Measurement uncertainty evaluation from correlated validation data: Determination of elemental impurities in pharmaceutical products by ICP-MS.
Pharmaceutical products as well as active pharmaceutical ingredients (APIs) are checked for levels of elemental contaminants to guarantee medicines administration will not involve the consumption of level of contaminants greater than their maximum admissible intake. However, the conformity decision is affected by the measurement uncertainty function of analytical steps performance, used standards quality and how measurement performance is assessed during method validation. When an ingredient is considered conform, since the measured concentration is lower than the maximum limit, the risk of a false acceptance depends on how close the measured concentration is from the limit and on the measurement uncertainty. The analytical methods used for pharmaceutical analysis should be validated by ICH and USP recommendations, in order to prove measurements are fit for purpose. The validation must also be economically feasible and have an acceptable duration. This work discusses how to evaluate the uncertainty of elemental analysis in pharmaceutical ingredients from data collected during the validation of the analytical method by following ICH guidelines and USP chapters. A top-down uncertainty evaluation based on results from the analysis of a model API intermediate, with the native analyte after spiking at three concentration levels, where analyses are performed by two analysts in two different days, is presented. The impact of the correlation of some uncertainty components of collected results on the uncertainty evaluation is discussed and considered in the calculations. The developed measurement model was checked by a cross-validation procedure where some validation data was randomly removed and used for an independent model control. The developed uncertainty evaluation methodology was successfully applied to the analysis of Pd in a model API intermediate by ICP-MS after a micro-wave assisted acid digestion, where the risk of a false acceptance of the pharmaceuticals is determined. The measurement performance data and used spreadsheet are made available as Supplementary Material.


Proteomic prediction and Renin angiotensin aldosterone system Inhibition prevention Of early diabetic nephRopathy in TYpe 2 diabetic patients with normoalbuminuria (PRIORITY): essential study design and rationale of a randomised clinical multicentre trial.
Diabetes mellitus affects 9% of the European population and accounts for 15% of healthcare expenditure, in particular, due to excess costs related to complications. Clinical trials aiming for earlier prevention of diabetic nephropathy by renin angiotensin system blocking treatment in normoalbumuric patients have given mixed results. This might reflect that the large fraction of normoalbuminuric patients are not at risk of progression, thereby reducing power in previous studies. A specific risk classifier based on urinary proteomics (chronic kidney disease (CKD)273) has been shown to identify normoalbuminuric diabetic patients who later progressed to overt kidney disease, and may hold the potential for selection of high-risk patients for early intervention. Combining the ability of CKD273 to identify patients at highest risk of progression with prescription of preventive aldosterone blockade only to this high-risk population will increase power. We aim to confirm performance of CKD273 in a prospective multicentre clinical trial and test the ability of spironolactone to delay progression of early diabetic nephropathy. Investigator-initiated, prospective multicentre clinical trial, with randomised double-masked placebo-controlled intervention and a prospective observational study. We aim to include 3280 type 2 diabetic participants with normoalbuminuria. The CKD273 classifier will be assessed in all participants. Participants with high-risk pattern are randomised to treatment with spironolactone 25 mg once daily, or placebo, whereas, those with low-risk pattern will be observed without intervention other than standard of care. Treatment or observational period is 3 years.The primary endpoint is development of confirmed microalbuminuria in 2 of 3 first morning voids urine samples. The study will be conducted under International Conference on Harmonisation - Good clinical practice (ICH-GCP) requirements, ethical principles of Declaration of Helsinki and national laws. This first new biomarker-directed intervention trial aiming at primary prevention of diabetic nephropathy may pave the way for personalised medicine approaches in treatment of diabetes complications. NCT02040441; Pre-results.


Mechanisms of atrial fibrillation: lessons from studies in patients.
Recent clinical studies performed in human subjects have provided important clues that improved our understanding of the mechanisms of atrial fibrillation (AF) and facilitated development of new treatment strategies. When analyzed together, these studies confirm the complexity and multifactorial nature of AF. Because a variety of mechanisms such as focal drivers within the thoracic veins, rotors in the left atrium, multiple reentrant circuits, and autonomic innervation may play a role in the initiation and maintenance of AF alone or in combination, the best strategy to eliminate AF may be the accurate identification of one or more of the mechanisms critical to the genesis of AF and to target the specific mechanism(s).


Preoperative risk factors for deep vein thrombosis in knee osteoarthritis patients undergoing total knee arthroplasty.
To analyze the risk factors for preoperative deep vein thrombosis (DVT) in knee osteoarthritis (OA) patients undergoing total knee arthroplasty (TKA). In this retrospective study, a total of 584 knee OA patients undergoing TKA were enrolled. General information, medical records and preoperative laboratory examination results of the patients were collected. According to the results of Doppler ultrasonography for the lower extremities, the patients were divided into DVT group and non-DVT group. Univariate and multivariate logistic regression were used to identify independent risk factors for preoperative DVT in knee OA patients undergoing TKA. The incidence of DVT before TKA was 6.85% (40 cases). The increase of ESR (Erythrocyte Sedimentation Rate), platelet crit, IL-6 (Interleukin-6), and PCT (Procalcitonin) were associated with the development of DVT before TKA. Factors as coronary heart disease (CHD), diabetes mellitus (DM), Chronic Kidney Disease (CKD), NLR (ratio of neutrophils to lymphocytes), lower limb venous blood stasis, the time from onset to admission, RBC, PLT were identified by multivariate logistic regression to be the independent risk factors for preoperative DVT in knee OA patients undergoing TKA. DM, CKD, CHD, increased in ESR, IL-6 and PCT, blood stasis of the lower extremities, increased in PLT, platelet crit and the time from onset to admission, decreased in RBC, were high risk factors for preoperative DVT in knee OA patients undergoing TKA.


Examiners' Perceptions in Surgical Education: The Blind Spot in the Assessment of OSCEs.
Interpreting objective structured clinical examination (OSCE) scores remains a challenging task.1 It remains unclear how examiners make a decision to score an OSCE the way they do. The examiners' thoughts and decision-making processes remain a hidden part of the OSCE assessment. One cannot assume that all examiners have a shared belief about what is important in an OSCE or what is the aim of an OSCE assessment. These differences are problematic, since they lead to students not getting the 'correct' score for their OSCE. Using a qualitative case study design,2 this study aims to gain in-depth insight into examiners' perceptions of OSCEs and their role as an examiner. For this purpose, semi-structured interviews with OSCE observers (n = 6) were conducted. The results show that the perception of examiners on the aim of OSCES and their motivation and task as examiner are crucial in understanding the differences among OSCE scores of examiners. Training and discussion among the examiners are needed to decrease the differences among the OSCE scores of examiners.


Effects of exogenous protease addition on fermentation and nutritive value of rehydrated corn and sorghum grains silages.
The study objective was to evaluate the effects of the addition of exogenous protease on the fermentation and nutritive value of rehydrated corn and sorghum grain silages during various storage periods. Treatments were applied using a 2 × 6 × 3 factorial combination, with 2 types of rehydrated grains (corn and sorghum), 6 doses of the enzyme (0, 0.3, 0.6, 0.9, 1.2, and 1.5%, based on natural matter) and 3 fermentation periods (0, 60, and 90 days) in a completely randomized design, with 4 replications. The protease aspergilopepsin I, of fungal origin, produced by Aspergillus niger, was used. The lactic acid concentration increased linearly as the enzyme dose increased in corn (CG) and sorghum (SG) grain silages, at 60 and 90 days of fermentation. There was an increase in the concentrations of ammonia nitrogen and soluble protein, as well as the in situ starch digestibility in rehydrated CG and SG silages, compared to the treatment without the addition of protease. The addition of 0.3% exogenous protease at the moment of CG ensiling and 0.5% in rehydrated SG increased the proteolytic activity during fermentation, providing an increase in in situ starch digestibility in a shorter storage time.


Adult wilms tumor in the renal pelvis: case report with review of the literature.
An 84-year-old man was referred to our hospital with gross hematuria. Abdominal computed tomography demonstrated a well-circumscribed enhanced mass in the right pelviureteral junction. Retroperitoneoscopic nephroureterectomy was performed because of a clinical diagnosis of renal pelvic carcinoma. Pathologic examination led to a final diagnosis of an adult Wilms tumor arising from the renal pelvis. This is the first report of a Wilms tumor in the renal pelvis of an adult patient.


Clinical trials. A pending subject.
Clinical trials are essential tools for the progress of clinical medicine in its diagnostic and therapeutic aspects. Since the first trial in 1948, which related tobacco use with lung cancer, there have been more than 150,000 clinical trials to date in various areas (paediatrics, cardiology, oncology, endocrinology, etc.). This article highlights the importance for all physicians to participate, over the course of their professional career, in a clinical trial, due to the inherent benefits for patients, the progress of medicine and for curricular prestige. The authors have created a synthesis of their experience with clinical trials on hypertension, diabetes, dyslipidaemia and ischaemic heart disease over the course of almost 3 decades. Furthermore, a brief reference has been made to the characteristics of a phase I unit, as well as to a number of research studies currently underway.


Tissue factor pathway inhibitor (TFPI) interferes with endothelial cell migration by inhibition of both the Erk pathway and focal adhesion proteins.
Tissue factor pathway inhibitor (TFPI) is a plasma Kunitz-type serine protease inhibitor that is mainly known for its inhibition of tissue factor-mediated coagulation. In addition to its anticoagulant properties, emerging data show that TFPI may also regulate endothelial cell functions via a non-haemostatic pathway. In this work we demonstrate that at concentrations within the physiological range, TFPI inhibits both endothelial cell migration and their differentiation into capillary-like structures in vitro. These effects were specific to endothelial cells since no inhibitory effect was observed on the migration of tumor (glioblastoma) cells. Inhibition of endothelial cell migration was correlated with a concomitant loss in cell adhesion, suggesting an alteration of focal adhesion complex integrity. Accordingly, we observed that TFPI inhibited the phosphorylation of focal adhesion kinase and paxillin, two key proteins involved in the scaffolding of these complexes, and that this effect was specific to endothelial cells. These results suggest that TFPI influences the angiogenic process via a non-haemostatic pathway, by downregulating the migratory mechanisms of endothelial cells.


Electrochemical behavior and determination of rutin on a pyridinium-based ionic liquid modified carbon paste electrode.
In this paper the electrochemical behavior of rutin on a pyridinium-typed ionic liquid modified carbon paste electrode (IL-CPE) was investigated and further used for rutin sample determination. The IL-CPE showed strong electrocatalytic effects to the oxidation of rutin. In phosphate buffer solution (PBS, pH 2.5; 0.1 M) a pair of well-defined cyclic voltammetric redox peaks of rutin appeared with the redox peak located at 512mV (Epa) and 448mV (Epc) (vs. SCE), respectively. The redox peak current was increased about 27.5 times more than that on traditional carbon paste electrode (CPE). The electrochemical parameters of rutin on the IL-CPE were calculated with the results of the charge transfer coefficient (alpha), the number of electron transfer (n) and the electrode reaction rate constant (k(s)) as 0.53, 1.80 and 2.39s(-1), respectively. The cathodic peak currents increased linearly with the concentration of rutin in the range from 5.0x10(-7) to 1.0x10(-4)M with the detection limit as 3.58x10(-7)M (3sigma). The relative standard deviation (RSD) of 10 successive detection of 5.0x10(-5)M rutin was 4.2%. The method was successfully applied to the determination of rutin content in tablets samples with good recovery. The modified electrode showed good stability and reproducibility without the influence of the coexisting substances.


Nitrogen sources alter ligninase and cellulase activities of thermophilic fungi isolated from compost and vermicompost.
Fungi harboring lignocellulolytic activity accelerate the composting process of agricultural wastes; however, using thermophilic fungal isolates for this process has been paid little attention. Moreover, exogenous nitrogen sources may differently affect fungal lignocellulolytic activity. A total of 250 thermophilic fungi were isolated from local compost and vermicompost samples. First, the isolates were qualitative assayed for ligninase and cellulase activities using Congo red (CR) and carboxymethyl cellulose (CMC) as substrates, respectively. Then, twenty superior isolates harboring higher ligninase and cellulase activities were selected and quantitatively assayed for both enzymes in basic mineral (BM) liquid medium supplemented with the relevant substrates and nitrogen sources including (NH4)2SO4 (AS), NH4NO3 (AN), urea (U), AS + U (1:1), or AN + U (1:1) with final nitrogen concentration of 0.3 g/L. The highest ligninase activities of 99.94, 89.82, 95.42, 96.25, and 98.34% of CR decolorization were recorded in isolates VC85, VC94, VC85, C145, and VC85 in the presence of AS, U, AS + U, AN, and AN + U, respectively. Mean ligninase activity of 63.75% in superior isolates was achieved in the presence of AS and ranked the highest among other N compounds. The isolates C200 and C184 exhibited the highest cellulolytic activity in the presence of AS and AN + U by 8.8 and 6.5 U/ml, respectively. Mean cellulase activity of 3.90 U/mL was achieved in AN + U and ranked the highest among other N compounds. Molecular identification of twenty superior isolates confirmed that all of them are belonging to Aspergillus fumigatus group. Focusing on the highest ligninase activity of the isolate VC85 in the presence of AS, the combination can be recommended as a potential bio-accelerator for compost production.


Design and performance of a high speed driver circuit for PIN diode switches used in microwave hyperthermia.
In many cancer treatment facilities where hyperthermia treatments are performed, there is a need to split a single channel microwave source into multiple, individually controllable channels. In the application reported here, microwave power is alternately switched between active and passive elements for each channel. The active element is either a microwave antenna inside a catheter in tumour tissue or a planar spiral applicator placed superficially, the total power to each channel being pulse width modulated from 1 to 99% of a 1 s duty cycle. The system is computer controlled and is capable of dividing and controlling power to 12 channels. PIN diode switch assemblies control the flow of power in each channel but they must be switched within a few microseconds to avoid failure at the high power levels used in the clinic. A high speed circuit was fabricated and tested to drive each PIN diode switch; the PIN diode switches are turned on in 1.5 microseconds and off in 3.0 microseconds, which meets the specifications for hot switching. The increase in speed over the former driver system is a factor of 5.1 for the on cycle and 1.5 x 10(5) for the off cycle and the maximum power tested in each channel was 80 W for 1 h at random duty cycles between 1 and 99%. The circuit was also tested with 40 W of power at duty cycles of 1, 50 and 99% for 1 h each; no failures or performance decrements were observed. The 12 channel circuit driver has been used for six months of clinical treatments without failure.


Sc-47 production from titanium targets using electron linacs.
In this work we have studied the feasibility of photonuclear production of (47)Sc from (48)Ti via (48)Ti(γ,p)(47)Sc reaction. Photon flux distribution for electron beams of different energies incident on tungsten converter was calculated using MCNPX radiation transport code. (47)Sc production rate dependence on electron beam energy was found and (47)Sc yields were estimated. It was shown that irradiating a natural Ti target results in numerous scandium isotopes which can reduce the specific activity of (47)Sc. Irradiating enriched (48)Ti targets with a 22MeV 1mA beam will result in hundreds of MBq/g activity of (47)Sc and no other isotopes of scandium. Decreasing the size of the target will result in much higher average photon flux through the target and tens of GBq/g levels of specific activity of (47)Sc. Increasing the beam energy will also result in higher yields, but as soon as the electron energy exceeds the (48)Ti(γ,np)(46)Sc reaction threshold, (46)Sc starts being produced and its fraction in total scandium atoms grows as beam energy increases. The results of the simulations were benchmarked by irradiating natural titanium foil with 22MeV electron beam incident on the tungsten converter. Measured (47)Sc activities were found to be in very good agreement with the predictions.


Optimizing and applying high-resolution, in-line laboratory phase-contrast X-ray imaging for low-density material samples.
In-line, or propagation-based phase-contrast X-ray imaging (PBI) is an attractive alternative to the attenuation-based modality for low-density, soft samples showing low attenuation contrast. With the growing availability of micro- and nano-focus X-ray tubes, the method is increasingly applied in the laboratory. Here, we discuss the technique and demonstrate its advantages for selected low-density, low attenuation material samples using a lab-based micro- and nano-computed tomography systems Easytom XL Ultra, providing micron and sub-micron range resolution PBI images. We demonstrate a multi-step optimization of the lab-based PBI technique on our scanner that includes choosing the optimal detector-source hardware combination available in the setup, then optimizing the imaging geometry and finally the phase retrieval process through a parametric study. We point out and elaborate on the effect of noise correlation and texturing due to phase retrieval. We demonstrate the overall benefits of using the phase image and the phase retrieval for the selected samples such as improved image quality, increased contrast-to-noise ratio while only marginally influencing the spatial resolution. The improvement in image quality also enables further image processing steps for detailed structural analysis of the samples, which would be much more complicated if not impossible based on the transmission image.


Sympathomimetic amines effectively control pain for interstitial cystitis that had not responded to other therapies.
To further investigate the efficacy of treatment of interstitial cystitis that had been refractory to standard treatment with sympathomimetic amines. Dextroamphetamine sulfate sustained release capsules up to 30 mg per day were prescribed in women with refractory painful bladder syndrome/interstitial cystitis in six new cases. The patients were carefully evaluated for relief of symptoms. All six women found marked relief in their painful bladder syndrome in a rather short length of time. The benefit persisted as long as the therapy was maintained. Temporary cessation resulted in prompt return of symptoms, but resumption of sympathomimetic amines again allowed good relief of bladder pain and related symptoms. Because of very few side-effects and no drug dependence in the dosage used, sympathomimetic amines should be considered for first-line therapy.


Molecular Docking as a Potential Approach in Repurposing Drugs Against COVID-19: a Systematic Review and Novel Pharmacophore Models.
This article provides a review of the recent literature related to the FDA-approved drugs that had been repurposed as potential drug candidates against COVID-19. Moreover, we performed a quality pharmacophore study for frequently studied targets, namely, the main protease, RNA-dependent RNA polymerase, and spike protein. Ever since the COVID-19 pandemic, the whole spectrum of scientific community is still unable to invent an absolute therapeutic agent for COVID-19. Considering such a fact, drug repurposing strategies seem a truly viable approach to develop novel therapeutic interventions. Drug repurposing explores previously approved drugs of known safety and pharmacokinetics profile for possible new effects, reducing the cost, time, and predicting prospective side effects and drug interactions. COVID-19 virulent machinery appeared similar to other viruses, making antiviral agents widely repurposed in pursuit for curative candidates. Our main protease pharmacophoric study revealed multiple features and could be a probable starting point for upcoming research.


Posttranscriptional repression of Escherichia coli OmpF protein in response to redox stress: positive control of the micF antisense RNA by the soxRS locus.
The soxRS regulon is a cornerstone of the adaptive defense systems of Escherichia coli against oxidative stress. Unexpectedly, activation of this regulon also enhances bacterial resistance to multiple antibiotics that seem unrelated to oxygen radicals. We previously correlated this multiple antibiotic resistance with a reduced rate of synthesis of the OmpF outer membrane porin that does not affect the OmpC or OmpA porins. Studies presented here, with operon and gene fusions of ompF to lacZ, show that the soxRS-dependent repression of OmpF is achieved posttranscriptionally. We also show posttranscriptional repression of OmpF mediated by the soxQ1 mutation, which maps to the marA locus. These repressions are dependent on the micF gene, which encodes a small RNA partially complementary to the 5' end of the ompF message. Northern (RNA) blotting experiments show that micF transcription is strongly inducible by the superoxide-generating agent paraquat in a manner that depends completely on the soxRS locus. The soxR-constitutive and soxQ1 mutations elevate the expression of micF in the absence of redox stress. However, the antibiotic resistance mediated by a soxR-constitutive mutation was only partially reversed upon deletion of micF. The soxRS regulon therefore includes other components that contribute to general antibiotic resistance, although the relation of this phenotype to oxidative stress remains to be established.


Anti-inflammatory, antimicrobial and antioxidant activities of Diospyros bipindensis (Gürke) extracts and its main constituents.
Diospyros bipindensis (Gürke) stem bark is used in Cameroon by Baka Pygmies for the treatment of respiratory disorders. To assess the anti-inflammatory, antibacterial and antioxidant properties of constituents from the bark extracts through bioassay-guided fractionation. The anti-inflammatory activity of extracts, fractions and pure compounds was assessed through the inhibition of the pro-inflammatory mediator nuclear factor-kappa B (NF-κB) transcriptional activity and nitric oxide (NO) production. DPPH, ABTS and ORAC assays were used for determining the antioxidant properties. The activity against Streptococcus pneumoniae, Staphylococcus aureus, Streptococcus pyogenes, Escherichia coli and Klebsiella pneumoniae, was evaluated on the basis of the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) by the macrodilution method. The water extract showed antimicrobial activity against S. pneumoniae (MIC: 300 μg/ml) and S. pyogenes (MIC: 300 μg/ml). The dichloromethane extract efficiently inhibited NF-κB transcriptional activity and NO production and exhibited significant antioxidant activity in the ORAC assay. An interesting activity was also found against S. pneumoniae (MIC: 200 μg/ml), S. aureus (MIC: 400 μg/ml) and S. pyogenes (MIC: 200 μg/ml). The phytochemical investigation of the dichloromethane extract afforded plumbagin, canaliculatin, ismailin, betulinic acid and 4-hydroxy-5-methyl-coumarin as the main constituents. Plumbagin and ismailin were found to be responsible for the main biological activities observed. These results may provide a rational support for the traditional use of Diospyros bipindensis stem bark in the treatment of respiratory disorders, since the anti-inflammatory, antimicrobial and antioxidant compounds isolated from the dichloromethane extract were also present in the traditional water extract.


Optimization of two-stage pretreatment for maximizing ethanol production in 1.5G technology.
Two-stage pretreatment conditions were optimized to convert corn fiber, separated from whole stillage in a corn dry grind ethanol plant, to fermentable sugars via hydrolysis. Liquid hot water pretreatment (25% solids) at 180 °C for 10 min, followed by three cycles of disk milling, provided maximum glucose, xylose, and arabinose yields of 88.5%, 41.0%, and 30.4% respectively after hydrolysis with Cellulase I. The glucose, xylose, and arabinose yields with Cellulase II at optimum conditions were 94.9%, 74.2%, and 66.3%, respectively. SSF of corn fiber using engineered yeast, with both Cellulase I and II, provided maximum ethanol concentrations of 2.13% and 2.73% (v/v). The protein content in the residual solid after fermentation was 47.95% and 52.05% for Cellulase I and II, respectively. This technology provides additional ethanol in a dry grind plant by converting corn fiber into ethanol and increases the protein content of DDGS, thereby improving the quality.


Soft tissue studies with 805 nm diode laser radiation: thermal effects with contact tips and comparison with effects of 1064 nm Nd:YAG laser radiation.
This study compares the perioperative depths of thermal coagulation, charring, and incision in rabbit liver, internal anterior abdominal wall skeletal muscle, and abdominal skin and in swine liver and abdominal skin obtained with 805 nm diode laser and 1,064 nm Nd:YAG laser radiation using 300-microns-diameter conical-tip and 1,200-microns-diameter spherical-tip contact fibers by hand. Additionally, the total depth of tissue necrosis surrounding incisions made with both lasers and contact tips is determined 48 hours postoperatively in the three tissues, and healing of the liver and abdominal wall muscle 21 and 35 days postoperatively is assessed histologically. Perioperatively determined charring, coagulation, and incision depth obtained in all tissues with either 805 nm or 1,064 nm laser radiation were sensibly equivalent at equal laser power values for each of the two contact tip shapes tested. At equal laser power values, coagulation depths obtained in rabbit abdominal skin using the 300-microns-diameter conical tip differed significantly (P < or = 0.01) from those values obtained with the 1,200-microns-diameter spherical tip. Incision depths obtained with the two different contact tip shapes at equal laser power settings in the different tissues studied differed in a few instances with no apparent pattern relating to tissue type or laser power. Depth of incisions obtained with both laser and tip types increases in the range of 6-12 w, but plateaus in the range 12-18 w in the tissues studied. Incisions obtained with both diode and Nd:YAG laser contact were essentially hemostatic, with self-limiting oozing at most.(ABSTRACT TRUNCATED AT 250 WORDS)


Effects of childhood adversity and resilience on Taiwanese youth health behaviors.
Adverse childhood experiences (ACEs) can leave negative impacts on one's health behaviors or social functioning later in life. Resilient characteristics have been shown to mitigate effects against risk behaviors in developing adolescents. However, clinical and research attention has rarely been given to jointly consider the effects of ACEs and resilient characteristics on health behaviors in Taiwanese youth. A total of 200 individuals aged 15-22 years were recruited from primary care settings, communities, and schools. Participants completed questionnaires assessing their ACEs, resilient characteristics, and health behaviors. Univariate analysis was firstly used to describe the correlates of ACEs and resilient characteristics. Further multivariate logistic regression analysis was used to examine the association of both factors with health behaviors. More than half (61.5%) of those surveyed had been exposed to at least one category of ACE. Verbal (37%) and physical (21%) abuses were the most common types of ACEs. The counts in the ACE categories were associated with being involved in physical fights (odds ratio 1.28 [confidence interval 1.01-1.63]), property damage (1.29 [1.03-1.61]), running away from home (1.30 [1.05-1.60]), bullying victimization (1.37 [1.16-1.61]), and sleep problems/tiredness (1.25 [1.03-1.52]). Meanwhile, resilience scores were associated with decreased odds of infrequent seatbelt use (0.47 [0.23-0.97]), low fruit and vegetable intake (0.42 [0.21-0.86]) unsatisfied body image (0.46 [0.22-0.97]), and sleep problems/tiredness (0.37 [0.18-0.79]). ACEs and resilience characteristics play a significant role in shaping youth health behaviors. Further research should be undertaken to identify ways to build resilience against health risks in youth with prior ACE exposure.


Angiographically proven cervical venous engorgement: a possible concurrent cause in the pathophysiology of Hirayama's myelopathy.
The objective of this study is to discuss the possible role of cervical posterior epidural plexus engorgement during cervical flexion in the pathogenesis of Hirayama myelopathy. In Hirayama disease, MRI during neck flexion often shows that the posterior dura detaches from the posterior arches compressing the spinal cord. Autopsies demonstrated asymmetric changes in the anterior horns consistent with chronic ischemic damage, attributed to arterial insufficiency during flexion or to microcirculatory changes due to compression by the tight dura. In a 15-year-old patient with 5-year history of distal upper limbs weakness, MRI demonstrated marked venous engorgement of the posterior epidural plexus in cervical flexion, confirmed by angiography. Laminectomy from C3 to C6 with duraplasty was performed. At one-year follow-up, the clinical condition of the patient remained stable. In Hirayama myelopathy, compression of the spinal cord by the tight dura is probably the most important pathogenetic factor. However, venous congestion in flexion might play an additional role in determining spinal cord ischemic changes.


A novel microtube culture system that enhances the in vitro development of parthenogenetic murine embryos.
Oil is an indispensable material in micro-droplet culture; it prevents medium from evaporation, and its transparency facilitates monitoring. However, lipophilic factors in the medium can be absorbed into the oil overlay, and conversely, deleterious materials can diffuse into the medium. In the present study, we describe a novel oil-free microtube culture (MTC) system. Parthenogenetic mouse embryos were placed into 0.2-mL thin-wall flat cap PCR tubes and cultured to the blastocyst stage. Conventional drop culture was used as a control. Embryos in MTC had a higher blastocyst formation rate (89.2%) and larger population of cells in the blastocysts (92.0+/-6.9; mean+/-S.E.M.) compared with drop culture (78.3% and 74.7+/-8.1; P<0.05 for each). The large blastocyst cell population in MTC was due to higher numbers of trophectoderm (TE) cells (70.5+/-5.9 versus 53.8+/-7.4; P<0.05) rather than inner cell mass cells. The presence of more TE cells was attributed to faster development in MTC. Embryos cultured in oil-covered MTC had fewer TE cells (61.5+/-5.6) than oil-free cultures (70.5+/-5.9; P<0.05). In conclusion, oil-free MTC was an alternative to conventional micro-drops, without the deleterious effects of oil.


[Synchronous bilateral breast cancer and pregnancy: about 3 cases at Joliot Curie institute of Dakar (Senegal)].
To describe our diagnostic and therapeutic strategy against synchronous, bilateral breast cancer during the pregnancy. Gestational age at diagnosis of cancer was 7; 21 and 25 weeks respectively. Patients had stage IV and IIIA of breast cancer in two and one case respectively. They all received chemotherapy, two cases during pregnancy (6TEC and 3AC) and one case after delivery. Bilateral mastectomy was performed in one case. One patient died. The others were alive but all metastatic. Fetal growth restriction was noted in one case. This association leads to delayed diagnosis of cancer. Surgery is feasible and the type of intervention is only determined by the cancer stage. Chemotherapy is feasible and is associated with less foetal complications in the last two quarters of pregnancy.


Histocompatibility reaction in tissue and cells of the marine sponge Suberites domuncula in vitro and in vivo: central role of the allograft inflammatory factor 1.
Sponges (Porifera) are the phylogenetically oldest still extant metazoan phylum. Recently elements of their immune system have been cloned and analyzed, primarily from the demosponges Suberites domuncula and Geodia cydonium. By differential display, two genes were identified in S. domuncula, whose translation products are involved in graft rejection/fusion: the allograft inflammatory factor (AIF-1) and the Tcf-like transcription factor (TCF). Since the AIF-1 and TCF genes are upregulated in vivo after tissue transplantation, especially in allografts, we investigated whether this reaction can be monitored in vitro. Therefore, the autogeneic and the allogeneic mixed sponge cell reaction (MSCR) system was applied for the first time to identify distinct factors in sponges in vitro. The results confirm that the two AIF-1 and TCF genes are induced during allogeneic MSCR. Furthermore, the recombinant sponge AIF-1 causes an upregulation of the expression of the TCF. We conclude that the AIF-1 and TCF genes are upregulated in sponges during histoincompatibility reactions; the data support the view that sponges have immune systems composed of highly complex elements related to those found in mammalian systems.


Associations between serum vitamin D status and the cardiometabolic profile of patients with obstructive sleep apnea.
Obstructive sleep apnea (OSA) and the metabolic syndrome (MetS) frequently coexist. Low serum vitamin D has been positively associated with OSA presence and severity; however, data on its link to cardiometabolic features in patients with OSA remain scarce. We aimed to assess serum 25-hydroxyvitamin D [25(OH)D] and explore its association with cardiometabolic parameters in OSA. This was a cross-sectional study among 262 patients (49 ± 9 years old, 73% men) with polysomnography-diagnosed OSA. Participants were evaluated in terms of anthropometric indices, lifestyle habits, blood pressure, biochemical, plasma inflammatory and urinary oxidative stress markers, and the presence of MetS. Serum 25(OH)D was assessed by chemiluminescence, and vitamin D deficiency (VDD) was defined as 25(OH)D < 20 ng/mL. Median (1st, 3rd quartile) serum 25(OH)D levels were 17.7 (13.4, 22.9) ng/mL and 63% of participants had VDD. Serum 25(OH)D correlated negatively with body mass index (BMI), homeostasis model of assessment of insulin resistance (HOMA-IR), total cholesterol, low-density lipoprotein cholesterol, triglycerides, high-sensitivity C-reactive protein (hsCRP), and urinary oxidized guanine species (oxG), and positively with high-density lipoprotein cholesterol (all P < 0.050). In logistic regression analysis, serum 25(OH)D was associated with lower odds of MetS [odds ratio (95% confidence interval): 0.94 (0.90-0.98)], after adjustment for age, sex, season of blood sampling, Mediterranean diet score, physical activity, smoking, apnea-hypopnea index, HOMA-IR, hsCRP, and oxG. In the same multivariate model, VDD was associated with ~ twofold greater odds of MetS [2.39 (1.15, 4.97)]. VDD is highly prevalent and is associated with a detrimental cardiometabolic profile among patients with OSA.


Intact mass spectrometry screening to optimize hydroxyl radical dose for protein footprinting.
Hydroxyl radical protein footprinting (HRPF) using synchrotron radiation is a well-validated method to assess protein structure in the native solution state. In this method, X-ray radiolysis of water generates hydroxyl radicals that can react with solvent accessible side chains of proteins, with mass spectrometry used to detect the resulting labeled products. An ideal footprinting dose provides sufficient labeling to measure the structure but not so much as to influence the results. The optimization of hydroxyl radical dose is typically performed using an indirect Alexa488 fluorescence assay sensitive to hydroxyl radical concentration, but full evaluation of the experiment's outcome relies upon bottom-up liquid chromatography mass spectrometry (LC-MS) measurements to directly determine sites and extent of oxidative labeling at the peptide and protein level. A direct evaluation of the extent of labeling to provide direct and absolute measurements of dose and "safe" dose ranges in terms of, for example, average numbers of labels per protein, would provide immediate feedback on experimental outcomes prior to embarking on detailed LC-MS analyses. To this end, we describe an approach to integrate intact MS screening of labeled samples immediately following exposure, along with metrics to quantify the extent of observed labeling from the intact mass spectra. Intact MS results on the model protein lysozyme were evaluated in the context of Alexa488 assay results and a bottom-up LC-MS analysis of the same samples. This approach provides a basis for placing delivered hydroxyl radical dose metrics on firmer technical grounds for synchrotron X-ray footprinting of proteins, with explicit parameters to increase the likelihood of a productive experimental outcome. Further, the method directs approaches to provide absolute and direct dosimetry for all types of labeling for protein footprinting.


The Transcriptome of Schistosoma mansoni Developing Eggs Reveals Key Mediators in Pathogenesis and Life Cycle Propagation.
Schistosomiasis, the most important helminthic disease of humanity, is caused by infection with parasitic flatworms of the genus Schistosoma. The disease is driven by parasite eggs becoming trapped in host tissues, followed by inflammation and granuloma formation. Despite abundant transcriptome data for most developmental stages of the three main human-infective schistosome species-Schistosoma mansoni, S. japonicum and S. haematobium-the transcriptomic profiles of developing eggs remain under unexplored. In this study, we performed RNAseq of S. mansoni eggs laid in vitro during early and late embryogenesis, days 1-3 and 3-6 post-oviposition, respectively. Analysis of the transcriptomes identified hundreds of up-regulated genes during the later stage, including venom allergen-like (VAL) proteins, well-established host immunomodulators, and genes involved in organogenesis of the miracidium larva. In addition, the transcriptomes of the in vitro laid eggs were compared with existing publicly available RNA-seq datasets from S. mansoni eggs collected from the livers of rodent hosts. Analysis of enriched GO terms and pathway annotations revealed cell division and protein synthesis processes associated with early embryogenesis, whereas cellular metabolic processes, microtubule-based movement, and microtubule cytoskeleton organization were enriched in the later developmental time point. This is the first transcriptomic analysis of S. mansoni embryonic development, and will facilitate our understanding of infection pathogenesis, miracidial development and life cycle progression of schistosomes.


On the use of mercury as a means of locating background sources in ultra low-background HPGe-detector systems.
In low-level gamma-ray spectrometry, it is common to measure large samples in order to obtain low detection limits for the massic activity (in mBq/kg). These samples have significant shielding effects. In order to study whether the background sources in three ultra low-background HPGe detectors were located in the detector or in the shield, Marinelli beakers filled with hyperpure mercury were measured. Although the measurements were hampered by the presence of cosmogenically produced (194)Hg, information regarding the major background location of (40)K, (60)Co, (137)Cs, (210)Pb, (226)Ra, (228)Ra and (228)Th could be obtained.


Vocal imprecision as a universal constraint on the structure of musical scales.
Theories of the origin of musical scales from the ancient Greeks to the present day have assumed that the intervals comprising scales are defined by specific mathematical ratios. Such theories are predicated on pre-tunable instruments, and yet the voice is almost certainly the original musical instrument. Therefore, the analysis of vocal scales offers a more naturalistic approach to understanding the origin of musical scales. In the present study, we conducted a large-scale computational analysis of vocal pitch-class properties and their implications for scale structure. We analyzed 418 field recordings of solo, unaccompanied songs from across 10 principal musical-style regions of the world. The results revealed a mean vocal pitch-class imprecision of approximately 1.5 semitones, and comparable results were obtained across all regions. These results suggest that vocal imprecision is universal and is mainly derived from the physiological limitations of the voice. Such vocal imprecision fundamentally constrains the formation of musical scale structure: it provides a lower limit on the spacing between adjacent scale tones and thus an upper limit on the number of scale tones that an octave can contain. We discuss these results in terms of an Interval Spacing model of the evolution of musical scales.


Radiology Malpractice Claims in the United States From 2008 to 2012: Characteristics and Implications.
The aim of this study was to compare the frequency and liability costs associated with radiology malpractice claims relative to other medical services and to evaluate the clinical context and case disposition associated with radiology malpractice claims. This HIPAA-compliant study was exempted from institutional review board approval. The Comparative Benchmarking System database, a repository of more than 300,000 medical malpractice cases in the United States, was queried for closed claims over a five-year period (2008-2012). Claims were categorized by the medical service primarily responsible for the claim and the paid total loss. For all cases in which radiology was the primary responsible service, the case abstracts were evaluated to determine injury severity, claimant type by setting, claim allegation, process of care involved, case disposition, modality involved, and body section. Intracategory comparisons were made on the basis of the frequency of indemnity payment and total indemnity payment for paid cases, using χ(2) and Wilcoxon rank-sum tests. Radiology was the eighth most likely responsible service to be implicated in a medical malpractice claim, with a median total paid loss (indemnity payment plus defense cost plus administrative expense) per closed case of $30,091 (mean, $205,619 ± $508,883). Radiology claims were most commonly associated with high- and medium-severity injuries (93.3% [820 of 879]; 95% confidence interval [CI], 91.7%-94.95%), the outpatient setting (66.3% [581 of 876]; 95% CI, 63.0%-69.2%), and diagnosis-related allegations (ie, failure to diagnose or delayed diagnosis) (57.3% [504 of 879]; 95% CI, 54.0%-60.6%). A high proportion of claims pertained to cancer diagnoses (44.0% [222 of 504]; 95% CI, 39.7%-48.3%). A total of 62.3% (548 of 879; 95% CI, 59.1%-65.5%) of radiology claims were closed without indemnity payments; 37.7% (331 of 879; 95% CI, 34.5%-40.9%) were closed with a median indemnity payment of $175,000 (range, $112-$6,691,762; mean $481,094 ± $727,636). Radiology malpractice claims most commonly involve diagnosis-related allegations in the outpatient setting, particularly cancer diagnoses, with approximately one-third of claims resulting in payouts to the claimants.


Outcome of Hamstring Autograft With Preserved Insertions Compared With Free Hamstring Autograft in Anterior Cruciate Ligament Surgery at 2-Year Follow-up.
To compare mechanical stability, functional outcome, and level of return to sports activity in patients undergoing anterior cruciate ligament (ACL) reconstruction with a free hamstring graft versus a graft with preserved insertions at 2-year follow-up. This study was a prospective, single-blind (the evaluator was blinded), randomized trial of 110 adult professional athletes who were randomly allocated into 2 groups. Group 1 consisted of 55 patients who underwent ACL reconstruction with hamstring tendon autograft with preserved insertions (technique 1), and group 2 consisted of 55 patients who underwent ACL reconstruction with free hamstring tendon autograft (technique 2). An anteromedial portal was used for drilling of the femoral tunnel in all cases. Patients were assessed for a minimum follow-up of 2 years with clinical tests, the Activities of Daily Living Function Scale and Sports Function Scale (Cincinnati knee score), knee arthrometer (KT-1000) testing, and the Tegner activity scale. The average age of the patients was 27.0 ± 7.5 years in group 1 and 27.2 ± 5.7 years in group 2. At 24 months, the mean side-to-side difference by KT-1000 testing was 1.4 in group 1 and 2.2 in group 2 (P < .0001); the mean Cincinnati knee score (Activities of Daily Living Function Scale and Sports Function Scale) was 418.5 (median, 420; range, 400-420) and 406.8 (median, 420; range, 350-420), respectively (P < .0001); and the mean difference between the preinjury and postsurgery Tegner level of sports activity was 0.3 and 1.08, respectively (P = .027). Although ACL reconstruction using hamstring autograft with preserved insertions resulted in statistically superior anterior stability, a better functional outcome, and a closer return to the preinjury level of sports activity as compared with free autograft, no clinically significant difference was proved. Level I, randomized controlled trial.


Efficacy and Safety of Lenvatinib for Patients With Advanced Hepatocellular Carcinoma: A Retrospective, Real-world Study Conducted in Japan.
We evaluated real-world efficacy and toxicity of lenvatinib in 142 patients with advanced hepatocellular carcinoma (HCC) at six tertiary referral centres. The patients with advanced HCC treated with lenvatinib were grouped into two categories based on REFLECT criteria for analysis of efficacy and safety. The primary endpoint was progression-free survival (PFS). The objective response rate (ORR) at week 12 of therapy was 41.5%, with a median PFS of 176 days. Child-Pugh score of 5 points, the presence of extrahepatic metastasis and adverse effects grade 2 or higher were considered independent factors associated with both better PFS and ORR. The ORR for patients who fulfilled the REFLECT inclusion criteria was significantly higher than that for those who did not. However, no significant differences in PFS were observed between the two groups. The incidence rate of adverse effects grade 3 or higher was 40.1%, which was similar for the two groups. Lenvatinib is safe and effective for patients, whether or not they satisfy REFLECT criteria. The result warrants replication in a larger study.


Anatomical and histological study of Liliequist's membrane: with emphasis on its nature and lateral attachments.
In previous studies, some disagreements regarding the nature (inner or outer arachnoid membrane) and lateral boundaries (temporal uncus or tentorial edge) of Liliequist's membrane remain. The aim was to clarify whether Liliequist's membrane is an inner or outer arachnoid membrane, and the distribution of Liliequist's membrane with emphasis on its lateral attachments. Liliequist's membrane was investigated by microsurgical dissection in 24 formalin-fixed adult cadaver heads and by histological sections of sellar-suprasellar specimens from another four formalin-fixed adult cadaver heads. The results obtained in the present study indicated that 1) Liliequist's membrane arises from the basal arachnoid membrane and has two components: a basal part comprising a folding inner layer of the arachnoid mater and an attaching part consisting of accumulated arachnoid trabeculae; 2) similar histological features are also present in other inner arachnoid membranes with attachments on basal arachnoid membrane, demonstrating Liliequist's membrane is an inner arachnoid membrane; 3) laterally, Liliequist's membrane attaches to the anterior tentorial edge constantly and to the mesial temporal uncus in more than half; 4) the oculomotor nerve courses above Liliequist's membrane and is fixed on Liliequist's membrane by the oculomotor membrane, which can also attach on temporal uncus and should be differentiated from the true temporal attachments of Liliequist's membrane. Liliequist's membrane is an inner rather than outer arachnoid membrane. Understanding of its individual variation and topographic relationships with surrounding neurovascular and arachnoid structures is important for neurosurgical practice.


Assessing Colorectal Cancer Screening Barriers by Two Methods.
Colorectal cancer (CRC) is commonly diagnosed in the USA despite screening tests that have decreased CRC incidence and mortality. Finding the best method to identify patient-level screening barriers is important to improve CRC screening rates. A group-randomized trial was conducted among ten primary-care clinics. Clinics were randomized to a multi-level (clinic, provider, patient) CRC screening intervention or usual care (2007-2013). Subsequent to clinic- and provider-level interventions, a three-step, patient-level intervention was conducted. One step of the patient-level intervention was a CRC screening barriers counseling call conducted by a lay health advisor (LHA). During the call, two methods were used to identify CRC screening barriers. An open-ended question was used first to determine why participants had not completed screening (without probes). Subsequently, the LHA read a list of additional potential screening barriers and asked participants whether each barrier was applicable (with probes). A generalized estimating equation approach was used to compare the two methods. Participants (n = 109) were female (59%), had a mean age of 57.2 years, and were white (67%) or black (31%). Most participants had some college education or a college degree (79%), annual household income $30,000+ (60%), and health insurance (80%). The number of CRC screening barriers increased with probing compared to the open-ended question format (OR 2.10, 95% CI 1.92-2.31; p < 0.01). The ranking of reported CRC screening barriers did not vary by assessment method. However, the methodology used to document CRC screening barriers may influence the content of patient-directed interventions.


Polyunsaturated fatty acids augment free radical generation in tumor cells in vitro.
Polyunsaturated fatty acids (PUFAs) have been shown to inhibit both normal and tumor cell growth in vitro. As PUFAs are known to induce a respiratory burst and free radical generation in polymorphonuclear leukocytes and since free radicals are toxic to cells, we investigated the effect of PUFAs on a measure of free radical generation (nitroblue tetrazolium reduction) in normal human fibroblasts and breast cancer cells in vitro. Results suggested that linoleate (LA), gamma-linolenate (GLA), arachidonate (AA) and eicosapentaenoate (EPA) can enhance nitroblue tetrazolium reduction in tumor cells but not in normal cells. GLA, AA and EPA were 1 1/2 to 2 times more effective than LA in inducing free radical generation. This difference was not due to increased uptake of LA, AA and EPA by tumor cells. In fact, the uptake of LA was the same both in normal and tumor cells whereas that of AA and EPA occurred at approximately half the rate in the tumor cells compared to normal cells. This indicates that PUFA induced growth inhibition and cytotoxicity to tumor cells may, at least in part, be due to enhanced free radical generation.


Safety of biologics approved for treating rheumatoid arthritis: analysis of spontaneous reports of adverse events.
Despite the effectiveness of biologics approved for the treatment of rheumatoid arthritis, they have been associated with serious adverse events (AEs). Biologics are used under close supervision of health care professionals. In Portugal, they are legally required to report AEs occurring during the treatment. This study aims at investigating post-marketing safety monitoring data of biologics in Portugal by comparing the frequency of spontaneously reported adverse events between 2009 and 2011 with the frequency of such events in the summary of the product characteristics of each biologic. Sales data for biologics were obtained from IMS Health and converted into defined daily doses/1,000 inhabitants/day in order to estimate a proportion of the population treated. The frequency of AEs was estimated as the percentage of patients in which an AE may have occurred. The use of each biologic was estimated for adalimumab at 1,439 patients/year, etanercept 1,944 patients/year, and infliximab 3,211 patients/year. A total of 992 AEs were reported: 207 for adalimumab, 199 for etanercept, and 586 for infliximab. Of the 515 different spontaneously reported AEs, 194 were included for comparisons with the SPCs. Of those, 31 (16 %) were similarly frequent, and 163 (84.0 %) occurred less frequently compared with SPCs' data. These results suggest an insufficient post-marketing safety monitoring of biologics in Portugal.


Decreased sialic acid content of erythrocytes in patients with aplastic anaemia.
The sialic acid content of erythrocytes from patients with aplastic anaemia was determined and compared with those in patients with several haematological disorders and healthy individuals. The sialic acid was released enzymatically with Vibrio cholerae sialidase and quantitated by the thiobarbituric acid method (Aminoff, 1961). The sialic acid content of normal erythrocytes was 538 +/- 31 nmol/ml of packed erythrocytes. That of erythrocytes from patients with aplastic anaemia was 480 +/- 35 nmol/ml of packed erythrocytes, which was significantly lower than normal (P less than 0.01). In contrast, erythrocytes from patients with myeloproliferative disorders showed significantly (P less than 0.05) higher sialic acid contents (564 +/- 45 nmol/ml of packed erythrocytes). These results suggest that some membrane changes occur in erythrocytes in patients with these disorders.


Assessment of the discriminative stimulus effects of cocaine in the rat: lack of interaction with opioids.
The present study examined the effects of several opioid agonists and antagonists in rats trained to discriminate cocaine (10 mg/kg) from saline in a two-lever, food-reinforced, discrimination task. Neither fentanyl, a mu agonist, nor the delta agonist BW 373U86 elicited cocaine-appropriate responding. Although pretreatment with fentanyl failed to alter the discriminative stimulus effects of low doses of cocaine, cocaine reversed the rate-suppressant effects of fentanyl. Although the kappa agonist U50,488H decreased response rates, it did not substitute for cocaine. Injection of U50,488H in combination with the training dose of cocaine (10 mg/kg) reversed the rate-suppressant effects of U50,488H but failed to affect the cocaine cue. Administration of U50,488H (3 mg/kg), in conjunction with several doses of cocaine, did not shift the cocaine dose-response curve. Naltrindole and naltrexone, delta and mu antagonists respectively, did not block the effects of cocaine. Further, naltrindole did not substitute for the cocaine cue. Complete generalization was observed to the dopamine uptake inhibitor bupropion (30 mg/kg). These results suggest that fentanyl and U50,488H, at doses that purportedly influence mesolimbic dopamine levels, do not alter the discriminative stimulus effects of cocaine. Moreover, activation of delta receptors and blockade of mu and delta receptors are similarly ineffective.


The influence of cellular amino acids and the Na+ : K+ pump on the membrane potential of the Ehrlich ascites tumor cell.
The membrane potential of the Ehrlich ascites tumor cell was shown to be influenced by its amino acid content and the activity of the Na+ :K+ pump. The membrane potential (monitored by the fluorescent dye, 3,3'-dipropylthiodicarbocyanine iodide) varied with the size of the endogenous amino acid pool and with the concentration of accumulated 2-aminoisobutyrate. When cellular amino acid content was high, the cells were hyperpolarized; as the pool declined in size, the cells were depolarized. The hyperpolarization seen with cellular amino acid required cellular Na+ but not cellular ATP. Na+ efflux was more rapid from cells containing 2-aminoisobutyrate than from cells low in internal amino acids. These observations indicate that the hyperpolarization recorded in cells with high cellular amino acid content resulted from the electrogenic co-efflux of Na+ and amino acids. Cellular ATP levels were found to decline rapidly in the presence of the dye and hence the influence of the pump was seen only if glucose was added to the cells. When the cells contained normal Na+ (approx. 30mM), the Na+ :K+ pump was shown to have little effect on the membrane potential (the addition of ouabain had little effect on the potential). When cellular Na+ was raised to 60mM, the activity of the pump changed the membrane potential from the range -25 to -30 mV to -44 to -63 mV. This hyperpolarization required external K+ and was inhibited by ouabain.


A sensitive staining method for detecting acidic polysaccharides in cellulose acetate and agarose gels.
A sensitive staining method has been developed for the detection of acidic polysaccharides in cellulose acetate and agarose gels. The method is based on the precipitation of bovine serum albumin by acidic polysaccharides at acidic pH values and the subsequent staining of precipitated protein with amido black or Coomassie brilliant blue R-250 stains. The detection limit of acidic polysaccharides is 15-40 ng on cellulose acetate strips and 50-150 ng on agarose plates. The sensitivity of the described staining technique is of the same order for a wide range of acidic polysaccharides of different origin in contrast to Alcian blue and toluidine blue stains, which detect only mucopolysaccharides of animal origin at comparable levels. The method was also applied to the colorimetric quantitative determination of acidic polysaccharides after electrophoretic separation.


Relation between centric slide and Angle's classification.
To compare the difference in sagittal cephalometric analysis in centric occlusion (CO) and in centric relation (CR), among Angle's classes i.e. class I, class II, and class III. Comparative, cross-sectional study. Orthodontic Outpatient Department, Karachi Medical and Dental College, Karachi, from July 2003 to December 2004. Eighty pre-treatment orthodontic patients were divided into 3 groups according to Angle's classification. The first lateral cephalogram was taken in CO. Then leaf gauges were placed in between incisors for deprogramming. The second cephalogram was taken with leaf gauges in place to obtain centric relation. Both the radiographs were traced. Angle ANB was measured for sagittal analysis. ANB angle compares the position of mandible with the maxilla relating it with the cranial base. Angles from both cephalograms were compared with each other. Significant difference (p<0.0001) was found in values of ANB in CO (2.93 degrees -/+3.7) and CR (4.88 degrees -/+3.4) cephalograms. Significant CO-CR discrepancy was found (p<0.05) in Angle's class III patients (3.05 degrees -/+0.55) in comparison to Angle's class I (1.96 degrees -/+0.94) and class II (1.65 degrees -/+0.65). CO-CR discrepancy can occur regardless of Angle's classification. Special emphasis should be given to Angle's class III patients for the centric slide.


Current postgraduate training in emergency medicine in the Nordic countries.
Emergency Medicine (EM) is an independent specialty in all five Nordic countries. This study aims to evaluate the structure of post-graduate EM training in the area. A leading hospital or hospitals in EM training in each country were identified. An e-survey was sent to each hospital to gather data on patient volume and physician staffing, curriculum, trainee supervision, and monitoring of progression in training. Data were collected from one center in Iceland and Norway, two in Finland and Sweden, and four centers in Denmark. The data from each country in Denmark, Finland, and Sweden, were pooled to represent that country. The percentage of consultants with EM specialist recognition ranged from 49-100% of all consultants working in the participating departments. The number of patients seen annually per each full time EM consultant was almost three times higher in Finland than in Sweden. In Iceland, Denmark, and Sweden a consultant was present 24/7 in the ED but not in all centers in the other countries. The level of trainee autonomy in clinical practice varied between countries. Requirements for completing standardized courses, completing final exams, scientific and quality improvement projects, and evaluation of trainee progression, varied between the countries. All Nordic countries have established EM training programs. Despite cultural similarities, there are significant differences in how the EM training is structured between the countries. Writing and implementing a standardized training curriculum and assessment system for EM training in the Nordic countries should be considered.


A common IGF1R gene variant predicts later life breast cancer risk in women with preeclampsia.
Preeclampsia has been inconsistently associated with altered later life risk of cancer. This study utilizes the Nurses' Health Study 2 (NHS2) to determine if the future risk of breast and non-breast cancers in women who experience preeclampsia is modified by carrying a protective variant of rs2016347, a functional insulin-like growth factor receptor-1 (IGF1R) single nucleotide polymorphism. This retrospective cohort study completed within the NHS2 evaluated participants enrolled in 1989 and followed them through 2015, with a study population of 86,751 after exclusions. Cox proportional hazards models both with and without the impact of rs2016347 genotype were used to assess the risk of invasive breast cancer, hormone receptor-positive (HR+) breast cancer, and non-breast cancers. Women with preeclampsia had no change in risk of all breast, HR+ breast, or non-breast cancers when not considering genotype. However, women carrying at least one T allele of rs2016347 had a lower risk of HR+ breast cancer, HR 0.67, 95% CI: 0.47-0.97, P = 0.04, with interaction term P = 0.06. For non-breast cancers as a group, women carrying a T allele had an HR 0.76, 95% CI: 0.53-1.08, P = 0.12, with interaction term P = 0.26. This retrospective cohort study found that women with preeclampsia who carry a T allele of IGF1R rs2016347 had a reduced future risk of developing HR+ breast cancer, and a reduced but not statistically significant decreased risk of non-breast cancers suggesting a possible role for the IGF-1 axis in the development of cancer in these women.


Plasma membrane-mediated leakage of liposomes induced by interaction with murine thymocytic leukemia cells.
The interaction of liposomes with BW 5147 murine thymocytic leukemia cells was studied using fluorescent probes (entrapped carboxyfluorescein and fluorescent phosphatidylethanolamine) in conjunction with a Ficoll-Paque discontinous gradient system for rapid separation of liposomes from cells. Reversible liposomal binding to discrete sites on the BW cell surface was found to represent the major form of interaction; uptake of intact liposomal contents by a process such as liposome-BW cell membrane fusion was found to apparently represent a minor pathway of interaction (2%). Liposomal lysis was found to be associated with the process of liposomal binding (perhaps as a result of the binding itself). Lysis was followed by release of the entrapped carboxyfluorescein into the media and its subsequent uptake by the cells. This lysis was shown to be dependent upon discrete membrane-associated sites that have some of the properties of proteins. The results of these studies suggest that liposomal binding to the cells, subsequent lysis of the liposomes and cellular uptake of their contents should be seriously considered in all studies of liposome-cell interactions as an alternate mode of interaction to the four modes (fusion, endocytosis, adsorption and lipid exchange) previously emphasized in the literature.


Two-dimensional hollow carbon skeleton decorated with ultrafine Co3O4 nanoparticles for enhanced lithium storage.
Transition metal oxides have shown high theoretical capacities as anode materials and have been considered as high potential materials to substitute graphite for composing new generations of lithium-ion batteries (LIBs). However, the considerable volume changes of transition metal oxide materials during practical processes have limited their applications. Herein, we report a simple approach to construct a two-dimensional (2D) hollow carbon skeleton decorated with ultrafine Co3O4 nanoparticles (Co3O4/C). This composite is derived from a leaf-like zeolitic imidazolate framework-L (ZIF-L (Co)) via etching coordination using tannic acid (TA). The Co3O4/C has a unique structure consisting of 2D carbon skeleton, ultrafine Co3O4 nanoparticle, and open channel, which can accelerate electron transport, alleviate volume change, and facilitate ion diffusion. Benefiting from these features, the LIBs assembled using Co3O4/C as anode material exhibits superior reversible cycle performance and impressive rate property. This study provides an efficient strategy for implementing transition metal oxide-based composites for energy storage applications.


Effects of excimer laser phototherapeutic keratectomy in limbal-conjunctival autograft transplantation for recurrent pterygium: a retrospective case control study.
Repeated surgery excisions could induce obvious irregular astigmatism in patients with recurrent pterygium. Our study is aimed to illustrate the effect of adjunct excimer laser phototherapeutic keratectomy (PTK) in limbal-conjunctival autograft transplantation on visual quality for patients with recurrent pterygium. Retrospective case-control study. Eyes that underwent pterygium excision with (PTK group) or without (control group) PTK from 2006 to 2017 were retrospectively included. Recurrence rate, preoperative and postoperative surface regularity index (SRI), surface asymmetry index (SAI), cylinder and LogMAR vision were collected. Postoperative anterior segment optical coherence topography and in vivo confocal microscopy were performed to monitor the cornea epithelium healing and cellular recovery process respectively. A total of 99 eyes of 99 patients were collected, of which 39 were treated with PTK and 60 without PTK. The mean follow-up time was 50.4 ± 38.1 months. The recurrence rate was 10.3% (4 eyes) in the PTK group and 13.3% (8 eyes) in the control group (p = 0.759). The SRI decreased 0.53 (range: - 0.88, 2.81), SAI decreased 0.53 (range: - 0.64, 2.94), and the cylinder decreased 2.08 (range:-0.16, 9.40) D in the PTK group, and the corresponding values were 0.48 (range:-0.45, 2.27), 0.27 (range:-1.06, 2.21) and 0.71 (range:-1.75, 3.55) D in the control group, respectively (Z = 1.76, 2.15, and 3.97, p = 0.005, 0.016, and 0.000 respectively). LogMAR vision improved in both groups after surgery, with an improvement of 0.18 (range: 0.00, 0.70) in the PTK group and 0.06 (range: - 0.12, 0.50) in the control group (Z = 4.08, p = 0.000). Besides, the eyes treated with PTK showed faster re-epithelization and better cellular recovery. For recurrent pterygium, surgical excision with adjunct PTK might be a better option with improved corneal surface and vision outcomes.


Detection of microscopic diffusion anisotropy in human cortical gray matter in vivo with double diffusion encoding.
To detect microscopic diffusion anisotropy in human cortical gray matter in vivo with double diffusion encoding experiments. Double diffusion encoding experiments were performed on a 3 T whole-body MR system using echo-planar imaging. Angular double diffusion encoding measurements were acquired with 8 × 8 and 12 × 12 planar direction combinations and were analyzed in three regions of interest containing white matter, mostly cortical gray matter, and one having significant contributions from cerebrospinal fluid. Inversion with variable recovery times served to estimate and eliminate white matter partial volume effects. To investigate the influence of magnetic field inhomogeneities, experiments with gradient offsets and cross-term compensated diffusion weightings were performed. The MA index, a rotationally invariant measure of the microscopic diffusion anisotropy, was determined from measurements with 96 direction combinations. The angular signal modulation in the gray matter region of interest has two components, one being consistent, inter alia, with cross terms with field inhomogeneities while the other represents a signal difference between parallel/antiparallel and orthogonal direction combinations, ie, the fingerprint of microscopic diffusion anisotropy. Based on the amplitudes and their dependency on the inversion time, white matter partial volumes can be excluded as the sole source for this modulation, providing strong evidence for the detection of microscopic diffusion anisotropy in cortical gray matter. MA maps of healthy volunteers show considerably lower values in cortical gray matter compared with white matter. Microscopic diffusion anisotropy can be measured in human cortical brain matter, which could help to characterize the microstructure of healthy and pathological tissue.


Effects of antibiotics and Saccharomyces boulardii on bacterial translocation in burn injury.
To investigate the effects of antibiotics and the probiotic, Saccharomyces boulardii, on indigenous microflora and bacterial translocation (BT) in burned rats. Twenty-three male albino rats were divided into a sham burn group (group 1, n = 7) exposed to 21 degrees C water, a burn + antibiotic group (group 2, n = 8), and a burn + antibiotic + S. boulardii group (group 3, n = 8) exposed to 95 degrees C water for 10 s, producing a full-thickness burn to 30% of the total body surface area. Ampicillin-sulbactam (1,000 mg/kg per day) was given as two doses via an orogastric feeding tube to groups 2 and 3. Saccharomyces boulardii (1 mg/g body weight per day) was given as two doses via the same route to group 3. All rats were killed on the fifth day postburn and cultures of the mesenteric lymph nodes, liver, spleen, blood, and cecal contents were done. The incidences of BT were 0% (0/7) in group 1, 87.5% (7/8) in group 2, and 37.5% (3/8) in group 3. A significant increase in the BT incidence was found in group 2 (P < 0.01), while a significant decrease was found in group 3 when compared with group 1. The total bacteria count of cecal flora was significantly lower in group 3 than in group 1 (P < 0.01). The decrease in Gram-negative bacteria in the cecal flora was significant in group 3. These results suggest that the incidence of BT in burn injury is enhanced by using an antibiotic, and that S. boulardii decreases the incidence of antibiotic-induced BT. Thus, we conclude that S. boulardii can effectively protect the intestinal ecologic equilibrium and prevent BT in burn injury victims.


Expression of T-cell receptor-γδ in normal human skin, inflammatory dermatoses and mycosis fungoides.
T cells expressing the γδ T-cell receptor (TCR) (γδ T cells) are found in normal epithelial tissues such as the skin. However, the proportions of γδ T cells that may be observed in commonly encountered cutaneous diseases with a prominent lymphocytic infiltrate have not been elucidated. Our pathology database was searched for cases of mycosis fungoides, erythema multiforme, graft-versus-host disease, lichen planus, lupus panniculitis and spongiotic dermatitis. Immunostaining for CD3, βF1 and the TCR γ chain was performed on formalin-fixed paraffin-embedded specimens retrieved from these cases to determine the normal range of γδ T cells in these diseases. In 100 of the 101 cases studied (99.0%), γδ T cells accounted for less than 10% of the T-cell infiltrate. Furthermore, γδ T cells were essentially absent in 74 cases (73.3%). These results suggest that γδ T cells very rarely account for more than 10% of the lymphocytic infiltrate in common inflammatory or infiltrative processes of the skin. Any case of suspected mycosis fungoides or interface dermatitis that possesses more than 10% γδ T cells should raise consideration for further investigation.


Two-dimensional bismuth oxybromide coupled with molybdenum disulphide for enhanced dye degradation using low power energy-saving light bulb.
In the present work, two-dimensional bismuth oxybromide (BiOBr) was synthesized and coupled with co-catalyst molybdenum disulphide (MoS2) via a simple hydrothermal process. The photoactivity of the resulting hybrid photocatalyst (MoS2/BiOBr) was evaluated under the irradiation of 15 W energy-saving light bulb at ambient condition using Reactive Black 5 (RB5) as model dye solution. The photo-degradation of RB5 by BiOBr loaded with 0.2 wt% MoS2 (MoBi-2) exhibited more than 1.4 and 5.0 folds of enhancement over pristine BiOBr and titanium dioxide (Degussa, P25), respectively. The increased photocatalytic performance was a result of an efficient migration of excited electrons from BiOBr to MoS2, prolonging the electron-hole pairs recombination rate. A possible charge transfer diagram of this hybrid composite photocatalyst, and the reaction mechanism for the photodegradation of RB5 were proposed.


Ammonium influx pathways into astrocytes and neurones of hippocampal slices.
Ammonium (NH(4) (+) ) is required to maintain pathways involved in shuttling metabolic precursors between astrocytes and neurones. Under hyperammonaemic conditions, increases in the cellular influx of NH(4) (+) , and accompanying changes in ion concentrations, may contribute to disruptions in metabolism and neurotransmission. We investigated mechanisms of cellular NH(4) (+) influx in hippocampal slices by measuring acute NH(4) (+) /NH(3) -evoked changes in intracellular pH (pH(i) ) and sodium ([Na(+) ](i) ). In both astrocytes and neurones, application of 5 mM NH(4) Cl for 30-45 min decreased pH(i) by 0.2-0.3 units, consistent with NH(4) (+) influx. In astrocytes, but not neurones, acidifications were accompanied by [Na(+) ](i) increases of 25-30 mM. Glial [Na(+) ](i) increases were blocked by bumetanide, suggesting that NH(4) (+) /NH(3) activated Na(+) -dependent, K(+) , Cl(-) cotransport. Bumetanide also reduced NH(4) (+) /NH(3) -evoked acidifications in astrocytes. Neuronal acidifications were insensitive to bumetanide and inhibition of Cl(-) -dependent transport and K(+) channels, but were prevented by inhibition of Na(+) ,K(+) -ATPase with ouabain. Furthermore, ouabain reduced astrocyte acidifications. Our results suggest that following rapid elevation of NH(4) (+) , Na(+) ,K(+) -ATPase is the major influx pathway for NH(4) (+) in neurones, whereas Na(+) ,K(+) -ATPase and Na(+) -dependent, K(+) , Cl(-) cotransport mediate NH(4) (+) transport into astrocytes. The different mechanisms of NH(4) (+) influx in astrocytes and neurones may contribute to the different susceptibility of both cell types to acute hyperammonaemic conditions.


Private sector involvement in the control of contagious bovine pleuropneumonia (CBPP) in the Kazungula district of Zambia benefitted the community and the control strategy.
Contagious bovine pleuropneumonia (CBPP) is a disease of economic importance that is widely distributed in sub-Saharan African and contributes significantly to cattle morbidity and mortality. Lack of resources to implement eradication measures has led to the disease becoming endemic in most areas in sub-Saharan Africa where governments have little resources and the majority of the people are poor. Usually, control and eradication of such diseases as CBPP is treated as a public good by governments and to achieve this, governments are usually assisted by nongovernment organisations, bilateral government programmes and international donors. The private sector, which usually is companies that run businesses to make profit, although not very well established in sub-Saharan Africa could play a big role in the eradication of CBPP in the region. This could play a dual role of promoting investment and also eradicate livestock diseases which have proved a menace in the livestock sector. This paper highlights the role played by the private sector in the control of CBPP in Zambia.


Heat shock of cultivated and wild wheats during early seedling stage: growth, cell viability and heat shock proteins.
Growth, cell viability and heat shock proteins (HSPs) in Bezostaya-1, Cukurova-86 and Diyarbakir-81 cultivated wheat cultivars and three Aegilops species were investigated. Etiolated seedlings were exposed to 23 degrees C, 32 degrees C, 35 degrees C, 37 degrees C and 38 degrees C for 24 h, and 35 degrees C (24 h) --> 50 degrees C (1 h) and 37 degrees C (24 h) --> (50 degrees C (1 h). At the end of recovery growth periods, the shoot lengths of the genotypes generally decreased significantly at 35, 37 and 38 degrees C. The acquired thermal tolerance (ATT) in intact seedlings was over 50% at 35 degrees C --> 50 degrees C and 37 degrees C --> 50 degrees C, but in cell viability test it ranged from 2.75% (Ae. triuncialis) to 32.87 (Bezostaya-1) at 35 degrees C, and from 2.82% (Ae. triuncialis) to 37.82 (Bezostaya-1) at 37 degrees C. Ae. triuncialis was most sensitive genotype in both ATT determination. In electrophoretic profiles of proteins, while some HSPs were newly synthesized, some normal cellular proteins disappeared at 37 degrees C and 37 degrees C -->50 degrees C compared to 23 degrees C. The number of low molecular weight (LMW) HSPs were more than intermediate- (IMW) and high- (HMW) HSPs. The genotypes had both common (12 HSPs between at least two genotypes) and genotype-specific (33 HSPs) LMW HSPs. The common HSP of 19.8 kDa (pI 6.5) was synthesized in Bezostaya-1, Cukurova-86, Diyarbakir-81, Ae. biuncialis and Ae. umbellulata. Bezostaya-1 is the only genotype that synthesized 12 IMW and 2 HMW HSPs at 37 degrees C --> 50 degrees C. Ae. triuncialis had only two common LMW HSPs [22.1 (pI 7.1) and 24.2 kDa (pI 6.5)].


Educational Interventions to Improve Advance Care Planning Discussions, Documentation and Billing.
To determine the impact of educational interventions, clinic workflow redesign, and quality improvement coaching on the frequency of advance care planning (ACP) activities for patients over the age of 65. Nonrandomized before-and-after study. 13 ambulatory care clinics with 81 primary care providers in eastern and central North Carolina. Patients across 13 primary care clinics staffed by 66 physicians, 8 physician assistants and 7 family nurse practitioners. Interprofessional, interactive ACP training for the entire interprofessional team and quality improvement project management with an emphasis on workflow redesign. From July 2017 through June 2018-number of ACP discussions, number of written ACP documents incorporated into the electronic medical record (EMR), number of ACP encounters billed. Following the interventions, healthcare providers were more than twice as likely to conduct ACP discussions with their patients. Patients were 1.4 times more likely to have an ACP document included in their electronic medical record. Providers were significantly (p < 0.05) more likely to bill for an ACP encounter in only one clinic. Implementing ACP education for all clinic staff, planning for workflow changes to involve the entire interprofessional team and supporting ACP activities with quality improvement coaching leads to statistically significant improvements in the frequency of ACP discussions, the number of ACP documents included in the electronic medical record and number of ACP encounters billed.


Essential fatty acids in serum lecithin of children with atopic dermatitis and in umbilical cord serum of infants with high or low IgE levels.
The fatty acid composition of serum lecithin from children with atopic dermatitis (AD) was found to be abnormal, characterized by significantly increased proportion of linoleic acid and reduced levels of metabolites of this fatty acid. The levels of linoleic acid in umbilical cord serum lecithin were significantly higher in babies with high serum IgE than in those with low or non-demonstrable serum IgE. Since elevated cord serum IgE is strongly associated with development of atopic disease the results suggest that fatty acid changes may be a basic feature of AD. Immunologic dysfunction in patients with AD may possibly partly be a consequence of the fatty acid abnormality.


Comparison of two prognostic models predicting survival in patients with malignant melanoma.
Accurate predictions of prognosis are important in the clinical management of patients with malignant melanoma. Primary lesions from 55 patients with cutaneous melanoma, having 10 or more years of clinical follow-up, were evaluated by two models predicting patient survival. One model was simple and relied solely on tumor thickness. The other model was complex and considered stage of tumor progression, and six clinical and histological variables. Accuracy of the two models was determined by retrospective review of the medical record, and the predictive power of each model was compared by receiver operating characteristic (ROC) curve analysis. The area under the ROC curve (a measure of the "goodness" of the model) for the single variable model was 0.70 +/- 0.0775 standard error (SE), and the area under the ROC curve for the multiple variable model was 0.77 +/- 0.0779 SE. Although a modest improvement in predictive power is suggested for the multiple variable model, the SEs for the two models overlap, and the difference is not statistically significant. Further study using a larger database may be required to determine definitively if the multiple variable model significantly increases the ability to predict patient survival or death, and make better clinical decisions.


Terson syndrome in reverse: intraventricular haemorrhage following primary intraocular haemorrhage.
Terson syndrome (TS) describes the presence of intraocular haemorrhage in patients with intracranial haemorrhage or traumatic brain injury. The aetiology of TS is controversial as an anatomical conduit between the vitreous humour and subarachnoid space remains contested. We herewith present a case of primary vitreous haemorrhage with secondary intracranial extension into the ventricles. Cranial CT demonstrates blood within the left optic nerve and chiasm but not within the subarachnoid space. This unusual phenomenon, which has not been reported before, may be described as 'Terson syndrome in reverse'. We explore mechanisms by which blood within the globe may track into the ventricular system, contextualising recent advances in the understanding of ocular-intracranial fluid transport.


N-Pyridinyl Sulfilimines as a Source for α-Imino Gold Carbenes: Access to 2-Amino-Substituted N-Fused Imidazoles.
Gold-catalyzed formal 1,3-dipolar annulation between readily accessible N-pyridinylsulfilimines and ynamides is reported. A diverse set of imidazole derivatives is prepared from the corresponding sulfilimines and ynamides. These functionalized cyclic products can undergo further transformations to afford diverse imidazole frameworks. Moreover, in situ synthesis is feasible and shows good potential in the synthesis of nucleoside analogues.


The MDM2 inhibitor Nutlin-3 modulates dendritic cell-induced T cell proliferation.
Nutlin-3, a small molecule inhibitor of the MDM2/p53 interaction, has been recently taken into consideration as a promising therapeutic tool for tumor treatment based on its ability to stabilize and activate the p53 transcription factor pathway. Since Nutlin-3 displays non cell-autonomous tumor-suppressor activities, we wanted to investigate its effect on dendritic cell functions, given the central role of these cells in the modulation of the immune response. We found that Nutlin-3 alone slightly affected the levels of major histocompatibility complex and costimulatory molecules and significantly promoted the ability of dendritic cells to stimulate T cells in the mixed lymphocyte reaction. Taken together, our findings suggest that the ability of Nutlin-3 to modulate dendritic cell functions and therefore lymphocyte proliferation might represent an additional important mechanism by which Nutlin-3 exerts its non cell-autonomous tumor-suppression function.


Differential expression of enhancer of zeste homolog 2 (EZH2) protein in small cell and aggressive B-cell non-Hodgkin lymphomas and differential regulation of EZH2 expression by p-ERK1/2 and MYC in aggressive B-cell lymphomas.
EZH2, a member of the polycomb protein group, is an important methyltransferase that is overexpressed in various neoplasms. We found that in small cell B-cell lymphomas, EZH2 is expressed in <40% of neoplastic cells, with heterogenous signal intensity. In aggressive B-cell lymphomas, 70-100% of tumor cells were positive for EZH2 expression with high signal intensity, which correlated with a high proliferation rate. We investigated the potential signaling molecules that regulate EZH2 overexpression in aggressive B-cell lymphomas and found that 80% of cases of EZH2-positive diffuse large B-cell lymphoma show high p-ERK1/2 expression (average ~57% tumor cell positivity). In contrast, only a small percentage of tumor cells (~10%) show p-ERK1/2 expression in Burkitt lymphoma and double hit lymphoma. On average, 91 and 76% of neoplastic cells were positive for MYC expression in Burkitt lymphoma and double hit lymphoma, respectively, while only 20% neoplastic cells were positive for MYC expression in diffuse large B-cell lymphoma. None of the aggressive B-cell lymphomas showed significant p-STAT3 expression in EZH2-overexpressed cases. The correlation of EZH2 expression with aggressive behavior and proliferation rate in B-cell neoplasms suggests that this molecule may function as an oncogenic protein in these neoplasms, with possible regulation by different signaling cascades in different types of aggressive B-cell lymphomas: p-ERK-related signaling in diffuse large B-cell lymphoma, and MYC-related signaling in Burkitt lymphoma and double hit lymphoma. Furthermore, EZH2 and associated signaling cascades may serve as therapeutic targets for the treatment of aggressive B-cell lymphomas.


5-HT3 receptor antagonist MDL 72222 attenuates cocaine- and mazindol-, but not methylphenidate-induced neurochemical and behavioral effects in the rat.
It has previously been demonstrated that the 5-HT(3) receptors located in the mesolimbic brain areas are able to modulate the dopaminergic effects of various abused drugs, including cocaine (COC). The present experiments investigated the role of 5-HT(3) receptors in the actions of selected monoamine uptake inhibitors. The ability of the 5-HT(3) receptor antagonist MDL 72222 (MDL; 0.1 and 1.0 mg/kg) to modify the neurochemical and behavioral changes induced by COC (20 mg/kg), mazindol (MAZ; 10 mg/kg), and methylphenidate (MP; 5.0 or 10, and 20 mg/kg) was assessed with an in vivo microdialysis technique, a conditioned place preference method, and motor activity measurements. MDL robustly attenuated the elevation of extracellular dopamine levels in the nucleus accumbens, acquisition of place preference, and motor activity induced by COC and MAZ, but not those induced by MP, the only drug with no significant effect on 5-HT. In contrast, expression of COC-induced place preference was not attenuated by MDL. These results show that COC- and MAZ-induced reward-related neurochemical and behavioral effects, preferentially those implicated in development of conditioned reward, are modified by the 5-HT(3) blockade. In contrast to COC and MAZ, the changes induced by MP, which has less effect on the serotonergic system, remain unchanged. Thus it appears that involvement of a serotonergic component in the mechanism of action of a drug could be a prerequisite for effective antagonism by 5-HT(3) receptor blockers.


Occupational hand dermatitis among cement workers in Taiwan.
Occupational dermatitis among cement workers is a major occupational health concern. The two most important occupational hazards for cement workers are irritant and allergic cement contact dermatitis. The objective of this study was to investigate the severity of occupational cement contact dermatitis and the common allergens among cement workers in Taiwan. A total of 97 cement workers from the Cement Workers' Association of Tainan City and County participated in this study. A structured questionnaire was used to evaluate the demographic data and work-related activities of these cement workers. A complete skin examination was conducted, and skin manifestations were assessed by a dermatologist. Allergens from European Standard Tray (Chemotechnique Diagnostic AB, Sweden) with a total of 25 substances were used for patch testing. Our results showed that 65 out of 97 cement workers were suffering from occupational cement hand contact dermatitis. The most affected skin area was the hand. Thickening of the dorsal surface of the hand, especially around the metacarpophalangeal joint area, and hyperkeratosis of the palm were the major skin manifestations. The results of the patch test showed that 24 out of 97 were allergic to potassium dichromate, nine were allergic to thiuram mix, nine were allergic to fragrance mix and seven were allergic to cobalt chloride. The final diagnosis, based on the results of the skin examination and the patch test, showed that 43 of 97 cement workers had irritant cement contact dermatitis and 22 had allergic cement contact dermatitis. We conclude that occupational cement hand dermatitis among cement workers is an important and severe issue in Taiwan, and the most common allergens among cement workers are potassium dichromate, thiuram mix, fragrance mix and cobalt chloride. The high positive rate of chromium hypersensitivity among cement workers reflects the urgency to regulate the addition of ferrous sulfate to cement in Taiwan.


The complete nucleotide sequence of two cold-adapted, temperature-sensitive attenuated mutant vaccine viruses (cp12 and cp45) derived from the JS strain of human parainfluenza virus type 3 (PIV3).
Two cold-passaged mutant vaccine viruses (cp12 and cp45) derived from the JS wild-type (wt) strain of human parainfluenza virus type 3 (PIV3) have been sequenced. These mutant viruses display the cold-adapted (ca), temperature-sensitive (ts), and attenuation (att) phenotypes. Sequence data indicate that both cp12 and cp45 sustained nucleotide substitutions during cold passage and subsequent cloning. Fifteen nucleotide changes were present in cp12 and 18 in cp45. Of these changes, some were present in the sequence of the prototype wt strain (Wash/47885/57) or were non-coding changes present in the open reading frames (ORFs). These were considered unlikely to be of significance in contributing to phenotypic differences between the mutants and the JS wt. There were nine remaining changes in cp12 and eight in cp45 that would most likely contribute to their phenotypes. For cp12, two were non-coding changes in regulatory regions, one in the 3' genome leader and one in the NP gene transcription start signal. The remaining seven changes resulted in amino acid substitutions in NP, F, HN, and L. For cp45, two mutations were in a non-coding regulatory region, the 3' genome leader. The remaining six changes resulted in amino acid substitutions in F, HN, and L. Only one amino acid substitution was conserved between cp12 and cp45 (a valine to alanine change at position 384 of the HN gene). These results should prove useful in the future in understanding the genetic basis of attenuation of the cold-passaged PIV3 candidate vaccine viruses.


Late, severe, noninfectious diarrhea after renal transplantation: high-risk factors, therapy, and prognosis.
Late severe noninfectious diarrhea in renal transplant recipients can lead to malnutrition and even graft loss. The purpose of this study was to evaluate risk factors associated with this condition and summarize therapy for these patients. For more than 36 months we observed a cohort of 541 recipients who underwent kidney transplantation from January 2001 to June 2007. They were provided a calcineurin inhibitor (CNI) combined with mycophenolate mofetil (MMF). The four group includes a continuous cyclosporine (CsA); a preconversion to tacrolimus and a postconversion group as well as a continuous tacrolimus group. The rate of severe late noninfectious diarrhea was compared among the four groups. Risk factors were analyzed between the diarrhea and nondiarrhea cohorts. Clinical characteristics, efficacy, and safety were observed after modifying the immunosuppressive protocol for late severe noninfectious diarrhea recipients. Twenty-eight recipients presented with late sever noninfectious diarrhea. No patients displayed chronic diarrhea in the CsA (n = 145) or preconversion group (n = 95). The rate of diarrhea was 7.31% in the postconversion and 7.35% in the tacrolimus group. Using multivariate Cox proportional hazards analysis, factors associated with an increased risk of noninfectious diarrhea were cytochrome P450(CYP)3A5 *3/*3 type, chronic renal allograft dysfunction, and patient ingestion of Tripterygium wilfordii Hook F. All diarrheal recipients experienced weight loss, hypoalbuminia, and an increased serum creatinine. All affected patients underwent adjustment of the immunosuppressive regimen to achieve remission. Renal allograft survival in recipients with diarrhea was worse than that in nondiarrheal recipients receiving tacrolimus combined with MMF. Tacrolimus with MMF increased the risk of late severe noninfectious diarrhea among renal transplant recipients compared with hosts treats with CsA plus MMF. The CYP3A5 *3/*3 type, chronic renal allograft dysfunction, and T. wilfordii supplementation were high-risk factors for late diarrhea. Prompt adjustment of immunosuppression was an effective, feasible therapy for these patients.


Short-term effects of prednisolone on neuromuscular transmission in normal rats and those with experimental autoimmune myasthenia gravis.
Electrophysiological investigations of the effects of bath-applied prednisolone at the neuromuscular junction were performed in muscles from normal rats and rats with experimental autoimmune myasthenia gravis (EAMG). In muscles from both groups, prednisolone reversible and significantly depressed the amplitudes of minature end-plate potentials (MEPPs), end-plate potentials (EPPs) and indirectly elicited action potentials (APs) without affecting resting membrane potentials. Prednisolone also caused a significant reduction in EPP rise time to peak and half-decay time while markedly increasing MEPP frequency and AP rise time to peak and duration. These effects were shown to be dose-dependent. The percentage decrease in amplitude after prednisolone perfusion was similar for EPPs and MEPPs, indicating that the depressive effect of prednisolone at the junction is postsynaptic. In all of the parameters studied, the percentage effect of prednisolone was the same in EAMG and normal preparations. No stimulus-linked repetitive EPPs or APs were observed after prednisolone. It is concluded that prednisolone has a depressive effect on neuromuscular transmission, but that this occurs only at high concentrations of the drug which are not achieved during the treatment of myasthenia gravis.


[Acanthamoeba isolated from child thymus: Acanthamoeba growth during cocultivation with human and murine eukaryotic cells and induction of cytopathogenic effect on these cells].
A cell line of the ameba assigned to the genus Acanthamoeba by morphological and cariotypic signs and by the specific features of its life cycle was isolated from the thymus of a child operated on for heart disease. The line received the name CDHT (Cells Derived from Human Thymus). Actively proliferating mammalian cells maintain the multiplication of Acanthamoeba cells in vitro. Using thymocytes as an example, it was shown that death of these cells occurred through the mechanism of apoptosis. The human thymocytes preincubated with the supernatant obtained through the cocultivation of CDHT and thymocytes may also undergo apoptosis.