Biophysical study of novel oligoelectrolyte-based nonviral gene delivery systems for mammalian cells.
Gene therapy is an important treatment for genetic and acquired diseases. The success of gene therapy is largely dependent on the development of suitable vectors for gene transfer. Vectors are needed to overcome cellular barriers and to achieve efficient DNA delivery with low cytotoxicity. In the present study, we synthesized and characterized a novel comb-like oligoelectrolyte nanocarrier, BG-2, as a nonviral gene delivery vector. A novel surface-active oligoelectrolyte of comb-like structure was synthesized via controlled radical copolymerization using oligoperoxide Cu(+2) coordinating complex as a multisite initiator of graft copolymerization. The critical micellar concentration was determined by Nile Red fluorescence. Complex formation of DNA with BG-2 was determined by YOYO-1 fluorescence. The physicochemical properties of DNA in complex with BG-2 have been investigated by electrophoresis, dynamic light scattering and fluorescence spectroscopy. The BG-2/DNA complex was demonstrated by scanning electron microscopy. Interactions between BG-2/DNA complex and model membranes were also studied. The sensitivity of the DNA molecule, complexed with BG-2, against deoxyribonuclease I and serum nucleases was assessed by agarose gel electrophoresis. BG-2 efficiency in the transfection of HeLa cells was determined by measuring luciferase gene expression using a luminometer and cytotoxicity was also evaluated. BG-2 oligoelectrolyte was successful in overcoming cellular barriers as a result of forming stable and small sized complexes with DNA, interacting with model membranes in a desirable manner and protecting DNA from nuclease. The transfection efficiency was quite high and cytotoxicity was low. BG-2 appears to be a promising nonviral vector with low cytotoxicity and efficient transfection properties.


Changes of the transcriptional and fatty acid profiles in response to n-3 fatty acids in SH-SY5Y neuroblastoma cells.
Synthesis of docosahexaenoic acid (DHA) from its metabolic precursors contributes to membrane incorporation of this FA within the central nervous system. Although cultured neural cells are able to produce DHA, the membrane DHA contents resulting from metabolic conversion do not match the high values of those resulting from supplementation with preformed DHA. We have examined whether the DHA precursors down-regulate the incorporation of newly formed DHA within human neuroblastoma cells. SH-SY5Y cells were incubated with gradual doses of alpha-linolenic acid (alpha-LNA), EPA, or docosapentaenoic acid (DPA), and the incorporation of DHA into ethanolamine glycerophospholipids was analyzed as a reflection of synthesizing activity. The incorporation of EPA, DPA, and preformed DHA followed a dose-response saturating curve, whereas that of DHA synthesized either from alpha-LNA, EPA, or DPA peaked at concentrations of precursors below 15-30 microM and sharply decreased with higher doses. The mRNA encoding for six FA metabolism genes were quantified using real-time PCR. Two enzymes of the peroxisomal beta-oxidation, L-bifunctional protein and peroxisomal acyl-CoA oxidase, were expressed at lower levels than fatty acyl-CoA ligase 3 (FACL3) and delta6-desaturase (delta6-D). The delta6-D mRNA slightly increased between 16 and 48 h of culture, and this effect was abolished in the presence of 70 microM EPA. In contrast, the EPA treatment resulted in a time-dependent increase of FACL3 mRNA. The terminal step of DHA synthesis seems to form a "metabolic bottleneck," resulting in accretion of EPA and DPA when the precursor concentration exceeds a specific threshold value. We conclude that the critical precursor- concentration window of responsiveness may originate from the low basal expression level of peroxisomal enzymes.


Interlaboratory comparison of a standardized toxicity test using the nematode Caenorhabditis elegans (ISO 10872).
A ring test was carried out within the standardization process of ISO 10872 to evaluate the precision of the toxicity test for the nematode Caenorhabditis elegans. Eight different laboratories tested aqueous solutions of the reference substance benzylcetyldimethylammonium chloride as well as native sediments and soils for toxic effects on the growth and reproduction of C. elegans. Validity criteria were met in all laboratories. Average median- and low-effect concentrations were determined to be 15.1 mg L(-1) (EC50) and 8.7 mg L(-1) (EC10) for growth and 7.5 mg L(-1) (EC50) and 3.8 mg L(-1) (EC10) for reproduction of C. elegans, with ECx values showing a high degree of reproducibility (CV(R) : <21% and <11% for EC10 and EC50, respectively) and repeatability (CV(r) : <20% and <7% for EC10 and EC50, respectively). The toxic effects of the sediments and soils revealed by the different laboratories were well related to each samples' degree of chemical contamination. Moreover, the effects showed an acceptable reproducibility (CV(R) : 5-33% and 0-28% for growth and reproduction, respectively) and repeatability (CV(r) : 3-13% and 0-12% for growth and reproduction, respectively). The present study confirms that the toxicity test with C. elegans according to ISO 10872 is a reliable and precise tool to assess the toxicity of aqueous media, freshwater sediments, and soils.


Use of an endoscopic suturing device (the "ESD") to treat patients with gastroesophageal reflux disease, after unsuccessful EndoCinch endoluminal gastroplication: another failure.
Endoluminal gastroplication, using the EndoCinch procedure, has emerged as a potential endoscopic antireflux therapy. Although initial results have been promising, the long-term durability of the treatment is uncertain due to suture loss. A new endoscopic suturing device, the "ESD," has been developed that promises excellent visibility and endoscopic control. The aim of this study was to evaluate prospectively the feasibility and efficacy of the ESD method after EndoCinch failure. The study involved 20 patients with gastroesophageal reflux disease (GERD), who had been initially treated with an EndoCinch procedure, but had relapsed after a median of 7.5 months, with lost or dysfunctional sutures and with reflux symptoms that required proton pump inhibitor (PPI) treatment. Using the ESD, at least three plications were created at the gastroesophageal junction. Patients underwent endoscopy, 24-hour pH monitoring and esophageal manometry before treatment and 6 months afterwards. In addition, reflux symptoms as well as quality-of-life scores were assessed (using the SF-6 and GERD-HRQL scales). The ESD procedure (median procedure time 45 min) was performed successfully in all patients without major complications. After 6 months only one patient (5 %) still had all sutures in situ, while no remaining sutures could be detected in 3/20 (15 %). No significant changes in reflux esophagitis; 24-hour pH monitoring results (median pH < 4/24 h9.9 % vs. 12.3 %; P = 0.60); manometry findings (median lower esophageal sphincter pressure 7.2 mm Hg vs. 9.9 mm Hg; P = 0.22); PPI use; or reflux esophagitis could be detected after 6 months. While reflux symptoms improved (heartburn severity score 30 vs. 48, P < 0,05), no changes in quality-of-life scores were detected. Endoluminal gastroplication using the ESD is an easy and safe, but unfortunately ineffective procedure for endoscopic GERD treatment. Endoluminal gastroplication techniques clearly need refinements before these therapies can evolve as a treatment option for GERD patients.


Relationships between thromboxane production, platelet aggregability, and serum concentrations of ibuprofen or flurbiprofen.
Arachidonate-induced ex vivo platelet aggregations were performed before and after subjects received various doses of either ibuprofen or flurbiprofen. Both ibuprofen and flurbiprofen reduced the initial rate of thromboxane B2 (TxB2) production, but about 10% of control TxB2 production was refractory to either drug. Aggregation was observed after TxB2 concentrations exceeded a threshold of about 40 ng/ml. Once the threshold was achieved, aggregation and TxB2 production occurred independently of either drug. Ibuprofen and flurbiprofen concentrations of about 3 and 0.2 micrograms/ml, respectively, inhibited aggregation during the 5-minute testing period. A pharmacodynamic model was developed that predicts TxB2 production, aggregation lag times, and percent aggregation as a function of the time course of drug concentrations in serum.


A novel dendritic cell-targeted lentiviral vector, encoding Ag85A-ESAT6 fusion gene of Mycobacterium tuberculosis, could elicit potent cell-mediated immune responses in mice.
Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis (Mtb), leading to high mortality worldwide. It is well-established that cellular immunity plays a critical role to control Mtb infection. Dendritic Cells (DCs) are potent antigen presenting cells, which play an important role to prime cell-mediated immune responses. In vivo targeting of DCs has been shown to induce both strong cellular immunity and protection against tumor challenges. The aim of the present study was not only to assess the immunizing potential of a novel DC-targeted recombinant lentivirus expressing fusion antigen Ag85A-ESAT6 of Mtb, but also to compare it with a recombinant lentivirus with broad cellular tropism expressing the same antigen in mice. The findings demonstrated that our novel recombinant DC-targeted lentivector was able to successfully transduce and express the fusion antigen Ag85A-E6 in vitro and in vivo. Moreover, a single footpad injection of targeted lentivectors could elicit strong T-helper 1 (Th1) immunity against the above mentioned antigen, as indicated by the specific high-level production of IFN-γ and IL-2 using spleen lymphocytes and lymphoproliferative responses. Despite of these promising results, more attempts are required to elucidate the protective and therapeutic efficacy of this approach in future.


The negative priming effect in cognitive conflict processing.
The present study used event-related potentials (ERPs) to investigate the specific physiological mechanisms underlying the negative nature of cognitive conflict and its influence on affective word evaluations. The present study used an affective priming paradigm where Stroop stimuli were presented for 200ms after which affective target words had to be evaluated as being positive or negative. Behavioral results showed that reaction times (RTs) were shorter for positive targets following congruent primes relative to incongruent primes, and for negative targets following incongruent primes relative to congruent primes. The ERP results showed that the N2 amplitude (200-300ms) for incongruent stimuli was significantly larger than for congruent stimuli in the Stroop task, which indicated a significant conflict effect. Moreover, the N400 amplitude (300-500ms) was smaller for negative words following incongruent primes relative to congruent primes, and for positive words following congruent primes relative to incongruent primes. The results demonstrated that cognitive conflict modulated both behavioral and electrophysiological correlates of subsequent emotional processing, consistent with its hypothesized registration as an aversive signal.


Computer vision syndrome among computer office workers in a developing country: an evaluation of prevalence and risk factors.
Computer vision syndrome (CVS) is a group of visual symptoms experienced in relation to the use of computers. Nearly 60 million people suffer from CVS globally, resulting in reduced productivity at work and reduced quality of life of the computer worker. The present study aims to describe the prevalence of CVS and its associated factors among a nationally-representative sample of Sri Lankan computer workers. Two thousand five hundred computer office workers were invited for the study from all nine provinces of Sri Lanka between May and December 2009. A self-administered questionnaire was used to collect socio-demographic data, symptoms of CVS and its associated factors. A binary logistic regression analysis was performed in all patients with 'presence of CVS' as the dichotomous dependent variable and age, gender, duration of occupation, daily computer usage, pre-existing eye disease, not using a visual display terminal (VDT) filter, adjusting brightness of screen, use of contact lenses, angle of gaze and ergonomic practices knowledge as the continuous/dichotomous independent variables. A similar binary logistic regression analysis was performed in all patients with 'severity of CVS' as the dichotomous dependent variable and other continuous/dichotomous independent variables. Sample size was 2210 (response rate-88.4%). Mean age was 30.8 ± 8.1 years and 50.8% of the sample were males. The 1-year prevalence of CVS in the study population was 67.4%. Female gender (OR: 1.28), duration of occupation (OR: 1.07), daily computer usage (1.10), pre-existing eye disease (OR: 4.49), not using a VDT filter (OR: 1.02), use of contact lenses (OR: 3.21) and ergonomics practices knowledge (OR: 1.24) all were associated with significantly presence of CVS. The duration of occupation (OR: 1.04) and presence of pre-existing eye disease (OR: 1.54) were significantly associated with the presence of 'severe CVS'. Sri Lankan computer workers had a high prevalence of CVS. Female gender, longer duration of occupation, higher daily computer usage, pre-existing eye disease, not using a VDT filter, use of contact lenses and higher ergonomics practices knowledge all were associated with significantly with the presence of CVS. The factors associated with the severity of CVS were the duration of occupation and presence of pre-existing eye disease.


Production of a monoclonal antibody-bleomycin conjugate utilizing dextran T-40 and the antigen-targeting cytotoxicity of the conjugate.
Bleomycin (BLM), a potent anticancer glycopeptide antibiotic, was linked covalently to murine monoclonal anti-HLA IgG1 antibody (H-1) with the use of dextran T-40. As determined spectrophotometrically, the conjugate was composed of 57.5 moles BLM per mole antibody. Of the substituted BLM, 18.4% (10.6 moles BLM per mole antibody) exhibited antimicrobial activity. The BLM-(H-1) conjugate showed stronger cytotoxicity than BLM alone against HLA-bearing cells in cultivation after a 30-min exposure to the drugs. In the same experiment, the conjugate was less toxic than BLM against cells lacking HLA.


The development and validation of a simple liquid chromatography-tandem mass spectrometry method for polymyxin B1 and B2 quantification in different matrices.
Polymyxin B has resurfaced as a last-line treatment for multi-drug resistant Gram-negative bacteria. Accurate characterization and quantification of polymyxin B components is necessary to optimize this therapy. We developed and validated a robust, straightforward LC-MS/MS method to quantify polymyxin B1 and B2, the primary polymyxin B components, in various matrices (cation-adjusted Mueller-Hinton broth (CAMHB), human and rat plasma). Of sample preparation approaches investigated, two protein precipitation/extraction methods were developed as part of an analytical strategy based upon reverse-phase LC-MS/MS using electrospray ionization in positive multiple-reaction monitoring mode. Both methods were validated over therapeutically and experimentally relevant concentration ranges (CAMHB: 0.1-8.0μg/mL, rat and human plasma: 0.05-4.0μg/mL). Quality control samples spanning a relevant concentration range were employed to assess intra- and inter-day accuracy (relative error (%RSD)) and precision (coefficient of variation (CV%)). For polymyxin B1 and B2 in CAMHB, inter-day standard deviations were 1.18-4.59% and 0.777-1.23%, respectively, and accuracies were 94.2-99.3% and 94.4-99.1%. For rat plasma, inter-day standard deviations were 1.53%-5.64% and 4.07%-8.26%. Accuracies were 100.6-108.9% and 96.1-108.1%. For human plasma, inter-day standard deviations were 2.77-7.32% and 1.55-7.29%. Accuracies were 89.6-96.4% and 92.9-102.0%. Extraction recoveries for all matrices were >93.5%. Adsorption, storage, and long-term stability were assessed and were acceptable. Accuracy, precision, and cost-efficiency make this an ideal approach for quantifying polymyxin B in in vitro and in vivo samples including those from rat and human subjects.


Improvement in left ventricular function following coronary stenting in patients with acute myocardial infarction: 6-month and 3-year follow-up.
This study assesses hemodynamic and angiographic changes in patients with a patent infarct-related artery (IRA) after acute myocardial infarction (AMI). One hundred and seventy-seven patients with first AMI, who received a predischarge stenting to the IRA and sustained a patent IRA over 3 years, were stratified into 3 groups according to the baseline left ventricular ejection fraction (LVEF): group A included 63 patients with a LVEF of >49%, group B 73 patients with a LVEF of 40%-49%, and group C 41 patients with a LVEF of <40%. The hemodynamic and angiographic parameters were compared at baseline, 6-month and 3-year follow-up. The LV end-diastolic volume index increased 1, 4 and 4 ml/m(2) at 6 months and 4, 5 and 10 ml/m(2) at 3 years, respectively in group A, B and C. The LVEF increased 4%, 7% and 12% at 6 months and 6%, 8% and 14% at 3 years, respectively in group A, B and C. The stroke volume index increased 3, 7 and 12 ml/m(2) at 6 months and 6, 8 and 15 ml/m(2) at 3 years, respectively in group A, B and C. The LV wall motion score decreased 2, 3 and 3 at 6 months and was unchanged at 3 years, respectively in group A, B and C. The LV end-diastolic pressure decreased 2, 3 and 4 mm Hg, respectively in group A, B and C, at 6-month follow-up and remained stable at 3 years. Long-term beneficial effects in patients receiving a late predischarge intracoronary stenting following first AMI were seen and these may be related to patent IRA. A progressive improvement in left ventricular remodeling occurs in all patients regardless of their initial left ventricular function and the improvement continues for at least 3 years.


A Neuronal Pathway that Commands Deceleration in Drosophila Larval Light-Avoidance.
When facing a sudden danger or aversive condition while engaged in on-going forward motion, animals transiently slow down and make a turn to escape. The neural mechanisms underlying stimulation-induced deceleration in avoidance behavior are largely unknown. Here, we report that in Drosophila larvae, light-induced deceleration was commanded by a continuous neural pathway that included prothoracicotropic hormone neurons, eclosion hormone neurons, and tyrosine decarboxylase 2 motor neurons (the PET pathway). Inhibiting neurons in the PET pathway led to defects in light-avoidance due to insufficient deceleration and head casting. On the other hand, activation of PET pathway neurons specifically caused immediate deceleration in larval locomotion. Our findings reveal a neural substrate for the emergent deceleration response and provide a new understanding of the relationship between behavioral modules in animal avoidance responses.


Correction of congenital hydronephrosis in utero. I. The model: fetal urethral obstruction produces hydronephrosis and pulmonary hypoplasia in fetal lambs.
In an attempt to simulate congenital hydronephrosis secondary to urethral obstruction, we evaluated the consequences of obstructing the urethra in fetal lambs by several techniques. When the urethra was ligated before 108 days gestation, the bladder decompressed through the urachus and the upper-urinary tract developed normally. Ligation of both the urethra and urachus produced gross bilateral hydronephrosis, hydroureter, and megacystis, as well as severe pulmonary hypoplasia, and was associated with a high perinatal mortality. Intermittent urethral obstruction produced with a balloon constrictor or by diverting urine externally and pressure-limited obstruction produced with a Pudenz valve both proved unsatisfactory for producing congenital hydronephrosis. Finally, ligation of the urachus and progressive gradual obstruction of the urethra with an ameroid constrictor produced hydronephrosis and pulmonary hypoplasia. This model simulates urethral obstruction in the human fetus and produces a clinical and pathologic picture similar to that seen in infants born with congenital hydronephrosis.


Promising results in a 3-year follow-up for adults undergoing a one-stage surgery for residual talipes equinovarus as part of a humanitarian mission in Vietnam.
Rigid talipes equinovarus (TEV) is a complex foot deformity in which the foot is fixed in a plantarflexed, inverted, and adducted position. This pathology has the potential to severely limit basic life activities, which can be devastating for patients in developing countries. The objective of this study was to present the outcomes of patients with mature bones presenting with severe rigid TEV deformity who were operated on during a humanitarian mission to Vietnam using a single lateral approach and a simple and inexpensive fixation technique. This is a retrospective analysis of prospectively collected data. We analyzed the outcomes of patients who underwent surgery for a severe rigid TEV that prevented them from walking minimal distances unaided. All feet were fixed in a non-plantigrade position. The surgeries were conducted as part of two International Extremity Project (IEP) missions in Can Tho, Vietnam (2013 and 2018). Pre- and post-operative AOFAS scores were compared using the paired sample t-test. We operated on 14 feet of 12 patients, 6 (50%) of whom were males, aged 34.42 ± 11.7 (range 12 to 58). Four patients were followed for three months, two patients were followed for 12 months, and eight patients were followed for three years. On the final follow-up visit of each patient, all 14 operated feet were plantigrade with good alignment, and patients reported an improvement in daily activity. After 3 years of follow-up, the mean AOFAS score of eight patients with available data improved by 42.88 ± 3.91 points (95% CI 39.61 to 46.14, P < 0.01). Our patients also reported an improvement in mobility. At the final follow-up examination, no recurrence of the deformity was observed in any of the patients. Using low-technical surgical modalities, we were able to achieve plantigrade and walkable feet in patients with mature bones who had fixed rigid equinovarus. Level IV- Case Series.


Risk factors and outcomes for cholangitis after hepatic resection.
There is a paucity in the literature in regard to the incidence, risk factors, and outcomes for post-operative cholangitis following hepatic resection. Retrospective review of the ACS NSQIP main and targeted hepatectomy registries for 2012-2016. A total of 11,243 cases met the selection criteria. The incidence of post-operative cholangitis was 0.64% (151 cases). Multivariate analysis identified several risk factors associated with the development of post-operative cholangitis, stratified out by pre-operative and operative factors. The most significant risk factors were biliary anastomosis and pre-operative biliary stenting with odds ratios (OR) of 32.39 (95% CI 22.91-45.79, P value < 0.0001) and 18.32 (95% CI 10.51-31.94, P value < 0.0001) respectively. Cholangitis was significantly associated with post-operative bile leaks, liver failure, renal failure, organ space infections, sepsis/septic shock, need for reoperation, longer length of stay, increased readmission rates, and death. Largest analysis of post-operative cholangitis following hepatic resection. While a rare occurrence, it is associated with significantly increased risk for severe morbidity and mortality. The most significant risk factors were biliary anastomosis and stenting.


The Impact of Phenotypic Heterogeneity on Chemotactic Self-Organisation.
The capacity to aggregate through chemosensitive movement forms a paradigm of self-organisation, with examples spanning cellular and animal systems. A basic mechanism assumes a phenotypically homogeneous population that secretes its own attractant, with the well known system introduced more than five decades ago by Keller and Segel proving resolutely popular in modelling studies. The typical assumption of population phenotypic homogeneity, however, often lies at odds with the heterogeneity of natural systems, where populations may comprise distinct phenotypes that vary according to their chemotactic ability, attractant secretion, etc. To initiate an understanding into how this diversity can impact on autoaggregation, we propose a simple extension to the classical Keller and Segel model, in which the population is divided into two distinct phenotypes: those performing chemotaxis and those producing attractant. Using a combination of linear stability analysis and numerical simulations, we demonstrate that switching between these phenotypic states alters the capacity of a population to self-aggregate. Further, we show that switching based on the local environment (population density or chemoattractant level) leads to diverse patterning and provides a route through which a population can effectively curb the size and density of an aggregate. We discuss the results in the context of real world examples of chemotactic aggregation, as well as theoretical aspects of the model such as global existence and blow-up of solutions.


Effects of dietary casein and soy protein on metabolism of radiolabelled low density apolipoprotein B in rabbits.
Rabbits fed semipurified diets containing casein have elevated plasma cholesterol levels compared to those fed soy protein. As part of continuing studies on the mechanism of casein-induced hypercholesterolemia, two groups of six rabbits were fed these diets for 14 to 16 weeks. Animals fed the casein diet were found to have significantly higher plasma concentrations of protein, cholesterol, triacylglycerol, phospholipid and apolipoprotein B (apo B) associated with low density lipoprotein (LDL) than those fed the soy protein diet. Kinetic studies showed that the fractional catabolic rate of LDL-apo B was significantly lower in animals fed casein than in those fed soy protein regardless of whether the tracer LDL was obtained from donors fed casein or soy protein. The production rate of LDL-apo B was higher in casein-fed animals but this was not statistically significant. These results show that the efficiency of removal of LDL is significantly reduced in animals fed casein compared to those fed soy protein, and that the source of LDL did not affect the efficiency of its subsequent removal. The accumulation of LDL in casein-fed animals is consistent with down-regulation of the LDL receptor.


Radon decay products equilibrium at different aerosol concentrations.
The influence of aerosol particle concentration on the activity of short-lived radon decay products (RPDs) was investigated inside a standard radon box. The 218Po:214Pb:214Bi activity ratio was determined for unattached and attached fractions of radon decay products at different aerosol concentrations. For the unattached fraction, this ratio was 1:0.31:0.06, while for the attached aerosol fraction the activity ratio was 1:1.4:0.91 at a low aerosol concentration (2000 cm-3) and with the unattached fraction fp = 0.45. In this case, there was no additional aerosol in the standard radon box and the equilibrium shift ratio for the total activity (attached and unattached) was 1:0.55:0.27 for 218Po:214Pb:214Bi. When the aerosol was injected, the unattached fraction nearly disappeared at a high aerosol concentration (80000-90000 cm-3). The equilibrium shift ratio for the total activity at a high aerosol concentration was 1:2:1.5 for 218Po:214Pb:214Bi. This ratio was the same for the attached fraction. For no more than 0.01 of the unattached fractions, this ratio was 1:0.85:0.7. Radioactive equilibrium factors and unattached fractions were also determined at different aerosol particles concentrations. A negative correlation between unattached fraction and the radioactive equilibrium factor, as well as a positive correlation coefficient between the radioactive equilibrium factor and aerosol particle concentration in a standard radon box, has been determined.


Characterization of human fibrocytes as circulating adipocyte progenitors and the formation of human adipose tissue in SCID mice.
An increase in fat mass associated with obesity results from recruitment and differentiation of adipocyte progenitor cells. The precise origin of these cells is unknown, although accumulating evidence suggests that circulating stem cells can differentiate into cells of mesenchymal lineage. It is currently unclear whether a progenitor adipocyte population exists in circulation. One potential candidate is the fibrocyte, which may represent a common progenitor cell for several mesenchymal lineages. We demonstrate that these circulating progenitors become adipocytes when cultured under adipogenic conditions, with intracellular lipids accumulation and up-regulation of proteins specific for adipocyte differentiation, including leptin, PPARgamma, and FABP4. cDNA microarray analysis revealed gene clusters that were differentially regulated during adipogenesis of fibrocytes, which were similar to visceral and subcutaneous adipose tissue preadipocyte-to-adipocyte differentiation. Moreover, these progenitors engrafted and formed human adipose tissue following injection into SCID mice. Although fibrocytes express an array of chemokine receptors, we observed an up-regulation of CCR2 expression following fibrocytes differentiation into adipocytes, which was associated with increased chemotactic response to CCL2. This paradigm supports the notion that elevated CCL2 levels in visceral adipose tissue associated with Metabolic Syndrome is a chemotactic niche, whereby fibrocytes can home to and differentiate into adipocytes to perpetuate its tissue formation.


Outcome of anaemic monochorionic single survivors following early intrauterine rescue transfusion in cases of feto-fetal transfusion syndrome.
To evaluate the outcome of severely anaemic monochorionic (MC) twins surviving the death of their co-twin following early intrauterine rescue transfusion in cases of feto-fetal transfusion syndrome (FFTS). We reviewed all MC pregnancies complicated with FFTS following primary management, in which a single intrauterine fetal death (IUFD) was diagnosed with certainty within 24 hours between January 1999 and December 2006. We included MC survivors who presented ultrasound or Doppler features of fetal anaemia following the death of their co-twin. Intrauterine transfusion (IUT) was given to all survivors who were anaemic. Nineteen MC twin pregnancies presented a single intrauterine death (IUD) associated with an anaemic co-twin. Median gestational age at IUD was 23 [20-28] weeks. The median interval between IUD and IUT was 12 [8-24] hours. There were 58% (11/19) healthy survivors. Perinatal death rate was 26% (5/19) including 16% (3/19) intrauterine and 10% (2/19) neonatal deaths. Abnormal prenatal cerebral findings developed in 21% (4/19) cases, always within 1 month after the death of the co-twin. Considering occlusive techniques and other management separately, there were 64% (7/11) and 50% (4/8) healthy survivors, respectively, and perinatal death occurred in 36% (4/11) and 12.5% (1/8) of fetuses, respectively. Prenatal fetal cerebral lesions developed in 9% (1/11) of cases following occlusive techniques and in 37.5% (3/8) of fetuses when managed differently. The median gestational age at delivery in the survivors was 31 [25-38] weeks. In cases of FFTS with single anaemic survivors, early IUT could be offered following extensive counselling and close follow up.


Maintenance of highly contractile tissue-cultured avian skeletal myotubes in collagen gel.
Highly contractile skeletal myotubes differentiated in tissue culture are normally difficult to maintain on collagen-coated tissue culture dishes for extended periods because of their propensity to detach as a sheet of cells from their substratum. This detachment results in the release of mechanical tension in the growing cell "sheet" and, consequently, loss of cellular protein. We developed a simple method of culturing high density contractile primary avian myotubes embedded in a collagen gel matrix (collagel) attached to either a stainless steel mesh or nylon support structure. With this system the cells are maintained in a highly contractile state for extended periods in vitro under tension. Structural integrity of the myotubes can be maintained for up to 10 d in basal medium without serum or embryo extract. Total cellular protein and myosin heavy chain accumulation in the cells can be maintained for weeks at levels which are two to three times those found in time-matched controls that are under little tension. Morphologically, the myotubes are well differentiated with structural characteristics of neonatal myofibers. This new collagel culture system should prove useful in the analysis of in vitro gene expression during myotube to myofiber differentiation and its regulation by various environmental factors such as medium growth factors, innervation, and mechanical activity.


Population-specific renal proteomes of marine and freshwater three-spined sticklebacks.
Quantitative proteomics was used to reveal biochemical differences in kidneys of marine and freshwater three-spined sticklebacks. More than 1500 unambiguous proteins were identified, 106 of which are robustly co-translationally modified. Amino-terminal acetylation sites for 94 and proline hydroxylation sites for 12 proteins, including 4 protein disulfide isomerases having the consensus motif APWCGHCK, were determined. More than 1500 proteins were quantified by LC-MS/MS yielding 120 proteins with consistent population-specific abundance differences. Twenty-five of these were selected for validation by data-independent acquisition (DIA) and spectral library based MS2 quantitation. A dense biochemical network was revealed, which promotes the synthesis of the organic osmolytes betaine, sorbitol, trimethylamine oxid (TMAO), and urea. It contains 33 of 49 proteins that are elevated in marine compared to freshwater sticklebacks, including the most highly elevated proteins (dimethylaniline monooxygenase, alanine-glyoxylate aminotransferase, glycine N-methyltransferase). Freshwater stickleback kidneys contain elevated levels of proteolytic, cytoskeletal, extracellular matrix, and calcium signaling proteins. Proteins that are most elevated in freshwater sticklebacks are ES1 protein homolog, apoptosis-associated speck-like protein containing a CARD and caspase 1. Protein-abundance network analysis demonstrates significantly higher levels of synchronized abundance control in marine sticklebacks. The significance of these findings for biochemical diversification of renal function in marine and FW sticklebacks is discussed.


Reexpression of HPRT activity following cell fusion with polyethylene glycol.
Polyethylene glycol-1000 (PEG-1000) induced fusion of HPRT (E.C. 18.104.22.168) deficient Chinese hamster cells with alpha-galactosidase A (E.C. 220.127.116.11) deficient cells from a patient with Fabry's disease yielded hybrids which contained both human and hamster HPRT, G6PD (E.C. 22.214.171.124), and APRT (E.C. 184.108.40.206) and Chinese hamster alpha-galactosidase B. Thus PEG-1000 mediated somatic cell fusion led to reexpression of Chinese hamster HPRT. It did not restore the expression of human alpha-galactosidase. Since PEG-1000 treatment of HPRT- Chinese hamster cells in the absence of human cells yielded no HPRT+ cells, it is concluded that the element responsible for the restoration of rodent HPRT was contributed by the human cells and not by the agent employed to promote fusion.


Lipid metabolism of rat adipocytes in culture.
Isolated differentiated rat adipocytes embedded in a collagen matrix were maintained in culture for 0, 4 and 14 days. Assessment of viability by either anabolic glucose metabolism or catabolic lipolytic metabolism can be misleading; both types of metabolism should be measured to establish the status of cells. Cells maintained morphological integrity even when not viable as judged by anabolic glucose metabolism.


Instrumentation and patient characteristics that influence postoperative haemorrhage rates following tonsil and adenoid surgery.
To investigate the effect of the type instrumentation used and the age and gender characteristics of patients on postoperative haemorrhage rates following tonsil and adenoid surgery. A retrospective analysis of 13 593 procedures was performed from The Patient Episode Database for Wales between 1 January 1999 and 31 March 2004. National health policy changes created four periods of different instrument usage (reusable, single-use with diathermy, single-use alone, specified single-use with diathermy). These and the age and gender distribution of the patients were examined against four categories of postoperative haemorrhage. Postoperative haemorrhage rates were expressed as the number of complications per operations performed. Primary postoperative haemorrhage that occurred during the initial admission either required a return to theatre [R1] or was managed conservatively [N1]; secondary postoperative haemorrhage that required a return to hospital either returned to theatre [R2] or was managed conservatively [N2], were compared. Primary haemorrhage with return to theatre doubled, from the baseline rate with reusable instruments, from 0.6% (CI 0.5-0.8) to 1.2% (CI 0.7-1.9) when single-use instruments were introduced and remained high at 1.4% (CI 0.9-2.1) after the withdrawal of single-use diathermy. This haemorrhage rate returned to the baseline rate (0.6% CI 0.3-1.0) when specified single-use instruments were introduced. None of the other haemorrhage rates changed significantly throughout the four observation periods. Adenotonsillectomy and tonsillectomy patients have different age and gender patterns. In a univariate analysis, males over the age of 12 years were twice as likely to have haemorrhage with return to theatre than girls of the same age, 3.8% (CI 3.0-4.7) versus 1.7% (CI 1.4-2.1). A significant rise in serious postoperative primary haemorrhage but not secondary haemorrhage was seen following the initial introduction of single-use instruments that reverted to baseline with the introduction of specified single-use instruments. Diathermy does not appear to have affected the haemorrhage rates. There is a distinct age and gender pattern for tonsil and adenoid surgery and risk of postoperative haemorrhage. The use of arbitrary divisions of age may be misleading in studies that examine post-tonsillectomy haemorrhage.


Mitral valve prolapse syndrome: interventions for symptom control.
Mitral valve prolapse syndrome, the most common valvular disorder, is associated with symptoms of anxiety, fatigue, palpitations, headaches, chest pain and more. Many non-drug interventions can be employed to alleviate symptoms.


Helicopter cooling of heatstroke victims.
Theoretical and clinical evidence suggests that traditional ice water immersion is not the best therapy for heatstroke. We treated three patients with serious heatstroke using the downdraft from a light utility helicopter to dissipate heat primarily by evaporation and convection. The speed of cooling was substantially better than that reported in prior clinical series.


A randomized trial of acceptance-based behavioral therapy to improve mental health outcomes for LGBTQ+ persons: Study protocol.
During the COVID-19 pandemic, public health measures limited social interactions as an effective and protective intervention for all. For many, however, this social isolation exacerbated mental health symptoms. People who identify as lesbian, gay, bisexual, transgender, and queer (LGBTQ+) were already at elevated risk of anxiety and depression, relative to cisgender and heterosexual populations, and pandemic-related social isolation likely heightened these disparities. In our prior work with sexual and gender minorities, we developed and established feasibility and acceptability of a novel acceptance-based behavioral therapy (ABBT) intervention for HIV treatment. ABBT showed promise in improving social support and reducing mental health symptoms. In the current study, we investigate the efficacy of ABBT, compared to a treatment-as-usual control condition, in a full-scale randomized controlled trial to improve social support for LGBTQ+ persons living with anxiety and depression. Two hundred forty LGBTQ+ adults with anxiety and/or depressive symptoms will be recruited and equally randomized to receive: (a) the ABBT intervention, consisting of two 30-40 min sessions plus treatment-as-usual (TAU), or (b) TAU only. Primary outcomes are interviewer-assessed anxiety and depressive symptoms. Secondary outcomes are self-reported anxiety and depressive symptoms. Experiential avoidance and social support are hypothesized mediators and presence of an anxiety and/or depressive disorder is a hypothesized moderator. ABBT represents a novel, identify-affirming real-world approach to promoting social support as a means of improving mental health among individuals who identify as LGBTQ+. This study will contribute actionable data establishing the impact, mediational mechanisms, and effect modifiers of ABBT. govregistration: NCT05540067.


Mosquito larvicidal activity of Aloe vera (Family: Liliaceae) leaf extract and Bacillus sphaericus, against Chikungunya vector, Aedes aegypti.
The bio-efficacy of Aloe vera leaf extract and bacterial insecticide, Bacillus sphaericus larvicidal activity was assessed against the first to fourth instars larvae of Aedes aegypti, under the laboratory conditions. The plant material was shade dried at room temperature and powdered coarsely. A. vera and B. sphaericus show varied degrees of larvicidal activity against various instars larvae of A. aegypti. The LC50 of A. vera against the first to fourth instars larvae were 162.74, 201.43, 253.30 and 300.05 ppm and the LC90 442.98, 518.86, 563.18 and 612.96 ppm, respectively. B. sphaericus against the first to fourth instars larvae the LC50 values were 68.21, 79.13, 93.48, and 107.05 ppm and the LC90 values 149.15, 164.67, 183.84, and 201.09 ppm, respectively. However, the combined treatment of A. vera + B. sphaericus (1:2) material shows highest larvicidal activity of the LC50 values 54.80, 63.11, 74.66 and 95.10 ppm; The LC90 values of 145.29, 160.14, 179.74 and 209.98 ppm, against A. aegypti in all the tested concentrations than the individuals and clearly established that there is a substantial amount of synergist act. The present investigation clearly exhibits that both A. vera and B. sphaericus materials could serve as a potential larvicidal agent. Since, A. aegypti is a container breeder vector mosquito this user and eco-friendly and low-cost vector control strategy could be a viable solution to the existing dengue disease burden. Therefore, this study provides first report on the mosquito larvicidal activity the combined effect of A. vera leaf extract and B. sphaericus against as target species of A. aegypti.


MRI assessment of post-traumatic spinal instability.
The imaging evaluation of patients with spinal trauma has evolved over the past decades, and there has been particular interest in the concept of instability, to predict which a series of criteria have been proposed. We retrospectively evaluated the magnetic resonance imaging (MRI) findings in 50 patients with post-traumatic spinal instability according to Denis's three-column method, Daffner et al.'s radiographic criteria, and Oner et al.'s categorization of MRI findings; additionally, we evaluated the cord, the prevertebral tissue, and the epidural space. We suggest that an integrated panel of MRI information might be standardized in order to provide a more complete evaluation of spinal injury in an individual case and help planning surgical treatment.


MxMPK6-2-bHLH104 interaction is involved in reactive oxygen species signaling in response to iron deficiency in apple rootstock.
Iron (Fe) is a trace element necessary for plant growth. Many land plants have evolved a set of mechanisms associated with the Fe absorption process to deal with the problem of insufficient Fe supply in the soil. During Fe absorption, reactive oxygen species (ROS) can be used as a signal to initiate a response to stress caused by Fe deficiency. However, the molecular mechanisms underlying the involvement of ROS in the Fe deficiency stress response remains unclear. In this study, we have identified a kinase, MxMPK6-2, from Malus xiaojinensis, an apple rootstock that is highly efficient at Fe absorption. MxMPK6-2 has been shown to be responsive to ROS signals during Fe deficiency, and MxMPK6-2 overexpression in apple calli enhanced its tolerance to Fe deficiency. We further screened for proteins in the Fe absorption pathway and identified MxbHLH104, a transcription factor which interacts with MxMPK6-2. MxbHLH104 can be phosphorylated by MxMPK6-2 in vivo, and we confirmed that its phosphorylation increased Fe absorption in apple calli under Fe deficiency, with the presence of ROS promoting this process. Overall, we have demonstrated that MxMPK6-2 is responsive to ROS signaling during Fe deficiency, and is able to control its response by regulating MxbHLH104.


First molecular identification of human Dobrava virus infection in central Europe.
Viral RNA was amplified by reverse transcription-PCR from a patient suffering from hemorrhagic fever with renal syndrome (HFRS) in Germany. The virus strain could be assigned to the Dobrava hantavirus (DOBV). This is the first molecular identification of human infection by DOBV in central Europe and the first proof that a virus strain related to the DOBV-Aa lineage, carried by Apodemus agrarius rodents, is able to cause HFRS.


Qualitative and quantitative analysis of portal Doppler waveform and a novel factor of portal pulsatility: Systolic spike wave.
We attempted to identify the physiologic circumstances associated with the portal Doppler waveform. The subjects were 98 patients: 42 had cirrhosis; 23, hepatitis; and 33, no liver disorder. We measured right portal venous peak velocity, minimum velocity, and hepatic arterial peak systolic velocity. Portal venous pulsatility was calculated as portal venous minimum velocity divided by portal venous peak velocity. We analyzed portal waveforms both qualitatively and quantitatively. Portal Doppler waveforms were classified as nonphasic, monophasic, biphasic, biphasic including spike wave, and triphasic. Portal waveforms were phasic in 84 subjects who had a mild systolic dip during the arterial systolic period. Markedly portal pulsatility (portal venous pulsatility<0.6) was present in only 7 subjects. A systolic spike wave corresponding to a hepatic arterial peak systolic wave was detected in 48 subjects. Hepatic arterial peak systolic velocity was significantly higher in the group with spike wave than in the group without it. Although portal pulsatility is generally attributed to multiple factors, the present study has pointed out a new factor; transmission via the hepatic artery through the portal vein vasa vasorum. Hepatic venous drainage and hepatic arterial blood flow influence portal waveform.


Social distance influences the outcome evaluation of cooperation and conflict: Evidence from event-related potentials.
Previous research shows that social distance plays an important role in promoting cooperation and that subtle cues that reduce social distance increase the tendency to cooperate. However, it is unclear how social distance influences our outcome evaluation of cooperative and conflict feedback. The present study investigated the influence of social distance on cooperative and conflict behavior and the evaluation process of the cooperative and conflict outcomes, using the event-related potentials (ERPs) technique. We recorded ERPs from 14 normal adults playing a social game task against a friend and a stranger. The results showed that the FRN (Feedback Related Negativity) and P300 were affected by the opponent's choice to cooperate or aggress; however, only the P300 was affected by social distance. Specifically, when the opponent chose to cooperate, the feedback elicited a smaller FRN and a larger P300 amplitude; and compared with playing against friends, the P300 had a larger amplitude when participants gaming with strangers. Our results indicate that at the early stage of the evaluation of cooperation and conflict outcomes, individuals may initially and quickly encode the valence of outcomes, judging whether an outcome is consistent with their expectations. However, at the late stage, which involves a top-down cognitive appraisal process, some social factors, such as social distance, may moderate processing of attention resource allocation of feedback about outcomes, and of higher-level motivation/affective appraisal.


Density-dependent habitat selection in muskrats: a test of the ideal free distribution model.
Two predictions of the ideal free distribution model, a null hypothesis of habitat selection, were examined using free-ranging muskrats. We rejected the prediction that the proportion of the animals found in each of five habitats was independent of population size. Data on over-winter occupancy of muskrat dwellings tend also to refute the prediction of equal fitness reward among habitats. Habitat type and water-level had a profound effect on the suitability of a site for settlement. We concluded that the observed pattern of muskrat distribution followed more closely an ideal despotic distribution where some individuals benefited from a higher fitness because of resource monopolization. Current theories of density-dependent habitat selection, which assume an ideal free distribution, would not apply to muskrats and possibly to many other mammal species.


Spermatogenesis in men exposed to polybrominated biphenyl (PBB).
Experience with other halogenated hydrocarbons, animal experiments, and individual complaints indicated that polybrominated biphenyl (PBB) might have an effect on human spermatogenesis. Analysis of semen from 52 PBB-exposed men as compared with analysis of semen from a control group of 52 men not exposed to PBB revealed no differences in the distribution of sperm counts, motility, or morphology.


Factors influencing the distribution pattern of porphyrins in cell membranes.
The mechanism of the sensitizer-membrane interactions has been studied by following the distribution properties of selected porphyrins, including haematoporphyrin (HP) and protoporphyrin (PP), into unilamellar liposomes of dipalmitoyl phosphatidylcholine (DPPC). The endomembrane distribution of HP and PP has been checked as a function of the membrane fluidity and composition by fluorescence polarization and quenching techniques. At porphyrin concentrations below 0.5 microM, HP and PP exclusively localize in the inner phospholipid monolayer; at higher concentrations, the outer monolayer also becomes populated. The porphyrin binding sites in liposomes, however, are different for HP and PP: HP preferentially distributes into water-accessible lipid regions, while PP localizes in the most hydrophobic loci of the lipid matrix. A porphyrin redistribution occurs when the fluidity properties of the liposomes are changed by addition of cholesterol or cardiolipin. In DPPC-cholesterol vesicles, all HP molecules dissolve in DPPC-rich regions while all PP molecules partition in cholesterol-rich environments. In DPPC-cardiolipin vesicles both porphyrins preferentially localize in regions accessible to the external medium. The effect of the nature of the carrier on porphyrin distribution in membranes has been studied by following the uptake and photosensitization properties of free and DPPC-incorporated PP and HP with rat liver mitochondria. The porphyrin photosensitizing efficiency has been checked by following the impairment of the respiratory function of mitochondria upon irradiation. Liposome-bound HP is less active than aqueous HP in determining membrane photodamage in mitochondria. On the contrary, aqueous PP is a very poor sensitizer as compared to a DPPC liposome-entrapped drug.(ABSTRACT TRUNCATED AT 250 WORDS)


Nitroxides are more efficient inhibitors of oxidative damage to calf skin collagen than antioxidant vitamins.
Reactive oxygen species generated upon UV-A exposure appear to play a major role in dermal connective tissue transformations including degradation of skin collagen. Here we investigate on oxidative damage to collagen achieved by exposure to (i) UV-A irradiation and to (ii) AAPH-derived radicals and on its possible prevention using synthetic and natural antioxidants. Oxidative damage was identified through SDS-PAGE, circular dichroism spectroscopy and quantification of protein carbonyl residues. Collagen (2 mg/ml) exposed to UV-A and to AAPH-derived radicals was degraded in a time- and dose-dependent manner. Upon UV-A exposure, maximum damage was observable at 730 kJ/m2 UV-A, found to be equivalent to roughly 2 h of sunshine, while exposure to 5 mM AAPH for 2 h at 50 degrees C lead to maximum collagen degradation. In both cases, dose-dependent protection was achieved by incubation with muM concentrations of nitroxide radicals, where the extent of protection was shown to be dictated by their structural differences whereas the vitamins E and C proved less efficient inhibitors of collagen damage. These results suggest that nitroxide radicals may be able to prevent oxidative injury to dermal tissues in vivo alternatively to commonly used natural antioxidants.


Toxicity associated to radiotherapy treatment in lung cancer patients.
Radiation therapy in combination with other treatments, such as chemotherapy, increases loco-regional control and survival in patients with lung cancer. Nevertheless, the subsequent toxicity of this treatment occurs in up to 37% of the irradiated patients. Some factors related to the patient, including performance status, pulmonary function tests (FEV1, DCLO), tumour site, as well as treatment-related factors such as radiation dose, fractionation and addition of chemotherapy, can be related to the risk of pulmonary toxicity. With the advent of tridimensional conformal radiotherapy (3DCRT), dose-volume histograms can be generated to assess the dose received by the organs at risk. Volume dose (Vdose), mean lung dose (MLD) and normal tissue complication probability (NTCP) are the dosimetric parameters most frequently used. The possible relationship between these parameters and clinical and anatomical factors has to be considered. Steroid treatment should be started soon in case of pneumonitis to avoid the development of late pulmonary fibrosis. Finally, some pharmacological agents to prevent radiation-related pneumonitis are under investigation.


Relevance of lymph node micrometastases in radically resected endobronchial carcinoid tumors.
Endobronchial carcinoids may recur even if radically resected. This retrospective study investigates the clinical and prognostic relevance of lymph node micrometastasis in these neoplasms. Fifty-five patients underwent standard anatomic resection (lobectomy or pneumonectomy) with systematic routine (hilar and mediastinal) lymphadenectomy. After an evaluation of conventional prognostic factors, we reanalyzed lymph nodes of pN0 patients for micrometastasis using immunohistochemistry (anti-cytokeratin AE1/AE3 and anti-chromogranin A antibodies). We performed 9 pneumonectomies, 41 lobectomies, and 5 bilobectomies. Histologic examination showed 47 (85%) typical carcinoid and 8 (14.5%) atypical. Twelve patients were pN1 (8 typical, 4 atypical); after reanalysis another 8 revealed micrometastasis, 6 N1 micrometastasis (5 typical, 1 atypical), and 2 N2 micrometastasis (2 atypical), increasing subjects with nodal involvement (pN1 and N micrometastasis) from 12 (21.8%) to 20 (36.4%; p = 0.01). Micrometastases were more frequent in atypical carcinoids (p = 0.002). Local recurrence developed in 3 (5.4%) patients: 2 pN1 (1 typical, 1 atypical) and 1 N1-micrometastasis (1 typical). Distant relapse occurred in 2 (3.6%) patients, both N2 micrometastasis (2 atypical). After reanalysis, recurrence rate in patients with nodal disease increased from 16.7% to 25% (p = 0.01). All patients with recurrence died: all had pN1 or N micrometastasis. No patient confirmed as N0 had recurrence. Only histologic pattern and node status significantly influenced disease-free (p = 0.002 and p = 0.05) and overall survivals (p = 0.02 and p < 0.001), respectively. Micrometastasis worsen both disease-free (p < 0.0001) and overall (p < 0.001) survival rates at 5 and 10 years. Routine systematic lymphadenectomy with immunohistochemical detection of lymph node micrometastasis contributes to identification of a larger population at risk with a higher recurrence rate, allowing a more accurate staging of endobronchial carcinoids.


Immediate early and functional AP-1 cis-response elements are involved in the transcriptional regulation of the large subunit of herpes simplex virus type 2 ribonucleotide reductase (ICP10).
Expression from the promoter of the herpes simplex virus type 2 (HSV-2) large subunit of ribonucleotide reductase (ICP10) is stimulated by co-transfection with DNA that encodes the virion protein Vmw65 previously shown to activate in trans the transcription of all IE genes (Wymer et al., 1989). Specific cis response elements involved in ICP10 transcriptional regulation were studied by chloramphenicol acetyltransferase analysis with hybrid ICP10 promoter/CAT structural gene constructions containing wild type or site-directed mutations of the promoter sequences. The data indicate that Vmw65 activation requires an intact TAAT-GARAT motif while complex formation requires an intact Oct-1 element, and the AP-1 consensus elements in the ICP10 promoter are functional in vitro. Thus, expression from the wild type and GA-rich mutant constructions was enhanced 10-20-fold by co-transfection with DNA encoding Vmw65. The GARAT and POU homeobox (PHB) binding motifs were required for Vmw65 mediated activation but the mutant in the POU specific box (PSB) binding motif was activated at higher concentrations of Vmw65 DNA (1.0-3.0 micrograms). The PHB and PSB binding motifs were necessary for complex formation as determined by gel retardation analysis with in vitro synthesized OTF-1 and Vmw65 proteins. The GARAT and GA-rich elements were not required. CAT expression from pICP10-cat was enhanced by co-transfection with jun and fos encoding DNA, and the ICP10 promoter complexed with in vitro synthesized jun protein.


The protective action of ketanserin against lipopolysaccharide-induced shock in mice is mediated by inhibiting inducible NO synthase expression via the MEK/ERK pathway.
Nitric oxide (NO) plays an important role in the pathogenesis of endotoxic shock. This work tested the hypothesis that ketanserin could attenuate endotoxic shock by inhibiting the expression of inducible NO synthase (iNOS). The results demonstrated that ketanserin could inhibit iNOS expression in the heart, lungs, liver, and kidneys and nitrate production in the serum upon endotoxic shock in mice. In RAW264.7 cells, ketanserin significantly inhibited the expression of iNOS and decreased the production of NO, TNFα, IL-6, and reactive oxygen species upon lipopolysaccharide (LPS) challenge. Ketanserin also increased the level of ATP and mitochondrial membrane potential in RAW264.7 cells upon LPS exposure. LPS-induced iNOS expression was inhibited by the 5-HT2A receptor antagonist ritanserin and not the α1 receptor antagonist prazosin. Knockdown of 5-HT2A receptor by siRNA abolished the inhibitory effect of ketanserin on the expression of iNOS. These results indicated that the inhibitory effect of ketanserin on the expression of iNOS is mediated by blocking the 5-HT2A receptor. Furthermore, ketanserin significantly inhibited the activation of ERK1/2 and NF-κB signal. Pretreatment with PD184352, a specific inhibitor of ERK1/2, blocked the inhibitory effect of ketanserin on the expression of iNOS and NO production, indicating a critical role for the MEK/ERK1/2 signaling pathway. Collectively, our findings indicate that inhibition of the expression of iNOS via the MEK/ERK pathway mediates the protective effects of ketanserin against LPS-induced shock in mice.


Fine-mapping butyrophilin family genes revealed several polymorphisms influencing viral genotype selection in hepatitis C infection.
Host-viral genetic interaction has a key role in hepatitis C infection (HCV) and maybe in the viral selection. In a preliminary GWAS analysis, we identified BTN3A2 rs9104 to be associated with HCV genotype 1. Therefore, our aim was to determine the influence of BTN family on the selection of HCV genotype. We performed a fine-mapping analysis of BTN gene region in a cohort of chronic HCV infection (N=841), validating significant results in another independent chronic HCV infection cohort (N=637), according to selection of viral genotype. BTN3A2 rs9104, BTN3A2 rs733528, BTN2A1 rs6929846, BTN2A1 rs7763910 and BTN3A3 rs13220495 were associated with viral genotype selection. Interestingly, BTN3A2 rs9104 GG genotype was closely related to genotype 1 infection (80.7% (394/488) compared with genotype 3 infection (53.5% (23/43); P=0.0001) in patients harboring IL28B-CT/TT genotype, although this effect was not observed in IL28B-CC genotype. Similarly, BTN3A3 rs13220495 CC genotype was linked to genotype 3 infection (100% (32/32)) compared to genotype 1 (87.3% (137/157); P=0.028) in patients harboring IL28B-CC genotype, but did not in IL28B-CT/TT genotype. Genetic variants in the butyrophilin family genes may alter susceptibility to infection, selecting HCV genotype and influencing disease progression. BTN3A2 rs9104 was strongly associated with genotype 1 infection and the haplotype BTN3A3 rs13220495 CC+IL28B genotype CC was universal in patients with hepatitis C genotype 3a.


Geographic and urban-rural disparities in dietary energy and macronutrient composition among women of childbearing age: findings from the China health and nutrition survey,1991-2015.
Understanding nutritional status among women of childbearing age (WCA) is of increasing concern, as nutrient intakes may affect the health of WCA and well-being of their offspring. This study aimed to investigate secular trends of dietary energy and macronutrients intakes and access longitudinally the urban-rural and geographic disparities among Chinese WCA. A total of 10,219 participants were involved in three rounds of the Chinese Health and Nutrition Survey (CHNS:1991, 2004, and 2015). Average macronutrients intakes were compared against the Chinese Dietary Reference Intakes Standard (DRIs) to better assess adequacy. Mixed effect models were used to estimate the secular trends of dietary intake. A total of 10,219 participants were involved. Dietary fat, the percentage of energy (%E) from fat, and the proportion with more than 30% of energy from fat and less than 50% from carbohydrates increased notably over time (p < 0.001). In 2015, urban western WCA had the most dietary fat (89.5 g/d), %E from fat (41.4%), with the highest proportion of energy from fat (81.7%) and carbohydrate (72.1%) out the range of DRIs. From 1991 to 2015, the average urban-rural differences in dietary fat decreased from 15.7 g/d to 3.2 g/d among eastern WCA. However, it increased to 16.4 g/d and 6.3 g/d among central and western WCA, respectively. WCA was experiencing a rapid transformation to a high-fat diet. Temporal variation with obvious urban-rural and geographic disparities in dietary. energy and macronutrient composition persistently existed among Chinese WCA.These findings have implications of future public strategies to strengthen the nutrition propaganda and education of balanced diet for WCA to help them to improve their nutritional status, especially for those living in western China.


Do blood plasma levels of oxytocin moderate the effect of nasally administered oxytocin on social orienting in high-functioning male adults with autism spectrum disorder?
The study investigated whether baseline plasma oxytocin (OXT) concentrations might moderate the effects of nasally administered OXT on social orienting. Thirty-one males with Autism spectrum disorder (ASD) and thirty healthy males participated in a double-blind placebo-controlled crossover trial. After administration of the compound, participants were viewing pictures from the International Affective Picture System that represented a systematic variation of pleasant, unpleasant and neutral scenes with and without humans. The outcome measures were a cardiac evoked response (ECR) and a cortical evoked long latency parietal positivity (LPP). Males with ASD had significantly higher plasma baseline levels than the controls. In the absence of general treatment effects, higher baseline concentrations were found to be associated with larger treatment effects, particularly in the group of males with ASD. Higher post-treatment plasma OXT concentrations were found to be associated with smaller treatment effects and larger orienting responses in the placebo situation in the group of controls. We interpret our findings as suggesting that it is the central availability of OXT determining how much of the nasally administered OXT will become centrally absorbed and how much of it will become released into the bloodstream.


Do older patients with stage IB non-small-cell lung cancer obtain survival benefits from surgery? A propensity score matching study using SEER data.
Surgical resection is the preferred treatment for Stage IB non-small cell lung cancer (NSCLC), but one-third of patients still do not receive surgery, which might be due to the lack of robust and dedicated studies on the outcomes of surgical treatment in older patients with stage IB NSCLC. This study aims to investigate whether older patients with stage IB NSCLC benefit from surgery. Data of patients with NSCLC were downloaded from the SEER (Surveillance, Epidemiology, and End Results) database classifications were converted from the 7th edition staging system to the 8th edition staging system, and older patients (aged ≥65 years at diagnosis) with stage IB NSCLC were included. The propensity score matching (PSM) method was used to balance the distribution proportions of clinical characteristics between the surgery and no surgery groups. After 1:1 propensity score matching, the distribution proportions of clinical characteristics were balanced between the surgery and no surgery groups (all P > 0.05). The overall and disease-specific survival rates of patients in the surgery group were significantly higher than those of patients in the no-surgery group (both P < 0.001). Furthermore, subgroup analysis showed that receiving surgery was a protective factor for overall survival and disease-specific survival of patients in all clinical character-related subgroups. Ultimately, univariate and multivariate Cox regression analyses showed that sex, tumor size, tumor grade, and tumor classification were independent prognostic factors for overall and disease-specific survival in patients undergoing surgery. Older patients with Stage IB NSCLC can benefit significantly from surgical treatment after eliminating confounding factors, which is expected to provide strong medical evidence for surgical treatment.


Ion exchange removal and resin regeneration to treat per- and polyfluoroalkyl ether acids and other emerging PFAS in drinking water.
Ion exchange (IX) is a promising technology to remove legacy anionic per- and polyfluoroalkyl substances (PFAS) from water. As increasing numbers of per- and polyfluoroalkyl ether acids (PFEA) and other emerging PFAS were detected in the environment, it is necessary to understand how well IX resins remove these emerging PFAS for drinking water treatment. In this study, nine commercially available IX resins were tested to treat a drinking water source spiked with 40 legacy and emerging PFAS at 600 ng/L, including PFEA, perfluoroalkyl carboxylic and sulfonic acids, fluorotelomer sulfonic acids, perfluoroalkane sulfonamides, perfluoroalkane sulfonamidoacetic acids, and zwitterionic species. With limited contact time (15 min), PFAS properties such as the fluorinated chain length, charge, and functional groups all affected PFAS adsorption to resins. However, the impact of PFAS properties on PFAS removal became less pronounced when the contact time increased beyond 2 h, while the resin polymer matrix became the critical factor for PFAS removal. All five tested polystyrene-divinylbenzene (PS-DVB) resins achieved more than 90% removal in 24 h of 35 PFAS compounds, while polymethacrylate and polyacrylic resins achieved >90% removal for less than half of the compounds. Regenerating PS-DVB resin was investigated using different salt species, regenerant pH, brine concentrations, and methanol contents. Sodium chloride and ammonium chloride were found the best brines for regenerating the tested resins. Increasing brine concentrations enhanced the regeneration efficiency, especially for short-chain PFAS. Using simple salt regenerants, up to 94% of selected short-chain PFAS was released from resins designed for general water treatment, but no meaningful regeneration was achieved for long-chain PFAS or PFAS-specific resins when the organic solvent content was less than 20%.


Chromosomal abnormalities and sister-chromatid exchange in bone marrow cells of mice and Chinese hamsters after inhalation and intraperitoneal administration. II. Cyclophosphamide.
The genotoxic effects of cyclophosphamide (CPP), a human and animal carcinogen requiring metabolic activation, were studied in bone marrow cells of mice and Chinese hamsters, analyzing chromosome abnormalities (CA) and sister-chromatid exchange (SCE) after a 2-h inhalation or a single intraperitoneal administration. In order to compare the genotoxicity after the different routes of administration in the dose range of 10-110 mg CPP/kg body weight, the systemic dose obtained by inhalation was calculated from blood concentrations and the inhalation duration after an analysis of the CPP blood kinetics. In NMRI mice the frequency of bone marrow cells with chromosome abnormalities was higher after aerosol exposure than after intraperitoneal administration of comparable CPP doses. In Chinese hamsters the CA frequency was similar with both exposure routes. Inhaled CPP was found to induce a higher frequency of CA and SCE in the bone marrow cells of mice compared to those of Chinese hamsters. The findings suggest that for genotoxins requiring metabolic activation species differences exist with respect to the influence of the route of entry and the sensitivity of bone marrow cells.


Long-term airway space changes after mandibular setback using bilateral sagittal split osteotomy.
Eleven patients were evaluated retrospectively for their long-term changes in hypopharyngeal airway space after surgical correction of mandibular hyperplasia. All patients had undergone mandibular setbacks by way of bilateral sagittal split osteotomies using rigid fixation. The cephalometric evaluation of hypopharyngeal airway space was based on stable craniofacial landmarks. The results of this study indicate that mandibular setback procedures create relative narrowing of the hypopharyngeal airway space, as seen on lateral cephalometric radiographs, which in isolated cases might contribute to the development of an obstructive sleep apnea syndrome. The 2-dimensional evaluation of hypopharyngeal airway space and the clinical implications of the results are discussed.


Quantitive cytokine mRNA expression profiles in the colonic mucosa of patients with steroid naïve ulcerative colitis during active and quiescent disease.
Cytokines have validated roles in the immunopathogenesis of inflammatory bowel disease (IBD). This study was to investigate the expressions of tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, IL-8, and IL-10 mRNAs in the colonic mucosa of patients with ulcerative colitis (UC) during active and quiescent UC. At colonoscopy, biopsies were taken from inflamed and non-inflamed mucosa of patients with steroid-naive UC (n = 15), non-IBD inflammatory colitis controls (ICC, n = 6), and non-colitis controls (NCC, n = 14). The presence of extensive mononuclear cells and neutrophils infiltrate in the lamina propria, cryptitis, and epithelial damage defined an inflammatory lesion in the mucosa. Quantitative cytokine mRNA expressions in biopsies were measured by real-time polymerase chain reaction (PCR). Of 15 UC patients, 3 remitted with 5-aminosalicylate and 11 received granulocytapheresis; of these, 10 remitted. At baseline, IL-6, IL-8, TNF-alpha, and IL-10 mRNAs were high in inflamed mucosa compared with NCC (P < 0.01). In active UC, IL-6, IL-8 and IL-10 mRNAs were high compared with non-inflamed mucosa (P = 0.03, P = 0.03, P < 0.05, respectively). Both TNF-alpha mRNA (P = 0.03) and IL-6 mRNA (P = 0.04) were higher in UC compared with ICC. Even in non-inflamed mucosa, IL-8 and TNF-alpha mRNA expressions were high compared with NCC. Both IL-6 and IL-8 mRNAs decreased to normal levels after granulocytapheresis. During active UC, all 4 cytokine mRNA levels were high; only IL-6 and IL-8 mRNAs decreased to normal levels during remission. IL-8 mRNA was high even at sites of endoscopically quiescent UC during active disease. Steroid naïve patients respond well to granulocytapheresis.


Polyester Urethane Implants for Orbital Trapdoor Fracture Repair in Children.
The objective of this study was to present the surgical and clinical outcomes of polyester urethane implants in orbital trapdoor fracture repair in children. This is the first study reporting on the use of polyester urethane implants to repair orbital fractures. A retrospective review was performed of children with orbital trapdoor fracture who underwent surgical repair in the authors' medical center over 6 years. The trapdoor fracture diagnosis was based on clinical and computed topographic findings. Eight patients with trapdoor fracture were identified. All cases were repaired by polyester urethane implant placement. Patients' mean age was 13.5 years and mean follow-up was 13.6 months. All patients had orbital floor fractures. Two fractures also involved the medial wall. Seven patients had inferior rectus muscle entrapment and 1 had medial rectus muscle entrapment. Three patients had residual diplopia on extreme upgaze after the surgical repair. No patient had enophthalmos or infraorbital paresthesia at the end of follow-up. No postoperative complications associated with the used material were reported. The polyester urethane implant is a reliable, safe, inexpensive, and effective implant for trapdoor fracture repair in children. It can serve as a promising alternative to implants fabricated from other synthetic materials for orbital floor fracture repair.


A UPLC-MS/MS method for the simultaneous plasma quantification of all isomeric forms of the new anti-HCV protease inhibitors boceprevir and telaprevir.
HCV infection affects over 170 milions people in the world. Up to now standard-of-care therapy consisted in ribavirin plus interferon-α 2a or 2b. Recently, two new Direct Acting Antivirals (DAA), inhibitors of NS3 HCV protease, have been developed and approved for the routine use on HCV genotype 1 infected patients: boceprevir and telaprevir. Protease inhibitors show complex pharmacokinetics (strong metabolism of both drugs by CYP3A and drug interactions), they require a TID dosage and, furthermore, they are present in plasma patients in two different isomeric forms. In this work, we developed and validated an UPLC-tandem mass spectrometry assay method capable of monitoring the telaprevir and boceprevir concentrations in plasma, discriminating each isomer of both drugs. Blank plasma used for Standards and QCs preparation, was obtained from blood of healthy donors. The calibration curves for each isomer of telaprevir and boceprevir were linear in a range from 46.87 ng/mL to 6000 ng/mL and from 23.43 to 3000 ng/mL, respectively (mean r(2)>0.998 for all analytes). QCs at three different concentration were prepared: High, Medium and Low. Each Standard, QC and patient sample was treated with a protein precipitation protocol with a solution containing acidified acetonitrile and Internal Standard (6,7-Dimethyl-2,3-di(2-pyridyl)quinoxaline). An aliquot of supernatant was diluted and then directly analyzed by reverse-phase UPLC-MS/MS. Accuracy (mean 98.47%), intra-day (mean <5.21%) and inter-day (mean <7.57%) precision for telaprevir and boceprevir quality controls fitted all FDA guidelines. All compounds resulted stable in our method conditions and the extraction procedure showed a recovery near to 100%. Finally, we tested this method by monitoring plasma concentrations in 9 HCV+ patients, for each triple combined therapy, with good results. The observed median ratio between S and R isomers for boceprevir and telaprevir were 1.22 (IQR 1.10-1.33) and 1.52 (IQR 1.21-1.67), respectively. The use of this simple assay method could be an important tool for management of HCV-1 DAAs treated patients.


The effect of coronary artery occlusion on the cardiovascular response to an aversive stress.
The cardiovascular response to a controlled aversive stress was investigated both before and during acute myocardial ischemia. Classical aversive conditioning (a 30-s tone reinforced with a 1-s shock) served as the model of stress while anterior wall myocardial ischemia was induced by the occlusion (hydraulic occluder) of the left anterior descending coronary artery. The conditional response consisted of significant increases (P less than 0.01) in mean arterial pressure (AP, 13.8 +/- 1.9 mm Hg, 14.3%), left ventricular (LV) dP/dtmax (1300 +/- 324 mm Hg/s, 34.7%) and heart rate (HR, 44 +/- 4 beats/min, 46.8%). Mean coronary vascular resistance significantly (P less than 0.01) increased first (CVR, 0.52 +/- 0.18 mm Hg/ml/min, 17.2%), then decreased (0.77 +/- 0.14 mm Hg/ml/min, 25.5%). In contrast, during acute myocardial ischemia both the HR and d(LVP)/dtmax conditional response were significantly reduced (P less than 0.01) by 58 and 54%, respectively (HR, 20.7 +/- 3.8 beats/min, d(LVP)/dt, 756 +/- 226 mm Hg/s). In addition, the initial CVR increase was virtually eliminated (0.13 +/- 0.10 mm Hg/ml/min, 74.0% reduction) while the CVR decrease was significantly reduced (P less than 0.01) by 48% (0.40 +/- 0.15 mm Hg/ml/min) during the coronary occlusion. The mean AP response, however, was not affected by myocardial ischemia. The pre-conditional stimulus cardiovascular variables were similarly unaffected by the coronary artery occlusion. Thus, coronary artery occlusion attenuated both the coronary and cardiac response elicited by an aversive stress. The mechanism mediating this inhibition of the conditional response remains to be determined but may involve the activation of vagal depressor reflexes during the acute ischemia.


Full-thickness skin grafting in preventing recurrence of Dupuytren's disease: A systematic review.
Dupuytren's disease is a progressive hand disorder characterized by the formation of pathologic nodules and cords in the palm, often leading to progressive flexion contractures. Hand surgery is currently the only efficient treatment to restore hand function, however, reported recurrence rates vary widely. Currently, it remains unclear which type of fasciectomy is associated with the lowest recurrence rate. Therefore, a systematic review was conducted to compare recurrence rates after limited fasciectomy, open palm technique, or dermofasciectomy with a full-thickness skin graft. 79 studies were included, and results suggest that a dermofasciectomy with a full-thickness skin graft is associated with a lower recurrence rate. However, most studies were case series in which a wide range of definitions of recurrence was used. Moreover, all studies had a high risk of bias. Therefore, additional comparative clinical trials are recommended that use an unambiguous definition of recurrence. LEVEL OF EVIDENCE III: .


Prognostic Significance of Incidental Deep Vein Thrombosis in Patients with Cancer Presenting with Incidental Pulmonary Embolism.
In symptomatic acute pulmonary embolism (PE), the presence of deep vein thrombosis (DVT) is a risk factor for 30- and 90-day mortality. In patients with cancer and incidental PE, the prognostic effect of concomitant incidental DVT is unknown. In this retrospective study, we examined the effect of incidental DVT on all-cause mortality in such patients. Adjusted Cox multivariate regression analysis was used for relevant covariates. From January 2010 to March 2018, we included 200 patients (mean age, 65.3 ± 12.4 years) who were followed up for 12.5 months (interquartile range 7.4-19.4 months). Of these patients, 62% had metastases, 31% had concomitant incidental DVT, and 40.1% (n = 81) died during follow-up. All-cause mortality did not increase in patients with DVT (hazard ratio [HR] 1.01, 95% confidence interval [CI] 0.43-2.75, p = 0.855). On multivariate analysis, weight (adjusted HR 0.96, 95% CI 0.92-0.99, p = 0.032), and metastasis (adjusted HR 10.26, 95% CI 2.35-44.9, p = 0.002) were predictors of all-cause mortality. In conclusion, low weight and presence of metastases were associated with all-cause mortality, while presence of concomitant DVT was unrelated to poorer survival.


The immune status of uraemic children/adolescents with chronic renal failure and renal replacement therapy.
In adults with chronic renal failure (CRF) and/or renal replacement therapy (RRT) various immunological abnormalities have been described, but few data are available for the paediatric age group. We performed basic in vitro immunological studies in 26 patients 10 months-19 years of age with advanced renal failure, 11 with CRF (creatinine clearance 16.8 +/- 5.2 ml/min per 1.73 m2), 15 on RRT with haemodialysis (HD; n = 9) and continuous ambulatory peritoneal dialysis (CAPD; n = 6) as well as in 16 healthy controls. None had clinical evidence of deranged immune function. No significant differences were found in the percentages of B- and T-cells, T-cell subsets CD3, CD4, CD8 and mitogenic responses to phytohaemagglutinin and concanavalin A (Con A) between RRT patients (HD = CAPD) and control children. Most parameters in CRF patients were also normal, although they had a low percentage of B-cells (12.1 +/- 4.1; RRT: 19.7 +/- 6.5; controls: 18.5 +/- 7.1; P less than 0.01), relatively low levels of serum immunoglobulin G (948.4 +/- 209.4 mg/dl; HD: 1374.7 +/- 235.2 mg/dl; P less than 0.01; CAPD: 966.3 +/- 430.2 mg/dl, NS) and a high normal response to Con A (34.3 +/- 13.6 cpm x 10(-3); RRT: 24.5 +/- 11.3 cpm x 10(-3); controls: 23.4 +/- 9.9 cpm x 10(-3), P less than 0.01). All these values were, however, well within the normal accepted range. These data indicate that children/adolescents with CRF and/or RRT have no significant basic in vitro immunological defects. This study did not test the functional immune status of the young uraemic patients.


Reducing the genotoxic potential of retroviral vectors.
The recent development of leukemia in gene therapy patients with X-linked severe combined immunodeficiency disease because of retroviral vector insertional mutagenesis has prompted reassessment of the genotoxic potential of integrating vector systems. In this chapter, various strategies are described to reduce the associated risks of retroviral genomic integration. These include deletion of strong transcriptional enhancer-promoter elements in the retroviral long terminal repeats, flanking the retroviral transcriptional unit with enhancer blocking sequences and designing vectors with improved RNA 3' end processing. Protocols are provided to evaluate the relative biosafety of the modified vectors based on their ability to immortalize hematopoietic progenitor cells and propensity to trigger clonal hematopoiesis or leukemogenesis following hematopoietic stem cell transplantation.


Incidence and malignancy rates classified by The Bethesda System for Reporting Thyroid Cytopathology (TBSRTC) - An 8-year tertiary center experience in Thailand.
Incidence and Malignancy Rates Classified by The Bethesda System for Reporting Thyroid Cytopathology (TBSRTC) - An 8-year Tertiary Center experience in Thailand. Fine-needle aspiration (FNA) of the thyroid is considered the best diagnostic tool for preoperative evaluation of thyroid nodules. The introduction of The Bethesda System for Reporting Thyroid Cytopathology (TBSRTC) in 2010 provided the opportunity to establish worldwide standard for reporting and terminology guidelines for diagnostic categories. It is recommended that every institution evaluates the risk of malignancy (ROM) in each category for quality improvement process. To assess the effectiveness of TBSRTC method at our institution using cyto-histological correlation. A retrospective 8-year (2010-2017) audit of thyroid FNA done by thyroid specialists at Theptarin hospital. The FNA results were classified according to TBSRTC. Histopathology reports for operated cases were used to correlate cytology and final histopathology. A total of 2735 thyroid FNA from 2115 patients (mean age 45.7 ± 13.1 years, female 89.8%) were examined. The rates of non-diagnostic, benign, atypia of undetermined significance (AUS), follicular neoplasm, suspected for malignancy, and malignant cases were 21.1%, 66.6%, 4.7%, 2.4%, 1.8%, and 3.3% respectively. There were 188 patients (9%) who underwent surgical resection with available histopathology. Malignancy rates in operated thyroid nodules were 20.0%, 4.2%, 9.4%, 23.5%, 57.1%, and 90.3% for categories 1 to 6, respectively. The sensitivity, specificity, negative predictive value, and positive predictive value were 96.6%, 88.5%, 95.8%, and 90.3, respectively. Preoperative diagnosis of thyroid nodules using TBSRTC in our hospital was comparable with other studies. The uniform diagnostic criteria of the Bethesda System help avoid misinterpretation while sharing local experience with international benchmarks.


Ultrasonography Technique in Abdominal Subcutaneous Adipose Tissue Measurement: A Systematic Review.
There are currently several reported techniques of sonographic subcutaneous adipose tissue (SAT) measurement described in the literature. This systematic review aims to report techniques of SAT measurement using ultrasonography. A systematic literature search was performed and identified 39 relevant articles using ultrasonography to quantify abdominal SAT. The following parameters were collected: type of measurement, ultrasound machine make/model, transducer frequency, external/internal landmarks, pressure applied on probe, special techniques and inter-/intraobserver reliability. Literature findings related to the above parameters were summarized. A summary of the most common techniques and parameters is provided, serving as a reference for a necessary standardized approach.


Antigenically distinct subpopulations of myeloid progenitor cells (CFU-GM) in human peripheral blood and marrow.
Two types of progenitor cells of the human granulocytic and monocytic lineages (CFU-GM) can be distinguished by using mouse monoclonal antibodies against human hemopoietic cells. Type 1 CFU-GM contribute all of the peripheral blood CFU-GM as well as a small fraction of bone marrow CFU-GM and express surface antigens recognized by "anti-lymphomonocytic" monoclonal antibodies S3-13 and S17-25 but not the antigens recognized by R1B19 and WGHS-29-1 (two monoclonal antibodies that react with all the cells of the granulocytic lineage). Type 2 CFU-GM are present only in the marrow and react with S3-13, R1B19, and WGHS-29-1. Partial reactivity with S17-25 was observed only in the complement-dependent cytotoxicity test. In vitro culture of type 1 CFU-GM in liquid medium in the presence of granulocyte-macrophage colony-stimulatory factor (GM-CSF) generates colony-forming cells that have the surface phenotype of type 2 CFU-GM. This finding supports the idea of two different stages of maturation of myelomonocytic progenitor cells represented by type 1 and type 2 CFU-GM.


Coronary artery bypass grafting in patients with left ventricular dysfunction: predictors of long-term survival and impact of surgical strategies.
In the surgical management of ischemic cardiomyopathy, factors associated with long-term prognosis after coronary artery bypass grafting (CABG) in patients with severe left ventricular (LV) dysfunction are poorly understood. This study aimed to determine predictors of clinical outcomes in patients with severe LV dysfunction undergoing CABG. Out of 6084 patients who underwent CABG between 1997 and 2011, 476 patients (aged 62.6 ± 9.3 years, 100 females) were identified as having severe LV dysfunction (ejection fraction ≤ 35%), preoperatively. All-cause mortality and adverse cardiac events (myocardial infarction, repeat revascularization, stroke and hospitalization due to cardiovascular causes) were evaluated during a median follow-up period of 55.2 months (inter-quartile range: 26.4-94.8 months). During the follow-up, 187 patients (39.3%) died and 126 cardiac events occurred in 104 patients (21.8%). Five-year survival and event-free survival rates were 72.1 ± 2.2% and 61.3 ± 2.4%, respectively. On Cox-regression analysis, old age (P < 0.001), recent MI (P < 0.001), history of coronary stenting (P = 0.023), decreased glomerular filtration rate (P < 0.001), and presence of mitral regurgitation (≥moderate) (P = 0.012) or LV wall thinning (P = 0.007) emerged as significant and independent predictors of death. After adjustment for important covariates affecting outcomes, none of the pump strategy (off-pump vs. on-pump), concomitant mitral surgery or surgical ventricular reconstruction (SVR) affected survival or event-free survival (P = 0.082 to >0.99). Long-term survival following CABG in patients with severe LV dysfunction was affected by age, renal function, recent MI, prior coronary stenting, and presence of mitral regurgitation or LV wall thinning. Neither concomitant mitral surgery nor SVR, however, had significant influence on clinical outcomes.


Beta-adrenergic receptors regulating vascular smooth muscle tone are only localized to the intraocular segment of the long posterior ciliary artery in bovine eye.
Beta-adrenergic drugs are important drugs in glaucoma treatment. Their exact mechanism of action is not yet fully understood but a decreased perfusion pressure in the ciliary body due to blockade of vasodilatory beta-adrenoceptors is thought to participate in the reduction of intraocular pressure. This study investigates the vasodilator action of beta adrenergic-agents in intra- and extraocular arteries from bovine and human eyes. Ring segments of retinal, choroidal, intraocular segment of long posterior ciliary artery, and segments of extraocular posterior ciliary arteries from bovine eyes and short posterior ciliary arteries from freshly enucleated human eyes were mounted on an isometric myograph and their reactivity to beta-adrenergic drugs were studied. Of all vessel types examined only the intraocular segment of long posterior ciliary artery precontracted with prostaglandin F2 alpha responded to 1-isoprenaline (1 nM-1 microM) with a graded concentration-dependent relaxation (32 +/- 7%, n = 10) and a pD2 of 7.5 +/- 0.2 concentrations of 1-isoprenaline greater than 1 microM induced contractions which were antagonized by a combined blockade of alpha1- and alpha2-adrenoceptors. Salbutamol (selective beta2-agonist) induced relaxations of similar magnitude as that of 1-isoprenaline but the sensitivity of the intraocular segment of long posterior ciliary arteries to salbutamol was 312 times less than that to 1-isoprenaline, pD2 7.2 +/- 0.2 and 4.4 +/- 0.1 (n = 6), respectively, whereas dobutamine (selective beta 1-agonist) had no effect between 1 nM and 10 microM. Betaxolol (selective beta 1-antagonist) and 1CI 115811 (selective beta 2-antagonist) competitively antagonized isoprenaline-induced relaxations giving pKB-values of 5.9 +/- 0.1 (n = 11) and 8.5 +/- 0.1 (n = 6), respectively. The slope of the Schild-plots were equal to unity. Human posterior ciliary arteries with spontaneous tone or tone induced by 10 microM PGF2 alpha did not react to isoprenaline (1 nM-10 microM). The bovine intraocular segment of long posterior ciliary arteries contracted to 1-noradrenaline without alpha-adrenoceptor blockade and addition of propranolol or cocaine did not change the concentration-response curve. 1-Phenylephrine and B-HT933 induced both potent and strong contractions indicating the presence of both alpha 1- and alpha 2-adrenoceptors in these arteries. Although beta-adrenoceptors have been identified in various parts of the ocular circulation by radio-ligand binding techniques our data show that beta-adrenoceptors linked to regulation of vascular tone are only found in intraocular branches of the intraocular segment of long posterior ciliary artery in bovine eyes.(ABSTRACT TRUNCATED AT 400 WORDS)


Quantitative surface EMG of pericranial muscles. Reproducibility and variability.
We present reproducibility data on a quantitative electromyographic investigation of the temporal and frontal muscles using surface electrodes. Such data have not been published before. They constitute a necessary basis for future evaluation of the importance of pericranial muscles in tension-type headache. Root mean squares and mean rectified values of the electrical activity were measured in 20 healthy subjects. Power spectra were calculated and the mean and median frequencies were extracted. The coefficient of intraindividual variation in RMS from the right temporal muscle within the same examination was 14%, between several examinations on the same day 18% and between measurements on 2 different days 18%. The corresponding coefficients of interindividual variation were 36, 31 and 38%. The left temporal and frontal muscles showed similar coefficients of variation. At rest, no differences were found between muscles, but during maximal voluntary contraction the frontal muscle revealed a 55% lower RMS values than the temporal muscles. The method used was reproducible and reliable in measuring quantitative electrical activity of pericranial muscles.


Laboratory-based testing for hepatitis C infection using dried blood spot samples: A systematic review and meta-analysis of diagnostic accuracy.
The use of dried blood spot (DBS) samples can facilitate the implementation of reflex testing by circumventing the need for centrifugation and freezing of venous blood samples. This systematic review assessed the accuracy of using DBS samples to diagnose chronic hepatitis C virus (HCV) infection. A comprehensive search was undertaken to identify articles published up to July 2020 evaluating the diagnostic accuracy of anti-HCV, HCV-RNA and HCV core antigen tests using DBS. Screening, data extraction, quality appraisal and Grading of Recommendations, Assessment, Development and Evaluations certainty of the evidence assessment were performed independently by two reviewers. Meta-analysis, meta-regression and sensitivity analyses were conducted. The evidence demonstrates that laboratory-based anti-HCV and HCV-RNA tests using DBS samples have high diagnostic accuracy. All comparisons were between DBS and venous samples. For the detection of anti-HCV, sensitivity was 95% (95% CI: 92%-97%) and specificity was 99% ([95% CI: 98%-99%]; n = 25; I2 = 81%; moderate certainty). For the detection of HCV-RNA, the sensitivity was 95% (95% CI: 93%-97%) and specificity was 97% ([95% CI: 94%-98%]; n = 20; I2 = 52%; moderate certainty). The sensitivity of HCV core antigen tests was 86% (95% CI: 79%-91%) and specificity was 98% ([95% CI: 94%-99%]; n = 5; I2 = 37%; low certainty) compared with HCV-RNA (the gold standard for detecting chronic HCV). DBS samples could facilitate diagnosis of chronic HCV infection as the necessary sequential tests (anti-HCV and then HCV-RNA or HCV core antigen) can be undertaken using the same blood sample. This could reduce loss of patient follow-up and support international efforts towards HCV elimination in both high and low prevalence settings.


The vitamin D status and its effects on life quality among the elderly in Jinan, China.
A challenge for researchers is that vitamin D has many functions in the body, and vitamin D status across the world is not optimistic. There was little data regarding insufficient vitamin D status and its adverse effect on health of elderly Chinese people. So we selected 686 subjects (310 men and 376 women) aged 60-89 years to explore the correlation between vitamin D status and their life quality in a community-based osteoporosis prevention study at Qianfoshan Hospital of Shandong University from 2009 to 2010. All subjects were divided into three groups: vitamin D adequacy [serum 25(OH)D ≧ 75 nmol/L], hypovitaminosis D (50-74.9 nmol/L), and vitamin D insufficiency (<50 nmol/L). Participants were requested to complete SF-36 health survey. The regression models were used to evaluate respectively the association between 25(OH)D levels and parameters of SF-36. As a result, serum 25(OH)D levels were 58.60 nmol/L in male [95% confidence interval (95% CI): 56.7-60.5 nmol/L] and 54.17 nmol/L in female (95% CI: 52.8-55.8 nmol/L). Vitamin D adequacy was in 19.4% of male and 14.4% of female. With aging, proportions of insufficient vitamin D increased significantly in male (P=0.004), and in female (P<0.001). The elderly with hypovitaminosis D got lower scores than that with vitamin D adequacy (almost P<0.01). Regression analysis showed that vitamin D appeared a positive association with parameters of SF-36 (almost P<0.001). In conclusion, vitamin D status in the elderly was severely inadequate, which decreased their life quality and cause adverse effect on their health.