Assessment of fatigue, monitor placement, and surgical experience during simulated laparoscopic surgery.
Laparoscopic surgery requires the surgeon to assume atypical postures for extended periods of time, potentially leading to fatigue and chronic injury. This study assessed the level of muscle activity and compared the effects of fatigue, monitor placement, and surgical experience during simulated laparoscopic surgery. Four attending and four resident surgeons repeated a series of four tasks with a View site and a standard operating room monitor. Electromyography (EMG) activity and muscular discomfort scores were obtained before and after a "fatigue session." Two variables, the EMG amplitudes and discomfort scores, were analyzed. The EMG amplitudes generally exceed the recommended threshold limits of acceptable muscular load. EMG data and discomfort scores demonstrated a fatigue response in several muscle groups. Minimal differences between the two monitor positions were seen. Overall, the EMG data and discomfort scores showed less muscle activity and discomfort in the attending surgeons. Laparoscopic surgery required a relatively high muscular load, putting surgeons at risk for fatigue and injury. Altering the monitor placement did not reduce the surgeon's risk of fatigue. Experience slightly reduced the level of fatigue, but not enough to reduce the surgeon's risk category.


Abnormal Changes of IL3/IL3R and Its Downstream Signaling Pathways in the Prion-Infected Cell Line and in the Brains of Scrapie-Infected Rodents.
Interleukin 3 (IL-3) plays an important role in hematopoiesis and immune regulation, brain IL-3/IL-3R signaling has been shown to involve in the physiological and pathological processes of a variety of neurodegenerative diseases, but its role in prion diseases is rarely described. Here, the changes of IL-3/IL-3R and its downstream signaling pathways in a scrapie-infected cell line and in the brains of several scrapie-infected rodent models were evaluated by various methods. Markedly decreased IL-3Rα were observed in the brains of scrapie-infected rodents at terminal stage and in the prion-infected cell model, which showed increased in the brain samples collected at early and middle stage of infection. The IL-3 levels were almost unchanged in the brains of scrapie-infected mice and in the prion-infected cell line. Morphological assays identified close co-localization of the increased IL-3Rα signals with NeuN- and Iba1-positive cells, whereas co-localization of IL-3 signals with NeuN- and GFAP-positive cells in the scrapie-infected brain tissues. Some downstream components of IL-3/IL-3R pathways, including JAK2-STAT5 and PI3K/AKT/mTOR pathways, were downregulated in the brains of scrapie-infected rodents at terminal stage and in the prion-infected cells. Stimulation of recombinant IL-3 on the cultured cells showed prion that the prion-infected cells displayed markedly more reluctant responses of JAK2-STAT5 and PI3K/AKT/mTOR pathways than the normal partner cells. These data suggest that although prion infection or PrPSc accumulation in brain tissues does not affect IL-3 expression, it significantly downregulates IL-3R levels, thereby inhibiting the downstream pathways of IL-3/IL-3R and blocking the neuroregulatory and neuroprotective activities of IL-3.


Cardiac Rehabilitation Prevents Recurrent Revascularization in Patients With Coronary Heart Disease: A POPULATION-BASED COHORT STUDY IN TAIWAN.
To evaluate the effects of cardiac rehabilitation (CR) provided within the first 3 months of revascularization on reducing recurrent revascularization in patients with coronary heart disease in Taiwan. In this population-based cohort study, we used the claims data of 1 million beneficiaries who were randomly selected from all beneficiaries enrolled in Taiwan's National Health Insurance program from 1996 to 2000. Between 2000 and 2007, 2838 patients underwent a first-event revascularization. Of these patients, 442 (15.6%) underwent CR within the first 3 months of admission for revascularization. The remaining 84.4% (n = 2396) served as the non-CR group. All the study patients were followed-up until the end of 2008 for any recurrent revascularization. A propensity score-adjusted Cox proportional hazard model was used to estimate the relative risk of recurrent revascularization associated with CR. During the 1- to 9-year follow-up, 69 patients (15.6%) in the CR group and 840 (35.1%) patients in the non-CR group experienced recurrent revascularization. The results of the propensity score-adjusted Cox proportional hazard regression analysis showed that CR was significantly associated with a reduced risk of recurrent revascularization with a hazard ratio of 0.48 (95% CI, 0.37 to -0.62). Cardiac rehabilitation within the first 3 months of revascularization is significantly associated with a reduced risk of recurrent revascularization. This preventive effect was more pronounced in men compared with other subgroups of patients.


Energy balance in children and young adults receiving haemodialysis for chronic renal failure.
This study was designed to determine the contribution of energy expenditure to the energy imbalance seen in uraemic children. Resting energy expenditure (REE) was measured using open-circuit indirect calorimetry in eight uraemic haemodialysed subjects aged 9.3-20.4 years and in 10 healthy children. Linear correlations between REE and both body weight and fat-free mass as measured by anthropometry were found in both controls and uraemic subjects (respectively: r = 0.76 and r = 0.88 for body weight and r = 0.73 and r = 0.90 for fat-free mass). Measured REE in uraemic patients was not different from the value predicted by using actual body weight and fat-free mass in the regression equation of REE on body weight and fat-free mass in controls (paired t test: p = 0.70 and p = 0.19 respectively). These data suggest that the energy imbalance seen in uraemic children is not due to increased energy expenditure and is therefore probably due to decreased food intake.


An improved method for separation of low-molecular-weight polypeptides by electrophoresis in sodium dodecyl sulfate-polyacrylamide gel.
An improved system for SDS-polyacrylamide gel electrophoresis, capable of analyzing polypeptides having molecular weights from 1500 to 100,000 (especially showing high resolving power in the 1500 to 25,000 molecular weight range) is described. The 10 to 18% linear gradient gel containing 7 M urea with an acrylamide:bisacrylamide ratio of 20:1 and the Laemmli discontinuous buffer was used. The use of the gel with a high crosslinkage ratio is shown to be effective in lowering the leakage of low-molecular-weight polypeptides from the gel. This method has facilitated rapid detection of small amounts of low-molecular-weight polypeptides in body fluids by the use of silver stain. A procedure is presented for the elimination of false bands on the gel frequently encountered during silver staining. The separation patterns of enzymatic cleavage products of proteins, uremic plasma, and urines from nephropathy patients are illustrated. This system is also applicable in the separation of lipopolysaccharides and also for the detection of phospholipids.


Epidemiology of adult atopic dermatitis.
Atopic dermatitis (AD) is driven by a complex gene-environment interaction. Many of the risk factors and genetic underpinning previously observed for pediatric AD may not apply to adult atopic dermatitis, suggesting that these may largely be different disorders. Whereas AD is classically thought of as a pediatric disease, recent studies have shown high rates of disease in adults as well. Risk factors for persistence of childhood-onset AD, as well as adult-onset AD, are reviewed. Adults with AD are particularly vulnerable to exogenous insults from the outside environment, including climate, ultraviolet exposure, pollution, irritants and pruritogens, and microbes. Finally, adult AD is associated with a substantial health care burden, with increased utilization, direct and indirect costs of care, and lost work productivity.


Potential role of nuclear factor-kappaB in the induction of nitric oxide and tumor necrosis factor-alpha by oligochitosan in macrophages.
Oligochitosan, having an average molecular weight of 1000 Da and a degree of N-acetylation below 15%, can be obtained by either chemical or enzymic hydrolysis of chitosan. The present investigation demonstrated that oligochitosan can significantly increase the activity of inducible nitric oxide synthase (iNOS) and induce the synthesis of nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha) in macrophages. Moreover, nuclear factor-kappaB (NF-kappaB) protein levels in nuclear extract are increased in response to oligochitosan. Blocking NF-kappaB with specific inhibitor results in decreased levels of NO and TNF-alpha. These results indicate that NF-kappaB plays a potential role in the induction of NO and TNF-alpha by oligochitosan in macrophages.


Systematic Review of Irreversible Electroporation Role in Management of Locally Advanced Pancreatic Cancer.
Ablative techniques provide in patients with locally advanced pancreatic cancer (LAPC) symptomatic relief, survival benefit and potential downsizing. Irreversible Electroporation (IRE) represents potentially an ideal solution as no thermal tissue damage occurs. The purpose of this review is to present an overview on safety, feasibility, oncological results, survival and quality of life improvement obtained by IRE. A systematic search was performed in PubMed, regarding the use of IRE on PC in humans for studies published in English up to March 2019. 15 original studies embodying 691 patients with unresectable LAPC who underwent IRE were included. As emerged, IRE works better on tumour sizes between 3-4 cm. Oncological results are promising: median OS from diagnosis or treatment up to 27 months. Two groups investigated borderline resectable tumours treated with IRE before resection with margin attenuation, whereas IRE has proved to be effective in pain control. Electroporation is bringing new hopes in LAPC management. The first aim of IRE is to offer a palliative treatment. Further efforts are needed for patient selection, as well as the use of IRE for 'margin accentuation' during surgical resection. Even if promising, IRE needs to be validated in large, randomized, prospective series.


In vivo real-time vessel imaging and ex vivo 3D reconstruction of atherosclerotic plaque in apolipoprotein E-knockout mice using synchrotron radiation microscopy.
To examine the pathophysiology of atherosclerosis, imaging the vascular wall and pathology without tissue damage is required. We used the unmonochromatized synchrotron X-ray to acquire in vivo real-time and ex vivo images of atherosclerotic lesions in apo E-knockout mice without contrast agents or staining. In the five apo E-knockout mice (apo E-/-, 12, 24, 32, 48, 62-week-old, 3 males) and age/sex matched five wild type mice on cow diets, we acquired in vivo real-time images of thoracic aorta without contrast agents and then, the central arterial trees were dissected intact. Ex vivo synchrotron images with tomographic reconstruction were done and compared with the corresponding pathology. For all living animals, in vivo real-time images of thoracic aorta could be acquired without contrast agents but could not identify the atherosclerotic lesions. Ex vivo images accurately determined aortic wall and atherosclerotic plaque without staining in comparison to histopathology according to the AHA classification (r=0.84, p<0.001). The volume rendered 3 D images of plaque showed central cholesterol clefts as matched with optical images. The combination of synchrotron enhanced X-ray microscopy and genetically engineered hyperlipidemic animals would be a useful tool to investigate the changes of advanced atherosclerotic lesions.


Solid phase enzyme immunoassay of cyclic adenosine 3',5'-monophosphate. Effect of coating strategy upon assay performance in comparison with radioimmunoassay.
We have evaluated two novel enzyme-linked immunosorbent assays (ELISAs) used to quantitate cyclic AMP. In one assay ELISA plates are coated with antigen consisting of a cyclic AMP-polylysine conjugate. Cyclic AMP samples added to plates are quantified by their ability to decrease the binding and anti-cyclic AMP antibodies to the coated antigen. A second ELISA utilizes a plated anti-immunoglobin technique in which plates are coated first with anti-goat IgG and then with goat anti-cyclic AMP antiserum. Cyclic AMP samples are quantified by their ability to compete with cyclic AMP-peroxidase conjugates for binding to the plated anti-cyclic AMP antibodies. The plated anti-immunoglobin ELISA proved to be somewhat more sensitive than the plated antigen ELISA and was comparable in sensitivity to an automated RIA for measuring cyclic AMP in standards and urine samples. Our data fit with the generalization that optimal ELISA sensitivity is obtained through the use of plated anti-immunoglobins rather than plated antigens. Further they demonstrate the practicality of utilizing small ligand-enzyme conjugates for ELISAs.


Design of biomimetic fibrillar interfaces: 2. Mechanics of enhanced adhesion.
This study addresses the strength and toughness of generic fibrillar structures. We show that the stress sigmac required to pull a fibril out of adhesive contact with a substrate has the form sigma(c) = sigma(0)Phi(chi). In this equation, sigma(0) is the interfacial strength, Phi(chi) is a dimensionless function satisfying 0 <or= Phi(chi) <or= 1 and chi is a dimensionless parameter that depends on the interfacial properties, as well as the fibril stiffness and radius. Pull-off is flaw sensitive for chi >> 1, but is flaw insensitive for chi < 1. The important parameter chi also controls the stability of a homogeneously deformed non-fibrillar (flat) interface. Using these results, we show that the work to fail a unit area of fibrillar surface can be much higher than the intrinsic work of adhesion for a flat interface of the same material. In addition, we show that cross-sectional fibril dimensions control the pull-off force, which increases with decreasing fibril radius. Finally, an increase in fibril length is shown to increase the work necessary to separate a fibrillar interface. Besides our calculations involving a single fibril, we study the concept of equal load sharing (ELS) for a perfect interface containing many fibrils. We obtain the practical work of adhesion for an idealized fibrillated interface under equal load sharing. We then analyse the peeling of a fibrillar surface from a rigid substrate and establish a criterion for ELS.


Phenotypic hiding: the carryover of mutations in RNA viruses as shown by detection of mar mutants in influenza virus.
When influenza virus monoclonal antibody-resistant (mar) mutants are selected by incubation in vitro with excess antibody, 90 to 99% of the mutants are not detectable. This observation may be explained by encapsidation of mar mutant RNAs within phenotypically wild-type envelopes. This phenotypic hiding can be revealed by selection of mar mutants in vivo after virus uncoating. Using experimental procedures appropriate to detect all viable mar mutants in a virus population, we determined precisely the mutation rates to the mar genotype by the fluctuation test for two nonoverlapping monoclonal antibodies.


Characteristics and trends of 3,4-methylenedioxymethamphetamine (MDMA) tablets found in Taiwan from 2002 to February 2005.
One hundred and eighty-one 3,4-methylenedioxymethamphetamine (MDMA) containing tablets were sampled from confiscated drugs received by the Taiwan National Bureau of Controlled Drugs for testing from 2002 to February 2005. Sample tablets demonstrated various colors and logos. The appearances, contents of MDMA and other components in these tablets were analyzed in order to understand the characteristics and trends of MDMA use. Samples were analyzed using GC-MS methodology. Deuterated internal standards were used for drug quantification. The MDMA contents varied from 16 to 193 mg/tablet. 66-71% of the tablets seized each year contained only MDMA, and the content of MDMA in MDMA only tablets varied from 89 to 133 mg/tablet. There was a decreasing trend in MDMA content in these tablets over time. Other components commonly found besides MDMA included caffeine (18%), methamphetamine (7%), 3,4-methylenedioxyethylamphetamine (MDEA) (7%) and amphetamine (4%). 3,4-Methylenedioxyamphetamine (MDA), ketamine, ephedrine, diazepam, chlorzoxazone and nicotinamide were also detected. During the study period, the number of other drugs found as well as the combinations of different drugs detected in these tablets increased.


Assessment of the psychometric properties of the Spanish version of EORTC QLQ-MY20 and evaluation of health-related quality of Life outcomes in patients with relapsed and/or refractory multiple myeloma in the real-world setting in Spain: results from the CharisMMa study.
We evaluated the psychometric properties of the Spanish version of the European Organization for Research and Treatment of Multiple Myeloma (MM) specific quality-of-life (QoL) questionnaire module (QLQ-MY20) in relapsed/refractory MM (RRMM) patients. This was an observational, cross-sectional, multicenter study using EORTC QLQ-C30 and QLQ-MY20 in RRMM patients (ClinicalTrials.gov ID NCT03188536). We assessed the non-response rate, ceiling/floor effects, internal consistency, test-retest reliability, and validity. The study included 276 patients (53.3% males, mean [SD] age of 67.4 [10.5] years). The EORTC QLQ-MY20 showed a low non-response rate, very low ceiling and floor effects, and good internal consistency. The test-retest reliability assessment revealed good temporary stability, the construct validity analysis stated four main factors similar to the ones of the original version, and the criterion validity assessment showed no differences between groups. In conclusion, the Spanish version of EORTC QLQ-MY20 is a reliable and valid tool for assessing QoL in RRMM patients.


Silencing of long non-coding RNA ZFAS1 alleviates LPS-induced acute lung injury by mediating the miR-96-5p/OXSR1 axis in sepsis.
Extensive studies have revealed that long non-coding RNAs (lncRNAs) are associated with sepsis-induced acute lung injury (ALI). This study focused on the function and potential mechanisms of lncRNA zinc finger antisense 1 (ZFAS1) in a cell model of sepsis-induced ALI. To induce sepsis-induced ALI in vitro and in vivo, mice were subjected to cecal ligation and puncture (CLP) operation, and human small airway epithelial cells (HSAECs) were stimulated with lipopolysaccharide (LPS) (10 μg/mL). Relative expression of oxidative stress-responsive 1 (OXSR1), lncRNA ZFAS1, and microRNA (miR)-96-5p was detected by quantitative real-time polymerase chain reaction (qRT-PCR). Relative protein expression of Bax, Bcl-2, and OXSR1 was determined by western blotting. Moreover, enzyme-linked immunosorbent assay was used to measure the levels of IL-6, IL-1β, and TNF-α. A dual-luciferase reporter assay was conducted to test the targeting interplay between ZFAS1/OXSR1 and miR-96-5p. Up-regulation of lncRNA ZFAS1 and OXSR1 and down-regulation of miR-96-5p was observed in lung tissues of CLP-induced mice and LPS-treated HSAECs. Decreased ZFAS1 expression or increased miR-96-5p expression repressed inflammation and apoptosis and promoted cell viability in LPS-treated HSAECs. The lncRNA ZFAS1 competitively binds to miR-96-5p and inversely modulates miR-96-5p expression. MiR-96-5p directly targets OXSR1 and inversely regulates OXSR1 expression. In addition, the protective effects of ZFAS1 knockdown on LPS-induced HSAECs were reversed by miR-96-5p inhibition or OXSR1 overexpression. Down-regulation of lncRNA ZFAS1 attenuated LPS-induced ALI in HSAECs by regulating the miR-96-5p/OXSR1 axis.


Automated remote transapical wound closure system study.
A readily applied and reliable means to create and close transapical wound access sites through a small intercostal opening could benefit many cardiac patients. Using relevant surgical tissue models, this research evaluated a new approach to enable single-port transapical wound site closure toward the eventual development of safe percutaneous access to the left ventricle. Novel techniques and technologies were developed and successfully tested for remote automated placement of 2 pledgeted horizontal mattress sutures delivered concentrically around a transmural apical guidewire, which provided a tract to enable therapeutic intervention. Mechanical pledgeted knots secured these sutures after cannula removal. Automated transapical closures were created in 47 ex vivo porcine hearts and in 10 human cadavers, 8 through a thoracotomy and 2 thoracoscopically. Automated apical closures achieved hemostasis in an acute beating heart thoracotomy model in 3 anesthetized pigs. An ex vivo porcine heart pressurized infusate model demonstrated the intraventricular pressure tolerated by automated closures (mean, 327 mm Hg) compared with hand-sutured closures (mean, 303 mm Hg). All automated apical closures were effective. Early results encourage further evaluation.


Identification of cbbBc as an additional distal gene of the chromosomal cbb CO2 fixation operon from Ralstonia eutropha.
Ralstonia eutropha (formerly Alcaligenes eutrophus) strain H16 possesses two highly homologous cbb operons encoding most of the Calvin cycle enzymes. One copy of the operon is located on the chromosome, the other on the megaplasmid pHG1 of the organism. Sequence analysis of the region downstream of the presumptive 3'-terminal gene (cbbAc) of the chromosomal operon revealed the presence of an open reading frame comprising 2,274 bp. Evidence is presented that this open reading frame is an additional distal gene (designated cbbBc) of the operon. In contrast to the other genes of the operon, cbbBc is not duplicated in the plasmid-borne operon. The deduced amino acid sequence of the cbbBc product (757 residues, molecular mass 83.17 kDa) showed the highest similarity to the large catalytic subunits of various bacterial formate dehydrogenases (FDH), suggesting that cbbBc might represent a structural FDH gene of R. eutropha. However, the properties of a cbbBc mutant strain indicated that the potential gene product is not related to known FDH of the organism. Transcriptional analysis in the homologous host and heterologous expression in Escherichia coli demonstrated that cbbBc is an active gene, which apparently has no essential function in the autotrophic metabolism of R. eutropha. The gene is a novel member of cbb operons in autotrophic bacteria.


A rapid method for capture and identification of immunogenic proteins in Bordetella pertussis enriched membranes fractions: a fast-track strategy applicable to other microorganisms.
Mass spectrometry (MS) coupled with 1-D and 2-D electrophoresis can be utilized to detect and identify immunogenic proteins, but these methods are laborious and time-consuming. We describe an alternative, simple, rapid gel-free strategy to identify multiple immunogenic proteins from Bordetella pertussis (Bp). It couples immunoprecipitation to nano liquid chromatography- tandem mass spectrometry (IP-nLC-MS/MS) and is significantly both time- and labor-saving. We developed a gel-free magnetic bead-based immunoprecipitation (IP) method using different NP-40/PBS concentrations in which solubilized proteins of Bp Tohama I membrane fractions were precipitated with polyclonal rabbit anti-Bp whole cell immune sera. Immune complexes were analyzed by MS and Scaffold analysis (>95% protein identification probability). Total immunoproteins identified were 50, 63 and 49 for 0.90%, 0.45% and 0.22% NP-40/PBS buffer concentrations respectively. Known Bp proteins identified included pertactin, serotype 2 fimbrial subunit and filamentous hemagglutinin. As proof of concept that this gel-free protein immunoprecipitation method enabled the capture of multiple immunogenic proteins, IP samples were also analyzed by SDS-PAGE and immunoblotting. Bypassing gels and subjecting immunoprecipitated proteins directly to MS is a simple and rapid antigen identification method with relatively high throughput. IP-nLC-MS/MS provides a novel alternative approach for current methods used for the identification of immunogenic proteins.


Flocculectomy and unit activity in the vestibular nuclei during visual-vestibular interactions.
Activity of neurons in the vestibular nuclei of alert monkeys was recorded extracellularly after total unilateral and bilateral flocculectomy and partial paraflocculectomy. Type 1 horizontal cells that were encountered after flocculectomy responded to visual and vestibular stimuli and to conflict stimulation, i.e., to rotation in a subject-stationary visual surround, as do vestibular neurons in the normal animal. The major difference between neurons in the normal and lesioned animals was that more time was needed to reach steady state firing levels during optokinetic stimulation at a constant velocity after operation. As shown previously (Waespe et al. 1983) the longer time course is related to increased initial retinal slip velocities that occur after flocculectomy as a result of an inability to change eye velocity rapidly in response to visual stimulation. It does not signify a change in the dynamics of neurons in the vestibular nuclei that mediate the vestibulo-ocular reflex (VOR). The similarity in modulation of horizontal Type 1 vestibular neurons in normal and flocculectomized monkeys makes it unlikely that floccular Purkinje cells suppress the horizontal VOR in the monkey during conflicting visual and vestibular stimuli by inhibiting or disfacilitating secondary or tertiary vestibular neurons. This is consistent with earlier findings that indicate that visual-oculomotor pathways responsible for ocular pursuit or for rapid changes in OKN do not go through the vestibular nuclei. Rather the point of interaction of the flocculus output with the VOR appears to be external to the vestibular nuclei. There was a close correspondence between the slow phase velocity of nystagmus and unit activity in the vestibular nuclei under a wide variety of test conditions after flocculectomy. This is consistent with the postulate that frequencies of vestibular nuclei neurons represent a summation of activity in direct vestibulo-oculomotor pathways and indirect pathways that include the velocity storage mechanism. These are the major remaining sources of activity that generate slow phases of nystagmus after the direct visual-oculomotor pathways have been partially interrupted by flocculectomy (Waespe et al. 1983). Horizontal Type 1 neurons which responded to vestibular and optokinetic stimulation with increases in frequency above 1 spike/s/degree/s were rarely encountered after flocculectomy. These cells were present on the normal side in a monkey after unilateral flocculectomy. We infer that vestibular nuclei neurons that project mossy fibers to the flocculus are inactivated or disappear as a result of surgical ablation of their axons. This could also contribute to the reduced gain of vestibular nystagmus, OKAN and off-vertical nystagmus that was observed in some of the animals after lesion.


Promoter-specific p53-dependent histone acetylation following DNA damage.
We have used chromatin immunoprecipitation (ChIP) to measure p53-dependent histone acetylation at the p21, MDM2, and PUMA promoters. The pattern of histone acetylation was different at each promoter. H3 and H4 acetylation increased at both the p21 and PUMA promoters in response to p53 activation, whereas there was only a minimal increase in H4 acetylation and no increase in H3 acetylation at the MDM2 promoter. The high p53 occupancy of the p21, MDM2 and PUMA promoters has been attributed to the presence of two p53 binding sites in these promoters, but mutation of the p53 binding sites in integrated p21 promoter constructs showed that the two sites in the p21 promoter do not cooperate to stabilize p53 binding. Despite 10-fold higher p53 binding to the proximal than the distal site in the p21 promoter, both sites showed similar patterns of H3 and H4 acetylation. Mutation of the binding sites showed that acetylation of the proximal, low-affinity site requires p53 binding to that site but not to the distal, high-affinity site. Since low-affinity p53 binding sites can confer strong acetylation, the DNA binding affinity in vitro is an unreliable guide to the likely importance of p53 in regulating candidate target genes in vivo.


The lncRNA HOTAIR via miR-17-5p is involved in arsenite-induced hepatic fibrosis through regulation of Th17 cell differentiation.
Arsenic compounds are toxins that are widely distributed in the environment. Chronic exposure to low levels of these compounds can cause hepatic fibrosis and other damage. Th17 differentiation of CD4+ T cells and the secretion of IL-17 activates hepatic stellate cells (HSCs), which are involved in hepatic fibrosis, but their mechanisms in arsenic-induced hepatic fibrosis are unclear. We found, in arsenite-induced fibrotic livers of mice, increases of CD4+ T cell infiltration, Th17 cell nuclear receptor retinoic acid receptor-related orphan receptor γt (RORγt), and secretion of the pro-inflammatory cytokine IL-17. There were also elevated levels of the lncRNA, HOTAIR. For Jurkat cells, arsenite elevated levels of HOTAIR and protein levels of RORγt and IL-17A, decreased miR-17-5p, promoted Th17 cell differentiation, and released IL-17. The culture medium of arsenite-treated Jurkat cells activated LX-2 cells. Down-regulation of HOTAIR or up-regulation of miR-17-5p blocked arsenite-induced Th17 cell differentiation, which inhibited the LX-2 cell activation. However, down-regulation of HOTAIR and miR-17-5p reversed this inhibitory effect. For mice, silencing of HOTAIR diminished the hepatic levels of RORγt and IL-17A and alleviated arsenite-induced hepatic fibrosis. These results demonstrate that, for CD4+ T cells, arsenite promotes RORγt-mediated Th17 cell differentiation through HOTAIR down-regulation of miR-17-5p, and increases the secretion of cytokine IL-17A, which activates HSCs; the activated HSCs facilitate hepatic fibrosis. The findings reveal a new mechanism and a potential therapeutic target for arsenite-induced hepatic fibrosis.


Severe adenovirus pneumonia with hemophagocytic syndrome and respiratory failure.
We report the case of an 18-month-old infant with severe serotype 3 adenovirus pneumonia, exceptionally associated with hemophagocytic syndrome. Treatment included cidofovir and mechanical ventilation for 13 days. The child developed chronic respiratory insufficiency due to bronchiectasis and bronchiolitis obliterans.


High-throughput sequencing of peripheral blood for minimal residual disease monitoring in childhood precursor B-cell acute lymphoblastic leukemia: A prospective feasibility study.
Minimal residual disease (MRD) measured on end-of-induction bone marrow (BM) is the most important biomarker for guiding therapy in pediatric acute lymphoblastic leukemia (ALL). Due to limited sensitivity of current approaches, peripheral blood (PB) is not a reliable source for identifying patients needing treatment changes. We sought to determine if high-throughput sequencing (HTS) (next-generation sequencing) of rearranged immunoglobulin and T-cell receptor genes can overcome this and be used to measure MRD in PB. We employed a quantitative HTS approach to accurately measure MRD from one million cell equivalents of DNA from 17 PB samples collected at day 29 after induction therapy in patients with precursor B-cell ALL. We compared these results to the gold-standard real-time PCR result obtained from their paired BM samples, median follow-up 49 months. With the increased sensitivity, detecting up to one abnormal cell in a million normal cells, we were able to detect MRD in the PB by HTS in all those patients requiring treatment intensification (MRD ≥ 0.005% in BM). This is proof of principle that using the increased sensitivity of HTS, PB can be used to measure MRD and stratify children with ALL. The method is cost effective, rapid, accurate, and reproducible, with inherent advantages in children. Importantly, increasing the frequency testing by PB as opposed to intermittent BM sampling may allow extension of the dynamic range of MRD, giving a more complete picture of the kinetics of disease remission while improving relapse prediction and speed of detection.


Per- and postoperative changes in the concentration of serum thyreotropin under general anaesthesia, compared to general anaesthesia with epidural analgesia.
In 18 patients scheduled for lower intraabdominal surgery (hysterectomy), changes in thyreotropin (TSH) thyroxine (T4), triiodothyronine (T3) binding of thyroid hormones to plasma proteins (T3-uptake) and glucose in serum were evaluated. In eight patients afferent neurogenic impulses from the surgical area were blocked (Th4-S5) with bupivacaine 0.5% infused continuously into the epidural space from the start of the operation until 6 h postoperatively. All patients received general anaesthesia with thiopentone, pethidine, pancuronium and nitrous-oxide plus oxygen. The patients receiving epidural analgesia had no increase in plasma-TSH, compared to the other group, which had a significant (P less than 0.05) increase peroperatively. The patients receiving epidural analgesia were pain-free and the normal stress-induced increase in plasma-glucose was abolished. Concerning T3 we found a significant decrease in both groups and a steady level of T4- and T3-uptake without significant fluctuations. Thus it can be concluded that the effects of surgical trauma on plasma-TSH concentration are markedly similar to the effects of other anterior pituitary hormones, i.e. HGH, prolactin and ACTH.


Ciliary neurotrophic factor prevents ischemia-induced learning disability and neuronal loss in gerbils.
Ciliary neurotrophic factor (CNTF) has been shown to have potent neurotrophic activity on peripheral and central neurons in vitro and in vivo. However, it remains to be determined whether or not CNTF rescues hippocampal CA1 neurons from lethal ischemia and prevents ischemia-induced learning disability. In the present in vivo study, we infused CNTF continuously for 7 days into the lateral ventricle of gerbil starting 2 h before 3-min forebrain ischemia. CNTF infusion prevented the occurrence of ischemia-induced learning disability in a dose-dependent manner as revealed by the step-down passive avoidance task. Subsequent light and electron microscopic examinations showed that pyramidal neurons in the CA1 region of the hippocampus as well as synapses within the strata moleculare, lacunosum/radiatum and oriens of the region were significantly more numerous in gerbils infused with CNTF than in those receiving vehicle infusion. These findings suggest that CNTF has a trophic effect on ischemic hippocampal neurons.


Early Intervention Services for Schizophrenia: Looking Back and Looking Ahead.
This invited commentary provides a conceptual history of modern early intervention services, briefly reviews the accomplishments of an international clinical and research community, and offers proposals for how such services might participate in the next generation of progress. In keeping with the theme of this column, we make the argument that such services should orient around bi-directional knowledge translation across basic, clinical and policy domains.


Symptoms without signs in outpatients attending ophthalmology clinics.
Ophthalmic outpatients without physical signs have been shown, as a group, to be significantly more anxious and depressed than a control group. They also have a higher incidence of somatic complaints of a kind commonly made by patients with psychiatric 'disorder'. There was a higher incidence of past psychiatric history in the proband group, and they also gave a greater subjective assessment of recent conscious life stress.


The metabolism of L-m-tyrosine: the use of a putative precursor to investigate the increased production of m-hydroxymandelic acid in phenylketonuria.
Several urinary meta-substituted phenols appear to be of endogenous origin. However, the production of these compounds is reduced in phenylketonuria with the exception of m-hydroxymandelic acid whose excretion is approximately doubled. This phenomenon has been investigated in two patients with phenylketonuria using the putative precursor L-m-tyrosine labelled with deuterium. Metabolism of this compound in these patients was comparable to that in healthy adults although much less was converted to m-hydroxymandelic acid and the excretion pattern of this metabolite was different. This apparent anomaly is attributed to smaller metabolic compartments in phenylketonuria and a lower threshold for the metabolism of m-tyramine via beta-hydroxylation. Incorporation into the natural pathway was shown by depletion of endogenous m-hydroxymandelic acid. The results are further support for the ideas that the amine precursors of m-hydroxymandelic acid, m-tyramine and m-octopamine, have a functional role and may be important in the pathogenesis of phenylketonuria.


Hepatic hemangioma with arterioportal shunt: Prevalence and lesion characteristics based on DSA, CT and MR imaging.
To evaluate the prevalence and lesion characteristics of hepatic hemangioma (HH) with arterioportal shunt (APS) based on digital subtraction angiography (DSA), computed tomography (CT) and magnetic resonance (MR) imaging. Ninety-eight consecutive patients with 189 HHs who underwent DSA, CT and/or MR imaging of the liver between January 2014 and December 2017 were included. The diagnosis of APS was established by DSA. APS was categorized as peripheral and central shunt based on imaging findings. The incidence and appearance of APS on DSA were compared with those on CT/MR images. Eleven possible lesion characteristics associated with APS were compared between HHs with and those without APS. Multiple logistic regression modeling was used to identify the independent lesion characteristics associated with APS. APS was diagnosed in 103 (103/189, 54.5%) HHs on DSA, of which 96 lesions appeared as peripheral APS and 7 appeared as central. In contrast, APS was detected only in 57 HHs (57/103, 55.34%) on CT/MR imaging, of which 50 (50/96, 52.08%) appeared as peripheral APS while 7 (7/7, 100%) appeared as central. Lesion size (P < .001), enhancement rapidity (P = .031), and vascularization degree (P < .001) were found to be significant independent imaging characteristics associated with APS. APS can occur in HH with high frequency. DSA was superior to CT/MR imaging in detection of APS, particularly for the peripheral APS. Lesion size, enhancement rapidity and vascularization degree were associated with the presence of APS.


Cardiovascular Imaging and Interventional Procedures in Patients with Novel Coronavirus Infection.
The coronavirus disease 2019 (COVID-19) pandemic is a huge challenge to the health system because of the exponential increase in the number of individuals affected. The rational use of resources and correct and judicious indication for imaging exams and interventional procedures are necessary, prioritizing patient, healthcare personnel, and environmental safety. This review was aimed at guiding health professionals in safely and effectively performing imaging exams and interventional procedures.


Primary amyloidosis of urethra.
Primary localized amyloidosis of the urethra is rare; only 17 cases reported to date. Its clinical importance is significant, however, in that its presentation mimics carcinoma and treatment should be conservative. We describe an additional case and briefly review the literature.


Dynamics of lamivudine-resistant hepatitis B virus strains in patients with entecavir rescue therapy.
Entecavir rescue therapy is frequently used in patients with lamivudine-resistant hepatitis B virus (HBV) strains. The aim of this study was to investigate evolutionary patterns of HBV quasispecies during entecavir rescue therapy and evaluate their impacts on therapeutic efficacy. We enrolled 21 chronic hepatitis B patients who failed to respond to lamivudine therapy and were switched to entecavir treatment. Measurement of serum HBV DNA and sequence analysis of HBV reverse transcriptase were done up to 144 weeks. Four patients of this series showed a reversion to wild-type HBV after entecavir treatment and in three of them, a complete viral response (<2.6 log10 copies/ml) was achieved. An additional five patients developed entecavir genotypic resistance, with prior occurrence of lamivudine-resistant mutation (L180 M ± M204 V/I). A viral breakthrough was observed in four of the five patients with entecavir-resistant mutants. The remaining 12 patients of this series showed dominance of lamivudine-resistant mutants throughout the entecavir rescue therapy, and five of them achieved a complete viral response at the end of follow-up. The average HBV DNA level was significantly lower in patients with a reversion to wild-type HBV than in those without it (P < 0.05). In conclusion, reversion to wild-type HBV is a favorable indicator for response to entecavir rescue therapy in lamivudine-refractory patients with chronic hepatitis B. The presence of lamivudine-resistant mutations contributes to the development of entecavir resistance.


Hydrogen exchange in Pseudomonas cytochrome c-551.
Hydrogen exchange rates were measured or estimated for 75 amide protons in in ferrocytochrome c-551 from Pseudomonas aeruginosa (82 residues total) at neutral pH and 300 K. Rate constants span at least eight orders of magnitude. Rate constants or limiting estimates were determined by a combination of methods relying upon 1H-NMR spectroscopy, including the direct observation in one- or two-dimensional spectra of the decrease in proton intensity for samples dissolved in deuterium oxide, or, in a few favorable cases, saturation transfer from the solvent protic water. The heme ligand residues and the thioether bridge residues were slowly exchanging backbone amides, but the slowest exchanging backbone amides were found in two clusters. One was composed of Ile-48 and Lys-49 in the last turn of what is termed the 40's helix in the protein. The second was composed of Leu-74, Ala-75, Lys-76 and Val-78 in the C-terminal alpha helix.


Ionic liquid-linked dual magnetic microextraction: a novel and facile procedure for the determination of pyrethroids in honey samples.
A novel and facile microextraction technique, termed as ionic liquid-linked dual magnetic microextraction (IL-DMME), was developed for the determination of pyrethroids in honey samples. The distinct advantage of the proposed method is that high recoveries can be readily achieved through the combination of dispersive liquid-liquid microextraction (DLLME) and dispersive microsolid-phase extraction (D-μ-SPE) with the aid of synthetic ionic liquid and non-modified magnetic nanoparticles (MNPs), respectively. In the first DLLME step, [C6MIM]NTf2 was used to extract the pyrethroids without the addition of any toxic dispersive solvent. In the following D-μ-SPE steps, non-modified MNPs were added to retrieve the ionic liquid. The effect of different variables on the extraction efficiency was studied simultaneously using the response surface methodology. The Plackett-Burman design was first employed to screen for the variables that significantly affected the extraction efficiency. Central composite design (CCD) was then introduced to optimize the significant factors using a polynomial fit. The optimal experimental conditions obtained from this statistical evaluation included: ionic liquid volume, 75 μL; S-BaFe quantity, 60 mg; sonication time, 4 min; vortex time, 100 s; desorption time, 150 s; and no addition of salt. Under the optimal conditions, good linearity in the range of 0.5 μg L(-1)-500 μg L(-1), repeatability (RSD 1.1-3.8%), low LODs (0.03-0.05 μg L(-1)) and good recovery (86.7-98.2%) were obtained. Finally, the developed method was evaluated for the extraction and determination of four pyrethroids in real honey samples.


Mitochondrial permeability transition induced by reactive oxygen species is independent of cholesterol-regulated membrane fluidity.
Cholesterol enrichment of rat liver mitochondria (CHM) impairs atractyloside-induced mitochondrial permeability transition (MPT) due to decreased membrane fluidity. In this study we addressed the effect of cholesterol enrichment on MPT induced by reactive oxygen species (ROS). Superoxide anion generated by xanthine plus xanthine oxidase triggered mitochondrial swelling and cytochrome c release in CHM, which was prevented by butylated hydroxytoluene, an anti-voltage-dependent anion channel antibody, or cyclosporin A. Furthermore, hydrogen peroxide generated by the combination of ganglioside GD3 and mitochondrial GSH depletion elicited mitochondrial swelling and release of cytochrome c, Smac/Diablo and apoptosis-inducing factor in control mitochondria and CHM. Thus, ROS induce MPT and apoptosome activation regardless of decreased mitochondrial membrane dynamics due to cholesterol enrichment.


Game of Drones: Multi-UAV Pursuit-Evasion Game With Online Motion Planning by Deep Reinforcement Learning.
As one of the tiniest flying objects, unmanned aerial vehicles (UAVs) are often expanded as the "swarm" to execute missions. In this article, we investigate the multiquadcopter and target pursuit-evasion game in the obstacles environment. For high-quality simulation of the urban environment, we propose the pursuit-evasion scenario (PES) framework to create the environment with a physics engine, which enables quadcopter agents to take actions and interact with the environment. On this basis, we construct multiagent coronal bidirectionally coordinated with target prediction network (CBC-TP Net) with a vectorized extension of multiagent deep deterministic policy gradient (MADDPG) formulation to ensure the effectiveness of the damaged "swarm" system in pursuit-evasion mission. Unlike traditional reinforcement learning, we design a target prediction network (TP Net) innovatively in the common framework to imitate the way of human thinking: situation prediction is always before decision-making. The experiments of the pursuit-evasion game are conducted to verify the state-of-the-art performance of the proposed strategy, both in the normal and antidamaged situations.


Heterogeneity of dermal OKT6+ cells in inflammatory and neoplastic skin diseases.
This immunopathologic study of both normal and pathologic skin specimens (contact dermatitis [CD], lichen planus [LP], cutaneous T cells lymphoma [CTCL], and histiocytosis X [HX]) allowed as to differentiate four types of dermal OKT6+ cells: (1) cells with the same morphologic features as epidermal Langerhans cells (LCs), rarely found in either normal or pathologic dermis; (2) cells structurally similar to LCs but lacking Birbeck granules (BGs), found mainly in CD and LP; (3) larger cells rich in cytoplasmic organelles, only 5% of which contained BGs. They were especially common CTCL; and (4) cells typical of HX.


Photoreceptor inner and outer segment defects in myopic foveoschisis.
To evaluate pathologic features of the photoreceptors in myopic foveoschisis with the Fourier-domain optical coherence tomography (FD-OCT). Observational case series. Seventeen eyes of 15 patients with myopic foveoschisis (foveal detachment type, six eyes; foveoschisis type, 11 eyes) were included. We observed the photoreceptor inner and outer segments (IS/OS) and evaluated the morphologic status using FD-OCT. Fundus photographs and time-domain OCT (TD-OCT) images also were obtained. IS/OS defects, which are uncommon in retinal detachments in eyes with myopia, were seen clearly in five eyes (three eyes [50%] with the foveal detachment type; two eyes [18%] with the foveal schisis type). Fundus photographs showed myopic chorioretinal atrophy in eight study eyes (47%), including diffuse atrophy in four eyes (24%) and patchy atrophy in four eyes (24%). The IS/OS defects on FD-OCT were accompanied by myopic chorioretinal atrophy in three (60%) of five eyes. FD-OCT enables observation of more detailed retinal structures in these myopic eyes. The IS/OS defects may be specific to, but are not rare, in myopic foveoschisis and may predict the postoperative visual recovery.


Osteochondroma of the proximal phalanx of toe presenting as widening of third web space- a rare case report.
Osteochondromas are the most common benign bone tumors, which usually affect the metaphyseal region of long bones. Foot is an unusual location for these tumors with phalanges being a much rare site. Only a few case reports have been made of phalangeal osteochondromas. A patient with proximal phalangeal osteochondroma of third toe, presenting clinically as widening of the third web space, is described. A simple extraperiosteal excision of the tumor was done which resolved his forefoot deformity. At five years followup the patient had no recurrence or symptoms.


A serial SPECT-CT study in a celiac disease patient with cerebellar ataxia and psychiatric symptoms.
Celiac disease (CD) is an immune-mediated systemic disorder triggered by gluten and related prolamins in genetically predisposed individuals. Here, we described a case of a 31-year-old Caucasian woman who exhibited cerebellar and psychiatric dysfunctions. The patient underwent single-photon emission computed tomography (SPECT-CT) before and after a gluten-free diet (GFD). There was an improvement in cerebellar perfusion accompanied by a remission of cerebellar manifestations. The maintenance of the psychiatric manifestations was related to the persistence of the hypoperfusion in the frontal lobes. The patient's psychiatric symptoms did not change after 4 months under a GFD in the hospital. To our knowledge, this is the first case that shows the relationship between improvement in cerebellar perfusion and remission of cerebellar clinical manifestations in a CD patient under a GFD.


Proteoform Analysis and Construction of Proteoform Families in Proteoform Suite.
Proteoform Suite is an interactive software program for the identification and quantification of intact proteoforms from mass spectrometry data. Proteoform Suite identifies proteoforms observed by intact-mass (MS1) analysis. In intact-mass analysis, unfragmented experimental proteoforms are compared to a database of known proteoform sequences and to one another, searching for mass differences corresponding to well-known post-translational modifications or amino acids. Intact-mass analysis enables proteoforms observed in the MS1 data without MS/MS (MS2) fragmentation to be identified. Proteoform Suite further facilitates the construction and visualization of proteoform families, which are the sets of proteoforms derived from individual genes. Bottom-up peptide identifications and top-down (MS2) proteoform identifications can be integrated into the Proteoform Suite analysis to increase the sensitivity and accuracy of the analysis. Proteoform Suite is open source and freely available at https://github.com/smith-chem-wisc/proteoform-suite .


Are family characteristics associated with attendance at family centered rounds in the PICU?.
The objective of this study was to identify if family characteristics or opinions affected participation in family centered rounds. Observational study of 431 patient encounters on daily work rounds, followed by 100 questionnaires completed by family members of patients in the unit during observation. PICU at a tertiary care, academic, free-standing children's hospital. Patients and families admitted to the PICU during the observation period. None. The most frequent family members present for rounds were mothers (40%). Race, educational level, age of the family member, age of the child, whether the admission was expected, and whether the family member was a medical professional had no association with whether the family member attended rounds. Both family members who were present and those who were not present felt being at rounds would improve the care of their child (87% vs. 100%, p = 0.57). A family's response that they preferred to attend rounds was the only factor associated with a higher likelihood of attending rounds (odds ratio 3.4, 95% confidence interval 1.1-10.8, p = 0.03). Families feel that participating in family centered rounds improves the care of their children. Those that like attending rounds are more likely to participate in family centered rounds, but family demographic characteristics were not associated with rounds attendance. Future studies are needed to identify barriers to family participation in family centered rounds.


Sexual dysfunctions in female university students and the correlation with body image: a cross-sectional study.
This study aims to correlate female sexual dysfunctions and dissatisfaction with female university students' body image. 276 female university students completed an online questionnaire, including sociodemographic data, the Female Sexual Function Index, and the Body Shape Questionnaire. 31.1% reported some degree of dissatisfaction with body image, and 39.5% had a risk of female sexual dysfunction. The majority of women with a risk of female sexual dysfunctions (69.4%) had no concerns with shape, and the majority of women without sexual problems (67.7%) had no concerns with shape. Pearson's coefficient underlined no correlation between Body Shape Questionnaire and Female Sexual Function Index domains. The body image perception did not seem to affect the sexual function of female university students in our sample. However, it is important to further investigate these variables to improve women's quality of life.


Determination of volatile components in fresh, frozen, and freeze-dried Padrón-type peppers by gas chromatography-mass spectrometry using dynamic headspace sampling and microwave desorption.
"Padrón-type" peppers are a small variety of Capsicum annuum cultivated mainly in Galicia, Spain. To compare the effects of freezing and freeze-drying on the volatile components of Padrón-type peppers, preserved samples are analyzed by means of dynamic headspace sampling on an adsorbent followed by microwave desorption into a gas chromatograph equipped with a mass spectrometric detector. Sixty-five compounds are identified, including hydrocarbons, terpenes, alcohols, phenols, ethers, aldehydes, ketones, esters, pyrroles, pyrazines, and sulfurous compounds. Fresh whole, homogenized, and freeze-dried peppers have characteristic volatile-component profiles, whereas frozen peppers have a highly variable volatile-component profile.


Characterization of cannabis strain-plant-derived extracellular vesicles as potential biomarkers.
The scientific interest in cannabis plants' beneficial properties has recently sparked certain interest in the possible functional characterization of plant-derived extracellular vesicles (PDEVs). Establishing the most appropriate and efficient isolation procedure for PDEVs remains a challenge due to vast differences in the physio-structural characteristics of different plants within the same genera and species. In this study, we employed a crude but standard isolation procedure for the extraction of apoplastic wash fluid (AWF) which is known to contain the PDEVs. This method includes a detailed stepwise process of PDEV extraction from five (5) cultivars of cannabis plants, namely: Citrus (C), Henola (HA), Bialobrezenski (BZ), Southern-Sunset (SS), and Cat-Daddy (CAD). Approximately, 150 leaves were collected from each plant strain. In order to collect PDEV pellets, apoplastic wash fluid (AWF) was extracted from plants via negative pressure permeabilization and infiltration followed by high-speed differential ultracentrifugation. Particle tracking analysis of PDEVs revealed particle size distribution in the range of 20 to 200 nm from all plant strains, while PDEV total protein concentration from HA was higher than that of SS. Although HA-PDEVs' total protein was higher than SS-PDEVs, SS-PDEVs' RNA yield was higher than that of HA-PDEVs. Our result suggests that the cannabis plant strains contain EVs, and PDEV concentration from the cannabis plant could be age or strain dependent. Overall, the results provide a guide for the selection and optimization of PDEV isolation methods for future studies.


The central insulin system and energy balance.
Insulin acts throughout the body to reduce circulating energy and to increase energy storage. Within the brain, insulin produces a net catabolic effect by reducing food intake and increasing energy expenditure; this is evidenced by the hypophagia and increased brown adipose tissue sympathetic nerve activity induced by central insulin infusion. Reducing the activity of the brain insulin system via administration of insulin antibodies, receptor antisense treatment, or receptor knockdown results in hyperphagia and increased adiposity. However, despite decades of research into the role of central insulin in food intake, many questions remain to be answered, including the underlying mechanism of action.


Management of primary congenital glaucoma by trabeculectomy in Nigeria.
To review the management of congenital glaucoma, bearing in mind the limited facilities available in developing countries. The medical records of children aged < or = 3 years with primary congenital glaucoma seen at two teaching hospitals and one private eye hospital in Enugu, Nigeria over a 5-year period were reviewed retrospectively. All the children underwent trabeculectomy without use of anti-metabolites. Fifty-seven eyes of 32 patients (5 unilateral, 26 bilateral) were reviewed. Pre-operatively, 63.2% of the eyes had a corneal diameter of 12-15 mm. After surgery, the number with corneal haziness decreased from 25 (43.8%) to 15 (26.4%) (p=0.05). The mean (SD) pre-operative intra-ocular pressure (IOP) was 28.3 (7.5) mmHg and the mean post-operative IOP was 17 (5.6) mmHg (p=0.001). When facilities for other treatment modalities are not available, primary trabeculectomy without the use of anti-metabolites can be employed to manage primary congenital glaucoma.


Genetic relationship between Streptococcus pneumoniae isolates from nasopharyngeal and cerebrospinal fluid of two infants with Pneumococcal Meningitis.
The molecular epidemiology of Streptococcus pneumoniae isolates from carriage and cerebrospinal fluid (CSF) concurrently recovered from the same individual has not yet been reported. By using pulsed-field gel electrophoresis, we demonstrated the genetic linkage among strains from CSF and nasopharynges of two children with pneumococcal meningitis.


Increased genetic gains in sheep, beef and dairy breeding programs from using female reproductive technologies combined with optimal contribution selection and genomic breeding values.
Female reproductive technologies such as multiple ovulation and embryo transfer (MOET) and juvenile in vitro embryo production and embryo transfer (JIVET) can boost rates of genetic gain but they can also increase rates of inbreeding. Inbreeding can be managed using the principles of optimal contribution selection (OCS), which maximizes genetic gain while placing a penalty on the rate of inbreeding. We evaluated the potential benefits and synergies that exist between genomic selection (GS) and reproductive technologies under OCS for sheep and cattle breeding programs. Various breeding program scenarios were simulated stochastically including: (1) a sheep breeding program for the selection of a single trait that could be measured either early or late in life; (2) a beef breeding program with an early or late trait; and (3) a dairy breeding program with a sex limited trait. OCS was applied using a range of penalties (severe to no penalty) on co-ancestry of selection candidates, with the possibility of using multiple ovulation and embryo transfer (MOET) and/or juvenile in vitro embryo production and embryo transfer (JIVET) for females. Each breeding program was simulated with and without genomic selection. All breeding programs could be penalized to result in an inbreeding rate of 1 % increase per generation. The addition of MOET to artificial insemination or natural breeding (AI/N), without the use of GS yielded an extra 25 to 60 % genetic gain. The further addition of JIVET did not yield an extra genetic gain. When GS was used, MOET and MOET + JIVET programs increased rates of genetic gain by 38 to 76 % and 51 to 81 % compared to AI/N, respectively. Large increases in genetic gain were found across species when female reproductive technologies combined with genomic selection were applied and inbreeding was managed, especially for breeding programs that focus on the selection of traits measured late in life or that are sex-limited. Optimal contribution selection was an effective tool to optimally allocate different combinations of reproductive technologies. Applying a range of penalties to co-ancestry of selection candidates allows a comprehensive exploration of the inbreeding vs. genetic gain space.


Surgical margins do not affect prognosis in high grade myxofibrosarcoma.
Myxofibrosarcoma (MFS) is one of the most common soft tissue sarcomas (STS) in elderly patients and it primarily affects the extremities. The aim of this retrospective analysis is to understand the natural history of MFS and whether adequate treatment influence prognosis. We reviewed 129 adult patients with primary, localized, FNCLCC grade 3 MFS of the extremities operated at Istituto Ortopedico Rizzoli, Bologna. Sarcoma specific survival (SS), local recurrence (LR) and distant metastasis (DM) were analyzed. Among excised MFS (119), 106 (89.9%) had R0 margins, 13 (10.1%) R1 margins. No significant correlation between margins adequacy and tumor depth, location and size was found. Estimated SS was 73.2% at 5 years and 66.3% at 10 years, with a better SS in superficial MFS (p = 0.011). Local recurred MFS had a worse SS (p = 0.049). Local recurrence-free rate was 74.3% at 5 and 10 years. Even if not significant, a better outcome in term of LR was observed in superficial MFS and R0 margins. Distant metastasis-free survival was 75.6% at 5 years and 72.9% at 10 years, with a better outcome in superficial MFS (p = 0.012). Myxofibrosarcoma remain a debated entity with specific behavior features. Myxofibrosarcoma tends to local recur due to its infiltrative grow pattern making difficult to achieve "safe margins". To date, surgical margins as classified for other STS are not predictive of LR and patients' survival. Tumor grade and depth are still the most important prognostic factors.


Spatial-Temporal Change Trend Analysis of Second-Hand House Price in Hefei Based on Spatial Network.
Spatial Markov chain can effectively explore the spatial evolution trend of housing price under the influence of lag factor. This paper uses spatial autocorrelation and spatial Markov to study 353 second-hand houses in Hefei. The results show that (1) the housing price of Hefei urban area presents a situation of "two points and one side," the high housing price is concentrated in the south and southwest of the urban area, and the price level gradually weakens from south to north, and the housing development shows a north-south differentiation. (2) There is a significant spatial autocorrelation between second-hand housing prices in Hefei. The "high-high" residential price clusters are mainly distributed in Shushan District and Binhu New Area, while the "low-low" residential price clusters are mostly in Yaohai district and its surrounding areas. The number of "low-high" agglomeration and "high-low" agglomeration is small, and the degree of change is not big. (3) Under the influence of different neighborhood environments, the housing prices in urban Area of Hefei show club convergence overall. At the same time, under the short-term influence of the policy, the housing prices of low level and middle and low level are promoted in the same neighborhood environment, while the housing prices of high level and middle and high level are negatively affected.


Is the Hitchcock Story Really True? Public Opinion on Hooded Crows in Cities as Input to Management.
In recent years, the Hooded crow (Corvus cornix) has become one of the most successful wild bird species in urban environments across Europe. Hooded crows can cause several problems in cities, including trash scattering, noise disturbance, and aggressive behavior toward humans or pets, and they can be potential vectors of pathogens. To find effective solutions, the public has to be involved in the decision-making process in urban planning management, managed by the city administration. In this study, we surveyed the attitude of people in Hungary towards crows and crow management by collecting information using an online questionnaire containing 65 questions published in 14 Facebook groups. We found that many people were familiar with corvid species and had personal experience with them. In most cases, these experiences were not negative, so the crows were not or only rarely perceived to cause problems to people, such as aggressive behavior, damage to cars or stealing something. Most respondents recognized that the presence of large numbers of hooded crows is a problem to be solved and acknowledged that they do not know how to resolve it. The majority of people expressed their interest in raising public awareness of crows but not in their management actions, which they believe should be implemented by experts. Most respondents preferred passive, harmless methods. More direct methods such as egg/chick removal from the nest, control by trapping, poisoned baits or firearms, or oral contraceptives were the least acceptable. These results express the difficulty in identifying a control method for managing hooded crow populations that is both acceptable to most people and effective at the same time. This study demonstrates the importance of involving public opinion in wildlife management and providing more information to citizens to reduce human-crow conflicts.


An interactive framework for an analysis of ECG signals.
In this study, we introduce and discuss a development of a highly interactive and user-friendly environment for an ECG signal analysis. The underlying neural architecture being a crux of this environment comes in the form of a self-organizing map. This map helps discover a structure in a set of ECG patterns and visualize a topology of the data. The role of the designer is to choose from some already visualized regions of the self-organizing map characterized by a significant level of data homogeneity and substantial difference from other regions. In the sequel, the regions are described by means of information granules-fuzzy sets that are essential in the characterization of the main relationships existing in the ECG data. The study introduces an original method of constructing membership functions that incorporates class membership as an important factor affecting changes in membership grades. The study includes a comprehensive descriptive modeling of highly dimensional ECG data.


Medical complications of psychiatric treatment.
Psychiatric medications are frequently an essential component of care for critically ill patients. Their use may lead to medical complications, however, as a result of (1) direct toxicity from psychotropic medications, (2) drug-drug interactions, or (3) intoxication or withdrawal states. These complications may be a nuisance (eg, dry mouth and nausea) or serious and life-threatening (eg, neuroleptic malignant syndrome and cardiac arrhythmias). This article addresses the most important medical complications (organized by organ systems) of psychiatric treatment.


A descriptive study of breast cancer worry.
Women with (n = 65) and without (n = 70) a family history of breast cancer reported on their thoughts and concern about the disease. Measures were taken across a 1-month interval and at a 1-year follow-up. Reported screening behaviors were also measured at baseline and the 1-year follow-up. Worry dissipated over time, suggesting that worry levels are affected by the measurement context. However, women with a family history of the disease maintained greater worry than those without such a history, suggesting that they may be chronically worried about the disease. Thinking and worrying about breast cancer were both modestly and positively related to the frequency of screening behaviors, suggesting that some kinds of worry can motivate self-protective behavior.


A new way of looking at environmental variables that may affect the age at menopause.
Environmental variables that might affect the age at menopause were analysed by means of the segmentation method AID. Marital status, occupation, smoking habits, age at last pregnancy and height were shown to be significant discriminating variables. The material and design of the study and the method of analysis are given and its results are discussed. A proposal is made concerning the nature of a 'secular trend'.


The Prevalence of Patient-Prosthesis Mismatch Can Be Reduced Using the Trifecta Aortic Prosthesis.
Some important studies have shown that patient-prosthesis mismatch is a frequent occurrence after surgical aortic valve replacement that impairs survival. The Trifecta valve (St. Jude Medical Inc, St. Paul, MN) has special architecture designed to achieve the best hemodynamic profile. The aim of this study was to determine the prevalence of mismatch when using this prosthesis. This study included 1,302 patients at 3 months postoperatively, 339 patients with a Trifecta prosthesis and 963 patients (the control group) with a Mitroflow aortic valve (Sorin Group Inc, Mitroflow Division, Vancouver, Canada). Multinomial multivariate logistic regression was calculated to estimate the association between the Trifecta prosthesis and moderate or severe patient-prosthesis mismatch. Any degree of mismatch was present in 5.9% of the Trifecta group and in 42.4% in the Mitroflow group. Moderate patient-prosthesis mismatch was present in 3.8% of the patients with a Trifecta valve and in 32.6% in the Mitroflow group. Severe mismatch was present in 2.1% of the patients with a Trifecta prosthesis and in 9.8% of the patients with a Mitroflow valve. All differences were statistically significant (p < 0.001). The odds ratio of the Trifecta prosthesis as protector against mismatch was 16.9 (95% confidence interval, 9.5 to 30.4) and 11.9 (95% confidence interval, 5.3 to 26.7) for moderate or severe mismatch, respectively. The prevalence of patient-prosthesis mismatch using the Trifecta aortic prosthesis is extraordinary low. This finding may have great clinical repercussions in patients undergoing surgical aortic valve replacement.


Etiology of Turkey "X" disease in retrospect: A case for the involvement of cyclopiazonic acid.
The etiology of the classical turkey "X" disease syndrome is reappraised based on original reports in conjunction with current information. The clinical signs described in those original reports cannot be totally explained as being typical for aflatoxicosis. The unexplained effects of the disease can be resolved by the proposed presence of the mycotoxin cyclopiazonic acid which is frequently produced byAspergillus flavus along with the aflatoxins.


The effect of oral absorption enhancers on the in vivo performance of insulin-loaded poly(ethylcyanoacrylate) nanospheres in diabetic rats.
Poly(ethylcyanoacrylate) (PECA) nanospheres have been employed as biodegradable polymeric carriers for oral (po) delivery of insulin. The main goal of this investigation was to screen various absorption enhancers, which were used to protect insulin-loaded PECA, by following their in vivo performance after oral administrations to streptozotocin-induced diabetic rats. The nanospheres were prepared by polymerization in a continuous aqueous phase at pH 2.5 and in the presence of Pluronic 68 (0.5%). This polymerization technique was able to hold 85 +/- 7.5% of insulin added, 30 min after initiation of polymerization. The drug loading was evaluated by HPLC. Insulin absorption after oral administration was evaluated by its hypoglycemic effect. The addition of protease inhibitor to insulin-loaded PECA nanospheres significantly reduced the blood glucose level after po administrations. Capric acid (0.5%) showed the maximum reduction in blood glucose level (bG max = 34.4 +/- 4.1 mg%), while cholic acid (0.5%) showed the fastest reduction in blood glucose level (Tmax, G = 6.0 +/- 0.2 h) among the tested oral absorption enhancers. Insulin associated with PECA nanospheres retained its biological activity up to 12 days in 50% of the rats in the presence of glycyrrhizic acid (1%). The per cent pharmacological availabilities (PA%) were in the order of capric acid (0.5%) > glycyrrhizic acid (1%) > deoxycholic acid (0.5%) > hydroxypropyl-beta-cyclodextrin (HPbetaCD) (1%) > cholic acid (0.5%). There was no significant difference in the PA% between capric acid (0.5%), glycyrrhizic acid (1%) and deoxycholic acid (0.5%). Although sodium cholate (0.5%) showed the lowest increase in PA% (3.1 +/- 3.11%), its Tmax G was the shortest (6 +/- 0.2 h) among the tested absorption enhancers. The reduction in blood glucose levels was maintained < 200 mg/dL in the order of glycyrrhizic acid (166 +/- 56 h) > capric acid (72 h) >; deoxycholic acid (66 +/- 6 h) > no enhancer (64 +/- 16 h) > HPbetaCI) (31 +/- 17.5) > cholic acid (22 +/- 5.7 h) > aprotenin (20 +/- 3.5 h). Therefore, capric acid and glycyrrhizic acid could be successfully used as oral absorption enhancers.


Cancer-derived exosomal circ_0038138 enhances glycolysis, growth, and metastasis of gastric adenocarcinoma via the miR-198/EZH2 axis.
This study aims to decipher the impact and downstream mechanisms of the bioinformatically identified circ_0038138 delivered by cancer-derived exosomes in gastric adenocarcinoma (GAC). Expression of circ_0038138 in clinical GAC tissues and exosomes (Exos) from clinical plasma samples (plasma-Exos) was predicted by bioinformatics analysis and validated by RT-qPCR. The binding affinity between circ_0038138, miR-198 and EZH2 was identified using luciferase activity, RIP, and RNA pull-down assays. GAC cells (AGS) were co-cultured with Exos isolated from GAC cell supernatant (GC9811-P). After co-culture, the behaviors of GAC cells including proliferation and glycolysis were assessed to identify the biological effect of exosomal circ_0038138. Also, in vivo effects of exosomal circ_0038138 on the tumorigenesis and lung metastasis of GAC cells were evaluated by developing nude mouse xenograft and metastatic models. circ_0038138 upregulation was detected in GAC tissues and plasma-Exos. Exos delivered circ_0038138 to GAC cells and potentiated the proliferative, migratory, invasive, and glycolytic potentials of GAC cells. Mechanistically, circ_0038138 competitively bound to miR-198, which in turn targeted EZH2 by binding to its 3'-UTR. Silencing of EZH2 promoted CXXC4 expression and inhibited Wnt/β-catenin pathway activation, thus repressing the malignancy and glycolysis of GAC cells. In vivo assay confirmed that exosomal circ_0038138 induced tumorigenesis and lung metastasis by regulating the miR-198/EZH2 axis. Collectively, our work suggests that the Exo-mediated transfer of circ_0038138 potentially facilitates the glycolysis, growth and metastasis of GAC cells via miR-198/EZH2 axis, which offers a potential prognostic marker and a therapeutic target for GAC.


The association between SYNTAX score and long-term outcomes in patients with unstable angina pectoris: a single-centre retrospective study.
The SYNTAX score affects clinical outcomes in early studies. However, the prognostic value of the SYNTAX Score for long-term outcomes and differences by SYNTAX score risk stratification in long-term prognosis between medical therapy and percutaneous coronary intervention (PCI) in patients with unstable angina pectoris (UAP) are not well known in the era of new generation drug-eluting stents and medication. In this single-centre retrospective study, a total of 2364 patients with UAP from January 2014 to June 2017 at Beijing Friendship Hospital were enrolled. The primary endpoint was a composite of major adverse cardiovascular events (MACEs), including all-cause death, cardiac death, nonfatal myocardial infarction and stroke at least 2 years after discharge. In this study, 1695 patients had low SYNTAX scores ([Formula: see text]), 432 patients had medium SYNTAX scores (23-32), 237 patients had high SYNTAX scores (≥ 33), 1018 received medical therapy, and 1346 patients underwent PCI. Long-term MACEs occurred in 95 patients during the 3.38 ± 0.99-year follow-up. Compared to the medical therapy group, the PCI group showed lower MACEs and cardiac death in patients with high SYNTAX scores (7.4% vs. 16.7%, P = 0.048; 3.7% vs. 14.6%, P = 0.004) but no reduction in patients with low and medium SYNTAX scores. Cox multivariate regression analysis showed that advanced age, diabetes mellitus, left ventricular ejection fraction (LVEF), hs-CRP and high SYNTAX score were independent predictors for MACEs in the medical therapy group (P < 0.05), whereas chronic kidney disease (CKD) and LVEF were predictors of MACEs in the PCI group. Compared to medical therapy, PCI could only significantly reduce long-term MACEs and cardiac death for patients with high SYNTAX scores but not for patients with low and medium SYNTAX scores. A high SYNTAX score could predict long-term MACEs for UAP patients in the medical therapy group but not in the PCI group.


Haem synthesis in sideroblastic anaemia.
Measurement of delta-aminolaevulinic acid (ALA) synthetase has been performed in bone marrow of 24 normal subjects and 20 patients with sideroblastic anaemia (five congenital, two pyridoxine-responsive, 12 primary acquired and one secondary form due to isoniazid therapy). ALA-synthetase activity without added pyridoxal-phosphate in vitro was decreased in four of the congenital cases, and in all 12 primary acquired cases. In two of these 16 cases with low ALA-synthetase activity and in a case with sideroblastic change secondary to isoniazid therapy, the enzyme activity was corrected to normal by pyridoxal-phosphate in vitro. ALA-synthetase activity was normal in both cases of pyridoxine-responsive anaemia while they were receiving pyridoxine therapy but in one of these cases, studied at repeated intervals after withdrawal of pyridoxine therapy, the bone marrow ALA-synthetase activity fell to very low levels unless pyridoxal-phosphate was added in vitro. In this case the Km of the enzyme for pyridoxal-phosphate was substantially greater than that of a control sample. Haem synthetase activity was reduced in the bone marrow of two of six cases of primary acquired sideroblastic anaemia and in one congenital case tested but was normal in the pyridoxine-responsive patient studied on pyridoxine therapy. Nevertheless, this patient showed reduced incorporation of 14C-labelled delta-aminolaevulinic acid into bone marrow haem. It is concluded that both congenital and primary acquired sideroblastic anaemia are a heterogenous group of disorders, with different defects in haem synthesis, reduced activity of delta-aminolaevulinic acid synthetase being common but not invariable in both types. Pyridoxine-responsive (congenital) sideroblastic anaemia may be due to the presence of an abnormal ALA-synthetase apoenzyme which requires excessive amounts of the coenzyme, pyridoxal-phosphate, to achieve normal activity; other abnormalities of haem synthesis may also be present.


Differences of liver membrane antibody frequency in alcoholic liver disease. Detection of IgG and IgA classes using radioimmunoassay.
The presence of liver membrane antibody in IgG and IgA was investigated by radioimmunoassay using isolated rabbit hepatocytes as target cells. This technique was more sensitive than the immunofluorescent method. IgG liver membrane antibodies were positive in 24% of patients with alcoholic liver disease. IgA liver membrane antibodies were detected in 58% of patients with alcoholic liver disease, whereas they were detected only in 21% of those with nonalcoholic liver disease, except for cases of autoimmune chronic active hepatitis. In alcoholic liver disease, IgA liver membrane antibodies were detected at a high frequency in a group of patients with alcoholic hepatitis and active cirrhosis (94%) as compared with that of fatty liver, hepatic fibrosis, and inactive cirrhosis (42%). These results suggest that alcoholic liver disease is characterized in part by a humoral immune response of IgA liver membrane antibodies.


Hierarchy structure and fracture mechanisms of the wild wolf tusk's enamel.
The self-growth and self-strengthening of natural biomaterials provided us strategies for new materials design. In this paper, the microstructure and fracture mechanisms of the wild wolf tusk's enamel were studied. The enamel included four-order hierarchies, which were the hydroxyapatite (HAP) fiber (first-order, nano-scale, ploy-crystals), enamel rod (second-order, micro-scale, rope-like), enamel type (third-order, meso-scale, mat-like) and the enamel patterns (forth-order, macro-scale), respectively. It was interesting to find that the numerous nano-grains distributed disorderly in a single HAP fiber. The thousands HAP fibers bundled together to form the rope-like enamel rod. The protein ligaments were discovered between adjacent enamel rods. The out enamel, inner enamel and P&D-zones showed a criss-cross type and ran through whole enamel pattern in three-dimensional space. The enamel of the wild wolf tusk exhibited an excellent fracture toughness based on the nanoindentation tests. The fracture morphology in transverse direction indicated that the cracks preferred to propagate along the weak interface (protein or interrod) and cut those enamel rods perpendicular to the propagation direction. However, the cracks extended obviously forward along the step-like paths from the outmost surface of the enamel to the enamel-dentin junction in the longitudinal direction. It was considered that the protein ligament was the main reason for the good fracture toughness of the bulk enamel. Our studies reveal that the design strategies of the natural material can be applied to guide the development of high-performance artificial materials.


Necrosis score as a prognostic factor in stage I-III colorectal cancer: a retrospective multicenter study.
Tumor necrosis results from failure to meet the requirement for rapid proliferation of tumor, related to unfavorable prognosis in colorectal cancer (CRC). However, previous studies used traditional microscopes to evaluate necrosis on slides, lacking a simultaneous phase and panoramic view for assessment. Therefore, we proposed a whole-slide images (WSIs)-based method to develop a necrosis score and validated its prognostic value in multicenter cohorts. Necrosis score was defined as the proportion of necrosis in the tumor area, semi-quantitatively classified into 3-level score groups by the cut-off of 10% and 30% on HE-stained WSIs. 768 patients from two centers were enrolled in this study, divided into a discovery (N = 445) and a validation (N = 323) cohort. The prognostic value of necrosis score was evaluated by Kaplan-Meier curves and the Cox model. Necrosis score was associated with overall survival, with hazard ratio for high vs. low in discovery and validation cohorts being 2.62 (95% confidence interval 1.59-4.32) and 2.51 (1.39-4.52), respectively. The 3-year disease free survival rates of necrosis-low, middle, and high were 83.6%, 80.2%, and 59.8% in discovery cohort, and 86.5%, 84.2%, and 66.5% in validation cohort. In necrosis middle plus high subgroup, there was a trend but no significant difference in overall survival between surgery alone and adjuvant chemotherapy group in stage II CRC (P = .075). As a stable prognostic factor, high-level necrosis evaluated by the proposed method on WSIs was associated with unfavorable outcomes. Additionally, adjuvant chemotherapy provide survival benefits for patients with high necrosis in stage II CRC.


The role of the upper sample size limit in two-stage bioequivalence designs.
Two-stage designs (TSDs) are currently recommended by the regulatory authorities for bioequivalence (BE) assessment. The TSDs presented until now rely on an assumed geometric mean ratio (GMR) value of the BE metric in stage I in order to avoid inflation of type I error. In contrast, this work proposes a more realistic TSD design where sample re-estimation relies not only on the variability of stage I, but also on the observed GMR. In these cases, an upper sample size limit (UL) is introduced in order to prevent inflation of type I error. The aim of this study is to unveil the impact of UL on two TSD bioequivalence approaches which are based entirely on the interim results. Monte Carlo simulations were used to investigate several different scenarios of UL levels, within-subject variability, different starting number of subjects, and GMR. The use of UL leads to no inflation of type I error. As UL values increase, the % probability of declaring BE becomes higher. The starting sample size and the variability of the study affect type I error. Increased UL levels result in higher total sample sizes of the TSD which are more pronounced for highly variable drugs.


Bringing breast cancer education to deaf women.
The Deaf community has not been adequately served by mainstream public health interventions. A breast cancer education program adapted for the needs of the Deaf community was evaluated by 123 deaf and hard-of-hearing women using pre- and post-surveys and focus groups. Among the findings were the difficulty of recruiting Deaf community members to education and research programs; low adherence to breast cancer screening guidelines; insufficient breast-health knowledge; endorsement of the program; and suggestions for strengthening it. Deaf women will benefit from breast cancer education programs that specifically address their language, culture, and preferred learning styles.


Immunohistochemical demonstration of prostaglandins in various tissues of the rat.
To demonstrate the tissue localization of prostaglandin (PG) E2, PGF2 alpha and 6-keto-PGF1 alpha (a stable metabolite of PGI2) various tissues, including decalcified periodontal tissue of 7-week-old male Wistar strain rats, were immunohistochemically examined using a streptavidin-biotin complex method. Besides tissue macrophages and endothelial cells in various tissues, hepatocytes, renal tubular cells, and parietal and chief cells in the gastric mucosa showed a positive reaction for the various PGs examined. PGs were demonstrated in the cytoplasm or in association with the cell membrane. We generally observed no difference between the localization patterns of PGE2-, PGF2 alpha-, and 6-keto-PGF1 alpha-positive cells in these tissues. However, in the periodontal ligament and alveolar bone, 6-keto-PGF1 alpha was localized in the cytoplasm of osteocytes, osteoblasts, cementocytes, and cementoblasts, while no reaction for PGE2 or PGF2 alpha was revealed in these cells. We demonstrated the immunohistochemical localization of PGs in various rat tissues including decalcified periodontal tissue and discuss the important roles of PGs in the modulation of their normal functions in these tissues.


Day-to-Day Variability in Subjective Age and Ageist Attitudes and Their Association With Depressive Symptoms.
Evidence of daily fluctuations in subjective age and their association with older adults' well-being was recently obtained. Yet, neither the simultaneous tracking of two daily views on aging (i.e., daily subjective age and daily ageist attitudes) nor their interactive effect on mental health (i.e., depressive symptoms) has been explored. We hypothesized that (a) at days on which older adults feel older or report high ageist attitudes they would report higher depressive symptoms, (b) combined older subjective age and high ageist attitudes will be associated with the highest daily depressive symptoms. Community-dwelling older adults (N = 134, mean age = 69.66) completed measures of subjective age, ageist attitudes, and depressive symptoms for 10 consecutive days. Daily older subjective age and higher ageist attitudes were related to higher depressive symptoms, but there was no combined effect of both on depressive symptoms. There was a significant three-way interaction between subjective age, ageist attitudes, and chronological age, demonstrating the interactive effect of subjective age and ageist attitudes on depressive symptoms only among the old-old respondents. Time-lagged analyses further showed that ageist attitudes during previous days predicted feeling older and more depressed on following days, but not vice versa. Results suggest that old-old individuals are more susceptible to a combination of negative views on aging on daily basis. Findings further support a daily assimilation process, whereby previous-day stereotypes are assimilated and manifested into one's identity and mental health, so that one feels older and more depressed on subsequent days.


UREKA: umbilical ring easy kannula access.
Standard techniques of laparoscopic access involve creating an abdominal wall defect and can result in complications. We describe the umbilical ring easy kannula access (UREKA) technique, evaluating safety and a decrease in complications related to port placement. UREKA is performed via a supra- or infraumbilical incision followed by circumferential dissection of the umbilical stalk. The umbilical skin is dissected free from the fascia, exposing the umbilical ring. Pneumoperitoneum is established either before or after placement of a dilating port through the open ring. We reviewed all laparoscopic procedures performed by one pediatric surgeon over 14 months using UREKA. Ninety-four patients underwent laparoscopic surgery with initial port placement via UREKA. Appendectomy (n=57) was the most common procedure, followed by fundoplication (15) and cholecystectomy (10). No intestinal, solid organ, vascular, or bladder injuries related to port placement occurred. The only postoperative complication was a superficial wound infection in a 135-kg patient following cholecystectomy, treated successfully with oral antibiotics alone. The umbilical ring persists to some degree in all pediatric patients and provides a safe portal of entry for laparoscopic surgery. UREKA has few complications and is a straightforward, reproducible technique for gaining initial laparoscopic access.


Culture, law and psychiatry in Hawaii.
Hawaii is a set of islands with a broad mix of cultures. The law is American, but it was predominantly derived from English Common Law. Thus, the law is derived from a very distant culture. Theoretically, this 'transplanted' law could cause difficulties for people from Pacific cultures. Indeed, it appears to create difficulties for individuals with mental illness. A substantial number of these people are arrested for disobedience of a law that is not sensible for their culture. This problem is an example of the need for cultural knowledge when a psychiatrist conducts an examination for the legal system. An individual may appear legally guilty or medically insane when acting within cultural norms.


Bax, Bak and mitochondrial oxidants are involved in hypoxia-reoxygenation-induced apoptosis in human placenta.
Although apoptosis is prominent in placental cells in pregnancy complications such as preeclampsia, the cause is unknown. We surmised that hypoxia-reoxygenation (HR) is the mechanism and hypothesized that mitochondrial oxidants and Bcl-2 proteins cause HR-induced placental apoptosis. Our goal was studying expression of five Bcl-2 proteins--Bcl-2, Bcl-xL, Bax, Bak, Bad--and testing effects of diazoxide and cyclosporine A on oxidative stress and apoptosis in villous tissues subjected to HR. Term human placentas were obtained from normal pregnancies following elective caesarean deliveries. Villous tissues were subjected to "repetitive HR" (one hour at 2% O(2) then one hour at 8% O(2), alternatively, for a total of 6h) or "prolonged HR" (3h at 2% O(2) then 3h of 8% O(2)). Samples maintained at 2% and 8% O(2) served as hypoxic and normoxic controls, respectively. Prolonged HR caused the most severe villous apoptotic changes, increased the expression of Bax and Bak mRNA and protein and reduced the expression of Bcl-2 mRNA. Pre-administration of diazoxide and cyclosporine A reduced TUNEL-positive nuclei and levels of nitrotyrosine and 4-hydroxy-2-nonenol after prolonged HR. Thus, duration of hypoxia and reoxygenation is important in determining severity of HR-induced apoptosis in placenta. These apoptotic changes are closely associated with Bax and Bak effects and oxidative stress in mitochondria.


Immunoblotting conditions for human hemoglobin chains.
Immunoblotting to analyze low-molecular-weight proteins like calmodulin and metallothioneins is challenging and requires modifications for reproducible detection. Human globin chains are 17-kDa proteins and are not detectable by conventional immunoblotting using nitrocellulose membranes. Here we describe an immunoblotting method using nitrocellulose membranes that allows quantitative analyses of globin chains. Although previous studies have demonstrated that the fixation of blotted membranes with glutaraldehyde improves immunodetection of low-molecular-weight proteins, we found that the detection sensitivity for human globins is increased markedly by fixation with paraformaldehyde, but not glutaraldehyde. This immunoblotting procedure facilitates studies of posttranscriptional mechanisms for globin gene expression.


Low sodium intake enhances sensitivity of 11-deoxycortisol and deoxycorticosterone to ACTH in ACTH-suppressed normal subjects.
Continued administration of ACTH to patients with hypopituitarism produced normal increases in steroids dependent on microsomal cytochrome P450(21) and P450(17 alpha) but reduced responses of steroids dependent on mitochondrial cytochrome P450(11 beta-18). To explore possible mechanisms and to determine whether this dissociation occurs with short-term ACTH suppression, we have examined the steroid responses to ACTH after 1 h in 12 normal subjects after equilibration on sodium intakes of 124 mmol/d [normal sodium diet (NSD)], 22 mmol/d [low sodium diet (LSD)], and 240 mmol/d [high sodium diet (HSD)] before and during continued ACTH suppression with dexamethasone (DEX). Two distinct patterns of steroid responses were observed. Deoxycorticosterone (DOC) responses were initially reduced during LSD-DEX but eventually returned to the NSD-control (NSD-CONT) values; in contrast 18-hydroxydeoxycorticosterone and corticosterone remained suppressed. 11-Deoxycortisol and 21-deoxycortisol showed patterns similar to DOC, with a return to normal ACTH responses on LSD-DEX. Basal cortisol levels were reduced and the ACTH response was unchanged by LSD. HSD-DEX reduced basal levels of all steroids as well as their ACTH responses. LSD and/or increased activity of the renin-angiotensin system have a significant impact on 17 alpha- and 21-hydroxylation functions in the zona fasciculata to maintain a normal ACTH response of microsomally dependent steroids under these conditions. In contrast, on HSD-DEX with the renin-angiotensin system suppressed, there is generalized impairment of steroid responses to ACTH.


Exposure to ebselen changes glutamate uptake and release by rat brain synaptosomes.
We investigated effects of Ebselen, diphenyl diselenide (PhSe)2 and diphenyl ditelluride (PhTe)2 on [3H]glutamate uptake and release by brain synaptosomes. Ebselen after acute exposure inhibited K+-stimulated [3H]glutamate release by brain synaptosomes. (PhSe)2 and (PhTe)2 did not change [3H]glutamate release by brain synaptosomes. Ebselen, (PhSe)2 and (PhTe)2 had no significantly effects on [3H]glutamate uptake after acute exposure. In vitro, Ebselen (100 microM) inhibited [3H]glutamate release and uptake. (PhSe)2 had no significant effect, while (PhTe)2 (100 microM) inhibited [3H]glutamate uptake by brain synaptosomes. In vitro, (PhSe)2, (PhTe)2 and Ebselen caused a significant inhibition of [3H]glutamate uptake by brain synaptic vesicles in vitro. The results demonstrated that organochalcogenides have a rather complex effect on glutamate homeostasis depending on the compound and the schedule of exposition. We propose that the neuroprotective action of Ebselen can be related, in addition to its glutathione peroxidase-like and antilipoperoxidative activity, to a direct interaction with the glutamatergic system by reducing K+-evoked glutamate release.


Stressed cybrids model demyelinated axons in multiple sclerosis.
Multiple sclerosis is likely caused by a complex interaction of multiple genes and environmental factors. The contribution of mitochondrial DNA genetic backgrounds has been frequently reported. To evaluate the effect of mitochondrial DNA haplogroups in the same genetic and environmental circumstances, we have built human transmitochondrial cell lines and simulated the effect of axon demyelination, one of the hallmarks of multiple sclerosis pathology, by altering the ionic gradients through the plasmalemma and increasing ATP consumption. In this model, mitochondrial biogenesis is observed. This process is larger in Uk cybrids, which mirrors their lower oxidative phosphorylation capacity in basal conditions. This model replicates a process occurring in both patients suffering from multiple sclerosis and several animal models of axon demyelination. Therefore, it can be used to analyze the contribution of various mitochondrial DNA genotypes to multiple sclerosis. In this sense, a longer or stronger energy stress, such as that associated with demyelinated axons in multiple sclerosis, could make Uk individuals more susceptible to this pathology. Finally, pharmacologic compounds targeted to mitochondrial biogenesis could be a potential therapy for multiple sclerosis.