000).


The La autoantigen contains a dimerization domain that is essential for enhancing translation.
The La autoantigen is an RNA-binding protein that is involved in initiation and termination of RNA polymerase III transcription. It also binds several viral RNAs, including those of poliovirus and human immunodeficiency virus (HIV). Binding of the La protein to these RNAs enhances their translation in vitro (K. Meerovitch, Y.V. Svitkin, H.S. Lee, F. Lejbkowicz, D.J. Kenan, E.K.L. Chan, V.L. Agol, J.D. Keene, and N. Sonenberg, J. Virol. 67:3798-3807, 1993, and Y.V. Svitkin, A. Pause, and N. Sonenberg, J. Virol. 68:7001-7007, 1994). Here, a functional domain in the carboxy-terminal half of La that is distinct from the RNA-binding domain is described. Deletion of this domain abrogated the ability of La protein to enhance translation of poliovirus RNA and a hybrid HIV trans-activation-response element-chloramphenicol acetyltransferase mRNA. Far-Western assays indicated that the La protein homodimerized in vitro, and the C-terminal deletions that caused a loss of activity in translation also abrogated the dimerization signal. Gel filtration chromatography of recombinant La protein confirmed that La protein exists as a dimer under native conditions. Addition of the purified dimerization domain resulted in a loss of translation stimulatory activity of La protein in cell-free-translation reactions.


A host-specific virulence protein of Erwinia herbicola pv. gypsophilae is translocated into human epithelial cells by the Type III secretion system of enteropathogenic Escherichia coli.
summary HsvG is a virulence factor that determines the host specificity of Erwinia herbicola pathovars gypsophilae and betae on gypsophila. We used the calmodulin adenylate cyclase reporter (CyaA) to demonstrate that HsvG is secreted and translocated into HeLa cells by the type III secretion system (TTSS) of the enteropathogenic Escherichia coli (EPEC). A fusion of HsvG-CyaA containing 271 amino acids of the N-terminus of HsvG were introduced into a wild-type EPEC, espB mutant deficient in translocation and an escV mutant deficient in secretion. A significant secretion was detected in EPEC/HsvG-CyaA and its espB mutant, but not with the escV mutant. Translocation was only observed with the wild-type EPEC, and not with the other two mutants. To localize the secretion and translocation signals of HsvG, fusions containing 39, 11 and 3 amino acids of the N-terminus of HsvG were constructed and expressed in EPEC. A fusion containing the first 39 N-terminal amino acids of HsvG was secreted and translocated at significant level (31-35%) as compared to the original fusion. In contrast, fusions containing the 3 and 11 amino acids failed to be secreted and translocated.


Pathogenic variants of AIPL1, MERTK, GUCY2D, and FOXE3 in Pakistani families with clinically heterogeneous eye diseases.
Significant number out of 2.2 billion vision impairments in the world can be attributed to genetics. The current study is aimed to decipher the genetic basis of Leber congenital Amaurosis (LCA), Anterior Segment dysgenesis (ASD), and Retinitis Pigmentosa (RP), segregating in four large consanguineous Pakistani families. The exome sequencing followed by segregation analysis via Sanger sequencing revealed the LCA phenotypes segregating in families GCUF01 and GCUF04 can be attributed to c.465G>T (p.(Gln155His)) missense and novel c.139_140delinsA p.(Pro47Trhfster38) frameshift variant of AIPL1 and GUCY2D, respectively. The c.1843A>T (p.(Lys615*) truncating allele of MERTK is homozygous in all the affected individuals, presumably suffering with RP, of the GCUF02 family. Meanwhile, co-segregation of the ASD phenotype and the c.289A>G (p.(Ile97Val)) variant of FOXE3 was found in the GCUF06 family. All the identified variants were either absent or present in very low frequencies in the control databases. Our in-silico analyses and 3D molecular modeling support the deleterious impact of these variants on the encoded proteins. Variants identified in MERTK, GUCY2D, and FOXE3 were categorized as "pathogenic" or "likely pathogenic", while the missense variant found in AIPL1 was deemed to have "uncertain significance" based upon the variant pathogenicity guidelines from the American College of Medical Genetics and Genomics (ACMG). This paper highlights the genetic diversity of vision disorders in the Pakistani population and reports the identification of four novel mutations in families who segregate clinically heterogeneous eye diseases. Our results give insight into the genotype-phenotype correlations of AIPL1, FOXE3, MERTK, and GUCY2D variants.


Lung ultrasound in infants with bronchiolitis.
Lung ultrasound (LUS) is nowadays a fast-growing field of study since the technique has been widely acknowledged as a cost-effective, radiation free, and ready available alternative to standard X-ray imaging. However, despite extensive acoustic characterization studies and documented medical evidences, a lot is still unknown about how ultrasounds interact with lung tissue. One of the most discussed lung artifacts are the B-lines [in all ages] and the subpleural consolidations (in young infants). Recently, LUS has been claimed to be able to detect pneumonia in infants with bronchiolitis, although this can be an overestimation due to the peculiar physiology of small peripheral airways of the pediatric lung (particularly in neonate/infants). Distinguishing consolidations from atelectasis in young infants with bronchiolitis can be challenging and those criteria well defined for adults and older children (size and bronchogram) cannot easily translated in this specific subset. Therefore, if decades of studies clearly defined the low risk of SBI in bronchiolitis, we need to be careful before stating that LUS may confirm pneumonia in such a high number of cases and, importantly, new and promising techniques such as LUS should give us new insights bringing us to improvements and not back to overuse of antibiotics. More studies are surely need on this topic.


Broadband room temperature strong coupling between quantum dots and metamaterials.
Herein, we report the first demonstration of room temperature enhanced light-matter coupling in the visible regime for metamaterials using cooperative coupled quasi two dimensional quantum dot assemblies located at precise distances from the hyperbolic metamaterial (HMM) templates. The non-monotonic variation of the magnitude of strong coupling, manifested in terms of strong splitting of the photoluminescence of quantum dots, can be explained in terms of enhanced LDOS near the surface of such metamaterials as well as the plasmon mediated super-radiance of closely spaced quantum dots (QDs). Our methodology of enhancing broadband, room temperature, light-matter coupling in the visible regime for metamaterials opens up new possibilities of utilising these materials for a wide range of applications including QD based thresholdless nanolasers and novel metamaterial based integrated photonic devices.