Upregulation of ERp57 promotes clear cell renal cell carcinoma progression by initiating a STAT3/ILF3 feedback loop.
ERp57 dysfunction has been shown to contribute to tumorigenesis in multiple malignances. However, the role of ERp57 in clear cell renal carcinoma (ccRCC) remains unclear. Cell proliferation ability was measured by MTT and colony forming assays. Western blotting and quantitative real-time PCR (qRT-PCR) were performed to measure protein and mRNA expression. Co-immunoprecipitation (CoIP) and proximity ligation assay (PLA) were performed to detect protein-protein interaction. Chromatin immunoprecipitation (ChIP), ribonucleoprotein immunoprecipitation (RIP), and oligo pull-down were used to confirm DNA-protein and RNA-protein interactions. Promoter luciferase analysis was used to detect transcription factor activity. Here we found ERp57 was overexpressed in ccRCC tissues, and the higher levels of ERp57 were correlated with poor survival in patients with ccRCC. In vivo and in vitro experiments showed that ccRCC cell proliferation was enhanced by ERp57 overexpression and inhibited by ERp57 deletion. Importantly, we found ERp57 positively regulated ILF3 expression in ccRCC cells. Mechanically, ERp57 was shown to bind to STAT3 protein and enhance the STAT3-mediated transcriptional activity of ILF3. Furthermore, ILF3 levels were increased in ccRCC tissues and associated with poor prognosis. Interestingly, we revealed that ILF3 could bind to ERp57 and positively regulate its expression by enhancing its mRNA stability. Furthermore, ccRCC cell proliferation was moderated via the ERp57/STAT3/ILF3 feedback loop. In summary, our results indicate that the ERp57/STAT3/ILF3 feedback loop plays a key role in the oncogenesis of ccRCC and provides a potential therapeutic target for ccRCC treatment.


Developing restoration planting mixes for active ski slopes: a multi-site reference community approach.
Downhill ski areas occupy large expanses of mountainous lands where restoration of ecosystem function is of increasing importance and interest. Establishing diverse native plant communities on ski runs should enhance sediment and water retention, wildlife habitat, biodiversity and aesthetics. Because ski slopes are managed for recreation, ski slope revegetation mixes must consist of low-stature or herbaceous plants that can tolerate typical environmental conditions on ski slopes (high elevation, disturbed soils, open, steep slopes). The most appropriate reference communities for selecting ski slope revegetation species are thus successional, or seral plant communities in similar environments (i.e., other ski slopes). Using results from a broad-scale reference community analysis, I evaluated plant communities naturally occurring on ski slopes from 21 active and abandoned ski areas throughout the northern Sierra Nevada to identify native plant species suitable for use in ski slope restoration. I constructed a baseline planting palette of regionally appropriate plant species (for restoration of either newly created or already existing ski runs) that is functionally diverse and is likely to succeed across a broad range of environments. I also identify a more comprehensive list of species for more specialized planting mixes based on site-specific goals and particular environmental settings. Establishing seral plant communities may be an appropriate restoration goal for many other types of managed lands, including roadsides, firebreaks and utility rights-of-way. This study describes an ecological (and potentially cost-effective) approach to developing restoration planting palettes for such managed lands.


Liquid-holding experiments with human lymphocytes. III. Experiments with G0 and G1 cells.
Liquid holding (LH) experiments were performed with human peripheral lymphocytes treated in the G0 (G0-LH) or the G1 (G1-LH) phase of the cell cycle with diepoxybutane (DEB) or methylnitrosourea (MNU). In the G0-LH system, treatment with DEB but not with MNU led to a lowering of the frequencies of sister-chromatid exchanges (SCE). In the G1-LH system treatment with both chemicals led to a lowering of the SCE frequencies during the LH. These results are concluded to mean that lesions induced by DEB but not by MNU can be repaired in G0 cells and that G1 cells can repair both DEB and MNU induced lesions.


Foveal cone thresholds.
The method of constant stimuli was used to estimate the psychometric functions for detection of one or two flashes when two light pulses were presented. The test stimulus consisted of two simultaneous 0.5 msec, 1' pulses separated by 17'. Observers reported seeing 0, 1 or 2 flashes. A computer-controlled direct-view apparatus allowed sampling of slightly different foveal locations on each trial. The data were analyzed assuming a binomial probability for sampling of L and M cones and Poisson distributed quantal fluctuation. Under these assumptions, the measurements imply that detection requires a minimum of 5-7 quanta absorbed per cone, and that the effective number of cones illuminated by the 1', 0.5 msec pulse is two. The estimated L/M cone ratio was 1.6 for one observer and 4.0 for the other; each observer's ratio was in general agreement with the value estimated independently by heterochromatic flicker photometry.


Short vs long-course antibiotic therapy in pyelonephritis: a comparison of systematic reviews and guidelines for the SIMI choosing wisely campaign.
The Italian Society of Internal Medicine (SIMI) Choosing Wisely Campaign has recently proposed, among its five items, to reduce the prescription of long-term intravenous antibiotics if not indicated. The aim of our study was to assess the available evidences on optimal duration of antibiotic treatment in pyelonephritis through a systematic review of secondary studies. We searched for all guidelines on pyelonephritis and systematic reviews assessing the optimal duration of antibiotic therapy in this type of infection. We compared the recommendations of the three most cited and recent guidelines on the topic of interest. We extracted data of non-duplicated RCT from the selected systematic reviews and performed meta-analyses for clinical and microbiological failure. A trial sequential analysis (TSA) was also achieved to identify the need for further evidence. We identified 4 systematic reviews, including data from 10 non-duplicated RCTs (1536 patients). The meta-analysis showed a higher rate of clinical cure for short-course antibiotic treatment (RR for clinical failure 0.70, 95% CI [0.53-0.94]). No significant difference in the rate of microbiological failure (RR 1.06, 95% CI [0.75-1.49]) was observed. In terms of clinical cure, the TSA suggests that current evidence is sufficient to consider short course at least as effective as long-course treatment. Selected guidelines recommend considering shorter courses, but do not cite most of the published RCTs. Short-course antibiotic treatment is at least as effective as longer courses for both microbiological and clinical success in the treatment of acute uncomplicated pyelonephritis.


Severe malaria in children: factors predictive of outcome and response to Quinine.
To identify clinical features of severe malaria and their association with adverse outcomes according to recently proposed WHO guidelines and observe treatment failure to Quinine. This descriptive study was performed at Civil Hospital Karachi from September 2007 to January 2008. Various clinical features and laboratory parameters were analyzed according to t WHO guidelines and treatment failure to anti-malarial drugs was recorded. Mean, frequencies, percentages and chi-square test were used for analysis. Statistical significance was defined as p-value < 0.05. Total of 81 patients were enrolled in the study. Mean age of children was 5.5 +/- 3.4 years. Type of malaria infections that were seen included falciparum 46 (57%), mixed infection 26 (32%) and vivax 9 (11%). Frequent clinical features included splenomegaly (74%), multiple organ dysfunction (MOD) (70%), cerebral malaria (31%) and malnutrition (27%). Thrombocytopenia (86%) and severe anaemia (42%) were the common laboratory findings. Shock (p < 0.001), renal failure (p < 0.001), hepatic involvement (p < 0.002) and cerebral malaria (p < 0.002) emerged as strong predictors of complications. Fourteen out of 81 cases showed early treatment failure to Quinine. Shock, renal failure, hepatic involvement and cerebral malaria are strongly associated with complications in severe malaria. MOD and malnutrition were identified as significant new clinical features present in severe malaria in this study.


Alterations in synaptic curvature in the dentate gyrus following induction of long-term potentiation, long-term depression, and treatment with the N-methyl-D-aspartate receptor antagonist CPP.
Alterations in curvature of the post synaptic density (PSD) and apposition zone (AZ), are believed to play an important role in determining synaptic efficacy. In the present study we have examined curvature of PSDs and AZs 24 h following homosynaptic long-term potentiation (LTP), and heterosynaptic long-term depression (LTD) in vivo, in awake adult rats. High frequency stimulation (HFS) applied to the medial perforant path to the dentate gyrus induced LTP while HFS stimulation of the lateral perforant path induced LTD in the middle molecular layer of the dentate gyrus (DG). Curvature changes were analysed in this area using three dimensional (3-D) reconstructions of electron microscope images of ultrathin serial sections. Very large and significant changes in 3-D measurements of AZ and PSD curvature occurred 24 h following both LTP and LTD, with a flattening of the normal concavity of mushroom spine heads and a change to convexity for thin spines. An N-methyl-D-aspartate (NMDA) receptor antagonist CPP (3-[(R)-2-Carboxypiperazin-4-yl]-propyl-1-phosphonic acid) blocked the changes in curvature of mushroom and thin spine PSDs and apposition zones, actually increasing the concavity of mushroom spines as the spine engulfed the presynaptic bouton. In order to establish whether these changes resulted from the effect of the NMDA antagonist or from its coincidence with synaptic activation during testing we examined the effects of CPP alone on PSD and apposition zone curvature. It was found that CPP alone also caused a small decrease in curvature of both PSD and apposition zone of mushroom and thin spines.


Estimation of parameters related to vaccine efficacy and dengue transmission from two large phase III studies.
A tetravalent dengue vaccine was shown to be efficacious against symptomatic dengue in two phase III efficacy studies performed in five Asian and five Latin American countries. The objective here was to estimate key parameters of a dengue transmission model using the data collected during these studies. Parameter estimation was based on a Sequential Monte Carlo approach and used a cohort version of the transmission model. Serotype-specific basic reproduction numbers were derived for each country. Parameters related to serotype interactions included duration of cross-protection and level of cross-enhancement characterized by differences in symptomaticity for primary, secondary and post-secondary infections. We tested several vaccine efficacy profiles and simulated the evolution of vaccine efficacy over time for the scenarios providing the best fit to the data. Two reference scenarios were identified. The first included temporary cross-protection and the second combined cross-protection and cross-enhancement upon wild-type infection and following vaccination. Both scenarios were associated with differences in efficacy by serotype, higher efficacy for pre-exposed subjects and against severe dengue, increase in efficacy with doses for naïve subjects and by a more important waning of vaccine protection for subjects when naïve than when pre-exposed. Over 20 years, the median reduction of dengue risk induced by the direct protection conferred by the vaccine ranged from 24% to 47% according to country for the first scenario and from 34% to 54% for the second. Our study is an important first step in deriving a general framework that combines disease dynamics and mechanisms of vaccine protection that could be used to assess the impact of vaccination at a population level.


Effect of His6-tag Position on the Expression and Properties of Phenylacetone Monooxygenase from Thermobifida fusca.
Phenylacetone monooxygenase (EC 18.104.22.168, PAMО) catalyzes oxidation of ketones with molecular oxygen and NADPH with the formation of esters. PAMО is a promising enzyme for biotechnological processes. In this work, we generated genetic constructs coding for PAMO from Thermobifida fusca, containing N- or C-terminal His6-tags (PAMO N and PAMO C, respectively), as well as PAMO L with the His6-tag attached to the enzyme C-terminus via a 19-a.a. spacer. All PAMO variants were expressed as catalytically active proteins in Escherichia coli BL21(DE3) cells; however, the expression level of PAMO N was 3 to 5 times higher than for the other two enzymes. The catalytic constants (kcat) of PAMO C and PAMO L were similar to that published for PAMO L produced in a different expression system; the catalytic constant for PAMO N was slightly lower (by 15%). The values of Michaelis constants with NADPH for all PAMО variants were in agreement within the published data for PAMO L (within the experimental error); however, the KM for benzylacetone was several times higher. Thermal inactivation studies and differential scanning calorimetry demonstrated that the thermal stability of PAMO N was 3 to 4 times higher compared to that of the enzymes with the C-terminal His6-tag.


Esophagovisceral anastomotic leak. A prospective statistical study of predisposing factors.
An esophagovisceral anastomotic leak is a life-threatening postoperative complication, especially in the mediastinum. Of the 242 patients who underwent intrathoracic esophagogastric anastomosis for esophageal carcinoma (182 patients) and adenocarcinoma of the cardia (60 patients) between January 1980 and June 1985, 14 (5.8%) had esophageal anastomotic leakage and two died (0.8%). Various clinical and biologic parameters and aspects of operative technique were studied prospectively and analyzed statistically to identify possible factors responsible for leaks. Both bivariate and multivariate statistical analysis with logistic regression showed that the following clinical and biologic factors do not influence anastomotic leakage: tumor stage, the curative or palliative purpose of resection, neoplastic permeation of anastomotic margins, total protein concentration below 5 gm/dl, albumin concentration below 3 gm/dl, patient's age, diabetes, high blood pressure, cirrhosis of the liver, and cardiac, respiratory, or renal diseases. Technical factors, on the contrary, were statistically significant and of great clinical importance: manual as opposed to mechanical suturing (chi 2 = 8.8, p = 0.013) and single-layer as opposed to double-layer suturing (chi 2 = 9.9, p = 0.043). The level of the anastomosis was found to be a further statistically significant factor: The incidence of leakage was greater when the anastomosis was located between the azygos vein and the lower pulmonary vein (chi 2 = 15.5, p = 0.004) than above the azygos vein or below the lower pulmonary vein.


A rapid method for measuring the phospholipid synthetic activity of incubated lymphocytes.
A rapid and simple method for estimating the phospholipid synthetic activity of incubated lymphocytes by a Florisil column technique following the uptake of 14C-oleic acid was established. Stimulation of phospholipid synthesis by PHA and inhibitions caused by Tween 20 or Tween 80 and heating were evaluated easily with this method.


Pressure-induced magnetic switching and linkage isomerism in K0.4Fe4[Cr(CN)6]2.8 x 16 H2O: X-ray absorption and magnetic circular dichroism studies.
The effect of applied pressure on the magnetic properties of the Prussian blue analogue K0.4Fe4[Cr(CN)6]2.8 x 16 H2O (1) has been analyzed by dc and ac magnetic susceptibility measurements. Under ambient conditions, 1 orders ferromagnetically at a critical temperature (T(C)) of 18.5 K. Under application of pressure in the 0-1200 MPa range, the magnetization of the material decreases and its critical temperature shifts to lower temperatures, reaching T(C) = 7.5 K at 1200 MPa. Pressure-dependent Raman and Mossbauer spectroscopy measurements show that this striking behavior is due to the isomerization of some Cr(III)-C[triple bond]N-Fe(II) linkages to the Cr(III)-N[triple bond]C-Fe(II) form. As a result, the ligand field around the iron(II) centers increases, and the diamagnetic low-spin state is populated. As the number of diamagnetic centers in the cubic lattice increases, the net magnetization and critical temperature of the material decrease considerably. The phenomenon is reversible: releasing the pressure restores the magnetic properties of the original material. However, we have found that under more severe pressure conditions, a metastable sample containing 22% Cr(III)-N[triple bond]C-Fe(II) linkages can be obtained. X-ray absorption spectroscopy and magnetic circular dichroism of this metastable sample confirm the linkage isomerization process.


Effects of calcium and magnesium ions upon fat absorption by sacs of everted hamster intestine.
Sacs of everted hamster jejunum were incubated with fatty acid and monoglyceride in bile salt solutions either with or without added salts of calcium and magnesium. At very low concentrations of Ca2+ and Mg2+, the intestine produced very few chylomicrons. However, at physiological concentrations of the ions, the gut-preparation released numerous chylomicron-like particles into the serosal fluid.


Role of Notch signaling during lipopolysaccharide-induced preterm labor.
Notch signaling pathways exert effects throughout pregnancy and are activated in response to TLR ligands. To investigate the role of Notch signaling in preterm labor, Notch receptors (Notch1-4), its ligand Delta-like protein-1, transcriptional repressor hairy and enhancer of split-1, and Notch deregulator Numb were assessed. Preterm labor was initiated on gestation d 14.5 by 1 of 2 methods: 1) inflammation-induced preterm labor: intrauterine injection of LPS (a TLR4 agonist) and 2) hormonally induced preterm labor: subcutaneous injection of mifepristone. Delta-like protein-1, Notch1, and hairy and enhancer of split-1 were elevated significantly, and Numb was decreased in the uterus and placenta of inflammation-induced preterm labor mice but remained unchanged in hormonally induced preterm labor compared with their respective controls. F4/80(+) macrophage polarization was skewed in the uterus of inflammation-induced preterm labor toward M1-positive (CD11c(+)) and double-positive [CD11c(+) (M1) and CD206(+) (M2)] cells. This process is dependent on activation of Notch signaling, as shown by suppression of M1 and M2 macrophage-associated cytokines in decidual macrophages in response to γ-secretase inhibitor (an inhibitor of Notch receptor processing) treatment ex vivo. γ-Secretase inhibitor treatment also diminished the LPS-induced secretion of proinflammatory cytokines and chemokines in decidual and placental cells cultured ex vivo. Furthermore, treatment with recombinant Delta-like protein-1 ligand enhanced the LPS-induced proinflammatory response. Notch ligands (Jagged 1 and 2 and Delta-like protein-4) and vascular endothelial growth factor and its receptor involved in angiogenesis were reduced significantly in the uterus and placenta during inflammation-induced preterm labor. These results suggest that up-regulation of Notch-related inflammation and down-regulation of angiogenesis factors may be associated with inflammation-induced preterm labor but not with hormonally induced preterm labor.


10 years of experience in adopting, implementing and evaluating progress testing for Saudi medical students.
The progress test (PT) is a comprehensive examination that is designed to match the knowledge acquisition necessary at graduation and monitors progress during the entire period of an undergraduate program. Qassim College of Medicine (QCM) began using the multi-institutional PT in the Kingdom of Saudi Arabia (KSA). This study aimed to determine if the PT can be utilized to assess the progress of medical students at different Saudi medical colleges with different educational approaches, as well as whether this testing modality could be accepted by other colleges. Beside the establishment of a PT committee, comprehensive blueprinting was crafted to sample 200 A-type multiple choice questions (MCQs) from different disciplines. The PT is a paper-and-pencil model and is answered in a 4-h period. All PT items followed a uniform design. In total, 13 rounds of the progress test have been conducted. The number of participating colleges increased from three (with 285 students) in the first test (May 2012) to more than 20 (with >6000 students) in the ninth round (February 2017). The average % scores for first-year students ranged from 3.0% to 7.9% while the average scores for fifth-year students ranged from 34.0% to 43.0%. The conduction of this meticulously crafted test to evaluate knowledge achievement at medical graduation is a fruitful tool and helps to provide constructive feedback for test-takers and other stakeholders relating to their relative positions among other fellows at the national level.


Cerebrospinal Fluid Diversion in Endoscopic Skull Base Reconstruction: An Evidence-Based Approach to the Use of Lumbar Drains.
Before the vascularized pedicled nasoseptal flap was popularized, lumbar drains (LDs) were routinely used for cerebral spinal fluid (CSF) diversion in endoscopic skull base reconstruction. LDs are not necessary in most CSF leaks encountered during skull base surgery. In this article, the use is considered of an LD in select high-risk settings in which a high-flow leak is anticipated and the patient has significant risk factors that make closure of the leak more challenging. Evidence for the use of LDs in preventing postoperative after endoscopic skull base reconstruction is reviewed and a rational framework for their use is proposed.


The 3D structure of a cyclosporin analogue in water is nearly identical to the cyclophilin-bound cyclosporin conformation.
The conformation of [D-MeSer3-D-Ser-(O-Gly)8]CS, a water soluble cyclosporin derivative, has been determined in (D6)DMSO and in water using NMR. In these polar solvents the conformation is identical and very similar to the structure found in the cyclophilin-cyclosporin complex. However, it differs significantly from its conformation in deuterated chloroform. This demonstrates unambiguously that the large structure change is induced primarily by the polar solvent rather than by complex formation with cyclophilin.


Efficacy and tolerability of telmisartan plus amlodipine in added-risk hypertensive patients.
Added-risk hypertensive patients with co-morbidities such as diabetes and metabolic syndrome often require two or more antihypertensives to achieve blood pressure (BP) targets. The aim of this sub-analysis was to determine the efficacy and safety of telmisartan 40 or 80 mg plus amlodipine 5 or 10 mg in patients with hypertension, stratified according to certain criteria such as type 2 diabetes mellitus and metabolic syndrome. Patients were treated for 8 weeks with telmisartan 20-80 mg plus amlodipine 2.5-10 mg. This post-hoc analysis included patients treated with higher doses, and stratified according to a number of sub-populations (age, race, diabetes, obesity, metabolic syndrome, elevated baseline systolic BP (SBP), renal impairment). Eight weeks' treatment with telmisartan plus amlodipine combinations provided consistent reductions in mean SBP/diastolic BP (DBP) across the different sub-populations, similar to the overall population. SBP/DBP reductions ranged from -13.5 to -34.7/-12.6 to -26.1 mmHg and BP goal rates (<140/90 mmHg) ranged from 29.8-100% for the four key dose combinations of telmisartan plus amlodipine. For the highest dose combination of telmisartan 80 mg plus amlodipine 10 mg, SBP/DBP reduction ranged from -19.1 to -34.7/-16.4 to -22.8 mmHg and goal attainment rate from 66.7% to 87.0%. Across the sub-populations, high SBP and DBP response rates were seen with combination treatment (83.3-97.7% and 75.0-95.7%, respectively, with telmisartan 80 mg plus amlodipine 10 mg). The combination was safe and well tolerated across all sub-populations and the incidence of peripheral oedema with telmisartan 40-80 mg plus amlodipine 10 mg was generally lower than with A10 monotherapy. Despite small patient numbers in some sub-populations and the post-hoc nature of the analysis, this does show that the combination of telmisartan plus amlodipine provides an effective, safe and well-tolerated antihypertensive treatment for added-risk hypertensive patients.


Self-concept and sexuality of spinal cord injured women.
Differences in perceived self-concept and sexual response before and after spinal cord injury were examined. Twenty-four women between the ages of 20 and 40 completed a questionnaire and participated in a brief taped interview. Most of the women viewed themselves as very or somewhat attractive and had been involved in a sexual relationship since injury. The majority viewed sexual relationships as very enjoyable, although many commented that changes in bowel and bladder function had inhibited sexual expression. The need for more effective sexual counseling was highlighted. A trend was noted for an interrelationship between sexuality and self-concept in adapting to acquired disability.


Characterization of immobilization methods for African swine fever virus protein and antibodies with a piezoelectric immunosensor.
A direct piezoelectric flow injection analysis immunoassay for the detection of African Swine Fever virus and antibodies is presented. The peptide-specific monoclonal antibody 18BG3 and the virus protein 73 were used for detection with a quartz crystal microbalance. Accumulation of the analyte on the surface of this mass-sensitive biosensor resulted in a shift of the resonant frequency. Highly selective receptor layers were applied on the sensing electrode of the quartz crystal for detection of the complementary analyte. Different immobilization methods proved to be appropriate for coating of the monoclonal antibody 18BG3. A quartz crystal covalently coated with the antibody 18BG3 detected virus protein VP73 samples more than 20 times and was stable for more than 30 days. The coating of virus protein was performed by physisorption. A sensor with a virus protein receptor layer detected antibody 18BG3 samples 10 times within one day. The sensor device was able to perform one measurement cycle including blocking and regeneration within 30 min. With the help of a suitable carrier liquid, measurements with serum samples were performed. The calibration curves for measurements in buffer and in serum could be determined and the detection limits for virus protein detection were 0.31 and 1 microgram/ml, and for antibody detection 0.1 and 0.2 microgram/ml, respectively.


The morphologic and biologic patterns of chemically induced pancreatic adenocarcinoma in Syrian golden hamsters after homologous transplantation.
A pancreatic adenocarcinoma induced by N-nitrosobis(2-oxopropyl)amine in the Syrian golden hamster was successfully transplanted to a homologous host by subcutaneous inoculation through 10 successive passages. The rate of 'tumor take' increased progressively with each generation from 60% to 100%, and the latency period after inoculation was reduced simultaneously from 6 weeks to 1 week in the second and following passages. The tumors grew rapidly, ulcerated the overlying skin, and metastasized to the regional lymph nodes and lungs. The animals usually died with multiple lung metastases between the 5th and 20th weeks. All transplanted tumors and their metastases retained the pattern of the original, well-differentiated adenocarcinomas.


Rarity of hypertension in adult patients with atopic dermatitis.
Patients with atopic dermatitis show a tendency for vasoconstriction of the small vessels in the skin. As peripheral vasoconstriction contributes to the cause of hypertension, it is natural to suppose that blood pressures might be on the high side in adult patients with atopic dermatitis. In the literature, however, there was little information on the subject. To study the incidence of hypertension in adult patients with atopic dermatitis. Blood pressure was measured in 521 adult patients with active atopic dermatitis (235 males; 286 females) aged 30-59 years, and 87 adults with "healed" atopic dermatitis (26 males; 61 females) aged 34-52 years. The blood pressures were classified as definite hypertension, borderline hypertension or normal blood pressure. In those patients aged 30-39 years with active atopic dermatitis, the incidence of definite hypertension in the male patients and the female patients was 1.1% and 1.6%, respectively. The incidence remained almost at a plateau for the 30-39-year-old age group through to the 50-59-year-old age group, in both the male and female patients. There was no difference in the incidence of definite hypertension between patients with severe dermatitis and patients with mild dermatitis. Adult patients with "healed" atopic dermatitis also showed a low incidence of definite hypertension. These findings indicate that hypertension is rare in adult patients with atopic dermatitis. It is most probable that the rarity of hypertension is a primary feature of the disease.


Relevance of anaesthesia for dofetilide-induced torsades de pointes in alpha1-adrenoceptor-stimulated rabbits.
No information is available concerning the effects of anaesthetics in the most frequently used in vivo pro-arrhythmia model. Accordingly, in this study we examined the effect of pentobarbital, propofol or alpha-chloralose anaesthesia on the pro-arrhythmic activity of the class III anti-arrhythmic dofetilide in alpha(1)-adrenoceptor-stimulated rabbits. Rabbits anaesthetized intravenously with pentobarbital, propofol or alpha-chloralose were infused simultaneously with the alpha(1)-adrenoceptor agonist phenylephrine (15 microg kg(-1) min(-1), i.v.) and dofetilide (0.04 mg kg(-1) min(-1), i.v.). The electrocardiographic QT interval, the T (peak)-T (end) interval and certain QT variability parameters were measured. The heart rate variability and the baroreflex sensitivity were utilized to assess the vagal nerve activity. The spectral power of the systolic arterial pressure was calculated in the frequency range 0.15-0.5 Hz to assess the sympathetic activity. Pentobarbital considerably reduced, whereas propofol did not significantly affect the incidence of dofetilide-induced torsades de pointes (TdP) as compared with the results with alpha-chloralose (40% (P=0.011) and 70% (P=0.211) vs 100%, respectively). In additional experiments, neither doubling of the rate of the dofetilide infusion nor tripling of the rate of phenylephrine infusion elevated the incidence of TdP to the level seen with alpha-chloralose. None of the repolarization-related parameters predicted TdP. The indices of the parasympathetic and sympathetic activity were significantly depressed in the alpha-chloralose and propofol anaesthesia groups. In rabbits, anaesthetics may affect drug-induced TdP genesis differently, which must be considered when results of different studies are compared.


Insights into interactions of biodegradable and non-biodegradable microplastics with heavy metals.
Biodegradable microplastics (BMPs) are considered to be environmentally friendly compared to non-biodegradable plastics (NMPs). However, BMPs are likely to become toxic during their transport because of the adsorption of pollutants (e.g., heavy metals) onto them. This study investigated the uptake of six heavy metals (Cd2+, Cu2+, Cr3+, Ni2+, Pb2+, and Zn2+) by a common BMPs (polylactic acid (PLA)) and compared their adsorption characteristics to those of three types of NMPs (polyethylene (PE), polypropylene (PP), and polyvinyl chloride (PVC)) for the first time. The order of heavy metal adsorption capacity among the four MPs was PE > PLA > PVC > PP. The findings suggest that BMPs contained more toxic heavy metals than some NMPs. Among the six heavy metals, Cr3+ showed considerably stronger adsorption than other heavy metals in both BMPS and NMPs. The adsorption of heavy metals on MPs can be well explained using the Langmuir isotherm model, while the adsorption kinetic curves showed the best fit to the pseudo-second-order kinetic equation. Desorption experiments revealed that BMPs released a higher percentage of heavy metals (54.6-62.6%) in the acidic environment in a shorter time (~ 6 h) compared to NMPs. Overall, this study provides insights into interactions of BMPs and NMPs with heavy metals and their removal mechanisms in aquatic environment.


Calcium sensors of ciliary outer arm dynein: functions and phylogenetic considerations for eukaryotic evolution.
The motility of eukaryotic cilia and flagella is modulated in response to several extracellular stimuli. Ca(2+) is the most critical intracellular factor for these changes in motility, directly acting on the axonemes and altering flagellar asymmetry. Calaxin is an opisthokont-specific neuronal calcium sensor protein first described in the sperm of the ascidian Ciona intestinalis. It binds to a heavy chain of two-headed outer arm dynein in a Ca(2+)-dependent manner and regulates 'asymmetric' wave propagation at high concentrations of Ca(2+). A Ca(2+)-binding subunit of outer arm dynein in Chlamydomonas reinhardtii, the light chain 4 (LC4), which is a Ca(2+)-sensor phylogenetically different from calaxin, shows Ca(2+)-dependent binding to a heavy chain of three-headed outer arm dynein. However, LC4 appears to participate in 'symmetric' wave propagation at high concentrations of Ca(2+). LC4-type dynein light chain is present in bikonts, except for some subclasses of the Excavata. Thus, flagellar asymmetry-symmetry conversion in response to Ca(2+) concentration represents a 'mirror image' relationship between Ciona and Chlamydomonas. Phylogenetic analyses indicate the duplication, divergence, and loss of heavy chain and Ca(2+)-sensors of outer arm dynein among excavate species. These features imply a divergence point with respect to Ca(2+)-dependent regulation of outer arm dynein in cilia and flagella during the evolution of eukaryotic supergroups.


Surveillance of concussion-related injuries using electronic medical records from the Canadian Primary Care Sentinel Surveillance Network (CPCSSN): a proof-of-concept.
Previous studies on traumatic brain injury trends in Canada have been restricted to hospitalization and emergency department visit data. However, many concussion patients may present first, or only, to family physicians. Therefore, the true burden of concussion in Canada is likely underestimated. The Canadian Primary Care Sentinel Surveillance Network (CPCSSN) collects information electronically from family physicians across Canada and can potentially be used for concussion surveillance. The objective of this study is to explore the feasibility of using data collated from CPCSSN for concussion surveillance purposes and examine trends over time. Electronic medical records housed by CPCSSN from 2010 to 2016 were analyzed. Case ascertainment was determined through a combination of International Classification of Diseases, Ninth Revision codes. Binomial regression models were used to calculate the prevalence ratio (PR) of concussion by age, sex, deprivation indices, body mass index, and comorbid conditions. Concussion prevalence rates increased from 2010 to 2016 (p < 0.001). Based on 2016 data, males had a higher prevalence of concussion compared with females (PR = 1.09; 95% CI 1.02, 1.18), and those aged 10-14 (PR = 8.52; 95% confidence interval [CI] 6.34, 11.44) and 15-19 (PR = 7.75; 95% CI 5.84, 10.28) had higher prevalence of concussion compared with those aged 0-4 years. This pilot study demonstrates the feasibility of using the CPCSSN system for surveillance of concussion in the Canadian population. The initial findings on prevalence are in agreement with previous studies that have used hospitalization or emergency department data.


Outcomes and risk factors for Kahook Dual Blade excisional goniotomy with concomitant phacoemulsification: a multicentre Canadian study.
To assess the outcomes and failure risk factors for Kahook Dual Blade (KDB) excisional goniotomy with cataract surgery (phaco-KDB) in eyes with various glaucoma subtypes and severities. This multisurgeon consecutive case series included glaucomatous eyes with cataract that underwent phaco-KDB and had a minimum follow-up of 12 months postoperatively. Efficacy was assessed by absolute and qualified surgical success (defined by different criteria) and changes in intraocular pressure (IOP) and antiglaucoma medication (AGM) at the last postoperative follow-up. Safety included best-corrected visual acuity, cup-to-disc ratio, visual field mean deviation, retinal nerve fibre layer thickness, and adverse events. A total of 108 eyes of 89 patients with a median follow-up of 18 months (range, 12-47 months) were included. IOP decreased by 26% from 19.1 ± 5.0 mm Hg to 14.1 ± 3.5 mm Hg (p < 0.001), AGM use decreased by 29% from 2.4 ± 1.3 medications to 1.7 ± 1.3 (p < 0.001), and 25% of eyes became free of AGMs (vs 3% at baseline). Qualified success rates achieved for IOP cutoffs of 18, 15, and 12 mm Hg were 87%, 68%, and 46%, respectively. Higher baseline IOP and postoperative incidence of IOP spikes were associated with a higher risk of surgical failure. Best-corrected visual acuity improved postoperatively (p < 0.001), and visual field mean deviation, cup-to-disc ratio, and retinal nerve fibre layer thickness remained stable. Overall, safety was favourable, and adverse events were transient and not sight threatening. This multicentre Canadian study provides real-world data that support the safety and efficacy of phaco-KDB in reducing IOP and AGM use with no evidence of disease progression during the follow-up period.


The yield of solid swallows in high resolution esophageal manometry among patients with dysphagia and gastro-esophageal reflux symptoms.
Swallowing disorders are frequently reported esophageal symptoms and represent a common cause for referrals to gastroenterology consultations. Esophageal high-resolution manometry (HRM) is considered the gold standard modality for the evaluation of esophageal peristalsis and esophageal sphincter. The standard protocol of HRM includes water swallows only, that may not assess appropriately esophageal function and symptoms. Hence, we investigated the diagnostic yield of combining solid food swallows (SFS) to standard HRM. We conducted a retrospective study at the Gastroenterology unit of EMMS Nazareth Hospital, Israel. Patients who underwent manometry mainly due to dysphagia or reflux symptoms between November 2019 and November 2020 were enrolled. All patients underwent routine HRM and completed ten 5 ml water swallows and 5 bread swallows. Analysis and reports were done by an expert gastroenterologist, based on the Chicago Classification version 3. Overall, 99 patients (45.5% men, mean age 52.1±16 years) who underwent HRM were included. The addition of solid swallows to the test changed manometry results in 43 patients (43.4%) (P<0.005). Furthermore, SFS improved the contractility in 26/42 cases of ineffective and/or absent motility (61%) (P< 0.005). Finally, SFS aggravated symptoms in 44 patients (44.6% more aggravation than in liquid only) (P<0.005). Combining SFS to HRM can improve the diagnostic yield of the study. Adding solid swallows is of special value in cases of ineffective esophageal motility but induces a high rate of symptoms aggravation.


Computational study of a transition state analog of phosphoryl transfer in the Ras-RasGAP complex: AlF(x) versus MgF3-.
The structures of the complexes between Ras*GDP bound to RasGAP in the presence of three probable gamma-phosphate analogs (AlF3, AlF4- and MgF3-) for the transition state (TS) of the hydrolysis of guanosine triphosphate (GTP) by the Ras-RasGAP enzymes have been modeled by quantum mechanical-molecular mechanical (QM/MM) calculations. These simulations contribute to the dispute on the nature of the TS in the hydrolysis reaction, since medium resolution X-ray crystallography cannot discern among stereochemically similar isoelectronic species (e.g., AlF3 or MgF3-). The optimized geometry for each structure has been found starting from experimental coordinates of one of them (PDBID: 1WQ1). Direct comparison of the experimental and computed geometry configurations in the immediate vicinity of the active site suggests that MgF3- is the most likely candidate for the phosphate analog in the experimental structure.


Cloning, expression pattern, chromosomal localization, and evolution analysis of Porcine gnaq, gna11, and gna14.
The Gq class is one subfamily of the G protein alpha subunits multigene family. It comprises four genes: Gnaq, Gna11, Gna14, and Gna15. In mice and humans, the alpha subunit is an essential component of G protein interaction with receptors and effectors. We report here the cloning and characterization of porcine Gnaq, Gna11, and Gna14. We have cloned the full-length coding sequences of porcine Gnaq, Gna11, and Gna14 (1,080, 1,080, and 1,068 bp, respectively) and then mapped them chromosomally to regions 1q21-27, 2q21-24, and 1q21-27 by radiation hybrid mapping. Tissue distribution analysis indicated that Gnaq and Gna11 were coexpressed in liver, heart, muscle, spleen, adipose tissue, brain, and uterus, but Gna14 mRNA was detected only in kidney and lung. The phylogenetic trees reveal that porcine Gnaq, Gna11, and Gna14 are evolutionarily closer to their human homologs. This is the first report of molecular cloning and characterization of porcine Gnaq, Gna11, and Gna14, which will be helpful for further understanding of the physiological role of Gq class genes in pigs.


Insulin receptor tyrosine kinase is defective in skeletal muscle of insulin-resistant obese mice.
Obese syndromes of genetic origin or experimentally induced are characterized by resistance to insulin both in vivo (association of hyperglycaemia and hyperinsulinaemia) and in vitro. Thus, skeletal muscle of obese mice, which is the most important target organ for the action of insulin, displays a reduced response to insulin. This hormonal resistance cannot be explained by the moderate decrease in the number of insulin receptors found in obese animals. In fact, it is generally believed that a biochemical event occurring very early after binding of insulin to its receptor, which is the first step in insulin action, is defective in obesity. One of the earliest post-binding events so far recognized, and which is thought to have a key role in cellular signalling by the insulin receptor, is the insulin-stimulated phosphorylation of its receptor. In an effort to localize the defect responsible for the insulin resistance in obesity, we have studied the insulin receptor protein kinase activity and we show here that insulin receptors from skeletal muscles of insulin-resistant obese mice have an altered kinase activity for phosphorylation of both the receptor itself and of exogeneous substrates.


First Dorsal Metatarsal Artery Flap for Coverage of Soft Tissue Defects of the Distal Foot: Delayed Technique, Proximal and Distally Based Fasciocutaneous and Adipofascial Variants.
Orthoplastic surgeons continue to encounter challenges for both proximal and distal soft issue defects in the foot. Patients with significant comorbidities often are nonideal surgical candidates. Distal free flaps historically have been utilized. The first dorsal metatarsal artery flap is a local flap with several variants, making it a versatile option in the orthoplastic surgeon's armamentarium for treatment of challenging soft tissue defects about the foot and ankle. The vascular tree is variable and preoperative vascular identification is required to determine flap design.


Survey of Ebola Preparedness in Washington State Emergency Departments.
The 2014 Ebola virus disease (EVD) outbreak in West Africa remains the most deadly in history. Emergency departments (EDs) are more likely to come into contact with potential EVD patients. It is important for EDs to be prepared to care for suspected EVD patients. Our objective was to understand the perceived challenges experienced by Washington State ED medical directors in EVD preparedness. An anonymous, electronic survey was sent to a convenience sample of ED medical directors across Washington State between November and February of 2014-2015. The perceived challenges of and attitudes toward EVD preparations were assessed and reported as stratified proportions. Of 85 medical directors contacted, 59 responses (69%) were received. This included EDs with annual patient volumes of 60,000 (12 hospitals, 20%). Among the perceived challenges in EVD preparations were spatial modifications (eg, building an anteroom for donning and doffing of personal protective equipment) and waste management planning. Ninety-five percent of respondents moderately or strongly agreed that it is important to have a predesignated hospital to care for EVD patients. Washington State ED medical directors have faced significant challenges in ensuring their EDs are prepared to safely care for suspected EVD patients. Attitudes toward EVD preparations are mixed. Varying levels of perceived importance may represent an additional barrier to statewide EVD preparedness. (Disaster Med Public Health Preparedness. 2016;10:662-668).


Effect of specific antibody and complement on the survival of Rochalimaea quintana in vitro.
No bactericidal effect was produced when Rochalimaea quintana was exposed for 1 h to a combination of high-titered anti-R. quintana rabbit serum and guinea pig complement.


Equimolar proportions of somatic-type core histones in chromatin of spermatogonia.
[3H]Leucine incorporation into histones of seminiferous epithelial cells of hypophysectomized rats was used to calculate the molar proportions of the core histones of spermatogonia. The molar proportions H3:H2B:(H2A + protein A24):H4 are 1:1:1:1, viz. identical with those reported by others for somatic cells. Similar results were obtained when molar proportions of histones of seminiferous epithelial cells from immature rat testis (predominantly populated with spermatogonia) were determined by the dye-binding method. These data are relevant to mechanisms for the replacement of some of the core histones by variants during the primary spermatocyte stages.


Turning walking pneumonia into recurrent abscesses: a curious case of CVID and review of the literature.
Common variable immunodeficiency (CVID) is a primary immunodeficiency disorder associated with a broad symptom presentation that is still being characterized. We report a rare case of recurrent mycoplasma skin abscesses in a patient with a history of autoimmune disorders and prolonged mycoplasma pneumonia who was diagnosed with CVID. A 34-year-old woman presented with a history of recurrent abscesses previously confirmed positive for Mycoplasma pneumoniae. Her past medical history of recurrent mycoplasma abscesses, prolonged mycoplasma pneumonia, and autoimmune disorders (mixed connective tissue disease and immune thrombocytopenia) raised suspicion of CVID. Workup included negative anti-mycoplasma antibody titers, hypogammaglobulinemia, and negative anti-pneumococcal antibody titers despite prior vaccination, solidifying the diagnosis of CVID. The patient was discharged on antibiotic and intravenous immunoglobulin therapy and now follows allergy and immunology long-term for treatment. Her diagnostic history underscores the importance of considering the various criteria of CVID for diagnosis, and her unique presentation of M. pneumoniae skin abscesses highlights the broad sequelae patients with CVID can manifest.


Trastuzumab-induced cardiotoxicity: the role of two-dimensional myocardial strain imaging in diagnosis and management.
A 45-year-old female breast cancer patient developed heart failure during adjuvant trastuzumab therapy. Her initial left ventricular ejection fraction (LVEF) was 39% and corresponding global longitudinal and circumferential systolic strain measurements were also significantly reduced. Trastuzumab was ceased and supportive cardiac therapy commenced. The ensuing LVEF and systolic strain measurements showed consistent improvement so that trastuzumab was recommenced (while supportive cardiac therapy continued). At this point, reduced circumferential systolic strain with preserved LVEF was observed. Subsequent echocardiograms revealed further reductions in circumferential and longitudinal systolic strain without reductions in LVEF.


Does maternal platelet-associated or platelet-bindable IgG correlate with levels in umbilical cord blood or colostrum during normal pregnancy?
Existing data regarding the ability to predict neonatal thrombocytopenia during maternal immune thrombocytopenia are confusing. We studied normal pregnancies (n = 20) to define normal values and the correlation between maternal and umbilical cord platelet counts, platelet-associated immunoglobulin G (IgG), and platelet-bindable IgG. The postpartum serum platelet-bindable IgG level was measured to evaluate peripartum changes and the correlation with colostrum platelet-bindable IgG (n = 6). The mean maternal platelet count was 181,500 cells/cm3 mm and the mean umbilical cord platelet count was 293,500 cells/mm3. The median maternal platelet-associated IgG was 803 molecules per platelet, umbilical cord platelet-associated IgG was 791 molecules per platelet, maternal platelet-bindable IgG was 92 molecules per platelet, and umbilical cord platelet-bindable IgG was 256 molecules per platelet. The postpartum median maternal platelet-bindable IgG was 333 molecules per platelet and for colostrum it was 297 molecules per platelet. No clinically useful correlations for predicting the neonatal platelet count during normal pregnancy were found. Normal pregnancies may have high levels of maternal- or umbilical cord platelet-associated IgG, perhaps due to nonspecific binding.


The chlamydial developmental cycle.
Intracellular parasitism by bacterial pathogens is a complex, multi-factorial process that has been exploited successfully by a wide variety of organisms. Members of the Order Chlamydiales are obligate intracellular bacteria that are transmitted as metabolically inactive particles and must differentiate, replicate, and re-differentiate within the host cell to carry out their life cycle. Understanding the developmental cycle has been greatly advanced by the availability of complete genome sequences, DNA microarrays, and advanced cell biology techniques. Measuring transcriptional changes throughout the cycle has allowed investigators to determine the nature of the temporal gene expression changes required for bacterial growth and development.


Pleuropulmonary amoebiasis in Ghana: a report of three cases.
Three cases of pleuropulmonary amoebiasis are reported for the first time from Ghana. It is suggested that where facilities for identification of the causal agent are not available a therapeutic trial should be given with drugs effective against Entamoeba histolytica.


Circulating autoantibodies against the NR2 peptide of the NMDA receptor are associated with subclinical brain damage in hypertensive patients with other pre-existing conditions for vascular risk.
Arterial hypertension (HT) and other vascular pre-existing conditions (PEC) generate asymptomatic brain damage which increases the occurrence of stroke and cognitive decline. The aim of this work was to explore if serum antibodies against the NR2 subunit of the NMDA receptor (NR2Ab) could predict subclinical brain damage (SBD) in hypertensive patients with PEC. Forty seven neurologically asymptomatic hypertensive subjects were classified according to the number of PEC (retinopathy, overweight/obesity, diabetes mellitus and dyslipidemia). NR2A/B Ab were measured in serum employing an ELISA method. 3.0-T Brain MRI imaging was performed, and visual ratings of white matter hyperintensities (WMH) and counts of dilated Virchow-Robin spaces (DVRS) and lacunes were obtained. Brain atrophy was evaluated with cortical thickness measurements and linear measures. Higher levels of NR2Ab were associated with more severe periventricular WMH (PWMH), more DVRS and more severe SBD; while greater frontal interhemispheric fissure width (IHFW), as a linear measure of frontal atrophy, was inversely related with NR2Ab. Overall and regional cortical thickness were not significantly associated with NR2 Ab. A multivariate analyses showed that IHFW and PWMH were the only variables independently associated with serum NR2Ab concentration. ROC analysis revealed that NR2Ab (cutoff: 1.7ng/ml) predicted PWMH with a sensitivity and specificity of 65% and 87% respectively. Serum NR2Ab levels may reflect SBD in HT subjects with PEC, especially in younger populations at risk, where age-related cortical atrophy has not yet been fully established.


Chemical and biological properties of diesel exhaust particles collected during selected segments of a simulated driving cycle.
Particle emissions, percentage of organic extractable materials, and mutagenicities of extracts from a diesel engine operating on a test stand have been determined for the full Federal Test Procedure driving cycle and several individual segments thereof. Particle samples were collected using a computer controlled high volume sampler. Extracts of the exhaust particles were screened for the potent mutagens nitropyrene/nitrofluoranthenes by mass spectrometry/mass spectrometry (MS/MS). Results indicate that a long acceleration from 0-55 mph produced approximately seven times more particles per second than the full cycle. Also, the 0- to 55-mph acceleration and a subsequent 55-mph cruise produced significantly higher amounts of mutagens than other segments or the full FTP cycle. A direct correlation of both NOx levels and temperature with mutagenicity was noted (r = 0.89 and r = 0.89). The specific activities of the extracts showed decreases or remained unchanged when assayed in TA-98 NR or TA-98 1,8 DNP6, nitroreductase deficient strains of TA-98. Three extracts were found to have high levels of nitropyrenes/nitrofluoranthenes, and two of the three had high specific activities in TA-98.


The Fragility of Significance in the Hip Arthroscopy Literature: A Systematic Review.
The purpose of the present study was to perform the first examination of the utility of p values and the degree of statistical fragility in the hip arthroscopy literature by applying both the Fragility Index (FI) and the Fragility Quotient (FQ) to dichotomous comparative trials. We hypothesized that dichotomous comparative trials evaluating categorical outcomes in the hip arthroscopy literature are statistically fragile. The PubMed and MEDLINE databases were queried from 2008-2018 for comparative studies evaluating dichotomous data in the hip arthroscopy literature. The present analysis included both randomized controlled trials (RCTs) and non-RCTs in which dichotomous data and associated p values were reported. Fragility analysis was performed with use of the Fisher exact test until an alteration of significance was determined. Of the 5,836 studies screened, 4,156 met the search criteria, with 52 comparative studies included for analysis. One hundred and fifty total outcome events with 33 significant (p < 0.05) outcomes and 117 nonsignificant (p ≥ 0.05) outcomes were identified. The final FI incorporating all 150 outcome events from 52 comparative studies was only 3.5 (interquartile range, 2 to 6), with an associated FQ of 0.032 (interquartile range, 0.017 to 0.063). Twenty-two studies (42.3%) either failed to report loss to follow-up (LTF) data or reported LTF greater than the overall FI of 3.5. The peer-reviewed hip arthroscopy literature may not be as stable as previously thought, as the sole reliance on a threshold p value has proven misleading. We therefore recommend reporting of the FI and FQ, in conjunction with p values, to aid in the evaluation and interpretation of statistical robustness and quantitative significance in future comparative hip arthroscopy studies.


Mre11 regulates CtIP-dependent double-strand break repair by interaction with CDK2.
Homologous recombination facilitates accurate repair of DNA double-strand breaks (DSBs) during the S and G2 phases of the cell cycle by using intact sister chromatids as sequence templates. Homologous recombination capacity is maximized in S and G2 by cyclin-dependent kinase (CDK) phosphorylation of CtIP, which subsequently interacts with BRCA1 and the Mre11-Rad50-NBS1 (MRN) complex. Here we show that, in human and mouse, Mre11 controls these events through a direct interaction with CDK2 that is required for CtIP phosphorylation and BRCA1 interaction in normally dividing cells. CDK2 binds the C terminus of Mre11, which is absent in an inherited allele causing ataxia telangiectasia-like disorder. This newly uncovered role for Mre11 does not require ATM activation or nuclease activities. Therefore, functions of MRN are not restricted to DNA damage responses but include regulating homologous recombination capacity during the normal mammalian cell cycle.


Proton magnetic resonance studies of alpha-keto acids.
Pulsed Fourier transform proton magnetic resonance was used to study alpha-ketoglutaramic, and several other alpha-keto acids in aqueous solutions as a function of pH. Most alpha-keto acids were found to exist in equilibrium with the hydrate (gem-doil). The equilibrium position favors the nonhydrated alphs-keto acid at neutral pH, but at low pH values (below the pKa of the alpha-carboxylic acid group) the hydrate predominates. We found evidence that alpha-ketoglutaric acid exists in a third equilibrium form which is assigned to the lactol. alpha-Ketoglutaramic acid (the alpha-keto acid analog of glutamine) which is known to exist predominantly in a cyclic form at pH 7.0 was shown to exist as a cyclic structure over a wide pH range. However, the cyclic form is an equilibrium mixture of 2-pyrrolidone-5-hydroxy-5-carboxylic and 1-pyrrolin-2-one-5-carboxylic acids.


Efficient Large-Scale 2D Culture System for Human Induced Pluripotent Stem Cells and Differentiated Cardiomyocytes.
Cardiac regenerative therapies utilizing human induced pluripotent stem cells (hiPSCs) are hampered by ineffective large-scale culture. hiPSCs were cultured in multilayer culture plates (CPs) with active gas ventilation (AGV), resulting in stable proliferation and pluripotency. Seeding of 1 × 106 hiPSCs per layer yielded 7.2 × 108 hiPSCs in 4-layer CPs and 1.7 × 109 hiPSCs in 10-layer CPs with pluripotency. hiPSCs were sequentially differentiated into cardiomyocytes (CMs) in a two-dimensional (2D) differentiation protocol. The efficiency of cardiac differentiation using 10-layer CPs with AGV was 66%-87%. Approximately 6.2-7.0 × 108 cells (4-layer) and 1.5-2.8 × 109 cells (10-layer) were obtained with AGV. After metabolic purification with glucose- and glutamine-depleted and lactate-supplemented media, a massive amount of purified CMs was prepared. Here, we present a scalable 2D culture system using multilayer CPs with AGV for hiPSC-derived CMs, which will facilitate clinical applications for severe heart failure in the near future.


A Copper-Based Metal-Organic Framework as an Efficient and Reusable Heterogeneous Catalyst for Ullmann and Goldberg Type C-N Coupling Reactions.
A highly porous metal-organic framework (Cu-TDPAT), constructed from a paddle-wheel type dinuclear copper cluster and 2,4,6-tris(3,5-dicarboxylphenylamino)-1,3,5-triazine (H₆TDPAT), has been tested in Ullmann and Goldberg type C-N coupling reactions of a wide range of primary and secondary amines with halobenzenes, affording the corresponding N-arylation compounds in moderate to excellent yields. The Cu-TDPAT catalyst could be easily separated from the reaction mixtures by simple filtration, and could be reused at least five times without any significant degradation in catalytic activity.


Synthesis and SAR of highly selective MMP-13 inhibitors.
The structure-based design and synthesis of a series of novel biphenyl sulfonamide carboxylic acids as potent MMP-13 inhibitors with selectivity over MMP-1, MMP-2, MMP-3, MMP-7, MMP-8, MMP-9, MMP-14, Aggrecanase 1, and TACE are described.


Transdermal nanolipoplex simultaneously inhibits subcutaneous melanoma growth and suppresses systemically metastatic melanoma by activating host immunity.
Benefit for clinical melanoma treatments, the transdermal neoadjuvant therapy could reduce surgery region and increase immunotherapy efficacy. Using lipoplex (Lipo-PEG-PEI-complex, LPPC) encapsulated doxorubicin (DOX) and carrying CpG oligodeoxynucleotide; the transdermally administered nano-liposomal drug complex (LPPC-DOX-CpG) would have high cytotoxicity and immunostimulatory activity to suppress systemic metastasis of melanoma. LPPC-DOX-CpG dramatically suppressed subcutaneous melanoma growth by inducing tumor cell apoptosis and recruiting immune cells into the tumor area. Animal studies further showed that the colonization and growth of spontaneously metastatic melanoma cells in the liver and lung were suppressed by transdermal LPPC-DOX-CpG. Furthermore, NGS analysis revealed IFN-γ and NF-κB pathways were triggered to recruit and activate the antigen-presenting-cells and effecter cells, which could activate the anti-tumor responses as the major mechanism responsible for the therapeutic effect of LPPC-DOX-CpG. Finally, we have successfully proved transdermal LPPC-DOX-CpG as a promising penetrative carrier to activate systemic anti-tumor immunity against subcutaneous and metastatic tumor.


Prediction of warmed-over flavour development in cooked chicken by colorimetric sensor array.
The aim of this study was to develop a simple and rapid method based on colorimetric sensor array (CSA) for evaluation of warmed-over flavour (WOF) in cooked chicken. All samples were classified according to storage time by CSA coupled with principle component analysis (PCA) or hierarchical cluster analysis (HCA). The CSA data were used to establish prediction models with thiobarbituric acid reactive substances (TBARS), pentanal, hexanal, or heptanal associated with WOF by partial least square regression (PLSR). For the TBARS model, the coefficient of determination (rp(2)) was 0.9997 in the prediction range of 0.28-0.69mg/kg. In each of the models for pentanal, hexanal, and heptanal, all rp(2) were higher than 0.960 in the range of 0.58-2.10mg/kg, 5.50-11.69mg/kg, and 0.09-0.16mg/kg, respectively. These results demonstrate that the CSA was able to predict WOF development and to distinguish between each storage time.


Usefulness of the mitral valve Z score in predicting the need to close moderate- to large-sized atrial septal defects in infants with aortic coarctation.
Left ventricular anatomic substrates predisposing the development of congestive heart failure in infants with atrial left-to-right shunting after aortic coarctation repair were characterized. Mitral valve, but not aortic or left ventricular, hypoplasia correlated with the development of heart failure and the need for atrial defect closure shortly after coarctation repair.


Genetic diversity in South African Nguni cattle ecotypes based on microsatellite markers.
The Nguni cattle breed is a landrace breed adapted to different ecological regions of South Africa. A number of ecotypes are recognised based on phenotype within the breed, but it is not known if they are genetically distinct. In this study, molecular characterisation was performed on Makhathini (MAK), Pedi (PED), Shangaan (SHA) and Venda (VEN) Nguni cattle ecotypes. Two Nguni cattle populations, not kept as separate ecotypes, from the University of Fort Hare (UFH) and Agricultural Research Council Loskop South farm (LOS) were also included. Genotypic data was generated for 189 unrelated Nguni cattle selected based on pedigree records using 22 microsatellite markers. The expected heterozygosity values varied from 69 % (UFH) to 72 % (PED) with a mean number of alleles ranging from 6.0 to 6.9. The F ST estimate demonstrated that 4.8 % of the total genetic variation was due to the genetic differentiation between the populations and 92.2 % accounted for differences within the populations. The genetic distances and structure analysis revealed the closest relationship between MAK, PEDI and SHA ecotypes, followed by SHA and VEN. The UFH population clustered with the MAK ecotype, indicating that they are more genetically similar, while the LOS cattle grouped as a distinct cluster. Results suggest that the genetic differentiation between the PED and SHA ecotypes is low and can be regarded as one ecotype based on limited genetic differences. The results of this study can be applied as a point of reference for further genetic studies towards conservation of Nguni cattle ecotypes.


What Triggers a Diagnosis of HIV Infection in the Tokyo Metropolitan Area? Implications for Preventing the Spread of HIV Infection in Japan.
Japan has not succeeded in reducing the annual number of new HIV-infected patients, although the prevalence of HIV infection is low (0.02%). A single-center observational study was conducted at the largest HIV clinic in Tokyo, which treats 15% of the total patients in Japan, to determine the reasons for having diagnostic tests in newly infected individuals. HIV-infected patients who visited our clinic for the first time between 2011 and 2014 were analyzed. The 598 study patients comprised one-third of the total reported number of new patients in Tokyo during the study period. 76% were Japanese MSM. The reasons for being tested which led to the diagnosis was voluntary testing in 32%, existing diseases in 53% (AIDS-defining diseases in 22%, sexually transmitted infections (STI) in 8%, diseases other than AIDS or STIs in 23%) and routine pre-surgery or on admission screening in 15%. 52% and 74% of the study patients and patients presented with AIDS, respectively, had never been tested. The median CD4 count in patients with history of previous testing (315/μL) was significantly higher than that of patients who had never been tested (203/μL, p<0.001). Only 32% of the newly HIV diagnosed patients were diagnosed because of voluntary testing, and 53% were diagnosed due to presence of other diseases. These results remain unchanged from our previous report 10 years earlier (2000-2004) on newly diagnosed patients at the same clinic. HIV testing has not been widely used by newly diagnosed patients in the Tokyo metropolitan area.


Label-free and quantitative analysis of C-reactive protein in human sera by tagged-internal standard assay on antibody arrays.
We have developed a new, high-throughput, competition-based tagged-internal standard (TIS) assay to measure the levels of blood proteins in human serum. In this assay, target proteins in the sample serum compete with tagged-internal standard proteins for binding to an antibody array. Antibody arrays are fabricated by immobilizing a target protein-specific antibody on the carboxylate-modified latex bead surface of well-type arrays. A solution of Alexa 546-conjugated target protein is added to a sample of human serum and applied to the well-type antibody array. The array is then analyzed with a fluorescence scanner and the level of unlabeled target protein in the human sera is inferred from the amount of tagged protein bound to the array. We successfully applied this assay to measure the level of C-reactive protein (CRP) in 92 unlabeled human sera. The TIS assay was found to be specific and reproducible for the quantitative analysis of CRP. The antibody array data from the TIS assay correlate well with clinical laboratory data obtained using the commercialized latex-enhanced turbidimetry immunoassay (n=3, r=0.967, CV=0.32%). Thus, the antibody array-based TIS assay system is high-throughput, quantitative, and label-free and may be useful in the rapid serodiagnosis of human disease.


Aminoglycoside-resistance mechanisms in multidrug-resistant Staphylococcus aureus clinical isolates.
Aminoglycoside resistance in six clinically isolated Staphylococcus aureus was evaluated. Genotypical examination revealed that three isolates (HLGR-10, HLGR-12, and MSSA-21) have aminoglycoside-modifying enzyme (AME) coding genes and another three (GRSA-2, GRSA-4, and GRSA-6) lacked these genes in their genome. Whereas isolates HLGR-10 and HLGR-14 possessed bifunctional AME coding gene aac(6')-aph(2''), and aph(3')-III and showed high-level resistance to gentamycin and streptomycin, MSSA-21 possessed aph(3')-III and exhibited low resistance to gentamycin, streptomycin, and kanamycin. The remaining three isolates (GRSA-2, GRSA-4, and GRSA-6) exhibited low resistance to all the aminoglycosides because they lack aminoglycoside-modifying enzyme coding genes in their genome. The transmission electron microscopy of the three isolates revealed changes in cell size, shape, and septa formation, supporting the view that the phenomenon of adaptive resistance is operative in these isolates.


Experience with MTBE as a solvent for common bile duct stones in patients with T-tube in situ.
The postcholecystectomy patients who have a T-tube in situ offer a convenient route through the T-tube to perfuse solvents into the common bile duct (CBD) for dissolving any retained common duct stones. If successful, this approach is much simpler and cheaper than the usual therapeutic modality used for CBD stones, namely, endoscopic papillotomy. Thus a most potent cholesterol solvent, methyl t-butyl ether (MTBE) was perfused through the T-tube into the CBD of five patients with retained common duct stones. The dose of the solvent varied, 1.5-5 mL 0.5-1 h, given 7-13 times amounting to a total of 20-66 mL. Instillation of MTBE in the T-tube was alternated with aspiration of the bile through T-tube. Only one patient showed complete disappearance of the bile duct stone following MTBE perfusion. Others did not show any appreciable response and had to be treated by endoscopic papillotomy (three patients) or mono-octanoin perfusion (one patient). Side-effects of MTBE perfusion included pain in the abdomen in all patients, somnolence and nausea/vertigo in two patients and the smell of ether on the breath in two patients. It is concluded that MTBE is not an effective agent for dissolution of retained CBD stones in patients with T-tube in situ.


Clinical investigation of the alteration in the middle ear impedance in patients with intracranial hypertension.
Marked pulsation occurring during middle ear impedance measurements may be caused by an intracranial hypertension sydnrome, with pulsations being transmitted to the middle ear by a variety of preformed natural pathways. Impedancemetry is a useful technique for diagnosing and following the syndrome.


Bladder papilloma in a 12-year-old girl.
Epithelial bladder tumours are rare in childhood. A case of bladder papilloma in a 12-year-old girl is reported. Emphasis is given to the important role of ultrasound in diagnosis. Endoscopic treatment was successful and no evidence of recurrence was found at 5-year follow-up. Exposure to a chemical environmental factor (volatile shoe adhesives) is suggested as a possible cause of the tumour.


Combination of alpha-1-acid glycoprotein and alpha-fetoprotein as an improved diagnostic tool for hepatocellular carcinoma.
To evaluate the diagnostic value of alpha-1-acid glycoprotein (AAG) and the combination with alpha-fetoprotein (AFP) in hepatocellular carcinoma (HCC) patients. AAG was measured in serum of 65 HCC patients and 54 chronic liver diseases (CLD) patients by using proteomic approach. Sensitivity and specificity of AAG and its combination with AFP were determined and compared with AFP alone for the diagnosis of HCC. The expression concentration of AAG was significantly higher in HCC patients than chronic liver disease with sensitivity (77%) and accuracy (83%). Receiver operating characteristic analysis yielded the following AUC: AFP 0.750 (CI 95% 0.663-0.837), AAG 0.907 (CI 95% 0.855-0.960) and AFP+AAG 0.943 (CI 95% 0.897-0.988). At a specificity of 90%, the combination of AFP+AAG had sensitivity 89% and accuracy 90%, which was higher than sensitivity (52.3%) and accuracy (70%) when using AFP alone. The combination of AAG and AFP shows high sensitivity and improves the accuracy of HCC diagnosis.


How does it feel to lack a sense of boundaries? A case study of a long-term mindfulness meditator.
This paper discusses the phenomenological nature of the sense of boundaries (SB), based on the case of S, who has practiced mindfulness in the Satipathana and Theravada Vipassana traditions for about 40years and accumulated around 20,000h of meditative practice. S's unique abilities enable him to describe his inner lived experience with great precision and clarity. S was asked to shift between three different stages: (a) the default state, (b) the dissolving of the SB, and (c) the disappearance of the SB. Based on his descriptions, we identified seven categories (with some overlap) that alter during the shifts between these stages, including the senses of: (1) internal versus external, (2) time, (3) location, (4) self, (5) agency (control), (6) ownership, and (7) center (first-person-egocentric-bodily perspective). Two other categories, the touching/touched structure and one's bodily feelings, do not fade away completely even when the sense-of-boundaries disappears.


Clinical Impact of Lesion Complexity on 2-Year Outcomes After Zotarolimus-Eluting Stents Implantation.
The clinical efficacy and safety of second-generation drug-eluting stents in complex percutaneous coronary interventions (PCIs) are not well established. The clinical influence of the lesion complexity after PCI with zotarolimus-eluting stents (ZES) was evaluated. From a prospective multicenter observational study, a total of 926 patients that underwent successful PCIs with ZES were included. Complex PCIs were defined as patients with ≥3 lesions treated, 3 vessels treated, severe calcified lesions, bifurcated lesions with 2 stents implanted, left main disease, chronic total occlusion lesions, and/or diffuse long (total stent length ≥60 mm) lesions and were compared to the noncomplex group. The primary outcome was incidence of target lesion failures at 2 years, defined as a composite of cardiac death, target lesion-myocardial infarctions, and target lesion revascularization. The patients were divided into complex PCI (n = 249) and noncomplex (n = 677) groups. In the complex PCI group, the 2-year risk of a target lesion failure was not significantly higher than in the noncomplex PCI group (4.8% vs 3.7%; adjusted hazard ratio: 1.373; 95% confidence interval: 0.689-2.738; P = 0.367). The same trend was observed for all composites of the clinical outcomes. Older age and advanced chronic kidney disease were independent predictors for the primary outcome. Up to 2 years after a ZES implantation, the clinical outcomes did not differ according to lesion complexity.


Map of physical interactions between extracellular domains of Arabidopsis leucine-rich repeat receptor kinases.
Plants use surface receptors to perceive information about many aspects of their local environment. These receptors physically interact to form both steady state and signalling competent complexes. The signalling events downstream of receptor activation impact both plant developmental and immune responses. Here, we present a comprehensive study of the physical interactions between the extracellular domains of leucine-rich repeat receptor kinases (LRR-RKs) in Arabidopsis. Using a sensitized assay, we tested reciprocal interactions among 200 of the 225 Arabidopsis LRR-RKs for a total search space of 40,000 interactions. Applying a stringent statistical cut-off and requiring that interactions performed well in both bait-prey and prey-bait orientations resulted in a high-confidence set of 567 bidirectional interactions. Additionally, we identified a total of 2,586 unidirectional interactions, which passed our stringent statistical cut-off in only one orientation. These datasets will guide further investigation into the regulatory roles of LRR-RKs in plant developmental and immune signalling decisions.


Lysozyme localization in human gastric and duodenal epithelium. An immunocytochemical study.
The distribution of lysozyme in normal gastric and duodenal mucosa was studied by light- and electron-microscopic immunocytochemical techniques (direct enzyme-labeled antibody method). In the duodenal mucosa, lysozyme was found in the Paneth cells and the epithelial cells of Brunner's glands. Electron-microscopically, lysozyme was found in rough endoplasmic reticulum and perinuclear spaces, which were assumed to be protein-synthesizing organelles, and also in the secretory granules of Paneth cells. Additionally, lysozyme was detected in the stomach in mucinous granules and in some parts of the rough endoplasmic reticulum within the epithelial cells of the pyloric glands, the mucous neck cells of the fundic glands, and in several surface epithelial cells of the plyoric and fundic regions. This suggests that some quantity of lysozyme in gastrointestinal secretion originates from the gastric and duodenal glands, and that it acts as a defense mechanism in the gastrointestinal tract.


Reverse apolipoprotein A-I mimetic peptide R-D4F inhibits neointimal formation following carotid artery ligation in mice.
The ApoA-I mimetic peptide D-4F has demonstrated potent atheroprotective actions in vivo and in vitro. We investigated the effect of R-D4F (ie, the D-4F peptide with reverse order of amino acids) on intimal hyperplasia after vascular injury in a mouse model of carotid artery ligation. Adult male C57BL/6J mice were pretreated intraperitoneally with vehicle, D-4F (1 mg/kg), or R-D4F (1 mg/kg or 5 mg/kg) daily for 3 days; the mice were then subjected to left carotid artery ligation. All treatments were continued for 28 days after surgery. Neither D-4F nor R-D4F treatment affected serum lipid levels. Morphometric analysis showed that the occluded vessels had significant neointimal formation, compared with the uninjured arteries in vehicle-treated mice. Like the D-4F treatment, R-D4F treatment significantly (P < 0.05) inhibited intimal hyperplasia (-42%), local neutrophil and macrophage infiltration, and mRNA expression of the proinflammatory mediator monocyte chemotactic protein 1 (-55%) and vascular cell adhesion protein 1 (-53%), compared with vehicle. Furthermore, the vasoprotective effect of high-dose R-D4F was significantly enhanced, compared with the low dose. In cultured mouse RAW 264.7 macrophages, pretreatment with R-D4F also effectively inhibited lipopolysaccharide-induced leukocyte integrin CD11b expression, a key molecule for leukocyte infiltration. Taken together, these results suggest that R-D4F has significant anti-inflammatory features and facilitates prevention of neointimal formation after vascular injury in mice.


Lipid Droplet-Related PLIN2 in CD68+ Tumor-Associated Macrophage of Oral Squamous Cell Carcinoma: Implications for Cancer Prognosis and Immunotherapy.
PLIN2 (adipose differentiation-related protein) belongs to the perilipin family and is a marker of lipid droplets (LDs). Numerous types of tumor exhibit a high PLIN2 level, but its tumorigenic or tumor-suppressive role has been in debate. Recently, LDs serve as innate immune hubs and show antimicrobial capacity. We here aimed to investigate the heterogeneous functions of PLIN2 in the tumor microenvironment and immune regulation. This retrospective study included 96 oral squamous cell carcinoma (OSCC) samples and analyzed the spatial distribution of PLIN2 by immunohistochemistry (IHC) and LD level by oil red O staining. A total of 21 serial sections were obtained to analyze the relationship between PLIN2 and immune cells by IHC and immunofluorescence (IF). Single-cell sequencing was used to analyze the cell locations of PLIN2. The values of diagnosis and prognosis of PLIN2 were also evaluated. Tumor Immune Estimation Resource (TIMER), cBioPortal databases, and IHC analysis were used to investigate the relationship between PLIN2 and OSCC immune microenvironment. PLIN2 was mainly expressed in tumor-infiltrating immunocytes (TIIs) of OSCC. Patients with high PLIN2 harbored more cytoplastic LDs. CD68+ tumor-associated macrophages (TAMs), instead of T cells and B cells, were found to be the main resource of PLIN2 in OSCC stroma and lung, pancreas, prostate, and testis. However, CD56+ NK cells also showed less extent of PLIN2 staining in OSCC. Moreover, patients with a high PLIN2 level in immune cells had a higher TNM stage and were susceptible to postoperative metastasis, but the escalated PLIN2 level in invasive tumor front independently predicted shorter metastasis-free survival. Furthermore, a high PLIN2 presentation in the microenvironment induced immune suppression which was featured as less infiltration of CD8+ T cells and more CD68+ TAMs and Foxp3+ Tregs, accompanied by more immune checkpoint molecules such as CSF1R, LGALS9, IL-10, CTLA-4, and TIGIT. CD68+ TAM-derived PLIN2 might participate in regulating immune balance of OSCC patients, which provides new insight into immune checkpoint therapy.


Psychological distress and quality of life in Iranian adolescents with overweight/obesity: mediating roles of weight bias internalization and insomnia.
To examine whether weight-related self-stigma (aka weight bias internalization) and insomnia are potential predictors of psychological distress and quality of life (QoL) among Iranian adolescents with overweight (OW)/obesity (OB). To examine whether weight-related self-stigma and insomnia are potential mediators in the relationship between excess weight and health outcomes of distress and QoL. All participants (n = 934; 444 males; mean age = 15.7 ± 1.2 years; zBMI = 2.8 ± 1.0) completed questionnaires on weight-related self-stigma and insomnia at baseline. Six months later, they completed questionnaires on psychological distress and QoL to assess health outcomes. Relationships among variables were tested using mediation analyses with bootstrapping method. Weight-related self-stigma significantly mediated the effects of zBMI on psychological distress (effect = 0.22; bootstrapping SE = 0.09; 95% CI = 0.08, 0.45), psychosocial QoL (effect = - 0.64; bootstrapping SE = 0.19; 95% CI = - 1.10, - 0.32), and physical QoL (effect = - 1.35; bootstrapping SE = 0.54; 95% CI = - 2.43, - 0.26). Insomnia also significantly mediated the effects of zBMI on psychological distress (effect = 2.18; bootstrapping SE = 0.31; 95% CI = 1.61, 2.81), psychosocial QoL (effect = - 0.89; bootstrapping SE = 0.33; 95% CI = - 1.60, - 0.28), and physical QoL (effect = - 0.83; bootstrapping SE = 0.42; 95% CI = - 1.69, - 0.02). Full mediations were found in psychosocial QoL; partial mediations were found in psychological distress and physical QoL. Weight-related self-stigma and insomnia were significant mediators in the effects of excess weight on health outcomes. Therefore, it is important to identify and treat weight-related self-stigma and insomnia for adolescents with OW/OB. Level V, cross-sectional descriptive study.


The role and regulation of the nuclear factor kappa B signalling pathway in human labour.
Within the discipline of reproductive biology, our understanding of one of the most fundamental biological processes is lacking--the cellular and molecular mechanisms that govern birth. This lack of understanding limits our ability to reduce the incidence of labour complications. The incidence of labour complications including: preterm labour; cervical incompetence; and post-date pregnancies has not diminished in decades. The key to improving the management of human labour and delivery is an understanding of how the multiple processes that are requisite for a successful labour and delivery are coordinated to achieve a timely birth. Processes of human labour include the formation of: contraction associated proteins; inflammatory mediators (e.g. cytokines); uterotonic phospholipid metabolites (e.g. prostaglandins); and the induction of extracellular matrix (ECM) remodelling. Increasingly, it is becoming evident that labour onset and birth are the result of cross-talk between multiple components of an integrated network. This hypothesis is supported by recent data implicating various upstream regulatory pathways in the control of key labour-associated processes, including the activity of enzymes involved in the formation of prostaglandins and extracellular matrix remodelling, and mediators of inflammation. Clearly, the biochemical pathways involved in the formation of these mediators represent potential sites for intervention that may translate to therapeutic interventions to delay or prevent preterm labour and delivery. Available data strongly implicate the nuclear factor-kappaB (NF-kappaB) family as candidate upstream regulators of multiple labour-associated processes. Not only do these data warrant further detailed analysis of the involvement of these pathways in the process of human labour but also promise new insights into the key mechanisms that trigger birth and the identification of new therapeutic interventions that will improve the management of labour.


Endoscopic treatment of benign esophageal tumors: case report of three patients.
Benign esophageal tumors are rare. Enucleation of the tumor is considered when the patient reports problems. The traditional approach is to use thoracotomy or laparotomy if the tumor is located in the distal esophagus. The use of minimally invasive techniques permits enucleation with all the concomitant advantages. Our experience with the minimally invasive management of three benign esophageal tumors is described. The methods and results of preoperative studies are reported. The surgical technique is described. Two patients could be managed using thoracoscopy, and one patient required conversion to laparoscopy. All the patients had complete and quick recoveries.


Potential study of basement membrane.
The membrane potential of a basement membrane (bovine-anterior-lens-capsule) was studied by use of the Gibbs-Donnan systems. The T.M.S. (Teorell, Meyer, Sievers) theory was applied to these systems for the treatment of experimental data. The results show that, in the basement membrane, the density of fixed charges such as ionogenic groups encountered in heparan sulfate proteoglycan is low. The ratios of the mobilities from the chloride anion and the alkaline cation are different to those in water. In this type of membrane, the mobility of Cl- is higher than the mobilities of K+, Na+ and Ca2+. This result leads us to investigate further the diffusion problem of electrolytes through basement membrane.


Effects on fracture healing of an antagonist of the vitamin K cycle.
The anticoagulant, dicumarol, inhibits the vitamin K cycle by blocking the conversion of the vitamin K epoxide. The effects of dicumarol on ossification have been tested by feeding it to rats in which a closed fracture of the metatarsals had been induced; the effects were studied up to 12 days postfracture. At 12 days, treatment with dicumarol caused a highly significant decrease in the amount of bone produced, without affecting the total size of the callus. Quantitative cytochemistry of unfixed, undemineralized sections showed that dicumarol also markedly affected the periosteal activities of glucose 6-phosphate dehydrogenase and of alkaline phosphatase in the first 2 mm from the fracture measured at 3 and 5 days postfracture when normally, new bone is first formed. In contrast, dicumarol had little effect on these activities in the fully formed callus.


Design of experiments guided multivariate calibration for the quantitation of injectable colistimethate sodium by ultra performance liquid chromatography - High resolution mass spectrometry.
Colistimethate sodium (CMS) is a widely administrated old-generation prodrug for the treatment of the life-threatening infections caused by multi-resistant Gram-negative bacteria. Until now, the quality control procedure of the CMS commercial products is based on microbiological assays. The aim of the study is the development of a chemical analysis methodology based on liquid chromatography - mass spectrometry (LC-MS) that could be used for the quality control of CMS products. The careful optimization of the LC and QToF-MS parameters was deemed crucial, as CMS is known to be a very complex mixture. Thus, a two stage Design of Experiments (DoE) pipeline has been followed, aiming towards the separation of the mixture components. According to the DoE results, a baseline-resolved chromatogram revealing more than 20 compounds was achieved. The separation was performed using a Waters Acquity BEH C8 column employing gradient elution. The mobile phase consisted of aq. ammonium formate 0.005 M (pH 6) (solvent A) and methanol/acetonitrile 79/21 (v/v) (solvent B). A second optimization experiment for the MS signal was employed in order to achieve maximum sensitivity. The singly charged signals were monitored for the validation in the positive ion mode. The calibration curve range was 50-110 μg mL-1, corresponding to the 80-120% of the nominal CMS amount in the commercial products. Due to the complexity of the CMS chromatograms and the corresponding spectrum of each chromatographic peak, untargeted and targeted approaches were performed employing the MZmine software. Furthermore, apart from the classical univariate statistical analysis, partial least squares regression (PLS-R) model was also employed, as the variables were more than the observations. The developed methodology has been employed to analyze several batches and inconsistences have been discovered.


Transient expression of the chloramphenicol acetyltransferase gene in cultured mosquito cells.
A recombinant plasmid in which the bacterial chloramphenicol acetyltransferase (CAT) gene is under the control of the Drosophila heat-shock protein (hsp) 70 promoter has been introduced into cultured mosquito (Aedes albopictus) cells using 1,5-dimethyl-1,5-diazaundecamethylene polymethobromide (polybrene) and dimethylsulfoxide (DMSO). CAT activity was induced by incubating transfected cells at 37 degrees C, and high levels of enzyme activity were maintained for more than 24 h after the temperature shock. Transfected DNA was maintained in the cells for at least 4 days. These experiments document an effective method for introducing purified DNA into cultured mosquito cells.


Pregnancy favors circulating IL-21-secreting TFH -like cell recovery in ARV-treated HIV-1-infected women.
Pregnancy appears to favor maternal antibody production. In contrast, by damaging follicular helper T cells (TFH ), HIV-1 infection compromises protective humoural immune response. Therefore, we aimed to investigate the frequency of different TFH -like cells in HIV-infected pregnant women (PW) before and after antiretroviral (ARV) therapy. Peripheral blood mononuclear cells, CD4+ T and B cells, were obtained from asymptomatic HIV-1-infected non-PW and PW just before and after ARV therapy. In some experiments, healthy HIV-1-negative PW were also tested. The frequency of different TFH -like cell subsets was determined by flow cytometry. The plasma titers of IgG anti-tetanus toxoid (TT), anti-HBsAg, and anti-gp41 were determined by ELISA. The in vitro production of total IgG, IL-21, and hormones (estrogen and progesterone) was quantified also by ELISA. Our results demonstrate that antiretroviral (ARV) therapy was more efficient in elevating the percentage of circulating IL-21-secreting TFH cells in HIV-1-infected pregnant women (PW) than in non-pregnant patients (nPW). Moreover, in co-culture systems, CD4+ T cells from ART-treated PW were more efficient in assisting B cells to produce IgG production. The in vivo anti-HBsAg IgG titers after ARV therapy were also significantly higher in PW, and their levels were directly associated with both IL-21+ TFH frequency and plasma concentration of estrogen. In summary, our results suggest that pregnancy favors the recovery of TFH -like cells after ARV therapy in HIV-1-infected women, which could help these mothers to protect their newborns from infectious diseases by transferring IgG across the placenta.


Impossibility of Consistent Distance Estimation from Sequence Lengths Under the TKF91 Model.
We consider the problem of distance estimation under the TKF91 model of sequence evolution by insertions, deletions and substitutions on a phylogeny. In an asymptotic regime where the expected sequence lengths tend to infinity, we show that no consistent distance estimation is possible from sequence lengths alone. More formally, we establish that the distributions of pairs of sequence lengths at different distances cannot be distinguished with probability going to one.


Antifungal activity of organobismuth compounds against the yeast Saccharomyces cerevisiae: structure-activity relationship.
Antifungal activity of organobismuth(III) and (V) compounds 1-9 was examined against the yeast, Saccharomyces cerevisiae. A clear structure-activity relationship was observed in these compounds. Thus, triarylbismuth dichlorides 2 [(4-YC6H4)3BiCl2: Y=MeO, F, Cl, CF3, CN, NO2] and halobismuthanes 6 [2-(t)BuSO2C6H4(4-YC6H4)BiX: Y=MeO, Me, H, Cl; X=Cl, Br, I], 7 [Bi(X)(C6H4-2-SO2C6H4-1'-): X=Cl, Br, I], 8 [2-Me2NCH2C6H4(Ph)BiX: X=Cl, Br] and 9 [4-MeC6H4(8-Me2NC10H6-1-)BiCl] showed the growth inhibition effect, while triarylbismuth difluorides 3 [(4-YC6H4)3BiF2] and triarylbismuthanes 1 [(4-YC6H4)3Bi], 4 [2-(t)BuSO2C6H4(4-YC6H4)2Bi] and 5 [4-YC6H4Bi(C6H4-2-SO2C6H4-1'-)] were not active at all irrespective of the nature of the substituents. Generation of the inhibition effect is governed by the facility of nucleophilic reaction at the bismuth center and the Lewis acidic bismuth center is an active site. Of all the bismuth compounds attempted, halobismuthanes 7 derived from diphenyl sulfone exhibited the highest activities. An X-ray crystallographic study of 7a [Bi(Cl)(C6H4-2-SO2C6H4-1'-)] revealed that the bismuth center adopts a seven-coordinated geometry, which is unusual in organobismuth(III) compounds, through the intramolecular and intermolecular coordination between the bismuth and oxygen atoms. The marked inhibition effect of 7 may be attributed to such a highly coordinated geometry, which allows the bismuth center to bind tightly with some biomolecules playing important roles in the growth of S. cerevisiae.


Front-runners for pharmacotherapeutic correction of the airway ion transport defect in cystic fibrosis.
Although cystic fibrosis (CF) patients display multiorgan dysfunction (e.g. pancreas, gut, and lung) it is lung disease that is the leading cause of premature death in these patients. CF lung disease is characterized by persistent pulmonary infection and mucus plugging of the airways initiated by the failure of solute transport across the airway epithelium. Many drug therapies aim to alleviate the secondary characteristics of CF lung disease; however, new therapies in development are targeted at correcting the ion transport deficiency of CF. The goal is to hydrate airway surfaces by stimulating secretion (through activation of the CF transmembrane conductance regulator and calcium-activated chloride channels), and/or inhibiting absorption (through the epithelial sodium channel) thereby stimulating healthy mucociliary clearance. If mucociliary clearance can be stimulated sufficiently from an early age, then there is the possibility that secondary lung infection may be eradicated from the syndrome of CF disease.


Surgical treatment for cervical aortic arch with aneurysm formation.
Cervical aortic arch is an unusual malformation. Cervical aortic arch with aneurysm formation is very rare. We report a case of cervical aortic arch associated with a saccular aneurysm in a 59-year-old Japanese man. The aneurysm protruded caudally and was located between the left common carotid and left subclavian arteries. Cardiopulmonary bypass and deep hypothermic circulatory arrest was applied as adjunct methods. A Dacron graft was sutured just distal to the left common carotid artery, with the patient in the Trendelenburg position. The proximal site was left open while oxygen-saturated venous blood was supplied in a retrograde manner to perfuse the lower body during occlusion of the descending aorta. Distal anastomosis to the descending aorta was performed during rewarming. The left subclavian artery was reconstructed by using a branch of the graft. This procedure is simple and useful for distal arch operations, especially in patients with Haughton D type aneurysms.