Ambient air pollution and respiratory emergency department visits.
A number of emergency department studies have corroborated findings from mortality and hospital admission studies regarding an association of ambient air pollution and respiratory outcomes. More refined assessment has been limited by study size and available air quality data. Measurements of 5 pollutants (particulate matter [PM10], ozone, nitrogen dioxide [NO2], carbon monoxide [CO], and sulfur dioxide [SO2]) were available for the entire study period (1 January 1993 to 31 August 2000); detailed measurements of particulate matter were available for 25 months. We obtained data on 4 million emergency department visits from 31 hospitals in Atlanta. Visits for asthma, chronic obstructive pulmonary disease, upper respiratory infection, and pneumonia were assessed in relation to air pollutants using Poisson generalized estimating equations. In single-pollutant models examining 3-day moving averages of pollutants (lags 0, 1, and 2): standard deviation increases of ozone, NO2, CO, and PM10 were associated with 1-3% increases in URI visits; a 2 microg/m increase of PM2.5 organic carbon was associated with a 3% increase in pneumonia visits; and standard deviation increases of NO2 and CO were associated with 2-3% increases in chronic obstructive pulmonary disease visits. Positive associations persisted beyond 3 days for several of the outcomes, and over a week for asthma. The results of this study contribute to the evidence of an association of several correlated gaseous and particulate pollutants, including ozone, NO2, CO, PM, and organic carbon, with specific respiratory conditions.


'LABNOTE', a laboratory notebook system designed for academic genomics groups.
We have developed a relational laboratory database system, adapted to the daily book-keeping needs of laboratories that must keep track of information acquired on hundreds or thousands of clones in an effective and user-friendly fashion. Data, whether final or related to experiments in progress, can be accessed in many different ways, e.g. by clone name, by gene, by experiment or through DNA sequence. Updating, import and export of results is made easier by specially developed tools. This system, in network version, serves several groups in our Institute and (over the Internet) elsewhere, and is instrumental in collaborative studies based on expression profiling. It can be used in many similar situations involving progressiveaccumulation of information on sets of clones or related objects.


Probiotics and Prebiotics: Present Status and Future Perspectives on Metabolic Disorders.
Metabolic disorders, including type 2 diabetes (T2DM) and cardiovascular disease (CVD), present an increasing public health concern and can significantly undermine an individual's quality of life. The relative risk of CVD, the primary cause of death in T2DM patients, is two to four times higher in people with T2DM compared with those who are non-diabetic. The prevalence of metabolic disorders has been associated with dynamic changes in dietary macronutrient intake and lifestyle changes over recent decades. Recently, the scientific community has considered alteration in gut microbiota composition to constitute one of the most probable factors in the development of metabolic disorders. The altered gut microbiota composition is strongly conducive to increased adiposity, β-cell dysfunction, metabolic endotoxemia, systemic inflammation, and oxidative stress. Probiotics and prebiotics can ameliorate T2DM and CVD through improvement of gut microbiota, which in turn leads to insulin-signaling stimulation and cholesterol-lowering effects. We analyze the currently available data to ascertain further potential benefits and limitations of probiotics and prebiotics in the treatment of metabolic disorders, including T2DM, CVD, and other disease (obesity). The current paper explores the relevant contemporary scientific literature to assist in the derivation of a general perspective of this broad area.


Nasal cancer in the textile and clothing industries.
A case-control study of 160 patients with cancers of the nasal cavity and paranasal sinuses and 290 controls showed an excess risk associated with employment in the textile or clothing industries, with the increase (relative risk [RR] = 2.1) found only among female workers. There was, however, no increasing trend in risk with years of employment or duration since first exposure. Both male and female workers were at an increased risk of adenocarcinoma (RR = 2.5), with further enhancement of risks for those experiencing dusty work conditions. Although aetiological inferences cannot be drawn from this study, the finding of raised risk supports some previous observations and the need for further investigation of the cancer experienced by textile and clothing workers.


The circadian clock regulator Bmal1 affects traumatic brain injury in rats through the p38 MAPK signalling pathway.
Traumatic brain injury (TBI) is still one of the main causes of death and disability worldwide. Bmal1 (brain and muscle Arnt-like protein-1) is the most central factor of the circadian rhythms that control life and cells. Studies have shown that Bmal1 is involved in inflammation, oxidative stress, vasodilation, glucose and lipid metabolism. This study explored the effect of Bmal1 on secondary brain injury after TBI in rats and the possible mechanism. We established a rat model of TBI induced by the free fall of a weight in rats. The Western blotting and immunofluorescence results showed that the Bmal1 levels decreased in the cerebral cortex after TBI, especially at 48 h. The effects of Bmal1 levels on rats after TBI were evaluated by brain oedema measurement, adhesive removal tests, behavioural tests, and TUNEL and FJC staining. We found that the recombinant Bmal1 protein increased Bmal1 levels after TBI and reduced brain oedema, neurobehavioural injury, somatosensory disturbances, and nerve cell necrosis and apoptosis. The ELISA results showed that Bmal1 overexpression could reduce the inflammatory factors IL-4 and TNF-α after TBI. In contrast, inhibiting Bmal1 expression had the opposite effect. The changes in Bmal1 levels were closely related to the phosphorylation of p38 MAPK after TBI. In conclusion, a decrease in Bmal1 after TBI may exacerbate pathological symptoms in vivo by activating p38 MAPK phosphorylation.


Depletion of Lubricant from Nanostructured Oil-Infused Surfaces by Pendant Condensate Droplets.
Due to recent advances in nanofabrication, phase-change condensation heat transfer has seen a renaissance. Compared to conventional heat transfer surfaces, nanostructured surfaces impregnated with chemically matched lubrication films (hereinafter referred to as "nanostructured lubricated surfaces") have been demonstrated to improve vapor-side phase-change condensation heat transfer by facilitating droplet nucleation, growth, and departure. While the presence of nanoscale roughness improves performance longevity by stabilizing the lubrication film via capillary forces, such enhancement is short-lived due to the eventual loss of lubrication oil by the departing droplets. The objective of this study is to characterize oil depletion caused by pendant droplets during condensation. For our study, we nanostructured, chemically functionalized, and lubricated horizontal copper tubes that are widely used in shell-and-tube heat exchangers in power plants and process industries. Using high-speed fluorescence imaging and thermogravimetric analysis, we show that shedding droplets exert a shear force on the oil in the wetting ridge at the water-oil interface. The viscous shear draws the lubrication film from the nanostructured surface onto the upper portion of the droplet and forms a ring-like oil skirt. Through detailed theoretical analysis, we show that the thickness of this oil skirt scales with the classical Landau-Levich-Derjaguin (LLD) theory for dip-coating. Our results reveal that droplets falling from horizontal tubes break unequally and leave behind small satellite droplets that retain the bulk of the oil in the wetting ridge. This observation is in stark contrast with the earlier description of droplets shedding from tilted flat plates where the entire oil-filled wetting ridge is demonstrated to leave the surface upon droplet departure. By selecting lubrication oils of varying viscosity and spreading coefficient, we provide evidence that the contribution of the wrapping layer to the rate of oil depletion is insignificant. Furthermore, we show that due to the nanoscale features on the tubes, nearly half of the lubrication film remains on the surface after 10 h of continuous steam condensation at ambient pressure, 23 °C, and 60% relative humidity, a 2-3-fold improvement over previous results.The insights gained from this work will provide guidelines for the rational design of long-lasting nanostructured lubricated surfaces for phase-change condensation.


Antinociceptive and anti-inflammatory activities of limonin isolated from the fruits of Evodia rutaecarpa var. bodinieri.
Antinociceptive and anti-inflammatory effects of limonin isolated from the dried fruits of Evodia rutaecarpa var. bodinieri were investigated. Oral administration of 30 or 100 mg/kg limonin significantly decreased the frequency of licking and biting behavior within a unit of time at the late phase without affecting that of the early phase in the formalin test. Limonin inhibited the rise of vascular permeability induced by acetic acid and the increase of paw edema induced by carrageenin. Limonin also showed inhibitory effects on bradykinin-induced paw edema and arachidonic acid-induced ear swelling. These results suggest that limonin possesses an antinociceptive effect and the effect may be accompanied by an anti-inflammatory action, and that the antinociceptive activity in Evodiae Fructus is partially attributable to limonin.


Interactions Between Silver Nanoparticles/Silver Ions and Liposomes: Evaluation of the Potential Passive Diffusion of Silver and Effects of Speciation.
Silver nanoparticles, used mainly for their antibacterial properties, are among the most common manufactured nanomaterials. How they interact with aquatic organisms, especially how they cross biological membranes, remains uncertain. Free Ag+ ions, released from these nanoparticles, are known to play an important role in their overall bioavailability. In this project, we have studied the uptake of dissolved and nanoparticulate silver by liposomes. These unilamellar vesicles, composed of phospholipids, have long been used as models for natural biological membranes, notably to study the potential uptake of solutes by passive diffusion through the phospholipid bilayer. The liposomes were synthesized using extrusion techniques and were exposed over time to dissolved silver under different conditions where Ag+, AgS2O3-, or AgCl0 were the dominant species. Similar experiments were conducted with the complexes HgCl 2 0 and Cd(DDC) 2 0 , both of which are hydrophobic and known to diffuse passively through biological membranes. The uptake kinetics of Ag+, HgCl 2 0 , and Cd(DDC) 2 0 show no increase in internalized concentrations over time, unlike AgS2O3- and AgCl0, which appear to pass through the phospholipid bilayer. These results are in contradiction with our initial hypothesis that lipophilic Hg and Cd complexes would be able to cross the membrane, whereas silver would not. Encapsulated tritiated water inside the liposomes was shown to rapidly diffuse through the lipid bilayer, suggesting a high permeability. We hypothesize that monovalent anions or complexes as well as small neutral complexes with a strong dipole can diffuse through our model membrane. Finally, liposomes were exposed to 5-nm polyvinylpyrrolidone-coated silver nanoparticles over time. No significant uptake of nanoparticulate silver was observed. Neither disruption of the membrane nor invagination of nanoparticles into the liposomes was observed. This suggests that the main risk caused by AgNPs for nonendocytotic biological cells would be the elevation of the free silver concentration near the membrane surface due to adsorption of AgNPs and subsequent oxidation/dissolution.


Controlled aggregation of adenine by sugars: physicochemical studies, molecular modelling simulations of sugar-aromatic CH-pi stacking interactions, and biological significance.
CH-Pi stacking interactions between carbohydrates and aromatic compounds play a central role in biomolecular recognition, especially in lectin-sugar and protein-glycolipid systems. In the present study, we have measured the solubility of the sparingly soluble aromatic base adenine in presence of various saccharides as an approach to investigate the interaction between adenine and sugars. Above 82.5 mM, adenine solutions gradually formed a crystalline precipitate which could be quantified by spectrophotometric turbidity measurements. Precipitation of adenine was increased by salts (NaCl and NaF) whereas it was prevented by DMSO, in agreement with the involvement of hydrophobic interactions (pi-pi stacking) in the vertical stacking of adenine molecules. Several monosaccharides and disaccharides were found to increase adenine solubility, with the following order: D-galactose = D-lactose > D-sucrose > D-glucose = D-maltose > D-ribose > D-fructose. Molecular mechanics simulations indicated that the potent cosolvent effect of beta-D-galactopyranose was probably mediated by CH-pi stacking interactions between its apolar surface and the aromatic structure of adenine. The polar OH groups of the sugars interacted with surrounding water molecules, ensuring the solubility of sugar-adenine complexes. In contrast, beta-D-fructofuranose, which has two polar faces, did not stack onto adenine and had a weak cosolvent effect. CH-pi stacking interactions were also demonstrated between 6-methylpurine and the sugar head group of glycolipids (glucosyl-, galactosyl- and lactosylceramide) but not with the charged head group of phosphatidylinositol-4,5-diphosphate. These data indicate that galactose-containing molecules have a high stacking propensity for aromatic compounds such as adenine, due to the specific structure of the galactose cycle.


Primary treatment for cleft lip and/or cleft palate in children in Japan.
To investigate current trends in primary treatment for children with cleft lip and/or cleft palate in Japan. Nationwide, retrospective study under the direction of the Academic Survey Committee of the Japanese Cleft Palate Association based on analysis of data obtained via a booklet-style questionnaire completed by institutions providing primary treatment for cleft lip and/or palate patients. PARTICIPANTS, PATIENTS: Patients were 4349 children undergoing primary repair for cleft lip and/or palate at 107 participating institutions between 1996 and 2000. Cleft type, laterality; use of infant palatal plate; and timing and technique of primary repair for cleft lip and/or palate were evaluated by cleft surgeons at 107 participating institutions. Of a total of 2874 patients with cleft lip and palate or cleft palate only, infant palatal plates were used with 1087 (37.8%) and were not used with 1787 (62.2%). Primary unilateral lip repair was performed at the age of 2 to 6 months in more than 90% of patients. Bilateral cleft lip was treated by one-stage repair in 285 patients (44.5%) and by two-stage repair in 258 (40.2%). Primary one-stage palatal repair was performed in 2212 (76.9%) and two-stage palatal repair in 262 (9.1%) cleft palate patients. Information on treatment of the remaining 400 (14%) patients was unavailable. This investigation clarified current trends in primary treatment for cleft lip and/or palate in Japan. The results suggest the need for an increase in regional core hospitals and greater variation in treatment options.


IL-22-Induced Ubiquitin-Specific Protease 15 Promotes Proliferation and Inflammation of Keratinocytes through Stabilization of Squamous Cell Carcinoma Antigen 2.
Ubiquitin-specific protease 15 (USP15) plays a significant role in regulating various biological processes in several autoimmune diseases and cancers. However, its role in psoriatic keratinocytes (KCs) has not been extensively studied. In this study, we described that USP15 promotes proliferation and inflammation in KCs by stabilizing squamous cell carcinoma antigen 2. We discovered that the expression of USP15 and squamous cell carcinoma antigen 2 was elevated in lesions from patients with clinical psoriasis and an imiquimod-induced psoriatic dermatitis mouse model. USP15 was able to bind, deubiquitinate, and stabilize squamous cell carcinoma antigen 2. Knocking down USP15 resulted in reduced KC inflammation and impaired KC viability and clonogenicity. Topically applying USP15 small interfering RNA significantly ameliorated imiquimod-induced psoriatic dermatitis and reduced the infiltration of T cells and neutrophils. In addition, we determined that IL-22 was a key cytokine that upregulated the expression of USP15. These findings provide insights regarding the mechanisms involved in the proliferation and inflammation of KCs mediated by IL-22, suggesting a potential IL-22-USP15-squamous cell carcinoma antigen 2 axis in the pathogenesis of psoriatic KCs.


Symptoms of Posttraumatic Stress During the COVID-19 Pandemic in the Governmental Public Health Workforce and General Population.
We aimed to estimate the prevalence of COVID-19-related posttraumatic stress symptoms (PTSS) in the governmental public health workforce and in US adults, assess differences in reporting PTSS within subgroups, and evaluate whether frontline workers reported higher levels of PTSS than persons in other jobs. We used data from 2 nationally representative studies: the 2021 Public Health Workforce Interests and Needs Survey (PH WINS) and the COVID-19 and Life Stressors Impact on Mental Health and Well-being (CLIMB) study. Our study found that the state and local governmental public health workforce was more likely to report PTSS than the general adult population. Almost a quarter of public health agency employees (24.7%) and 21.1% of adults reported at least 3 symptoms of posttraumatic stress. Differences in levels of PTSS appeared within demographic groups for both samples. Personal care and service frontline workers had 4.3 times the odds of reporting symptoms of posttraumatic stress than non-frontline workers.


How GRAIL controls Treg function to maintain self-tolerance.
Regulatory T cells (Tregs) normally maintain self-tolerance. Tregs recognize "self" such that when they are not working properly, such as in autoimmunity, the immune system can attack and destroy one's own tissues. Current therapies for autoimmunity rely on relatively ineffective and too often toxic therapies to "treat" the destructive inflammation. Restoring defective endogenous immune regulation (self-tolerance) would represent a paradigm shift in the therapy of these diseases. One recent approach to restore self-tolerance is to use "low dose IL-2" as a therapy to increase the number of circulating Tregs. However, studies to-date have not demonstrated that low-dose IL-2 therapy can restore concomitant Treg function, and phase 2 studies in low dose IL-2 treated patients with autoimmune diseases have failed to demonstrate significant clinical benefit. We hypothesize that the defect in self-tolerance seen in autoimmunity is not due to an insufficient number of available Tregs, but rather, due to defects in second messengers downstream of the IL-2R that normally control Treg function and stability. Previous studies from our lab and others have demonstrated that GRAIL (a ubiquitin E3 ligase) is important in Treg function. GRAIL expression is markedly diminished in Tregs from patients with autoimmune diseases and allergic asthma and is also diminished in Tregs of mice that are considered autoimmune prone. In the relevant pathway in Tregs, GRAIL normally blocks cullin ring ligase activity, which inhibits IL-2R desensitization in Tregs and consequently promotes Treg function. As a result of this defect in GRAIL expression, the Tregs of patients with autoimmune diseases and allergic asthma degrade IL-2R-associated pJAK1 following activation with low dose IL-2, and thus cannot maintain pSTAT5 expression. pSTAT5 controls the transcription of genes required for Treg function. Additionally, the GRAIL-mediated defect may also allow the degradation of the mTOR inhibitor, DEP domain-containing mTOR interacting protein (Deptor). This can lead to IL-2R activation of mTOR and loss of Treg stability in autoimmune patients. Using a monoclonal antibody to the remnant di-glycine tag on ubiquitinated proteins after trypsin digestion, we identified a protein that was ubiquitinated by GRAIL that is important in Treg function, cullin5. Our data demonstrate that GRAIL acts a negative regulator of IL-2R desensitization by ubiquitinating a lysine on cullin5 that must be neddylated to allow cullin5 cullin ring ligase activity. We hypothesize that a neddylation inhibitor in combination with low dose IL-2 activation could be used to substitute for GRAIL and restore Treg function and stability in the Tregs of autoimmune and allergic asthma patients. However, the neddylation activating enzyme inhibitors (NAEi) are toxic when given systemically. By generating a protein drug conjugate (PDC) consisting of a NAEi bound, via cleavable linkers, to a fusion protein of murine IL-2 (to target the drug to Tregs), we were able to use 1000-fold less of the neddylation inhibitor drug than the amount required for therapeutically effective systemic delivery. The PDC was effective in blocking the onset or the progression of disease in several mouse models of autoimmunity (type 1 diabetes, systemic lupus erythematosus, and multiple sclerosis) and a mouse model of allergic asthma in the absence of detectable toxicity. This PDC strategy represents targeted drug delivery at its best where the defect causing the disease was identified, a drug was designed and developed to correct the defect, and the drug was targeted and delivered only to cells that needed it, maximizing safety and efficacy.


Evidence for horizontal gene transfer from bacteroidetes bacteria to dinoflagellate minicircles.
Dinoflagellate protists harbor a characteristic peridinin-containing plastid that evolved from a red or haptophyte alga. In contrast to typical plastids that have ∼100-200 kb circular genomes, the dinoflagellate plastid genome is composed of minicircles that each encode 0-5 genes. It is commonly assumed that dinoflagellate minicircles are derived from a standard plastid genome through drastic reduction and fragmentation. However, we demonstrate that the ycf16 and ycf24 genes (encoded on the Ceratium AF490364 minicircle), as well as rpl28 and rpl33 (encoded on the Pyrocystis AF490367 minicircle), are related to sequences from Algoriphagus and/or Cytophaga bacteria belonging to the Bacteroidetes clade. Moreover, we identified a new open reading frame on the Pyrocystis minicircle encoding a SRP54 N domain, which is typical of FtsY proteins. Because neither of these minicircles share sequence similarity with any other dinoflagellate minicircles, and their genes resemble bacterial operons, we propose that these Ceratium and Pyrocystis minicircles resulted from a horizontal gene transfer (HGT) from a Bacteroidetes donor. Our findings are the first indication of HGT to dinoflagellate minicircles, highlighting yet another peculiar aspect of this plastid genome.


Tracking changes in behavioural dynamics using prediction error.
Automated analysis of video can now generate extensive time series of pose and motion in freely-moving organisms. This requires new quantitative tools to characterise behavioural dynamics. For the model roundworm Caenorhabditis elegans, body pose can be accurately quantified from video as coordinates in a single low-dimensional space. We focus on this well-established case as an illustrative example and propose a method to reveal subtle variations in behaviour at high time resolution. Our data-driven method, based on empirical dynamic modeling, quantifies behavioural change as prediction error with respect to a time-delay-embedded 'attractor' of behavioural dynamics. Because this attractor is constructed from a user-specified reference data set, the approach can be tailored to specific behaviours of interest at the individual or group level. We validate the approach by detecting small changes in the movement dynamics of C. elegans at the initiation and completion of delta turns. We then examine an escape response initiated by an aversive stimulus and find that the method can track return to baseline behaviour in individual worms and reveal variations in the escape response between worms. We suggest that this general approach-defining dynamic behaviours using reference attractors and quantifying dynamic changes using prediction error-may be of broad interest and relevance to behavioural researchers working with video-derived time series.


Integration of dosimetric parameters, clinical factors, and radiomics to predict symptomatic radiation pneumonitis in lung cancer patients undergoing combined immunotherapy and radiotherapy.
This study aimed to combine clinical/dosimetric factors and handcrafted/deep learning radiomic features to establish a predictive model for symptomatic (grade ≥2) radiation pneumonitis (RP) in lung cancer patients who received immunotherapy followed by radiotherapy. This study retrospectively collected data of 73 lung cancer patients with prior receipt of ICIs who underwent thoracic radiotherapy (TRT). Of these 73 patients, 41 (56.2%) developed symptomatic grade ≥2 RP. RP was defined per multidisciplinary clinician consensus using CTCAE v5.0. Regions of interest (ROIs) (from radiotherapy planning CT images) utilized herein were gross tumor volume (GTV), planning tumor volume (PTV), and PTV-GTV. Clinical/dosimetric (mean lung dose and V5-V30) parameters were collected, and 107 handcrafted radiomic (HCR) features were extracted from each ROI. Deep learning-based radiomic (DLR) features were also extracted based on pre-trained 3D residual network models. HCR models, Fusion HCR model, Fusion HCR + ResNet models, and Fusion HCR+ ResNet + Clinical models were built and compared using the receiver operating characteristic (ROC) curve with measurement of the area under the curve (AUC). Five-fold cross-validation was performed to avoid model overfitting. HCR models across various ROIs and the Fusion HCR model showed good predictive ability with AUCs from 0.740-0.808 and 0.740-0.802 in the training and testing cohorts, respectively. The addition of DLR features improved the effectiveness of HCR models (AUCs from 0.826-0.898 and 0.821-0.898 in both respective cohorts). The best performing prediction model (HCR+ ResNet + Clinical) combined HCR & DLR features with 7 clinical/dosimetric characteristics and achieved an average AUC of 0.936 and 0.946 in both respective cohorts. In patients undergoing combined immunotherapy/RT for lung cancer, integrating clinical/dosimetric factors and handcrafted/deep learning radiomic features can offer a high predictive capacity for RP, and merits further prospective validation.


Polypharmacy in the treatment of people diagnosed with borderline personality disorder: repeated cross-sectional study using New Zealand's national databases.
There is insufficient evidence to support the pharmacological treatment of borderline personality disorder. However, previous out-patient cohorts have described high rates of polypharmacy in this group. So far, there have been no national studies that have considered polypharmacy in borderline personality disorder. To describe psychotropic polypharmacy in people with borderline personality disorder in New Zealand. New Zealand's national databases have been used to link psychotropic medication dispensing data and diagnostic data for borderline personality disorder. Annual dispensing data for 2014 and 2019 have been compared. Fifty percent of people with borderline personality disorder who were dispensed medications had three or more psychotropic medications in 2014. This increased to 55.9% in 2019 (P < 0.001). Those on seven or more psychotropics increased from 8.4 to 10.7% (P < 0.023). Quetiapine was the most dispensed psychotropic medication, being given to 53.8% of people dispensed medication with borderline personality disorder in 2019. Lorazepam dispensing showed the largest increase, going from 15.5 to 26.7% between 2014 and 2019 (P < 0.001). There is a large burden of psychotropic polypharmacy in people with borderline personality disorder. This is concerning because of the lack of evidence regarding the efficacy of these medications in this group.


Dynamin-related protein 1 and mitochondrial fragmentation in neurodegenerative diseases.
The purpose of this article is to review the recent developments of abnormal mitochondrial dynamics, mitochondrial fragmentation, and neuronal damage in neurodegenerative diseases, including Alzheimer's, Parkinson's, Huntington's, and amyotrophic lateral sclerosis. The GTPase family of proteins, including fission proteins, dynamin related protein 1 (Drp1), mitochondrial fission 1 (Fis1), and fusion proteins (Mfn1, Mfn2 and Opa1) are essential to maintain mitochondrial fission and fusion balance, and to provide necessary adenosine triphosphate to neurons. Among these, Drp1 is involved in several important aspects of mitochondria, including shape, size, distribution, remodeling, and maintenance of mitochondria in mammalian cells. In addition, recent advancements in molecular, cellular, electron microscopy, and confocal imaging studies revealed that Drp1 is associated with several cellular functions, including mitochondrial and peroxisomal fragmentation, phosphorylation, SUMOylation, ubiquitination, and cell death. In the last two decades, tremendous progress has been made in researching mitochondrial dynamics, in yeast, worms, and mammalian cells; and this research has provided evidence linking Drp1 to neurodegenerative diseases. Researchers in the neurodegenerative disease field are beginning to recognize the possible involvement of Drp1 in causing mitochondrial fragmentation and abnormal mitochondrial dynamics in neurodegenerative diseases. This article summarizes research findings relating Drp1 to mitochondrial fission and fusion, in yeast, worms, and mammals. Based on findings from the Reddy laboratory and others', we propose that mutant proteins of neurodegenerative diseases, including AD, PD, HD, and ALS, interact with Drp1, activate mitochondrial fission machinery, fragment mitochondria excessively, and impair mitochondrial transport and mitochondrial dynamics, ultimately causing mitochondrial dysfunction and neuronal damage.


Role played by narcotics laboratories in the campaign against drug abuse and drug trafficking: a view from a developing country.
The narcotics laboratory at the national level identifies drugs for abuse and their accompanying substances in suspected samples, determines the purity and the possible origin of illicit drugs, carries out drug-related research, particularly on new sources of drugs liable to abuse, and, when required by the police or courts of law, provides supportive expertise in drug trafficking cases. Precaution must be taken to ensure that samples to be examined are representative. The university is a particularly appropriate setting for the location of a narcotics laboratory, especially if such a laboratory carries out complex work requiring assistance from other professional disciplines. Before new laboratory equipment is purchased, a careful study of requirements and financial resources should be made to ensure economical and optimum utilization of such equipment. In some situations the use of simple techniques, such as thin-layer chromatography, can be sufficient, while in others more sophisticated techniques may be required. Appropriate training of personnel is of particular importance for the effective functioning of a narcotics laboratory. The laboratory of the Department of Toxicology and Forensic Chemistry, University of Buenos Aires, provides for the training of personnel at three levels: The first level consists of basic training, which includes the use of kits for rapid identification of drugs in field conditions, for personnel from the police, gendarmerie, prefecture, customs and other agencies which deal with drug problems, but which have no previous skills in laboratory techniques; The second level is provided for professional laboratory personnel and usually lasts six months; The third level consists of two years' postgraduate university training for students who are expected to carry out complex laboratory work; an additional year is provided for trainees who are expected to assume responsibility in a laboratory unit.


Use of a self-designed bladder controller for restoring bladder function in paraplegic dogs.
To evaluate the results of a self-designed bladder controller for restoring micturition function in paraplegic dogs. The spinal cords of 4 dogs were transected above the cone. Electrodes were implanted in S2 bilaterally and connected to the subcutaneous receivers for external activation. Microsurgical technique was employed to perform dorsal rhizotomy of S1-3 intradurally. The dogs were stimulated daily to observe micturition. Urodynamic testing and vesicography were performed. All the dogs acquired micturition under the control of electric stimulation, with urine volume 80-140 ml per time. The mode of micturition was post-stimulus voiding. Vesicography revealed that the bladder was filled well and the bladder neck was open in the micturition course of electric stimulation. Residual urine volume was 15-20 ml. Urodynamic testing found that the bladder pressure and intraurethral pressure increased simultaneously, but when the intraurethral pressure was greater than the bladder pressure, no micturition occurred. The pressure decreased to baseline rapidly and the bladder pressure decreased slowly between two bursts. Micturition occurred when the bladder pressure was greater than the intraurethral pressure. The self-designed bladder controller together with a sacral deafferentation procedure can restore micturition function of paraplegic dogs.


Genetic control of single lumen formation in the zebrafish gut.
Most organs consist of networks of interconnected tubes that serve as conduits to transport fluid and cells and act as physiological barriers between compartments. Biological tubes are assembled through very diverse developmental processes that generate structures of different shapes and sizes. Nevertheless, all biological tubes invariably possess one single lumen. The mechanisms responsible for single lumen specification are not known. Here we show that zebrafish mutants for the MODY5 and familial GCKD gene tcf2 (also known as vhnf1) fail to specify a single lumen in their gut tube and instead develop multiple lumens. We show that Tcf2 controls single lumen formation by regulating claudin15 and Na+/K+-ATPase expression. Our in vivo and in vitro results indicate that Claudin15 functions in paracellular ion transport to specify single lumen formation. This work shows that single lumen formation is genetically controlled and appears to be driven by the accumulation of fluid.


Hepatic transcriptomic alterations for N,N-dimethyl-p-toluidine (DMPT) and p-toluidine after 5-day exposure in rats.
N,N-dimethyl-p-toluidine (DMPT), an accelerant for methyl methacrylate monomers in medical devices, was a liver carcinogen in male and female F344/N rats and B6C3F1 mice in a 2-year oral exposure study. p-Toluidine, a structurally related chemical, was a liver carcinogen in mice but not in rats in an 18-month feed exposure study. In this current study, liver transcriptomic data were used to characterize mechanisms in DMPT and p-toluidine liver toxicity and for conducting benchmark dose (BMD) analysis. Male F344/N rats were exposed orally to DMPT or p-toluidine (0, 1, 6, 20, 60 or 120 mg/kg/day) for 5 days. The liver was examined for lesions and transcriptomic alterations. Both chemicals caused mild hepatic toxicity at 60 and 120 mg/kg and dose-related transcriptomic alterations in the liver. There were 511 liver transcripts differentially expressed for DMPT and 354 for p-toluidine at 120 mg/kg/day (false discovery rate threshold of 5 %). The liver transcriptomic alterations were characteristic of an anti-oxidative damage response (activation of the Nrf2 pathway) and hepatic toxicity. The top cellular processes in gene ontology (GO) categories altered in livers exposed to DMPT or p-toluidine were used for BMD calculations. The lower confidence bound benchmark doses for these chemicals were 2 mg/kg/day for DMPT and 7 mg/kg/day for p-toluidine. These studies show the promise of using 5-day target organ transcriptomic data to identify chemical-induced molecular changes that can serve as markers for preliminary toxicity risk assessment.


The dimension of hyoid bone is independently associated with the severity of obstructive sleep apnea.
We hypothesized that the size of the hyoid bone itself may affect the severity of sleep apnea. The aim of this study was to identify the relationship between hyoid bone dimensions and the severity of sleep apnea using computerized tomography (CT) axial images. We retrospectively measured the hyoid bone in axial images of neck CTs and correlated these measurements with results of polysomnography in a total of 106 male patients. The new hyoid bone parameters studied in this study were as follows: distance between bilateral lesser horns (LH-d), distance between bilateral greater horns (GH-d), distance from the most anterior end of the hyoid arch to GH-d (AP), distance from the greater to the lesser horn on right and left sides (GH-LH), and the anterior angle between bilateral extensive lines from the greater to the lesser horn (H-angle). Data was analyzed using univariate and multivariate logistic regression, and Pearson correlation tests. We found a significant inverse correlation between the apnea-hypopnea index (AHI) and GH-d or AP. Neither the LH-d, GH-LH, nor H-angle were associated with the AHI. The patient group that met the criteria of both GH-d<45.4 and AP<33.4 demonstrated the most severe AHI. The lateral width or antero-posterior length of hyoid bone was associated with AHI and predicted the severity of sleep apnea in male patients. This finding supports the role of expansion hyoidplasty for treatment of sleep apnea. Pre-operative consideration of these parameters may improve surgical outcomes in male patients with sleep apnea.


Birth dates of men with cancer of the testis.
A recent rise in the incidence of cancer of the testis in affluent countries and affluent social strata suggests either an iatrogenic effect analogous with adenocarcinoma of the vagina or a prenatal infection analogous with congenital rubella syndrome. Three samples were assembled, two based on registration and one on death certification, and dates of birth were examined for the presence of steps, cycles, and irregular clusters. There was no evidence of a sudden step, and only equivocal evidence of clustering, but there was strong evidence of a temporal cycle. The cycle has a four-month period, analogous with a school-term-related cyclical component identified in other diseases. A prenatal infection is strongly suggested, but the pattern enables us to exclude many of the common childhood infectious fevers.


Manipulations of the mouse femoral nerve influence the accuracy of pathway reinnervation by motor neurons.
Previous studies using the femoral nerve model in both mice and rats have shown that regenerating motor axons prefer to reinnervate the terminal nerve branch to muscle versus a terminal nerve branch to skin, a process that has been termed preferential motor reinnervation (PMR). If end organ contact with muscle and skin is prevented, this preferential motor reinnervation still occurs in the rat. To better understand the process of preferential motor reinnervation in the mouse, we examined motor neuron reinnervation of muscle and cutaneous pathways without any end organ contact as well as with only cutaneous end organ contact. Surprisingly, there was no preferential motor reinnervation: Motor neurons preferred the cutaneous pathway over the muscle pathway when all end organ contact was prevented and showed an even greater preference for the cutaneous pathway when it was attached to skin.


Do transfusions cause necrotizing enterocolitis? The complementary role of randomized trials and observational studies.
A systematic review and a meta-analysis of the published literature on the association between transfusions in newborns and the occurrence of transfusion-associated necrotizing enterocolitis were performed. We discuss the differences between findings in randomized trials, and the results of observational studies that first explored this putative link. We suggest the following framework: where observational studies play a hypothesis generating- role for therapies and harm, and randomized studies allow an acid test of that hypothesis. It is acknowledged that not all questions can be subject to a randomized evaluation, but argued that this particular association is amenable to such a test.


[Expression of epidermal growth factor receptor and p53 protein in superficial cancer of the urinary bladder].
To determine the relationship of the immunohistochemical expression of p53 and EGF-r (epidermal growth factor receptor) and the recurrence rate and disease-free interval in superficial bladder cancer. 144 patients with superficial transitional cell carcinoma of the bladder were studied over a period of three years. Direct dilution anti-EGF-r monoclonal antibody (Biogenex, San Ramon, CA 94583, USA) was utilized for EGF-r determination. Anti-p53 mouse monoclonal antibody (DO7, Novocastra, Newcastle, 24 Claremont Place, UK) at a concentration of 1:100 was utilized for protein p53 determination and was considered negative if less than 10% of the tumor cells were stained and positive if 10% to 100% of the cells stained. EGF-r was determined only as either positive or negative regardless of percent of expression. 55 patients (38%) showed EGF-r and 14 (9.7%) showed p53 expression. The disease free survival was 54.08 months in the patients that showed EGF-r expression vs 30 months for those that did not, the difference being statistically significant (p = 0.027). However, no differences were found in this regard for p53 expression. Tumors that expressed EGF-r recurred in the same site as that of the primary tumor. By contrast, those that did not express EGF-r recurred in another or in multiple sites. The risk of recurrence is lower in bladder tumors that express EGF-r than those that do not, and when they recur, this generally occurs in the same site as the primary tumor. However, determination of p53 expression was not useful in determining the risk of recurrence or progression of superficial bladder tumors.


Marine-Inspired Enzymatic Mineralization of Dairy-Derived Whey Protein Isolate (WPI) Hydrogels for Bone Tissue Regeneration.
Whey protein isolate (WPI) is a by-product from the production of cheese and Greek yoghurt comprising β-lactoglobulin (β-lg) (75%). Hydrogels can be produced from WPI solutions through heating; hydrogels can be sterilized by autoclaving. WPI hydrogels have shown cytocompatibility and ability to enhance proliferation and osteogenic differentiation of bone-forming cells. Hence, they have promise in the area of bone tissue regeneration. In contrast to commonly used ceramic minerals for bone regeneration, a major advantage of hydrogels is the ease of their modification by incorporating biologically active substances such as enzymes. Calcium carbonate (CaCO3) is the main inorganic component of the exoskeletons of marine invertebrates. Two polymorphs of CaCO3, calcite and aragonite, have shown the ability to promote bone regeneration. Other authors have reported that the addition of magnesium to inorganic phases has a beneficial effect on bone-forming cell growth. In this study, we employed a biomimetic, marine-inspired approach to mineralize WPI hydrogels with an inorganic phase consisting of CaCO3 (mainly calcite) and CaCO3 enriched with magnesium using the calcifying enzyme urease. The novelty of this study lies in both the enzymatic mineralization of WPI hydrogels and enrichment of the mineral with magnesium. Calcium was incorporated into the mineral formed to a greater extent than magnesium. Increasing the concentration of magnesium in the mineralization medium led to a reduction in the amount and crystallinity of the mineral formed. Biological studies revealed that mineralized and unmineralized hydrogels were not cytotoxic and promoted cell viability to comparable extents (approximately 74% of standard tissue culture polystyrene). The presence of magnesium in the mineral formed had no adverse effect on cell viability. In short, WPI hydrogels, both unmineralized and mineralized with CaCO3 and magnesium-enriched CaCO3, show potential as biomaterials for bone regeneration.


Release of atrial natriuretic factor. Effects of repetitive stretch and temperature.
The effect of continuous and repetitive mechanical stretch on release of immunoreactive atrial natriuretic factor (IR-ANF) was studied in isolated rat atria distended by inflation of miniature balloon catheters. After 10 min incubation, the concentration of IR-ANF in the medium increased in proportion to the degree of continuous atrial stretch. Repetitive atrial expansion induced a rate dependent increase in release of IR-ANF which at the highest rate tested (89 inflations/min) was sixfold greater than that observed with continuous maximal expansion. Comparison of the amount of IR-ANF released from the right or left atrium separately revealed that during continuous and repetitive stretch, both atria contribute equally to the total amount of release IR-ANF. A reduction of bath temperature (37 degrees to 21.5 degrees C) decreased both basal- and stretch-induced IR-ANF release approximately 85%. It was concluded that release of atrial natriuretic factor is temperature-dependent and induced by both the extent and rate of atrial stretch.


Kinetic characterization of bile salt transport by human NTCP (SLC10A1).
The transport of bile acids facilitated by NTCP is an important factor in establishing bile flow. In this study, we examine the kinetics associated with human NTCP-dependent transport of two quantitatively important bile acids comprising the human bile acid pool, chenodeoxycholic acid and glycine-chenodeoxycholate, and secondary bile salt, 3-sulfo-glycolithocholate of potential toxicological significance. The study employed human NTCP overexpressing Chinese Hamster Ovary cells and results compared with taurocholate, a prototypical bile salt commonly used in transporter studies. GCDC and 3S-GLC but not CDCA were transported by NTCP. The efficient uptake of GCDC, TCA and 3S-GLC by NTCP enabled the determination of kinetics. GCDC displayed a lower KM (0.569±0.318μM) than TCA (6.44±3.83μM) and 3S-GLC (3.78±1.17μM). The apparent CLint value for GCDC was 20-fold greater (153±53μl/mg protein/min) than the apparent CLint for TCA (6.92±4.72μl/mg protein/min) and apparent CLint for 3S-GLC (8.05±1.33μl/mg protein/min). These kinetic results provide important complementary data on the substrate selectivity and specificity of NTCP to transport bile acids. NTCP transports GCDC with greater efficiency than TCA and has the same efficacy for 3S-GLC and TCA.


Use of a rapid, efficient inoculation method to induce papillomas by cottontail rabbit papillomavirus DNA shows that the E7 gene is required.
A simple inoculation method to induce papillomas efficiently with cottontail rabbit papillomavirus (CRPV) DNA is described. Using a jet injector, recombinant CRPV DNA is easily delivered to 100 or more sites per rabbit and induces typical epithelial papillomas in approximately 50% of those sites. Papillomas begin to form by 3 weeks and continue to develop for up to 7 weeks, a pattern similar to that reported following infection with intact virus. This system readily lends itself to investigation of viral gene function by delivering mutant viral genomes into an immunologically intact host. Two mutations in the E7 open reading frame were introduced into the complete CRPV genome and analyzed by this method. One was a frameshift mutation encoding just nine amino-terminal amino acids of the E7 protein; the other was an in-frame insertion mutation at position 9. Both E7 mutations were in a region of homology to the 300-kDa protein binding domain of adenovirus E1A protein. Neither mutant construct was able to induce papillomas, thereby demonstrating that the E7 gene participates in this biologic function. Exploitation of this approach, which demonstrates that a papillomavirus E7 gene is involved in the induction of papillomas in vivo, should permit detailed studies into molecular mechanisms involved in papilloma induction, malignant conversion, and host immune response. The high efficiency of papilloma induction with recombinant CRPV DNA suggests that the jet injector can also be used to study the biologic effects of other genetic elements in rabbits or in other species.


Undergraduate curriculum in palliative medicine at Tampere University increases students' knowledge.
Education in palliative medicine (PM) at medical schools reveals wide variation despite the increasing importance of palliative care. Many universities present poor description of the benefits and detailed content of the total curriculum in PM. Using the recommendations of European Association for Palliative Care (EAPC) as a reference, we evaluated the content and outcomes of the curriculum in PM at the University of Tampere, Finland. We searched for a PM curriculum by examining the teaching offered by every specialty and compared it to EAPC recommendations. Students' knowledge was evaluated using a progress test over three consecutive years. We found 53.5 teaching hours addressing PM issues, which exceeds the recommendation of the EAPC. Basics, symptom management, ethics, and communication skills were well established, while education in psychosocial/spiritual aspects, teamwork and self-reflection failed to reach the recommendations. Out of the maximum of 4.0, the progress test mean scores in PM among the third, fourth, fifth and sixth year students were 0.1 (SD 0.71), 0.69 (SD 1.28), 1.38 (SD 1.46) and 2.53 (SD 1.26), respectively (p < 0.001). This growing knowledge was associated with the timely increase in teaching provided through the PM discipline. In addition, the students who completed the optional PM course achieved better mean scores (2.66; SD 1.27) than the others (1.33; SD 1.43) (p < 0.001). The curriculum in PM at the University of Tampere is integrated into the teaching of many disciplines and complied well with the EAPC recommendations. This education led to increasing knowledge in PM among medical students.


MRSA surveillance programmes worldwide: moving towards a harmonised international approach.
Multinational surveillance programmes for methicillin-resistant Staphylococcus aureus (MRSA) are dependent on national structures for data collection. This study aimed to capture the diversity of national MRSA surveillance programmes and to propose a framework for harmonisation of MRSA surveillance. The International Society of Antimicrobial Chemotherapy (ISAC) MRSA Working Group conducted a structured survey on MRSA surveillance programmes and organised a webinar to discuss the programmes' strengths and challenges as well as guidelines for harmonisation. Completed surveys represented 24 MRSA surveillance programmes in 16 countries. Several countries reported separate epidemiological and microbiological surveillance. Informing clinicians and national policy-makers were the most common purposes of surveillance. Surveillance of bloodstream infections (BSIs) was present in all programmes. Other invasive infections were often included. Three countries reported active surveillance of MRSA carriage. Methodology and reporting of antimicrobial susceptibility, virulence factors, molecular genotyping and epidemiological metadata varied greatly. Current MRSA surveillance programmes rely upon heterogeneous data collection systems, which hampers international epidemiological monitoring and research. To harmonise MRSA surveillance, we suggest improving the integration of microbiological and epidemiological data, implementation of central biobanks for MRSA isolate collection, and inclusion of a representative sample of skin and soft-tissue infection cases in addition to all BSI cases.


Adiposity and risk of decline in glomerular filtration rate: meta-analysis of individual participant data in a global consortium.
To evaluate the associations between adiposity measures (body mass index, waist circumference, and waist-to-height ratio) with decline in glomerular filtration rate (GFR) and with all cause mortality. Individual participant data meta-analysis. Cohorts from 40 countries with data collected between 1970 and 2017. Adults in 39 general population cohorts (n=5 459 014), of which 21 (n=594 496) had data on waist circumference; six cohorts with high cardiovascular risk (n=84 417); and 18 cohorts with chronic kidney disease (n=91 607). GFR decline (estimated GFR decline ≥40%, initiation of kidney replacement therapy or estimated GFR <10 mL/min/1.73 m2) and all cause mortality. Over a mean follow-up of eight years, 246 607 (5.6%) individuals in the general population cohorts had GFR decline (18 118 (0.4%) end stage kidney disease events) and 782 329 (14.7%) died. Adjusting for age, sex, race, and current smoking, the hazard ratios for GFR decline comparing body mass indices 30, 35, and 40 with body mass index 25 were 1.18 (95% confidence interval 1.09 to 1.27), 1.69 (1.51 to 1.89), and 2.02 (1.80 to 2.27), respectively. Results were similar in all subgroups of estimated GFR. Associations weakened after adjustment for additional comorbidities, with respective hazard ratios of 1.03 (0.95 to 1.11), 1.28 (1.14 to 1.44), and 1.46 (1.28 to 1.67). The association between body mass index and death was J shaped, with the lowest risk at body mass index of 25. In the cohorts with high cardiovascular risk and chronic kidney disease (mean follow-up of six and four years, respectively), risk associations between higher body mass index and GFR decline were weaker than in the general population, and the association between body mass index and death was also J shaped, with the lowest risk between body mass index 25 and 30. In all cohort types, associations between higher waist circumference and higher waist-to-height ratio with GFR decline were similar to that of body mass index; however, increased risk of death was not associated with lower waist circumference or waist-to-height ratio, as was seen with body mass index. Elevated body mass index, waist circumference, and waist-to-height ratio are independent risk factors for GFR decline and death in individuals who have normal or reduced levels of estimated GFR.


Effects of Combining a Brief Cognitive Intervention with Transcranial Direct Current Stimulation on Pain Tolerance: A Randomized Controlled Pilot Study.
Cognitive behavioral therapy has been shown to be effective for treating chronic pain, and a growing literature shows the potential analgesic effects of minimally invasive brain stimulation. However, few studies have systematically investigated the potential benefits associated with combining approaches. The goal of this pilot laboratory study was to investigate the combination of a brief cognitive restructuring intervention and transcranial direct current stimulation (tDCS) over the left dorsolateral prefrontal cortex in affecting pain tolerance. Randomized, double-blind, placebo-controlled laboratory pilot. Medical University of South Carolina. A total of 79 healthy adult volunteers. Subjects were randomized into one of six groups: 1) anodal tDCS plus a brief cognitive intervention (BCI); 2) anodal tDCS plus pain education; 3) cathodal tDCS plus BCI; 4) cathodal tDCS plus pain education; 5) sham tDCS plus BCI; and 6) sham tDCS plus pain education. Participants underwent thermal pain tolerance testing pre- and postintervention using the Method of Limits. A significant main effect for time (pre-post intervention) was found, as well as for baseline thermal pain tolerance (covariate) in the model. A significant time × group interaction effect was found on thermal pain tolerance. Each of the five groups that received at least one active intervention outperformed the group receiving sham tDCS and pain education only (i.e., control group), with the exception of the anodal tDCS + education-only group. Cathodal tDCS combined with the BCI produced the largest analgesic effect. Combining cathodal tDCS with BCI yielded the largest analgesic effect of all the conditions tested. Future research might find stronger interactive effects of combined tDCS and a cognitive intervention with larger doses of each intervention. Because this controlled laboratory pilot employed an acute pain analogue and the cognitive intervention did not authentically represent cognitive behavioral therapy per se, the implications of the findings on chronic pain management remain unclear.


Capillary hemangioma of the testis: A rare benign tumour.
Testicular capillary hemangioma is a rare benign vascular tumour. We report a case of a 66-year-old man who underwent an uncomplicated radical orchiectomy for a painless left testicular mass. Pathology showed capillary hemangioma of the testis. There are only 22 cases reported in the English literature, including the presented case. Appropriate intra-operative recognition of this entity is vital to assess for potential testicular-sparing surgery.


Supramolecular polymerization: a coarse grained molecular dynamics study.
A coarse-grained (CG) force field to model the self-assembly of benzene-1,3,5-tricarboxamide (BTA) class of compounds in nonpolar solvents has been developed. The model includes an intrinsic point dipole embedded on one of the CG beads so as to impart a macrodipole moment to the oligomer, one of its characteristic feature. Chemical specificity has been preserved by benchmarking against results, including dimerization and solvation free energies, obtained from an all-atom representation. Starting from a well-dispersed configuration in n-nonane, BTA molecules self-assemble to form one-dimensional stacks. Free energy (FE) changes for the various manner in which short oligomers can exchange between the assembled and the dispersed states have been calculated. These calculations show BTA to self-assemble via a downhill cooperative mechanism with a nucleus size of three.


Phase noise measurements in dual-mode SC-cut crystal oscillators.
This paper describes the phase-noise characteristics and the analysis model of an SC-cut dual-mode oscillator. The C mode phase-noise sideband levels of -124 dBc at 10 Hz and -154 dBc at 10 kHz have been demonstrated using a dual-mode oscillator that simultaneously excited the C and B mode of a 10-MHz, third overtone, SC-cut crystal resonator. Based on Leeson's model, a phase-noise analysis model for dual-mode oscillators has been proposed also. Actual phase-noise levels of the C mode in dual-mode oscillation corresponded well to results calculated from the proposed model.


Decision analysis of dutasteride use for patients with negative prostate biopsy.
To determine whether the additional benefits of improved prostate cancer detection associated with 5α-reductase inhibitors are sufficient to warrant chemoprevention in the case where the degree of prostate cancer risk reduction is deemed inadequate. We reanalyzed data from REDUCE, a randomized trial of dutasteride for prostate cancer chemoprevention in men with prior negative biopsy. We evaluated whether statistical models using prostate-specific antigen (PSA) and PSA velocity could help predict the result of repeat prostate biopsy separately for dutasteride and placebo groups. Area under the curve was evaluated by 10-fold cross-validation. PSA velocity improved discrimination at 4 years in the dutasteride group but not at 2 years nor in the placebo group. At 2 years, dutasteride improved discrimination of PSA slightly (0.616 vs. 0.603 for any grade cancer; 0.681 vs. 0.676 for high-grade disease). Between-group differences in cancer rates at 4 years were small. Clinicians who are willing to treat at least 23 patients with dutasteride for 2 years to avoid 1 prostate cancer diagnosis should offer dutasteride after initial negative biopsy. Clinicians not willing to do so might consider dutasteride for its additional benefit of reducing unnecessary biopsy, although this benefit is apparent only under very restrictive conditions. It is difficult to justify extending treatment with dutasteride for >2 years.


Microsatellite analysis of genetic variation among and within Alpine marmot populations in the French Alps.
The genetic structure of the Alpine marmot, Marmota marmota, was studied by an analysis of five polymorphic microsatellite loci. Eight locations were sampled in the French Alps, one from Les Ecrins valley (n = 160), another from La Sassière valley (n = 289) and the six others from the Maurienne valley (n = 139). Information on social group structure was available for both Les Ecrins and La Sassière but not for the other samples. The high levels of genetic diversity observed are at odds with the results obtained using microsatellites, minisatellites and allozymes on Alpine marmots from Germany, Austria and Switzerland. Strong deficits in heterozygotes were found in Les Ecrins and La Sassière. They are caused by a Wahlund effect due to the family structure (i.e. differentiation between the family groups). The family groups exhibit excess of heterozygotes rather than deficits. This may be caused by outbreeding and this is compatible with recent results from the genetics of related social species when information on the social structure is taken into account. The observed outbreeding could be the result of females mating with transient males or males coming from neighbouring colonies. Both indicate that the species may not be as monogamous as is usually believed. The results are also compatible with a male-biased dispersal but do not allow us to exclude some female migration. We also found a significant correlation between geographical and genetic distance indicating that isolation by distance could be an issue in marmots. This study is the first that analysed populations of marmots taking into account the social structure within populations and assessing inbreeding at different levels (region, valley, population, and family groups). Our study clearly demonstrated that the sampling strategy and behavioural information can have dramatic effects on both the results and interpretation of the genetic data.


The evaluation of left ventricular functions with tissue doppler echocardiography in adults with celiac disease.
The aim of this study was to investigate the effects of celiac disease on cardiac functions using tissue Doppler echocardiography (TDE). The study included 30 patients with celiac disease (CD) and 30 healthy volunteers. Echocardiographic examinations were assessed by conventional echocardiography and tissue Doppler imaging. The peak systolic velocity (S'm), early diastolic myocardial peak velocity (E'm), late diastolic myocardial peak velocity (A'm), E'm/A'm ratio, myocardial precontraction time (PCT'm), myocardial contraction time (CT'm), and myocardial isovolumetric relaxation time (IVRT'm), E to E'm ratio were measured. In pulsed wave Doppler echocardiography, mitral late diastolic flow (A) velocity and E to E'm ratio were significantly higher (P = 0.02 and P = 0,017), E/A ratio was significantly lower (P = 0.008) and IVRT was significantly prolonged (P = 0.014) in patients with CD. In TDE, S'm, E'm, and E'm/A'm ratio were significantly lower, IVRT'm was longer (P = 0.009) from septal mitral annulus and S'm, E'm, E'm/A'm ratio were significantly lower, PCT'm, PCT/ET ratio, IVRT'm were longer, and MPI was higher from lateral mitral annulus in celiac group than controls. Our study confirms that patients with CD have impaired diastolic function. More importantly, we also demonstrated an impairment of myocardial systolic function in patients with CD by TDE. We recommend using TDE in addition to conventional echocardiography parameters for the cardiovascular risk assessment of patients with CD.


Effects of short-term sediment nutrient enrichment and grazer (Neritina Reclivata) removal on sediment microalgae in a shallow eutrophic estuary (Alabama, USA).
The olive snail (Neritina reclivata) is ubiquitous in tropical and sub-tropical systems of the Gulf of Mexico, however its impacts on sediment microalgae have been little studied. Many coastal systems around the world are being eutrophied due to human activities, and seemingly they will continue to be eutrophied to a further extent in the future. Exploring the single and combined impacts of further nutrient enrichment and grazing by the olive snail on sediment microalgae in such eutrophic systems is an important question for our understanding and management of these systems. Here we examine the effects of short-term nutrient enrichment and grazing by the olive snail N. reclivata on sediment microalgal biomass and composition in a shallow eutrophic estuary (Weeks Bay, Alabama, USA) of the Northern Gulf of Mexico. For this, we performed a series of factorial experiments adding or not nutrients and removing or not the snail, for a total of four treatments in each experiment: ambient grazing, ambient nutrients; ambient grazing, increased nutrients; no grazing, ambient nutrients; and no grazing, increased nutrients. We did not find any significant impact of nutrient addition in any of the eight short-term (i.e. four days) experiments carried out. Impacts by the snail were minor; we only found a decrease in biomass due to snail grazing in one of the eight experiments, and no impacts on microalgal (i.e. diatom) composition. High ambient nutrient concentrations in the sediment porewater and low snail abundances on the sediment could explain these findings. Our results suggest that ephemeral, short-term nutrient pulses into eutrophic coastal systems of the Northern Gulf of Mexico, such as Weeks Bay (Alabama, USA), should not greatly affect the abundance of sediment microalgae, even though those pulses occur in well-lit areas. The results further suggest the snail N. reclivata is not a major control of sediment microalgal populations in the subtidal sedimentary areas studied. Our findings contrast with the results of past work in sediments with well-lit and nutrient poor conditions, or sediments with high densities of other snail grazers. In conjunction this and other investigations indicate that the response of sediment microalgae to nutrient enrichment and modified grazer abundance depends to a large extent on the initial levels of nutrient availability and grazing before the system is altered.


Confocal reflectance imaging of head and neck surgical specimens. A comparison with histologic analysis.
Confocal reflectance microscopy (CRM) is an optical method of imaging tissue noninvasively without the need for fixation, sectioning, and staining as in standard histopathologic analysis. Image contrast is determined by natural differences in refractive indices of organelles and other subcellular structures within the tissues. Gray-scale images are displayed in real time on a video monitor and represent horizontal (en face) optical sections through the tissue. We hypothesized that CRM is capable of discerning histologic characteristics of different tissues in the head and neck. To examine the microscopic anatomy of freshly excised head and neck surgical specimens en bloc using CRM and to compare the findings with those generated by conventional histologic analysis. This was a pilot observational cohort study. Bone, muscle, nerve, thyroid, parotid, and ethmoid mucosa from human surgical specimens were imaged immediately after excision. Confocal images were compared with corresponding routine paraffin-embedded, hematoxylin-eosin-stained sections obtained from the same tissue. Characteristic histologic features of various tissues and cell types were readily discernible by CRM and correlated well with permanent sections. However, in all tissues examined, there was less microscopic detail visible in the CRM images than was appreciated in paraffin-embedded histologic sections. The CRM images revealed cytologic features without the artifacts of histologic processing and thus may have the potential for use as an adjunct to frozen-section analysis in intraoperative consultation.


Evaluation of the US Food and Drug Administration sentinel analysis tools in confirming previously observed drug-outcome associations: The case of clindamycin and Clostridium difficile infection.
The Food and Drug Administration's Sentinel System developed parameterized, reusable analytic programs for evaluation of medical product safety. Research on outpatient antibiotic exposures, and Clostridium difficile infection (CDI) with non-user reference groups led us to expect a higher rate of CDI among outpatient clindamycin users vs penicillin users. We evaluated the ability of the Cohort Identification and Descriptive Analysis and Propensity Score Matching tools to identify a higher rate of CDI among clindamycin users. We matched new users of outpatient dispensings of oral clindamycin or penicillin from 13 Data Partners 1:1 on propensity score and followed them for up to 60 days for development of CDI. We used Cox proportional hazards regression stratified by Data Partner and matched pair to compare CDI incidence. Propensity score models at 3 Data Partners had convergence warnings and a limited range of predicted values. We excluded these Data Partners despite adequate covariate balance after matching. From the 10 Data Partners where these models converged without warnings, we identified 807 919 new clindamycin users and 8 815 441 new penicillin users eligible for the analysis. The stratified analysis of 807 769 matched pairs included 840 events among clindamycin users and 290 among penicillin users (hazard ratio 2.90, 95% confidence interval 2.53, 3.31). This evaluation produced an expected result and identified several potential enhancements to the Propensity Score Matching tool. This study has important limitations. CDI risk may have been related to factors other than the inherent properties of the drugs, such as duration of use or subsequent exposures.


Epidemic dynamics on information-driven adaptive networks.
Research on the interplay between the dynamics on the network and the dynamics of the network has attracted much attention in recent years. In this work, we propose an information-driven adaptive model, where disease and disease information can evolve simultaneously. For the information-driven adaptive process, susceptible (infected) individuals who have abilities to recognize the disease would break the links of their infected (susceptible) neighbors to prevent the epidemic from further spreading. Simulation results and numerical analyses based on the pairwise approach indicate that the information-driven adaptive process can not only slow down the speed of epidemic spreading, but can also diminish the epidemic prevalence at the final state significantly. In addition, the disease spreading and information diffusion pattern on the lattice as well as on a real-world network give visual representations about how the disease is trapped into an isolated field with the information-driven adaptive process. Furthermore, we perform the local bifurcation analysis on four types of dynamical regions, including healthy, a continuous dynamic behavior, bistable and endemic, to understand the evolution of the observed dynamical behaviors. This work may shed some lights on understanding how information affects human activities on responding to epidemic spreading.


Prevention of central venous catheter-related infection in the neonatal unit: a literature review.
Central venous catheter infections are the leading cause of healthcare-associated bloodstream infections and contribute significantly to mortality and morbidity in neonatal intensive care units. Moreover, infection poses significant economic consequence which increased hospital costs and increased length of hospital stay. Prevention strategies are detailed in guidelines published by the Centers for Disease Control and Prevention (CDC) in the United States; nevertheless, recent surveys in neonatal units in the United States, and Australia and New Zealand demonstrate these are not always followed. This review discusses the numerous evidence-based strategies to prevent catheter infections including hand hygiene, maximal sterile barriers during insertion, skin disinfection, selection of insertion site, dressings, aseptic non-touch technique, disinfection of catheter hubs/ports, administration set management, prompt removal of catheter, antibiotic locks, systemic antibiotic prophylaxis and chlorhexidine bathing. Furthermore, it will describe different strategies that can be implemented into clinical practice to reduce infection rates. These include the use of care bundles including checklists, education and the use of CVC teams.


Overview of zinc absorption and excretion in the human gastrointestinal tract.
Zinc homeostasis is primarily maintained via the gastrointestinal system by the processes of absorption of exogenous zinc and gastrointestinal secretion and excretion of endogenous zinc. Although these processes modulate net absorption and the size of the readily exchangeable zinc pools, there are limits to the effectiveness of the homeostatic mechanisms of these and other systems. As a result of the interplay of the subcellular regulation of these mechanisms and host, dietary and environmental factors, zinc deficiency is not uncommon, especially on a global basis. This overview briefly reviews current understanding about the subcellular mechanisms of zinc absorption and transport. Factors recognized to affect zinc absorption at the whole body level are reviewed and include the amount and form of zinc consumed; dietary promoters, such as animal protein and low-molecular-weight organic compounds; dietary inhibitors, such as phytate and possibly iron and calcium when consumed as supplements; and physiologic states, such as pregnancy, lactation and early infancy, all of which increase the demand for absorbed zinc. The control of endogenously secreted zinc is less well understood. Available data suggest that the quantity of secreted zinc with each meal may be considerable and that efficient reabsorption is critical to the maintenance of normal zinc balance. Factors that have been proposed to interfere with the normal reabsorption of endogenous zinc include phytate and unabsorbed fat. Understanding of the dietary, physiologic, pathologic and environmental factors that may adversely affect these processes, and therefore zinc homeostasis, will be critical to preventing and treating zinc deficiency in human populations.


Steroidogenic factor 1/adrenal 4 binding protein transforms human bone marrow mesenchymal cells into steroidogenic cells.
Steroidogenic factor 1/adrenal 4 binding protein (SF-1/Ad4BP) is an essential nuclear receptor for steroidogenesis as well as for adrenal and gonadal gland development. Mesenchymal bone marrow cells (BMCs) contain pluripotent progenitor cells, which differentiate into multiple lineages. In a previous study, we reported that adenovirus-mediated forced expression of SF-1 could transform mouse primary long-term cultured BMCs into steroidogenic cells. For future clinical application, trials using human BMCs would be indispensable. In this study, we examined whether SF-1 could transform human BMCs into steroidogenic cells and compared the steroid profile of these cells with that of mouse steroidogenic BMCs. Primary cultured human BMCs infected with adenovirus containing bovine SF-1 cDNA could produce progesterone, corticosterone, cortisol, dehydroepiandrosterone, testosterone, and estradiol. Such a mixed character of adrenal and gonadal steroid production in human BMCs was supported by the expressions of P450scc, 3beta-hydroxysteroid dehydrogenase (3beta-HSD), P450c21, P450c11, P450c17, 17beta-HSD, and P450arom mRNAs. Unlike mouse steroidogenic BMCs, introduction of SF-1 into human BMCs caused dramatic inductions of both ACTH and LH receptors, thus leading to good responsiveness of the cells to ACTH and LH respectively. Importantly, among several factors that are known to be closely associated with adrenal and/or gonadal development, introduction of only SF-1 enabled the human BMCs to express P450scc and to produce cortisol and testosterone, suggesting that SF-1 is truly a master regulator for the production of steroidogenic cells from human BMCs.


Causal Analysis of Physiological Sleep Data Using Granger Causality and Score-Based Structure Learning.
Understanding how the human body works during sleep and how this varies in the population is a task with significant implications for medicine. Polysomnographic studies, or sleep studies, are a common diagnostic method that produces a significant quantity of time-series sensor data. This study seeks to learn the causal structure from data from polysomnographic studies carried out on 600 adult volunteers in the United States. Two methods are used to learn the causal structure of these data: the well-established Granger causality and "DYNOTEARS", a modern approach that uses continuous optimisation to learn dynamic Bayesian networks (DBNs). The results from the two methods are then compared. Both methods produce graphs that have a number of similarities, including the mutual causation between electrooculogram (EOG) and electroencephelogram (EEG) signals and between sleeping position and SpO2 (blood oxygen level). However, DYNOTEARS, unlike Granger causality, frequently finds a causal link to sleeping position from the other variables. Following the creation of these causal graphs, the relationship between the discovered causal structure and the characteristics of the participants is explored. It is found that there is an association between the waist size of a participant and whether a causal link is found between the electrocardiogram (ECG) measurement and the EOG and EEG measurements. It is concluded that a person's body shape appears to impact the relationship between their heart and brain during sleep and that Granger causality and DYNOTEARS can produce differing results on real-world data.


Emergence delirium in children is not related to intraoperative burst suppression - prospective, observational electrography study.
Emergence-delirium is the most frequent brain dysfunction in children recovering from general anaesthesia, though the pathophysiological background remains unclear. The presented study analysed an association between emergence delirium and intraoperative Burst Suppression activity in the electroencephalogram, a period of very deep hypnosis during general anaesthesia. In this prospective, observational cohort study at the Charité - university hospital in Berlin / Germany children aged 0.5 to 8 years, undergoing planned surgery, were included between September 2015 and February 2017. Intraoperative bi-frontal electroencephalograms were recorded. Occurrence and duration of Burst Suppression periods were visually analysed. Emergence delirium was assessed using the Pediatric Assessment of Emergence Delirium Score. From 97 children being analysed within this study, 40 children developed emergence delirium, and 57 children did not. Overall 52% of the children displayed intraoperative Burst Suppression periods; however, occurrence and duration of Burst Suppression (Emergence delirium group 55% / 261 + 462 s vs. Non-emergence delirium group 49% / 318 + 531 s) did not differ significantly between both groups. Our data reveal no correlation between the occurrence and duration of intraoperative Burst Suppression activity and the incidence of emergence delirium. Burst Suppression occurrence is frequent; however, it does not seem to have an unfavourable impact on cerebral function at emergence from general anaesthesia in children. NCT02481999, June 25, 2015.


Generation of envelope and defective interfering RNA mutants of tomato spotted wilt virus by mechanical passage.
During a series of mechanical transfers of tomato spotted wilt virus, two distinct types of mutants were generated. Firstly, a morphologically defective isolate was obtained which had lost the ability to produce the membrane glycoproteins and, as a consequence, was not able to form enveloped particles. Analysis of the genomic RNAs of this isolate suggested that this defect was caused by either point mutations or very small deletions in the medium genomic RNA segment. Secondly, isolates were obtained which had accumulated truncated forms of the large (L) RNA segment. These shortened L RNA molecules most likely represented defective interfering RNAs, since they replicated more rapidly than full-length L RNA and their appearance was often associated with symptom attenuation. Defective L RNAs of different sizes were generated after repeated transfers, and hybridization analysis using L RNA-specific cDNA probes showed that the internal regions deleted varied in length. The presence of defective L RNAs in nucleocapsid fractions as well as in enveloped virus particles indicates that all defective molecules retained the sequences required for replication, encapsidation by nucleocapsid proteins and packaging of the nucleocapsid into virus particles.


Capsule endoscopy in real life: a four-centre experience of 160 consecutive patients in Israel.
The Given Capsule procedure is a new system designed to aid the gastroenterologist in diagnosing small-bowel disease. Capsule endoscopy has provided gastroenterologists with easy access to small-bowel mucosa for examination by direct visualisation. The aim of our prospective study was to determine the indication for capsule endoscopy and to describe the identified pathology within the gastrointestinal tract. Patients with unexplained gastrointestinal bleeding from suspected small-bowel pathology of an undisclosed nature or with unexplained abdominal pain for more than 6 months were eligible to undergo capsule endoscopy. Patients with suspected intestinal obstruction or with a history of major abdominal surgery were excluded. One hundred and sixty patients (70 women, 90 men; mean age 49.0 +/- 19.8 years, range 13-91 years) ingested the capsule easily. They had undergone 6 +/- 2.9 investigative procedures before capsule endoscopy. Two (1.25%) patients who had non-natural excretions underwent surgery: one was found to have Meckel's diverticulum and the other had stricturing Crohn's disease. The mean haemoglobin in iron-deficiency anaemia patients was 9.1 +/- 2.2 g%. The diagnostic yield of capsule endoscopy was 57.7% in iron-deficiency anaemia and 52.4% in suspected Crohn's disease. Capsule endoscopy detected the more clinically significant intestinal lesions. It was of diagnostic value in patients with obscure bleeding, suspected Crohn's disease and malabsorption, but the diagnostic yield in patients with chronic pain or irritable bowel syndrome-type symptoms was low.


Circulating Extracellular DNA: Cause or Consequence of Thrombosis?
Thrombosis leads to ischemic organ damage in cardiovascular and thromboembolic diseases. Neutrophils promote thrombosis in vitro and in vivo by releasing neutrophil extracellular traps (NETs). NETs are composed of DNA filaments coated with histones and neutrophil enzymes such as myeloperoxidase (MPO). Circulating extracellular DNA (ceDNA) is widely used as a surrogate marker to monitor NET formation in thrombosis. This narrative review summarizes the association of ceDNA with human thrombosis. Levels of ceDNA indicate the extent and outcome of several cardiovascular and thromboembolic diseases, including myocardial infarction, stroke, and venous thromboembolism. ceDNA correlates with markers of coagulation and platelet consumption, thus supporting the hypothesis that ceDNA may be a surrogate marker of thrombus formation. In addition, ceDNA levels correlate with markers of cell injury and size of ischemic lesions, suggesting that ceDNA does not derive from NETs but is probably released from damaged organs. Few studies identified NET-specific biomarkers such as DNA-MPO complexes in the blood of patients with thrombosis. In conclusion, it remains to be established whether ceDNA in patients derives from NETs and is a cause or consequence of thrombosis.


Endoplasmic reticulum stress, diabetes mellitus, and tissue injury.
Endoplasmic reticulum (ER) stress is characterized by the accumulation of unfolded and misfolded proteins in the ER lumen. Unfolded and misfolded protein accumulation interferes with the ER function and triggers ER stress response. Thus, ER stress response, also called unfolded protein response (UPR), is an adaptive process that controls the protein amount in the ER lumen and the downstream protein demand. In normal conditions, the role of ER stress is to maintain ER homeostasis, restore ER function, and protect stressed cells from apoptosis, by coordinating gene expression, protein synthesis, and accelerating protein degradation through several molecular pathways. However, prolonged ER stress response plays a paradoxical role, which leads to cell damage, apoptosis, and concomitant tissue injuries. A number of tissue alterations are involved with diabetes mellitus progress and its comorbidities via ER stress. However, certain pharmacological agents affecting ER stress have been identified. In this review, we summarized the relationship between ER stress and insulin resistance development. Moreover, we aim to explain how ER stress influences type 2 diabetes mellitus (T2DM) development. In addition, we reviewed the literature on ER stress and UPR in three kinds of tissue injuries induced by T2DM. Finally, a retrospective analysis of the effects of anti-diabetes medications on ER stress is presented.


Biofunctionality of Enzymatically Derived Peptides from Codfish (Gadus morhua) Frame: Bulk In Vitro Properties, Quantitative Proteomics, and Bioinformatic Prediction.
Protein hydrolysates show great promise as bioactive food and feed ingredients and for valorization of side-streams from e.g., the fish processing industry. We present a novel approach for hydrolysate characterization that utilizes proteomics data for calculation of weighted mean peptide properties (length, molecular weight, and charge) and peptide-level abundance estimation. Using a novel bioinformatic approach for subsequent prediction of biofunctional properties of identified peptides, we are able to provide an unprecedented, in-depth characterization. The study further characterizes bulk emulsifying, foaming, and in vitro antioxidative properties of enzymatic hydrolysates derived from cod frame by application of Alcalase and Neutrase, individually and sequentially, as well as the influence of heat pre-treatment. All hydrolysates displayed comparable or higher emulsifying activity and stability than sodium caseinate. Heat-treatment significantly increased stability but showed a negative effect on the activity and degree of hydrolysis. Lower degrees of hydrolysis resulted in significantly higher chelating activity, while the opposite was observed for radical scavenging activity. Combining peptide abundance with bioinformatic prediction, we identified several peptides that are likely linked to the observed differences in bulk emulsifying properties. The study highlights the prospects of applying proteomics and bioinformatics for hydrolysate characterization and in food protein science.


Evolution of developmentally regulated genome rearrangements in eukaryotes.
Developmentally regulated genome rearrangements (DRGR)--processes that alter genomes either in specific cells or during specific life cycle stages--are widespread throughout eukaryotes. This contrasts with the view that genome structure and content remain essentially constant throughout an organism's life cycle. Here we review three categories of developmentally regulated genome processing in eukaryotes: genome-wide rearrangements, targeted rearrangements, and a special case of amplification of ribosomal DNA genes. Mapping these types of DRGR onto eukaryotic phylogeny indicates that each type of processing is found in multiple independent lineages. We propose that such genome rearrangements were present within the last common ancestor of extant eukaryotes, and that future research will yield evidence of homologous epigenetic mechanisms underlying genome processing among diverse eukaryotes.


Optical properties of human dermis in vitro and in vivo.
Condensed Monte Carlo simulation results have been used for calculating absorption and reduced scattering coefficients from the literature data on the measured total transmittance and total reflectance of samples of the human skin in vitro. The results of several measuring methods have been compared. We have also estimated the range for absorption coefficients and reduced scattering coefficients at 660 and 940 nm from measured intensities at the skin surface as a function of the distance from the location where the light enters the skin by using condensed Monte Carlo simulations for a homogeneous semi-infinite medium. The in vivo values for the absorption coefficients and the reduced scattering coefficients appear to be much smaller than the values from the in vitro measurements, that have been assumed until now. The discrepancies have been discussed in detail. Our in vivo results are in agreement with other in vivo measurements that are available in the literature.


EDTA dependent red cell neutrophil rosetting in autoimmune haemolytic anaemia.
Disodium ethylenediaminetetraacetate (EDTA) dependent platelet-neutrophil rosetting is a well-known but infrequently reported in vitro blood abnormality. We describe a patient with autoimmune haemolytic anaemia in whom EDTA dependent red cell-neutrophil rosettes were a persistent blood film feature.


Stress attenuates the flexible updating of aversive value.
In a dynamic environment, sources of threat or safety can unexpectedly change, requiring the flexible updating of stimulus-outcome associations that promote adaptive behavior. However, aversive contexts in which we are required to update predictions of threat are often marked by stress. Acute stress is thought to reduce behavioral flexibility, yet its influence on the modulation of aversive value has not been well characterized. Given that stress exposure is a prominent risk factor for anxiety and trauma-related disorders marked by persistent, inflexible responses to threat, here we examined how acute stress affects the flexible updating of threat responses. Participants completed an aversive learning task, in which one stimulus was probabilistically associated with an electric shock, while the other stimulus signaled safety. A day later, participants underwent an acute stress or control manipulation before completing a reversal learning task during which the original stimulus-outcome contingencies switched. Skin conductance and neuroendocrine responses provided indices of sympathetic arousal and stress responses, respectively. Despite equivalent initial learning, stressed participants showed marked impairments in reversal learning relative to controls. Additionally, reversal learning deficits across participants were related to heightened levels of alpha-amylase, a marker of noradrenergic activity. Finally, fitting arousal data to a computational reinforcement learning model revealed that stress-induced reversal learning deficits emerged from stress-specific changes in the weight assigned to prediction error signals, disrupting the adaptive adjustment of learning rates. Our findings provide insight into how stress renders individuals less sensitive to changes in aversive reinforcement and have implications for understanding clinical conditions marked by stress-related psychopathology.


Caspase-8 promotes c-Rel-dependent inflammatory cytokine expression and resistance against Toxoplasma gondii.
Caspase-8 is a key integrator of cell survival and cell death decisions during infection and inflammation. Following engagement of tumor necrosis factor superfamily receptors or certain Toll-like receptors (TLRs), caspase-8 initiates cell-extrinsic apoptosis while inhibiting RIPK3-dependent programmed necrosis. In addition, caspase-8 has an important, albeit less well understood, role in cell-intrinsic inflammatory gene expression. Macrophages lacking caspase-8 or the adaptor FADD have defective inflammatory cytokine expression and inflammasome priming in response to bacterial infection or TLR stimulation. How caspase-8 regulates cytokine gene expression, and whether caspase-8-mediated gene regulation has a physiological role during infection, remain poorly defined. Here we demonstrate that both caspase-8 enzymatic activity and scaffolding functions contribute to inflammatory cytokine gene expression. Caspase-8 enzymatic activity was necessary for maximal expression of Il1b and Il12b, but caspase-8 deficient cells exhibited a further decrease in expression of these genes. Furthermore, the ability of TLR stimuli to induce optimal IκB kinase phosphorylation and nuclear translocation of the nuclear factor kappa light chain enhancer of activated B cells family member c-Rel required caspase activity. Interestingly, overexpression of c-Rel was sufficient to restore expression of IL-12 and IL-1β in caspase-8-deficient cells. Moreover, Ripk3-/-Casp8-/- mice were unable to control infection by the intracellular parasite Toxoplasma gondii, which corresponded to defects in monocyte recruitment to the peritoneal cavity, and exogenous IL-12 restored monocyte recruitment and protection of caspase-8-deficient mice during acute toxoplasmosis. These findings provide insight into how caspase-8 controls inflammatory gene expression and identify a critical role for caspase-8 in host defense against eukaryotic pathogens.


Epigenomic Assessment of Cardiovascular Disease Risk and Interactions With Traditional Risk Metrics.
Background Epigenome-wide association studies for cardiometabolic risk factors have discovered multiple loci associated with incident cardiovascular disease (CVD). However, few studies have sought to directly optimize a predictor of CVD risk. Furthermore, it is challenging to train multivariate models across multiple studies in the presence of study- or batch effects. Methods and Results Here, we analyzed existing DNA methylation data collected using the Illumina HumanMethylation450 microarray to create a predictor of CVD risk across 3 cohorts: Women's Health Initiative, Framingham Heart Study Offspring Cohort, and Lothian Birth Cohorts. We trained Cox proportional hazards-based elastic net regressions for incident CVD separately in each cohort and used a recently introduced cross-study learning approach to integrate these individual scores into an ensemble predictor. The methylation-based risk score was associated with CVD time-to-event in a held-out fraction of the Framingham data set (hazard ratio per SD=1.28, 95% CI, 1.10-1.50) and predicted myocardial infarction status in the independent REGICOR (Girona Heart Registry) data set (odds ratio per SD=2.14, 95% CI, 1.58-2.89). These associations remained after adjustment for traditional cardiovascular risk factors and were similar to those from elastic net models trained on a directly merged data set. Additionally, we investigated interactions between the methylation-based risk score and both genetic and biochemical CVD risk, showing preliminary evidence of an enhanced performance in those with less traditional risk factor elevation. Conclusions This investigation provides proof-of-concept for a genome-wide, CVD-specific epigenomic risk score and suggests that DNA methylation data may enable the discovery of high-risk individuals who would be missed by alternative risk metrics.


Problem based learning.
Problem based learning (PBL) has been the subject of considerable interest and debate in medical undergraduate and, increasingly, postgraduate education in recent years. Its supporters maintain that PBL enhances learning by providing a highly motivational environment for acquisition of knowledge, which is well received by those who take part in it. Critics argue that PBL is a time consuming exercise, often undertaken by people with a limited appreciation of its complexities, and the lack of evidence that PBL translates into better clinical competence brings into question the relevance of such intensive learning methods in everyday practice. This article outlines the background to PBL, explains its mechanics in action, and considers the potential disadvantages of its use as a clinical learning tool in the emergency department.