Scanning transmission electron microscopy through-focal tilt-series on biological specimens.
Since scanning transmission electron microscopy can produce high signal-to-noise ratio bright-field images of thick (≥500 nm) specimens, this tool is emerging as the method of choice to study thick biological samples via tomographic approaches. However, in a convergent-beam configuration, the depth of field is limited because only a thin portion of the specimen (from a few nanometres to tens of nanometres depending on the convergence angle) can be imaged in focus. A method known as through-focal imaging enables recovery of the full depth of information by combining images acquired at different levels of focus. In this work, we compare tomographic reconstruction with the through-focal tilt-series approach (a multifocal series of images per tilt angle) with reconstruction with the classic tilt-series acquisition scheme (one single-focus image per tilt angle). We visualised the base of the flagellum in the protist Trypanosoma brucei via an acquisition and image-processing method tailored to obtain quantitative and qualitative descriptors of reconstruction volumes. Reconstructions using through-focal imaging contained more contrast and more details for thick (≥500 nm) biological samples.


Molecular evidence for deep evolutionary roots of bilaterality in animal development.
Nearly all metazoans show signs of bilaterality, yet it is believed the bilaterians arose from radially symmetric forms hundreds of millions of years ago. Cnidarians (corals, sea anemones, and "jellyfish") diverged from other animals before the radiation of the Bilateria. They are diploblastic and are often characterized as being radially symmetrical around their longitudinal (oral-aboral) axis. We have studied the deployment of orthologs of a number of family members of developmental regulatory genes that are expressed asymmetrically during bilaterian embryogenesis from the sea anemone, Nematostella vectensis. The secreted TGF-beta genes Nv-dpp, Nv-BMP5-8, six TGF-beta antagonists (NvChordin, NvNoggin1, NvNoggin2, NvGremlin, NvFollistatin, and NvFollistatin-like), the homeodomain proteins NvGoosecoid (NvGsc) and NvGbx, and the secreted guidance factor, NvNetrin, were studied. NvDpp, NvChordin, NvNoggin1, NvGsc, and NvNetrin are expressed asymmetrically along the axis perpendicular to the oral-aboral axis, the directive axis. Furthermore, NvGbx, and NvChordin are expressed in restricted domains on the left and right sides of the body, suggesting that the directive axis is homologous with the bilaterian dorsal-ventral axis. The asymmetric expression of NvNoggin1 and NvGsc appear to be maintained by the canonical Wnt signaling pathway. The asymmetric expression of NvNoggin1, NvNetrin, and Hox orthologs NvAnthox7, NvAnthox8, NvAnthox1a, and NvAnthox6, in conjunction with the observation that NvNoggin1 is able to induce a secondary axis in Xenopus embryos argues that N. vectensis could possess antecedents of the organization of the bilaterian central nervous system.


Cognitive impairment and dementia in Parkinson's disease: practical issues and management.
Cognitive impairment and dementia pose particular challenges in the management of patients with Parkinson's disease (PD). Decision-making capacity can render patients vulnerable in a way that requires careful ethical considerations by clinicians with respect to medical decision making, research participation, and public safety. Clinicians should discuss how future decisions will be made as early in the disease course as possible. Because of cognitive, visual, and motor impairments, PD may be associated with unsafe driving, leading to early driving cessation in many. DBS of the STN and, to a lesser degree, globus pallidus interna (GPi) has consistently been associated with decreased verbal fluency, but significant global cognitive decline is usually not observed in patients who undergo rigorous selection. There are some observations suggesting lesser cognitive decline in GPi DBS than STN DBS, but further research is required. Management of PD dementia (PDD) patients involves both pharmacological and nonpharmacological measures. Patients with PDD should be offered treatment with a cholinesterase inhibitor taking into account expected benefits and potential risks. Treatment with neuroleptics may be necessary to treat psychosis; classical neuroleptics, as well as risperidone and olanzapine, should be avoided. Quetiapine might be considered first-line treatment because it does not need special monitoring, although the strongest evidence for efficacy exists for clozapine. Evidence from randomized, controlled studies in the PDD population is lacking; selective serotonin reuptake inhibitors or serotonin-norepinephrine reuptake inhibitors may be used to treat depressive features. Clonazepam or melatonin may be useful in the treatment of rapid eye movement behavior disorder.


The hemolysin of Escherichia coli.
Many strains of E. coli elaborate a hemolysin which is responsible for the zone of beta-hemolysis surrounding bacterial colonies on blood agar. The significance of this cytolysin as a determinant of bacterial pathogenicity has been established in animal models with the use of genetically engineered, isogenic bacterial strains. An analogous role in human infections has been inferred from the high association of hemolysin production with disease. Studies at a molecular genetical level have defined 4 genes that are required for the synthesis, post-translational modification and secretion of the hemolysin. The structural gene hlyA encodes for a 107-110,000 polypeptide, which must be modified in an unknown manner to its active form by the product of the neighboring hlyC gene. Genes hlyB and hlyD encode for proteins that export the molecule to the extracellular medium. The signal for secretion is contained in the C-terminal portion of the toxin molecule. The secreted hemolysin attacks plasma membranes of target mammalian cells by inserting as a monomer into the bilayer and generating hydrophilic transmembrane pores of approximately 2 nm effective diameter. The pores display a marked selectivity for cations over anions and pore-opening is dependent on the presence of a correct transmembrane potential. Binding to a membrane target does not require the presence of a specific receptor, and pores may be generated in planar lipid membranes consisting solely of phosphatidylcholine. Pore formation in nucleated cells can trigger secondary reactions such as stimulation of arachidonate metabolism with release of lipid mediators, probably initiated by passive influx of extracellular Ca2+.(ABSTRACT TRUNCATED AT 250 WORDS)


Paroxysmal atrial fibrillation. A common but neglected entity.
Paroxysmal atrial fibrillation is an extremely common form of atrial fibrillation seen by most physicians. Despite making up more than 40% of the cases of atrial fibrillation, its management continues to be neglected in the medical literature. Many of the recent large-scale prospective trials have excluded these patients. Limited data suggest that paroxysmal atrial fibrillation should be treated in a similar fashion as chronic atrial fibrillation.


Effects of noise and spectral resolution on vowel and consonant recognition: acoustic and electric hearing.
Current multichannel cochlear implant devices provide high levels of speech performance in quiet. However, performance deteriorates rapidly with increasing levels of background noise. The goal of this study was to investigate whether the noise susceptibility of cochlear implant users is primarily due to the loss of fine spectral information. Recognition of vowels and consonants was measured as a function of signal-to-noise ratio in four normal-hearing listeners in conditions simulating cochlear implants with both CIS and SPEAK-like strategies. Six conditions were evaluated: 3-, 4-, 8-, and 16-band processors (CIS-like), a 6/20 band processor (SPEAK-like), and unprocessed speech. Recognition scores for vowels and consonants decreased as the S/N level worsened in all conditions, as expected. Phoneme recognition threshold (PRT) was defined as the S/N at which the recognition score fell to 50% of its level in quiet. The unprocessed speech had the best PRT, which worsened as the number of bands decreased. Recognition of vowels and consonants was further measured in three Nucleus-22 cochlear implant users using either their normal SPEAK speech processor or a custom processor with a four-channel CIS strategy. The best cochlear implant user showed similar performance with the CIS strategy in quiet and in noise to that of normal-hearing listeners when listening to correspondingly spectrally degraded speech. These findings suggest that the noise susceptibility of cochlear implant users is at least partly due to the loss of spectral resolution. Efforts to improve the effective number of spectral information channels should improve implant performance in noise.


Enhanced LTP of primary afferent neurotransmission in AMPA receptor GluR2-deficient mice.
Ca(2+)-permeable-AMPA receptors (AMPARs) are expressed in the superficial dorsal horn (SDH, laminae I/II) of the spinal cord, the area involved in transmission and modulation of sensory information, including nociception. A possible role of Ca(2+)-permeable-AMPARs in synaptic strengthening has been suggested in postnatal DH cultures, but their role in the long-lasting activity-dependent synaptic plasticity of primary afferent neurotransmission in the adult mouse SDH has not been investigated. In the present study the role of Ca(2+)-permeable-AMPARs in the regulation of long-lasting synaptic plasticity, specifically long-term potentiation (LTP) and long-term depression (LTD) in the SDH, was investigated using mice deficient in AMPAR GluR2 subunit. We show here that the GluR2 mutants exhibited no changes in passive membrane properties, but a significant increase in rectification of excitatory postsynaptic currents, the finding suggesting increased expression of Ca(2+)-permeable-AMPARs. In the absence of GluR2, high-frequency stimulation (HFS) of small-diameter primary afferent fibers induced LTP that is enhanced and non-saturating in the SDH at both primary afferent Adelta- and/or C-fibers monosynaptic and polysynaptic pathways, whereas neuronal excitability and paired-pulse depression were normal. The LTP could be induced in the presence of the NMDA receptor antagonist d-AP5, and L-type Ca(2+) channel blockers, suggesting that Ca(2+)-permeable-AMPARs are sufficient to induce LTP in the SDH neurons of adult mouse spinal cord. In contrast, the induction of HFS-LTD is reduced in the SDH of GluR2 mutants. These results suggest an important role for AMPAR GluR2 subunit in regulating synaptic plasticity with potential relevance for long-lasting hypersensitivity in pathological states.


Antibiotics and antibiotic-resistant bacteria in waters associated with a hospital in Ujjain, India.
Concerns have been raised about the public health implications of the presence of antibiotic residues in the aquatic environment and their effect on the development of bacterial resistance. While there is information on antibiotic residue levels in hospital effluent from some other countries, information on antibiotic residue levels in effluent from Indian hospitals is not available. Also, concurrent studies on antibiotic prescription quantity in a hospital and antibiotic residue levels and resistant bacteria in the effluent of the same hospital are few. Therefore, we quantified antibiotic residues in waters associated with a hospital in India and assessed their association, if any, with quantities of antibiotic prescribed in the hospital and the susceptibility of Escherichia coli found in the hospital effluent. This cross-sectional study was conducted in a teaching hospital outside the city of Ujjain in India. Seven antibiotics--amoxicillin, ceftriaxone, amikacin, ofloxacin, ciprofloxacin, norfloxacin and levofloxacin--were selected. Prescribed quantities were obtained from hospital records. The samples of the hospital associated water were analysed for the above mentioned antibiotics using well developed and validated liquid chromatography/tandem mass spectrometry technique after selectively isolating the analytes from the matrix using solid phase extraction. Escherichia coli isolates from these waters were tested for antibiotic susceptibility, by standard Kirby Bauer disc diffusion method using Clinical and Laboratory Standard Institute breakpoints. Ciprofloxacin was the highest prescribed antibiotic in the hospital and its residue levels in the hospital wastewater were also the highest. In samples of the municipal water supply and the groundwater, no antibiotics were detected. There was a positive correlation between the quantity of antibiotics prescribed in the hospital and antibiotic residue levels in the hospital wastewater. Wastewater samples collected in the afternoon contained both a higher number and higher levels of antibiotics compared to samples collected in the morning hours. No amikacin was found in the wastewater, but E.coli isolates from all wastewater samples were resistant to amikacin. Although ciprofloxacin was the most prevalent antibiotic detected in the wastewater, E.coli was not resistant to it. Antibiotics are entering the aquatic environment of countries like India through hospital effluent. In-depth studies are needed to establish the correlation, if any, between the quantities of antibiotics prescribed in hospitals and the levels of antibiotic residues found in hospital effluent. Further, the effect of this on the development of bacterial resistance in the environment and its subsequent public health impact need thorough assessment.


Evaluation of guided bone regeneration in rabbit femur using collagen membranes.
The aim of the study was to evaluate the mechanical performance and the structure of neoformed bone around hydroxyapatite-coated titanium fixtures according to guided bone regeneration techniques. Ten hydroxyapatite-coated titanium fixtures were inserted in the femurs of five rabbits, in which a cortical defect was created and after the insertion of the fixture, covered with a resorbable membrane obtained from bovine Achilles tendon collagen Type I (A implant). In the same femur, a second fixture was inserted in similar cavities without application of the membrane (B implant). After 60 days, the animals were sacrificed, and block sections of the femoral bone containing the implants were embedded in polymethylmetacrylate and subjected to tensile shear-stress at break testing. After the detachment of the implants from the bone, their surfaces were examined with a scanning electron microscope. Tensile shear-stress values for A and B implant specimens were comparable to some extent, but the former had a lower performance. In this regard, scanning electron microscope observations showed that the neoformed cortical bone present cervically around implant A was much thicker than around implant B.


Genetic heterogeneity in ADHD: DAT1 gene only affects probands without CD.
Previous studies have found heterogeneous association between DAT1-3'-UTR-VNTR and attention deficit hyperactivity disorder (ADHD). Various proportions of conduct disorder (CD) comorbidity in their ADHD samples may partially explain the observational discrepancies. Evidence for this comes from family and twin studies which found ADHD probands with CD (ADHD + CD) are genetically different from those without CD (ADHD - CD). Genotypes of 20 DAT1 markers were analyzed in 576 trios, consisting of 141 ADHD + CD and 435 ADHD - CD. In addition to the classical TDT test, a specific genetic heterogeneity test was performed to identify variants that have different transmission patterns in the two phenotypic subgroups. After multiple-test correction, rs40184 and rs2652511 were significant in TDT tests. Further heterogeneity test found the two SNPs had a significant transmission pattern difference between ADHD + CD and ADHD - CD children, indicating that DAT1 has a significantly greater genetic influence on ADHD without CD. Although the result needs further replications, it does highlight the importance of selecting genetically homogeneous samples for molecular genetic analyses of ADHD.


Theoretical Study of Microgel Functional Groups' Mobility.
Polymer microgels, micrometer-sized cross-linked polymer particles, are considered to be a promising type of advanced materials for a wide range of applications. To enhance the microgels' applicability, it is essential to incorporate various functional groups into a microparticle polymer network. Yet, the availability of functional groups for the interaction with surroundings depends strongly on the properties of the polymer network and has a great impact on further effective usage. In this theoretical study, we address this question and, with the help of coarse-grained molecular dynamics computer simulations, assess the segmental mobility and accessibility of functional groups bound to polymer network depending on microgel architecture and solvent quality. Additionally, we evaluate the minimum number of functional groups needed to facilitate the hopping mechanism between the functional groups (i.e., charge transfer). As an example of practical implementation of the obtained results, we estimate the optimal network topology for redox-active microgels to provide the maximum charge capacity for the dispersion electrolyte in redox-flow batteries.


The p53 protein induces stable miRNAs that have the potential to modify subsequent p53 responses.
The p53 tumour suppressor is a transcription factor that can increase the expression of mRNAs and microRNAs (miRNAs). HT29-tsp53 cells expressing a temperature sensitive variant of p53 have provided a useful model to rapidly and reversibly control p53 activity. In this model, the majority of p53-responsive mRNAs were upregulated rapidly but they were short-lived leading to rapid decay of the p53 response at the restrictive temperature. Here we used oligonucleotide microarrays and reverse transcriptase PCR to show that p53-induced miRNAs exhibited a distinct temporal pattern of expression. Whereas p53-induced miRNAs like miR-143-3p, miR-145-5p, miR-34a-5p and miR-139-5p increased as fast as mRNAs, they were extremely stable persisting long after p53 induced mRNAs and even their corresponding primary miRNAs had decayed to baseline levels. Three p53-induced mRNAs (MDM2, BTG2 and CDKN1A) are experimentally verified targets of one or more of these specific miRNAs so we hypothesized that the sustained expression of p53-induced miRNAs could be explained by a post-transcriptional feedback loop. Activation of consecutive p53 responses separated by a period of recovery led to the selective attenuation of a subset of p53 regulated mRNAs corresponding to those targeted by one or more of the p53-responsive miRNAs. Our results indicate that the long term expression of p53 responsive miRNAs leads to an excess of miRNAs during the second response and this likely prevents the induction of MDM2, BTG2 and CDKN1A mRNA and/or protein. These observations are likely to have important implications for daily cancer therapies that activate p53 in normal tissues and/or tumour cells.


Effect of Different Irrigation Solutions on the Colour Stability of Three Calcium Silicate-Based Materials.
Previous studies have shown discoloration of mineral trioxide aggregate (MTA) in contact with root canal irrigation solutions. However, there are limited data on colour stability of other calcium silicate-based materials (CSMs). This in vitro study aimed to evaluate the colour stability of three CSMs in contact with different irrigation solutions. Three CSMs including White MTA (wMTA) Angelus, calcium enriched mixture (CEM), and Biodentine were assessed in this study. Forty five samples of each material were mixed according to the manufactures' instructions and then placed in silicone tubes. After 24 hours, the materials were removed from the moulds and 9 samples of each material left dry or immersed in normal saline, 5% sodium hypochlorite (NaOCL), 2% chlorhexidinegluconate (CHX), or 17%EDTA for 24 hours. Colour changes were measured with a spectrophotometer. Data were evaluated with 2-way analysis of variance, one way analysis of variance and Tukey post hoc tests. The highest discoloration of all materials was observed after contact with CHX. In the MTA Angelus and CEM cement groups, significant differences were observed between CHX and NaOCl and also between these two irrigants with the other three irrigants (p < 0.05). In the Biodentine group, CHX created statistically significant discoloration compared to other irrigants (p < 0.05). Only wMTA Angelus showed a significantly higher discoloration in contact with EDTA compared to normal saline and dry condition (p < 0.05). wMTA Angelus showed a significantly higher colour change compared with CEM cement and Biodentine after contact with NaOCl, CHX, and EDTA (p < 0.05). The contact of wMTA, CEM cement, and Biodentine with CHX should be avoided because this leads to severe discoloration. Contact with sodium hypochlorite also leads to discoloration of wMTA and CEM cements. Among of the three tested materials, wMTA showed the highest discoloration after contact with NaOCl, CHX, and EDTA.


Fibroblasts from chronic wounds show altered TGF-beta-signaling and decreased TGF-beta Type II receptor expression.
Chronic wounds are characterized by failure to heal in a defined time frame. However, the pathogenic steps leading from the etiological factors to failure to heal are unknown. Recently, increasing evidence suggests that resident cells in chronic wounds display a number of critical abnormalities, including senescence and unresponsiveness to the stimulatory action of transforming growth factor-beta1 (TGF-beta1). In this study, we have determined some of the mechanisms that might be responsible for unresponsiveness to TGF-beta1. Using Northern analysis and affinity labeling, we show that venous ulcer fibroblasts have decreased TGF-beta Type II receptor expression. This finding is not the result of genetic mutation, as shown by experiments with Type II receptor satellite instability. Decreased Type II receptor expression was accompanied by failure of ulcer fibroblasts to phosphorylate Smad 2, Smad 3, and p42/44 mitogen activating protein kinase (MAPK), and was associated with a slower proliferative rate in response to TGF-beta1. We conclude that venous ulcer fibroblasts show decreased Type II receptor expression and display abnormalities in the downstream signaling pathway involving MAPK and the early Smad pathway. These findings suggest ways to address and treat the abnormal cellular phenotype of cells in chronic wounds.


Common bile duct stricture without obvious cause.
Common bile duct strictures without apparent cause are unusual, yet probably occur more often than has been recognized. Whether this condition represents a different cause of biliary stricture or is just a forme fruste of recognized injuries or causes is not clear. Nonetheless, the surgeon should be aware that such isolated strictures without obvious cause may occur. The common causes of biliary stricture should be ruled out for diagnostic and therapeutic reasons. The management of such strictures is the same as for strictures due to the generally recognized causes.


Effects of botulinum toxin-induced paralysis on postnatal development of the supraspinatus muscle.
The mechanical environment plays an important role in musculoskeletal tissue development. The present study characterized changes in supraspinatus muscle due to removal of mechanical cues during postnatal development. An intramuscular injection of botulinum toxin type A (BTX) was used to induce and maintain paralysis in the left shoulders of mice since birth while the right shoulders received saline and served as contralateral controls. A separate group of animals was allowed to develop normally without any injections. Muscles were examined postnatally at various time points. The maximum isometric tetanic force generated by the muscle was significantly reduced in the BTX group compared to saline and normal groups. The paralyzed muscles were smaller and showed significant muscle atrophy and fat accumulation on histologic evaluation. Myogenic genes myogenin, myoD1, myf5, myf6, and fast type II myosin heavy chain (MHC) isoform were significantly upregulated while slow type I MHC isoform was significantly downregulated in the BTX group. Adipogenic genes C/EBPα, PPARγ2, leptin, and lipoprotein lipase were significantly upregulated in the BTX group. Results indicate that reduced muscle loading secondary to BTX-induced paralysis leads to fat accumulation and muscle degeneration in the developing muscle. Understanding the molecular and compositional changes in developing supraspinatus muscles may be useful for identifying and addressing the pathological changes that occur in shoulder injuries such as neonatal brachial plexus palsy.


Paper-based chemiresistor for detection of ultralow concentrations of protein.
A new paper-based chemiresistor composed of a network of single-wall carbon nanotubes (SWCNTs) and anti-human immunoglobulin G (anti-HIgG) is reported herein. SWCNTs act as outstanding transducers because they provide high sensitivity in terms of resistance changes due to immunoreaction. As a result, the resistance-based biosensor reaches concentration detection as low as picomolar. The resulting paper-based biosensor is sensitive, selective and employs low-cost substrate and simple manufacturing stages. Since chemiresistors require low-power equipment and are able to detect low concentrations with inexpensive materials, the present approach may pave the way for the development of resistive biosensors at very low-cost with high performances.


[Characterization and nutritional value of a food artisan: the meat pie of Murcia].
The main aims of this study are to describe the characteristics of the meat pie, a typical product of the regional gastronomy of Murcia and to determine its nutritional and energy values, fatty acid profile and fat quality. There were studied 24 samples of Murcia's meat pie from the six best-selling retail establishments in this Region (four units per establishment).The moisture, protein, fat, carbohydrates, fibre and minerals, contents and the energy value, fatty acid profile and fat quality were analyzed using the Official Analysis Methods of Foods. All analyses were performed by triplicate. The average weight of this product was 192.3 ± 11.8 g, with three differentiated parts (base, filling and pastry lid). All ingredients were natural raw materials: wheat flour, lard, ground beef, sliced boiled egg and chorizo, water and spices (salt, pepper, garlic, paprika and nutmeg). Most of its organoleptic attributes are due to the type and amount of fat or lard. The combination of the other ingredients and the particular formulation of the spices are also responsible of other attributes. Due to its protein content (11.0%), this meat pie can replace other meat dishes, and be incorporated into a balanced diet. However, it is necessary to take into account its fat and energy contents (17.3 g and 317 kcal/100 g, respectively). Unlike many common pastry products, it contains no trans fatty acids. The results of the research show that the studied product remains, at present, an artisan food, and offer reliable information that it is representative of the energy and nutritional values of the Murcia's meat pie, a typical product of the gastronomy of the Region of Murcia.


[The occurrence of destructive hydrocephalus in cases with neonatal intraventricular hemorrhage].
A total of 21 neonates with intraventricular hemorrhage (IVH) were studied; 12 (57.1%) survived, and 7 (58.3%) of these 12 cases suffered from hydrocephalus. Hydrocephalus occurred only in patients with IVH of grades III and IV on Papile's classification, and its occurrence rate was 87.5% as the sum total for the two groups. Destructive hydrocephalus (DHC) occurred in 5 cases of grade III and grade IV, and its occurrence rate was 62.5% as the sum total for the two groups. It could be conjectured that the prognosis of the IVH depends upon the extent of the primary hemorrhagic lesions. However, in cases with DHC, the resistance of the cerebral parenchyma to the hydrodynamic force is decreased, so that secondary cerebral destruction yielding rapid and intense ventricular dilatation, which is responsible for the development of neurological disorders, might occur more easily than in cases with IVH of grades III and IV, and to the secondary parenchymal destruction that may arise in association with hydrocephalus. It is essential that a shunt operation should be performed as soon as possible after the diagnosis of DHC has been established so as to allow repair of the primary lesion and to minimize the secondary brain damage.


Catechol-tetraethylenepentamine co-deposition modified cellulose filter paper for α-glucosidase immobilization and inhibitor screening from traditional Chinese medicine.
Cellulose filter paper (CFP) is expected to be an ideal carrier for enzyme immobilization due to its sustainability and biocompatibility. However, the interaction between the carrier and enzyme might change the spatial conformation of the enzyme and its microenvironment, and thus the flexibility of the enzyme molecule or the transport of the substrate to the active site would be hampered. In this work, a two-component system of catechol and tetraethylene pentamine was used to replace dopamine, and a polydopamine-like composite layer was deposited on the surface of CFP to introduce amino groups, which was similar to the self-polymerization-adhesion behavior of dopamine. Using polyethylene glycol diglycidyl ether with flexible spacer arms as the cross-linking agent, α-glucosidase was covalently bonded to amino-modified CFP through an epoxy ring-opening reaction. The immobilized α-glucosidase exhibited greater tolerance to pH and high temperature. After 10 repeated uses, the immobilized α-glucosidase maintained relatively high enzyme activity. Its kinetic behavior was investigated to illustrate the reliability for enzyme inhibitor screening. Finally, a screening method combining an immobilized enzyme and capillary electrophoresis analysis was proposed and applied to screening inhibitors from 11 kinds of traditional Chinese medicines, among which Chebulae Fructus, Phyllanthi Fructus and Terminaliae Relliricae Fructus exhibited strong enzyme inhibitory activities.


TRAP1, a novel mitochondrial chaperone responsible for multi-drug resistance and protection from apoptotis in human colorectal carcinoma cells.
TRAP1 is a component of a pro-survival mitochondrial pathway up-regulated in tumor cells. The evaluation of TRAP1 expression in 26 human colorectal carcinomas showed up-regulation in 17/26 tumors. Accordingly, TRAP1 levels were increased in HT-29 colorectal carcinoma cells resistant to 5-fluorouracil, oxaliplatin and irinotecan. Thus, we investigated the role of TRAP1 in multi-drug resistance in human colorectal cancer. Interestingly, TRAP1 overexpression leads to 5-fluorouracil-, oxaliplatin- and irinotecan-resistant phenotypes in different neoplastic cells. Conversely, the inhibition of TRAP1 activity by TRAP1 ATPase antagonist, shepherdin, increased the sensitivity to oxaliplatin and irinotecan in colorectal carcinoma cells resistant to the single agents. These results suggest that the increased expression of TRAP1 could be part of a pro-survival pathway responsible for multi-drug resistance.


Mutagenesis in mice of nuclear hormone receptor binding sites in the Igf2/H19 imprinting control region.
The H19/Igf2 imprinting control region (ICR) is a DNA methylation-dependent chromatin insulator in somatic cells. The hypomethylated maternally inherited ICR binds the insulator protein CTCF at four sites, and blocks activity of the proximal Igf2 promoter by insulating it from the shared distal enhancers. The hypermethylated paternally inherited ICR lacks CTCF binding and insulator activity, but induces methylation-silencing of the paternal H19 promoter. The paternal-specific methylation of the ICR is established in the male germ cells, while the ICR emerges from the female germ line in an unmethylated form. Despite several attempts to find cis-regulatory elements, it is still unknown what determines these male and female germ cell-specific epigenetic modifications. We recently proposed that five in vivo footprints spanning fifteen half nuclear hormone receptor (NHR) binding sites within the ICR might be involved, and here we report on the effects of mutagenizing all of these half sites in mice. No effect was obtained--in the female and male germ lines the mutant ICR remained hypomethylated and hypermethylated, respectively. The ICR imprinting mechanism remains undefined.


Adrenocortical carcinoma: clinicopathological features, prognostic factors and outcome.
The purpose of this study was to investigate the clinicopathological characteristics and treatment outcomes of patients with adrenocortical carcinoma (AC). Twenty-four patients (10 females and 14 males) diagnosed with AC between 1998 and 2009 were evaluated. Clinical features and outcomes were reviewed. Median age was 46.5 years. One (4%) patient was classified as stage I, 10 (42%) were classified as stage II, 8 (33%) were classified as stage III and 5 (21%) were classified as stage IV. Tumor sizes ranged from 3 to 22 cm with a mean diameter of 11 cm. Five patients were locally inoperable at initial diagnosis. In addition to surgery, 2 of 19 patients were treated with an adjuvant cisplatin plus etoposide regimen. Sixteen patients were treated with chemotherapy after recurrence. Median survival time was 18 months. The 1- and 5-year overall survival estimates were 73 and 48%, respectively. Mean survival times for male and female patients were 58 and 12 months, respectively (p = 0.046). Early T stage (p = 0.04), lymph node negativity (p < 0.001), the absence of distant metastases (p < 0.001) and early stage (p < 0.001) were correlated with overall survival. AC is a rare disease with a poor prognosis. There are correlations between gender, stage and survival.


Cervical, mandibular, and parotid lymph nodes of dogs naturally infected with Leishmania infantum: a histopathologic and immunohistochemistry study and its correlation with facial skin lesions.
The parasite load in cervical, mandibular, and parotid lymph nodes and in the skin of the nose and the pinna from dogs infected with Leishmania infantum were investigated by histologic and immunohistochemical studies. Twenty-two infected dogs with and without signs of infection were examined to demonstrate correlation of signs with parasite load and the correlation of facial skin lesions with parasites in regional lymph nodes. Chronic inflammation of the skin was demonstrated in infected dogs that had no gross skin lesions, confirming that normal-appearing skin can harbor the parasite, likely playing a role in transmission. Dogs with facial skin lesions showed a higher parasite load in parotid lymph nodes than dogs without lesions of the facial skin, based on Leishman-Donovan unit analysis. Based on immunohistochemical analysis, parasite load in parotid and cervical nodes was correlated with that of skin of the nose and pinna, as was the parasite load in mandibular lymph nodes and skin of the external nose. We demonstrated a logical involvement of the lymphatic vessels and their specific anatomic draining sites.


Analysis of heart rate variability in the presence of ectopic beats using the heart timing signal.
The time-domain signals representing the heart rate variability (HRV) in the presence of an ectopic beat exhibit a sharp transient at the position of the ectopic beat, which corrupts the signal, particularly the power spectral density (PSD) of the HRV. Consequently, there is a need for correction of this type of beat prior to any HRV analysis. This paper deals with the PSD estimation of the HRV by means of the heart timing (HT) signal when ectopic beats are present. These beat occurrence times are modeled from a generalized, continuous time integral pulse frequency modulation model and, from this point of view, a specific method for minimizing the effect of the presence of ectopic beats is presented to work together with the HT signal. By using both, a white noise driven autoregressive model of the HRV signal with artificially introduced ectopic beats and actual heart rate series including ectopic beats, the more usual methods of HRV spectral estimation are compared. Results of the PSD estimation error function of the number of ectopic beats are presented. These results demonstrate that the proposed method has one order of magnitude lower error than usual ectopic beats removal strategies in preserving PSD, thus, this strategy better recovers the original clinical indexes of interest.


Exercise management in type 1 diabetes: a consensus statement.
Type 1 diabetes is a challenging condition to manage for various physiological and behavioural reasons. Regular exercise is important, but management of different forms of physical activity is particularly difficult for both the individual with type 1 diabetes and the health-care provider. People with type 1 diabetes tend to be at least as inactive as the general population, with a large percentage of individuals not maintaining a healthy body mass nor achieving the minimum amount of moderate to vigorous aerobic activity per week. Regular exercise can improve health and wellbeing, and can help individuals to achieve their target lipid profile, body composition, and fitness and glycaemic goals. However, several additional barriers to exercise can exist for a person with diabetes, including fear of hypoglycaemia, loss of glycaemic control, and inadequate knowledge around exercise management. This Review provides an up-to-date consensus on exercise management for individuals with type 1 diabetes who exercise regularly, including glucose targets for safe and effective exercise, and nutritional and insulin dose adjustments to protect against exercise-related glucose excursions.


Rapid establishment of a national surveillance of COVID-19 hospitalizations in Belgium.
In response to the COVID-19 epidemic, caused by a novel coronavirus, it was of great importance to rapidly collect as much accurate information as possible in order to characterize the public health threat and support the health authorities in its management. Hospital-based surveillance is paramount to monitor the severity of a disease in the population. Two separate surveillance systems, a Surge Capacity survey and a Clinical survey, were set up to collect complementary data on COVID-19 from Belgium's hospitals. The Surge Capacity survey collects aggregated data to monitor the hospital capacity through occupancy rates of beds and medical devices, and to follow a set of key epidemiological indicators over time. Participation is mandatory and the daily data collection includes prevalence and incidence figures on the number of COVID-19 patients in the hospital. The Clinical survey is strongly recommended by health authorities, focusses on specific patient characteristics and relies on individual patient data provided by the hospitals at admission and discharge. This national double-level hospital surveillance was implemented very rapidly after the first COVID-19 patients were hospitalized and revealed to be crucial to monitor hospital capacity over time and to better understand the disease in terms of risk groups and outcomes. The two approaches are complementary and serve different needs.


Political attitudes develop independently of personality traits.
The primary assumption within the recent personality and political orientations literature is that personality traits cause people to develop political attitudes. In contrast, research relying on traditional psychological and developmental theories suggests the relationship between most personality dimensions and political orientations are either not significant or weak. Research from behavioral genetics suggests the covariance between personality and political preferences is not causal, but due to a common, latent genetic factor that mutually influences both. The contradictory assumptions and findings from these research streams have yet to be resolved. This is in part due to the reliance on cross-sectional data and the lack of longitudinal genetically informative data. Here, using two independent longitudinal genetically informative samples, we examine the joint development of personality traits and attitude dimensions to explore the underlying causal mechanisms that drive the relationship between these features and provide a first step in resolving the causal question. We find change in personality over a ten-year period does not predict change in political attitudes, which does not support a causal relationship between personality traits and political attitudes as is frequently assumed. Rather, political attitudes are often more stable than the key personality traits assumed to be predicting them. Finally, the results from our genetic models find that no additional variance is accounted for by the causal pathway from personality traits to political attitudes. Our findings remain consistent with the original construction of the five-factor model of personality and developmental theories on attitude formation, but challenge recent work in this area.


pH-dependent structural transitions in cationic ionizable lipid mesophases are critical for lipid nanoparticle function.
Lipid nanoparticles (LNPs) are advanced core-shell particles for messenger RNA (mRNA) based therapies that are made of polyethylene glycol (PEG) lipid, distearoylphosphatidylcholine (DSPC), cationic ionizable lipid (CIL), cholesterol (chol), and mRNA. Yet the mechanism of pH-dependent response that is believed to cause endosomal release of LNPs is not well understood. Here, we show that eGFP (enhanced green fluorescent protein) protein expression in the mouse liver mediated by the ionizable lipids DLin-MC3-DMA (MC3), DLin-KC2-DMA (KC2), and DLinDMA (DD) ranks MC3 ≥ KC2 > DD despite similar delivery of mRNA per cell in all cell fractions isolated. We hypothesize that the three CIL-LNPs react differently to pH changes and hence study the structure of CIL/chol bulk phases in water. Using synchrotron X-ray scattering a sequence of ordered CIL/chol mesophases with lowering pH values are observed. These phases show isotropic inverse micellar, cubic Fd3m inverse micellar, inverse hexagonal [Formula: see text] and bicontinuous cubic Pn3m symmetry. If polyadenylic acid, as mRNA surrogate, is added to CIL/chol, excess lipid coexists with a condensed nucleic acid lipid [Formula: see text] phase. The next-neighbor distance in the excess phase shows a discontinuity at the Fd3m inverse micellar to inverse hexagonal [Formula: see text] transition occurring at pH 6 with distinctly larger spacing and hydration for DD vs. MC3 and KC2. In mRNA LNPs, DD showed larger internal spacing, as well as retarded onset and reduced level of DD-LNP-mediated eGFP expression in vitro compared to MC3 and KC2. Our data suggest that the pH-driven Fd3m-[Formula: see text] transition in bulk phases is a hallmark of CIL-specific differences in mRNA LNP efficacy.


Regulation of vitamin D metabolism: factors influencing the rate of formation of 1,25-dihydroxycholecalciferol by kidney homogenates.
The formation of 1,25-dihydroxycholecalciferol from 25-hydroxycholecalciferol by chick kidney homogenates is inhibited by increasing concentrations of Ca(2+). The apparent K(m) for the hydroxylation reaction is 1x10(-7)m, significantly lower than that reported for isolated mitochondria. Separated cytoplasmic and particulate fractions are inactive, but on recombination activity is restored, possibly because of the presence in the soluble fraction of a factor with a high affinity for 25-hydroxycholecalciferol.


Factors influencing the entry of dye into neutrophil leucocytes in the Nitroblue Tetrazolium test.
1. The Nitroblue Tetrazolium (NBT) test is a measure of the phagocytosis of particulate complexes of NBT and heparin and/or fibrinogen by neutrophil leucocytes. 2. Humoral factors in the plasma of ill patients stimulate neutrophil leucocytes of normal subjects to ingest these complexes. 3. The acute phase protein, orosomucoid and endotoxin stimulate reduction of NBT in vitro and could be responsible for some positive NBT tests in vivo. 4. The effect of factors promoting phagocytosis may be masked by an inability of neutrophil leucocytes to respond to stimulation. 5. This defective neutrophil function could result from the replacement of circulating neutrophil leucocytes by less mature, less actively phagocytic cells or by the presence of circulating immune complexes.


Developing an optimum computer-designed multispectral system comprising a monochrome CCD camera and a liquid-crystal tunable filter.
In a previous work [J. Opt. Soc. Am. A24, 942 (2007)JOAOD60740-323210.1364/JOSAA.24.000942] we made a complete theoretical and computational study of the influence of several parameters on the behavior of a planned multispectral system for imaging skylight, including the number of sensors and the spectral estimation algorithm. Here we follow up this study by using all the information obtained in the computational simulations to implement a real multispectral imaging system based on a monochrome CCD camera and a liquid-crystal tunable filter (LCTF). We were able to construct the optimum Gaussian sensors found in the simulations by adjusting the exposure times of some of the transmittance modes of the LCTF, hence obtaining really accurate spectral estimations of skylight with only a few optimum sensors.


Ultra-thin ultra-smooth and low-loss silver films on a germanium wetting layer.
We demonstrate a method to fabricate ultra-thin, ultra-smooth and low-loss silver (Ag) films using a very thin germanium (Ge) layer as a wetting material and a rapid post-annealing treatment. The addition of a Ge wetting layer greatly reduces the surface roughness of Ag films deposited on a glass substrate by electron-beam evaporation. The percolation threshold of Ag films and the minimal thickness of a uniformly continuous Ag film were significantly reduced using a Ge wetting layer in the fabrication. A rapid post-annealing treatment is demonstrated to reduce the loss of the ultra-thin Ag film to the ideal values allowed by the quantum size effect in smaller grains. Using the same wetting method, we have also extended our studies to ultra-smooth silver-silica lamellar composite films with ultra-thin Ag sublayers.


Psychological determinants of COVID-19 vaccine acceptance among urban slum dwellers of Bangladesh.
Coronavirus disease 2019 (COVID-19) vaccination has emerged as a promising approach to counter the harmful impacts of the pandemic. Understanding the psychological components that may impact an individual's attitude toward COVID-19 vaccination is crucial for generating evidence-based ways to minimize vaccine hesitancy. This study determined the psychological antecedents regarding vaccine acceptance among urban slum people of Bangladesh. From 5 July to 5 August 5, 2021, a face-to-face survey was conducted in the urban slum of two large cities in Bangladesh. The questionnaire considered socio-demographics, health-related characteristics, psychological determinants, sources of information, and conspiracy beliefs regarding COVID-19. The 5C sub-scales were used to assess psychological antecedents. Five stepwise binary logistic regression models evaluated significant predictors for confidence, complacency, calculation, constraints, and collective responsibility. Multinomial logistic regression was used to determine the relationship between psychological antecedents and vaccine acceptability. The study revealed that the slum residents with a high level of confident (89.94%), complacent (72.73%), having constraints (82.31%), calculative (84.80%), and responsible (93.30%) showed a higher vaccine acceptance rate. Higher vaccine acceptance was related to the believer in natural-made origin (85.96%) and those who rejected anti-vaccination (88.44%). The information acquired from newspapers differed significantly (p < 0.05), though TV or radio was the most common primary information source about COVID-19 vaccines (74.75%). The regression result revealed that marital status, education, family income, and perceived health condition were significantly associated with the 5C domains. Two psychological antecedents including complacency (OR = 3.97; p < 0.001) and collective responsibility (OR = 0.23; p < 0.001) were significantly associated with vaccine acceptance. Different predictors significantly affect psychological antecedents related to COVID-19 vaccine uptake. Therefore, considering the factors, targeted actions based on the findings may help to lower vaccine reluctance and boost vaccination rates.


A method for detecting long non-coding RNAs with tiled RNA expression microarrays.
Long non-coding ribonucleic acids (lncRNAs) have been proposed as biomarkers in prostate cancer. This paper proposes a selection method which uses data from tiled microarrays to identify relatively long regions of moderate expression independent of the microarray platform and probe design. The method is used to search for candidate long non-coding ribonucleic acids (lncRNAs) at locus 8q24 and is run on three independent experiments which all use samples from prostate cancer patients. The robustness of the method is tested by utilizing repeated copies of tiled probes. The method shows high consistency between experiments that used the same samples, but different probe layout. There also is statistically significant consistency when comparing experiments with different samples. The method selected the long non-coding ribonucleic acid PCNCR1 in all three experiments.


Phage Inhibit Pathogen Dissemination by Targeting Bacterial Migrants in a Chronic Infection Model.
The microbial communities inhabiting chronic infections are often composed of spatially organized micrometer-sized, highly dense aggregates. It has recently been hypothesized that aggregates are responsible for the high tolerance of chronic infections to host immune functions and antimicrobial therapies. Little is currently known regarding the mechanisms controlling aggregate formation and antimicrobial tolerance primarily because of the lack of robust, biologically relevant experimental systems that promote natural aggregate formation. Here, we developed an in vitro model based on chronic Pseudomonas aeruginosa infection of the cystic fibrosis (CF) lung. This model utilizes a synthetic sputum medium that readily promotes the formation of P. aeruginosa aggregates with sizes similar to those observed in human CF lung tissue. Using high-resolution imaging, we exploited this model to elucidate the life history of P. aeruginosa and the mechanisms that this bacterium utilizes to tolerate antimicrobials, specifically, bacteriophage. In the early stages of growth in synthetic sputum, planktonic cells form aggregates that increase in size over time by expansion. In later growth, migrant cells disperse from aggregates and colonize new areas, seeding new aggregates. When added simultaneously with phage, P. aeruginosa was readily killed and aggregates were unable to form. When added after initial aggregate formation, phage were unable to eliminate all of the aggregates because of exopolysaccharide production; however, seeding of new aggregates by dispersed migrants was inhibited. We propose a model in which aggregates provide a mechanism that allows P. aeruginosa to tolerate phage therapy during chronic infection without the need for genetic mutation.IMPORTANCE Bacteria in chronic infections often reside in communities composed of micrometer-sized, highly dense aggregates. A primary challenge for studying aggregates has been the lack of laboratory systems that promote natural aggregate formation in relevant environments. Here, we developed a growth medium that mimics chronic lung infection and promotes natural aggregate formation by the bacterium Pseudomonas aeruginosa High-resolution, single-cell microscopy allowed us to characterize P. aeruginosa's life history-seeding, aggregate formation, and dispersal-in this medium. Our results reveal that this bacterium readily forms aggregates that release migrants to colonize new areas. We also show that aggregates allow P. aeruginosa to tolerate therapeutic bacteriophage addition, although this treatment limits P. aeruginosa dissemination by targeting migrants.


The challenge of small-scale repeats for indel discovery.
Repetitive sequences are abundant in the human genome. Different classes of repetitive DNA sequences, including simple repeats, tandem repeats, segmental duplications, interspersed repeats, and other elements, collectively span more than 50% of the genome. Because repeat sequences occur in the genome at different scales they can cause various types of sequence analysis errors, including in alignment, de novo assembly, and annotation, among others. This mini-review highlights the challenges introduced by small-scale repeat sequences, especially near-identical tandem or closely located repeats and short tandem repeats, for discovering DNA insertion and deletion (indel) mutations from next-generation sequencing data. We also discuss the de Bruijn graph sequence assembly paradigm that is emerging as the most popular and promising approach for detecting indels. The human exome is taken as an example and highlights how these repetitive elements can obscure or introduce errors while detecting these types of mutations.


Pathogenesis of Interstitial Lung Disease in Children and Adults.
Interstitial lung diseases (ILDs) occur across the lifespan, from birth to advanced age. However, the causes, clinical manifestations, histopathology, and management of ILD differ greatly among infants, older children, and adults. The historical approach of classifying childhood ILD (chILD) using adult classification schemes may therefore have done more harm than good. Nevertheless, identification of novel forms of chILD in the past decade, such as surfactant metabolism dysfunction disorders and neuroendocrine cell hyperplasia of infancy (NEHI), as well as genomic analysis of adult ILDs, has taught us that identical genotypes may result in distinct phenotypes at different ages and developmental stages, and that lung developmental pathways and cellular phenotypes are often recapitulated in adult ILDs. Thus comparison of the pathophysiology of ILD in children and adults in the context of lung development is useful in understanding the pathogenesis of these disorders, and may lead to novel therapeutic interventions for ILDs at all ages.


Visible Light-Assisted Degradation of Sulfamethoxazole on 2D/0D Sulfur-Doped Bi2O3/MnO2 Z-Scheme Heterojunction Immobilized Photocatalysts.
Retrieving the spent photocatalysts from the reaction system is always a challenging task. Therefore, the present work is focused on immobilizing sulfur-doped-Bi2O3/MnO2 (S-BOMO) heterojunction photocatalysts over different support matrices and evaluating their performance for the removal of sulfamethoxazole (SMX) in water under visible light. Our findings revealed S-BOMO coated clay beads (S-BOMO CCB) achieving more than 86% (240 min) SMX degradation ∼3, ∼1.3, and ∼2 times higher compared to S-BOMO coated on the different substrates, including glass beads, floating stones, and polymer material substrates, respectively. Mott-Schottky measurements confirmed the construction of the Z-scheme heterojunction involving MnO2 and 2S-Bi2O3. This Z-scheme mechanism, along with its narrow band gap of 1.58 eV, resulted in a rapid spatial transfer of the photogenerated charge carriers between the semiconductors and is believed to enhance the overall photocatalytic activity of the nanocomposite. Radical trapping and electron paramagnetic resonance results clearly established the active role of hydroxyl radicals and hydrogen peroxide in the degradation of SMX. Further, the 2S-BOMO CCB demonstrated excellent stability and photocatalytic activity over multiple runs. According to the sensitivity analysis and the results of anion effect experiments, phosphate and sulfate ions exhibit a significant impact on sulfamethoxazole degradation. Toxicity analysis revealed that 2S-BOMO CCB and sulfamethoxazole degradation byproducts were apparently innocuous. Additionally, the practical applicability of 2S-BOMO CCB was examined in various real water matrices, with the degradation efficiency followed the order: tap water < groundwater < surface water < hospital wastewater < municipal wastewater < pharmaceutical industry wastewater. The economic assessment revealed the reduction in the overall cost of the immobilized 2S-BOMO following the recovery process. Overall, the findings of this work provided critical insights into the synthesis and performance of incredibly effective and stable immobilized photocatalysts for the degradation of pharmaceutical pollutants.


2021 European Thyroid Association Guideline on Thyroid Disorders prior to and during Assisted Reproduction.
Severe thyroid dysfunction may lead to menstrual disorders and subfertility. Fertility problems may persist even after restoring normal thyroid function, and then an assisted reproductive technology (ART) may be a solution. Prior to an ART treatment, ovarian stimulation is performed, leading to high oestradiol levels, which may lead to hypothyroidism in women with thyroid autoimmunity (TAI), necessitating levothyroxine (LT4) supplements before pregnancy. Moreover, women with the polycystic ovarian syndrome and idiopathic subfertility have a higher prevalence of TAI. Women with hypothyroidism treated with LT4 prior to ART should have a serum TSH level <2.5 mIU/L. Subfertile women with hyperthyroidism planning an ART procedure should be informed of the increased risk of maternal and foetal complications, and euthyroidism should be restored and maintained for several months prior to an ART treatment. Fertilisation rates and embryo quality may be impaired in women with TSH >4.0 mIU/L and improved with LT4 therapy. In meta-analyses that mainly included women with TSH levels >4.0 mIU/L, LT4 treatment increased live birth rates, but that was not the case in 2 recent interventional studies in euthyroid women with TAI. The importance of the increased use of intracytoplasmic sperm injection as a type of ART on pregnancy outcomes in women with TAI deserves more investigation. For all of the above reasons, women of subfertile couples should be screened routinely for the presence of thyroid disorders.


Replication-defective viruses modulate immune responses.
By immunizing inbred mice with purified replication-competent, defective virus particles, or an admixture of the two, differential effects on the cellular immune system have been uncovered. Defective virus, exemplified by the vesicular stomatitis virus (VSV) defective interfering particle (DI 0.33), induced in BALB/c mice low levels of proliferating, IL-2 secreting, and cytolytic Ag-specific T lymphocytes. This was not caused by a dominant suppressor cell response, or by a failure to stimulate lymphokine-secreting cells, but appeared to reflect a reduced efficiency of priming as compared with standard virus. Mice primed with a mixture of wt and DI virus showed reduced proliferation compared with mice primed with wt virus. When histocompatible target cells were sensitized by pure DI particles, they were neither recognized nor lysed by CD8+ CTL. Cells co-infected with wt and DI particles were not as readily lysed by CD8+ CTL as cells infected by VSV alone. The extent of this reduction was dependent on the concentration of DI particles. This suggests that DI particles may have prevented the proper presentation of endogenously synthesized Ag for recognition by CD8+ CTL. Metabolic labeling studies indicated that the presence of DI particles suppressed the synthesis of viral proteins in dually infected cells. However, CD4+ T lymphocyte clones recognized and efficiently lysed histocompatible Ia+ cells infected with DI particles alone or co-infected with replication-competent and defective virus.


[Composition and degree of sulfation of glycosaminoglycans from tissues of different animal species: heterogeneity and tissue specificity of heparan sulfates].
The composition and degree of sulfation of glycosaminoglycans (GAG) in proteoglycans from various animal tissues were studied. It was shown that sulfated GAG contain chondroitin sulfates AC and B as well as heparan sulfates. The bulk of GAG in the majority of tissues under study is represented by 2-3 types of heparan sulfate molecules differing in the degree of sulfation. According to the degree of sulfation heparan sulfates from all tissues studied can be classified into three groups. Homologous tissues of various animal species are characterized by a similar composition and the degree of sulfation The data obtained are discussed in terms of the feasible role of proteoheparan sulfates in specific cell-to-cell interactions.


Percutaneous antegrade transarterial treatment of iatrogenic radial arteriovenous fistula.
Site of access vascular complications is infrequent after transradial interventions. We report the case of a 66-year-old man referred to our hospital because of right forearm swelling, oedema and pain with functional forearm disability, 1 year after a transradial primary percutaneous coronary intervention (PCI). The diagnostic and procedural issues are discussed. This is the first description of a successful and well-tolerated radial arteriovenous fistula (AVF) treatment by means of percutaneous antegrade approach with the use of a short introducer and a biocompatible covered stent.


Role of the gut-associated and secondary lymphoid tissue in the induction of chronic colitis.
It is well known that enteric bacterial antigens drive the development of chronic colitis in a variety of different mouse models of the inflammatory bowel diseases (IBD). The objective of this study was to evaluate the role of gut-associated lymphoid tissue (GALT; Peyer's patches, isolated lymphoid follicles), mesenteric lymph nodes (MLNs) and spleen in the pathogenesis of chronic colitis in mice. Surgical as well as genetic approaches were used to generate lymphopenic mice devoid of one or more of these lymphoid tissues. For the first series of studies, we subjected recombinase activating gene-1-deficient mice (RAG(-/-) ) to sham surgery (Sham), mesenteric lymphadenectomy (MLNx), splenectomy (Splx) or both (MLNx/Splx). In a second series of studies we intercrossed lymphotoxinβ-deficient (LTβ(-/-) ) mice with RAG(-/-) animals to generate LTβ(-/-) x RAG(-/-) offspring that were anticipated to contain functional MLNs but be devoid of GALT and most peripheral lymph nodes. Flow purified naïve (CD4(+) CD45RB(high) ) T-cells were adoptively transferred into the different groups of RAG(-/-) recipients to induce chronic colitis. We found that at 3-5 wks following T-cell transfer, all four of the surgically-manipulated RAG(-/-) groups (Sham, MLNx, Splx and MLNx/Splx) developed chronic colitis that was similar in onset and severity. Flow cytometric analysis revealed no differences among the different groups with respect to surface expression of different gut-homing markers nor were there any differences noted in IFN-γ and IL-17 generation by mononuclear cells isolated among these surgically-manipulated mice. Although we anticipated that LTβ(-/-) x RAG(-/-) mice would contain functional MLNs but be devoid of GALT and peripheral lymph nodes (PLNs), we found that LTβ(-/-) x RAG(-/-) mice were in fact devoid of MLNs as well as GALT and PLNs. Adoptive transfer of CD45RB(high) T-cells into LTβ(-/-) x RAG(-/-) mice or their littermate controls (LTβ(+/+) x RAG(-/-) ) induced rapid and severe colitis in both groups. Taken together, our data demonstrate that: a) neither the GALT, MLNs nor PLNs are required for induction of chronic gut inflammation in this model of IBD and b) T-and/or B-cells may be required for the development of MLNs in LTβ(-/-) mice.


C20-D3-vitamin A slows lipofuscin accumulation and electrophysiological retinal degeneration in a mouse model of Stargardt disease.
Stargardt disease, also known as juvenile macular degeneration, occurs in approximately one in 10,000 people and results from genetic defects in the ABCA4 gene. The disease is characterized by premature accumulation of lipofuscin in the retinal pigment epithelium (RPE) of the eye and by vision loss. No cure or treatment is available. Although lipofuscin is considered a hallmark of Stargardt disease, its mechanism of formation and its role in disease pathogenesis are poorly understood. In this work we investigated the effects of long-term administration of deuterium-enriched vitamin A, C20-D(3)-vitamin A, on RPE lipofuscin deposition and eye function in a mouse model of Stargardt's disease. Results support the notion that lipofuscin forms partly as a result of the aberrant reactivity of vitamin A through the formation of vitamin A dimers, provide evidence that preventing vitamin A dimerization may slow disease related, retinal physiological changes and perhaps vision loss and suggest that administration of C20-D(3)-vitamin A may be a potential clinical strategy to ameliorate clinical symptoms resulting from ABCA4 genetic defects.


Randomized controlled multicenter international clinical trial of self-gripping Parietex™ ProGrip™ polyester mesh versus lightweight polypropylene mesh in open inguinal hernia repair: interim results at 3 months.
To compare clinical outcomes following sutureless Parietex™ ProGrip™ mesh repair to traditional Lichtenstein repair with lightweight polypropylene mesh secured with sutures. This is a 3-month interim report of a 1-year multicenter international study. Three hundred and two patients were randomized; 153 were treated with Lichtenstein repair (L group) and 149 with Parietex™ ProGrip™ precut mesh (P group) with or without fixation. The primary outcome measure was postoperative pain using the visual analog scale (VAS, 0-150 mm); other outcomes were assessed prior to surgery and up to 3 months postoperatively. Compared to baseline, pain score was lower in the P group at discharge (-10%) and at 7 days (-13%), while pain increased in the L group at discharge (+39%) and at 7 days (+21%). The difference between groups was significant at both time points (P = 0.007 and P = 0.039, respectively). In the P group, patients without fixation suffered less pain compared to those with single-suture fixation (1 month: -20.9 vs. -6.15%, P = 0.02; 3 months: -24.3 vs. -7.7%, P = 0.01). The infection rate was significantly lower in the P group during the 3-month follow-up (2.0 vs. 7.2%, P = 0.032). Surgery duration was significantly shorter in the P group (32.4 vs. 39.1 min; P < 0.001). No recurrence was observed at 3 months in both groups. Surgery duration, early postoperative, pain and infection rates were significantly reduced with self-gripping polyester mesh compared to Lichtenstein repair with polypropylene mesh. The use of fixation increased postoperative pain in the P group. The absence of early recurrence highlights the gripping efficiency effect.


Endocannabinoid signaling as a synaptic circuit breaker in neurological disease.
Cannabis sativa is one of the oldest herbal plants in the history of medicine. It was used in various therapeutic applications from pain to epilepsy, but its psychotropic effect has reduced its usage in recent medical practice. However, renewed interest has been fueled by major discoveries revealing that cannabis-derived compounds act through a signaling pathway in the human body. Here we review recent advances showing that endocannabinoid signaling is a key regulator of synaptic communication throughout the central nervous system. Its underlying molecular architecture is highly conserved in synapses from the spinal cord to the neocortex, and as a negative feed-back signal, it provides protection against excess presynaptic activity. The endocannabinoid signaling machinery operates on demand in a synapse-specific manner; therefore, its modulation offers new therapeutic opportunities for the selective control of deleterious neuronal activity in several neurological disorders.


Underlying structure of the National Institutes of Health Stroke Scale: results of a factor analysis. NINDS tPA Stroke Trial Investigators.
No stroke scale has been validated as an outcome measure using data from a clinical trial demonstrating a positive therapeutic effect. Therefore, we proposed to use data from the National Institute of Neurological Disorders and Stroke (NINDS) tPA Stroke Trial to determine whether the National Institutes of Health Stroke Scale (NIHSS) was valid in patients treated with tissue plasminogen activator (tPA) and to explore the underlying clinimetric structure of the NIHSS. We performed an exploratory factor analysis of NIHSS data from Part 1 (n=291) of the NINDS tPA Stroke Trial to derive a hypothesized underlying factor structure. We then performed a confirmatory factor analysis of this structure using NIHSS data from Part 2 of the same trial (n=333). We then tested whether this final factor structure could be found in tPA- and placebo-treated patients serially over time after stroke treatment. Using 3-month outcome data, we tested for an association between the NIHSS and other measures of stroke outcome. The exploratory analysis suggested that there were 2 factors underlying the NIHSS, representing left and right brain function, confirming the content validity of the scale. An alternative structure composed of 4 factors could be derived, with a better goodness of fit: the first 2 factors could represent left brain cortical and motor function, respectively, and the second 2 factors could represent right brain cortical and motor function, respectively. The same factor structures were then found in tPA and placebo patient groups studied serially over time, confirming the exploratory analysis. All 3-month clinical outcomes were associated with each other at subsequent time points, confirming predictive validity. This is the first study of the validity of a stroke scale in patients treated with effective stroke therapy. The NIHSS appeared to be valid in patients with acute stroke and for finding treatment-related differences. The scale was valid when used serially over time after stroke, up to 3 months, and showed good agreement with other measures of outcome.


Mini-Addenbrooke's Cognitive Examination (MACE): a Useful Cognitive Screening Instrument in Older People?
The Mini-Addenbrooke's Cognitive Examination (MACE) is a recently described brief cognitive screening instrument. To examine the test accuracy of MACE for the identification of dementia and mild cognitive impairment (MCI) in a cohort of older patients assessed in a neurology-led dedicated cognitive disorders clinic. Cross-sectional assessment of consecutive patients with MACE was performed independent of the reference standard diagnosis based on clinical interview of patient and, where possible, informant and structural brain imaging, and applying standard clinical diagnostic criteria for dementia and MCI. Various test accuracy metrics were examined at two MACE cut-offs ( ≤ 25/30 and ≤ 21/30), comparing the whole patient cohort with those aged ≥ 65 or ≥ 75 years, hence at different disease prevalences. Dependent upon the chosen cut-off, MACE was either very sensitive or very specific for the identification of any cognitive impairment in the older patient cohorts with increased disease prevalence. However, at both cut-offs the positive predictive values and post-test odds increased in the older patient cohorts. At the more sensitive cut-off, improvements in some new unitary test metrics were also seen. MACE is a valid instrument for identification of cognitive impairment in older people. Test accuracy metrics may differ with disease prevalence.


[Pharmacotherapy of Alzheimer dementia: therapy of cognitive symptoms--new results of clinical studies].
Recent investigations have given new insights into pathogenetical determinants of Alzheimer's disease. Amyloid deposition and neurofibrillary tangles are no longer considered to be primary pathological changes. Neurobiological research tries to work out the etiopathogenital cascade that finally causes Alzheimer's disease. So far, several relevant pathogenetical factors have been detected, e.g. pertubated control of glucose breakdown, impairment of oxidative metabolism, impaired neuroprotection due to increased oxidative stress and non-enzymatic protein glycation as well as immunological disturbances. Thus, new strategies for the development of cognition-enhancing drugs are emerging. The authors review reports on agents, that are under investigation for the treatment of cognitive symptomatology in Alzheimer's disease. Some of these agents have already been used for treatment of other medical conditions, e.g. nimodipine, memantine as well as selegiline. Many of them are still experimental. Promising strategies include antioxidative agents (e.g. vitamin E, vitamin C, beta-carotin), acetylcholinesterase-inhibitors with central selectivity (e.g. ENA 713), M1- and M4-muscarinic receptor agonists (milameline) as well as sabeluzole, a benzothazide derivative that shows neurotrophic activities and anti-inflammatory substances like indomethacin.


Thermoring basis for the TRPV3 bio-thermometer.
The thermosensitive transient receptor potential (TRP) channels are well-known as bio-thermometers with specific temperature thresholds and sensitivity. However, their precise structural origins are still mysterious. Here, graph theory was used to test how the temperature-dependent non-covalent interactions as identified in the 3D structures of thermo-gated TRPV3 could form a systematic fluidic grid-like mesh network with the constrained thermo-rings from the biggest grids to the smallest ones as necessary structural motifs for the variable temperature thresholds and sensitivity. The results showed that the heat-evoked melting of the biggest grids may control the specific temperature thresholds to initiate channel gating while the smaller grids may be required to secure heat efficacy. Together, all the grids along the lipid-dependent minimal gating pathway may be necessary to change with molar heat capacity for the specific temperature sensitivity. Therefore, this graph theory-based grid thermodynamic model may provide an extensive structural basis for the thermo-gated TRP channels.


Combining information from 3 anatomic regions in the diagnosis of glaucoma with time-domain optical coherence tomography.
To improve the diagnosis of glaucoma by combining time-domain optical coherence tomography (TD-OCT) measurements of the optic disc, circumpapillary retinal nerve fiber layer (RNFL), and macular retinal thickness. Ninety-six age-matched normal and 96 perimetric glaucoma participants were included in this observational, cross-sectional study. Or-logic, support vector machine, relevance vector machine, and linear discrimination function were used to analyze the performances of combined TD-OCT diagnostic variables. The area under the receiver-operating curve (AROC) was used to evaluate the diagnostic accuracy and to compare the diagnostic performance of single and combined anatomic variables. The best RNFL thickness variables were the inferior (AROC=0.900), overall (AROC=0.892), and superior quadrants (AROC=0.850). The best optic disc variables were horizontal integrated rim width (AROC=0.909), vertical integrated rim area (AROC=0.908), and cup/disc vertical ratio (AROC=0.890). All macular retinal thickness variables had AROCs of 0.829 or less. Combining the top 3 RNFL and optic disc variables in optimizing glaucoma diagnosis, support vector machine had the highest AROC, 0.954, followed by or-logic (AROC=0.946), linear discrimination function (AROC=0.946), and relevance vector machine (AROC=0.943). All combination diagnostic variables had significantly larger AROCs than any single diagnostic variable. There are no significant differences among the combination diagnostic indices. With TD-OCT, RNFL and optic disc variables had better diagnostic accuracy than macular retinal variables. Combining top RNFL and optic disc variables significantly improved diagnostic performance. Clinically, or-logic classification was the most practical analytical tool with sufficient accuracy to diagnose early glaucoma.


Transitioning the Adult with Type 2 Diabetes From the Acute to Chronic Care Setting: Strategies to Support Pragmatic Implementation Success.
Scientific evidence is available to guide the how to of medications management when patients with diabetes are hospitalized or present to the Emergency Department. However, few clinical trials in the diabetes field have addressed the execution, coupled with established implementation effectiveness evaluation frameworks to help inform and assess implementation practices to support the transition in care. These deficiencies may be overcome by (1) applying the principles of implementation and delivery systems science; (2) engaging the principles of human factors (HF) throughout the design, development, and evaluation planning activities; and (3) utilizing mixed methods to design the intervention, workflow processes, and evaluate the intervention for sustainability within existing care delivery models. This article provides a discussion of implementation science and human factors science including an overview of commonly used frameworks which can be applied to structure design and implementation of sustainable and generalizable interventions.


Insulin-like growth factors 1 and 2 (IGF-1 and IGF-2) mRNA levels in relation to the gastrointestinal tract (GIT) development in newborn calves.
The study was aimed at determining the age-depended changes in insulin-like growth factors 1 and 2 (IGF-1 and IGF-2) mRNA levels in relation to the gastrointestinal tract (GIT) development in newborn calves. Twenty four male calves (5-day-old) were randomly allocated to 4 groups which were slaughtered at day 5, 12, 19 and 26 of their life, respectively. Tissue samples from the rumen, abomasum, duodenum, jejunum and liver were taken for analysis, and the level of IGF-1 and IGF-2 mRNA was determined using RT semi-quantitative PCR method. Both IGF-1 and IGF-2 mRNA level was the biggest on 5 day of life, significantly decreased up to day 12-19 in most of analyzed GIT parts, and started to increase thereafter. In the rumen the reduction in the amount of IGFs transcripts was associated with the reduction in papillae length and tunica muscularis thickness. The abomasum weight and tunica mucosa thickness increased from the 5 day up to 19 day of life, whereas abomasal IGF-1 mRNA level decreased together with calf's age up to 19 day of life, and the level of IGF-2 mRNA did not change. The reduction in IGFs mRNA level after 5 day of life in the duodenum (IGF-1 and IGF-2) and in the jejunum (IGF-1) was associated with reduction in villi length (duodenum and jejunum), and the increase of crypt depth (duodenum).


Cycloserine Induced Late Onset Psychosis and Ethambutol Induced Peripheral Neuropathy Associated with MDR-TB Treatment in an Indian Patient- A Rare Case Report.
Adverse reactions and toxicity inevitably accompany all treatment courses for drug-resistant TB. Our case underscores the importance of awareness regarding neuropsychiatric adverse reactions due to MDR-TB therapy and reversible nature of it. Cycloserine induced psychosis is most life threatening complication and sometimes could be fatal. A 42-year-old male on MDR-TB therapy got admitted for his persistent psychotic complaints like hallucinations, delusions and suicidal ideations, despite being treated with quetiapine/olanzapine. Eventually patient was rehabilitated, cycloserine was stopped and psychotic events regressed slowly. Other culprit drugs like ethambutol and levofloxacin causing psychosis was ruled out because there was no relapse of psychotic events despite being continued with these drugs. He also complained of tingling, numbness, swaying, pain and weakness. On examination, he had distal motor weakness in lower limbs, tandem gait positive, altered position sense, and tenderness over toes and positive Romberg's sign with ataxia. He was diagnosed to have drug induced sensorimotor peripheral neuropathy. All these symptoms persisted after stopping cycloserine and patient continued to have neuropathy with ethambutol and ethionamide. Considering the nature of neuropathy which was mild, mixed sensorimotor and resolved completely after 2-3 weeks of stopping, it was more in favour of ethambutol. However, we could not rule out the possibility of ethionamide or (ethionamide + ethambutol) causing neuropathy or both could have accelerated the neurotoxic effects of cycloserine which remained elusive.


Multiple sites for interaction of prostaglandin and vasopressin in toad urinary bladder.
The interaction of vasopressin with prostaglandins were examined in the toad bladder by determining water flows, cAMP levels, and cAMP-dependent protein kinase activity. Both water flow and activation of cAMP-kinase in response to vasopressin were enhanced after prostaglandin inhibition, consistent with inhibition of vasopressin-induced cAMP generation by endogenous prostaglandins. On the other hand exogeneous PGE stimulated cAMP generation. PGE1 (10(-7) M) alone did not increase water flow but activated kinase more than vasopressin only. Addition of PGE1 (10(-7) M) and vasopressin inhibited water flow as compared with vasopressin along but increased the kinase ratio above that with vasopressin only. PGE2 (10(-5) M) increased the cAMP content and kinase ratio even more than vasopressin but again resulted in no water flow. Addition of vasopressin and PGE2 (10(-5) M) increased water flow but did not alter cAMP content or the kinase ratio compared with PGE2 alone. Similar results were obtained with PGE1. Accordingly, prostaglandin dissociates cAMP levels and kinase ratio from the hydroosmotic response, suggesting that PGE2 inhibits steps distal to cAMP. Consistent with this, in bladders pretreated with naproxen or meclofenamate, PGE2 (10(-8) to 10(-6) M) inhibited the response to submaximal doses of cAMP (5 mM) or 8-bromo-cAMP (0.03 mM). Furthermore, pretreatment with naproxen significantly enhanced the response to cAMP (5 mM). These studies provide evidence for vasopressin-PGE interaction at the site of cAMP generation and also at a step(s) unrelated to cAMP generation.


Multiple metastatic malignant meningioma: a case report.
We report a case of a 27-year-old man who presented with a clinical history of swelling on the left side of his head. MRI scan showed a left frontal meningioma. The patient underwent a macroscopically complete resection. The histological diagnosis was malignant meningioma. Ten months after the initial diagnosis, multiple distant metastases were detected. Risk factors leading to metastasis in this case are discussed with a review of the literature.


Autonomous hyperparathyroidism not associated with increased aluminum absorption.
A single 6-g dose of aluminum hydroxide was given to eight patients on regular hemodialysis before and 3 days after parathyroidectomy (PTX). Serum aluminum levels were measured before and after 5 h after ingestion of aluminum hydroxide to examine indirectly the effects on absorption of aluminum from the gastrointestinal tract of both high and low parathyroid hormone (PTH). Plasma PTH levels fell significantly from 2,724 +/- 1,830 ng/l (mean +/- 1 SD) before PTX to 352 +/- 63 ng/l after PTX. However, no significant change was detected in serum aluminum levels before or after PTX (1.45 +/- 0.96 to 1.92 +/- 0.88 mumol/l before PTX; 2.02 +/- 1.41 to 2.89 +/- 1.19 mol/l after). These findings indicate that in patients with chronic renal failure, high plasma levels of PTH are not associated with significant increases in serum aluminum concentrations after ingestion of a large quantity of aluminum hydroxide. Parathyroid hormone may not enhance intestinal absorption of aluminum in humans.


Of mice and men (and women) and the acylation-stimulating protein pathway.
The storage and release of energy by adipocytes is of fundamental biologic importance. Not surprisingly, therefore, the rate at which these processes occur can be modulated by a variety of physiologic molecules. A newly recognized participant is produced by adipocytes themselves: acylation-stimulating protein (ASP). This article focuses on the most recent in-vivo evidence regarding how the ASP pathway may influence energy storage and release. In brief, the rate at which triglycerides are cleared from plasma (i.e. the rate at which they are hydrolysed) is determined by lipoprotein lipase and insulin, which is the principal hormone that regulates lipoprotein lipase. By contrast, the ASP pathway modulates the rate at which fatty acids are taken up and converted to triglycerides by adipocytes. Under certain circumstances, however, reduction of activity of the ASP pathway may negatively impact on the first step of the process. ASP also influences the rate at which fatty acids are released by adipocytes, and it is clear that insulin and ASP interact in a variety of ways that involve energy storage and release. Accordingly, to understand the impact of any intervention on energy storage and release by adipocytes, the effects of both insulin and ASP must be taken into account.


Persistence, Discordance and Diversity of Staphylococcus aureus Nasal and Oropharyngeal Colonization in School-aged Children.
The anterior nares are regarded as the primary site for Staphylococcus aureus colonization, although studies have highlighted the potential importance of colonization at extra-nasal sites, including the oropharynx. Accordingly, the aims of this study were to assess the prevalence, persistence and molecular epidemiology of S. aureus colonization in the nares and oropharynx of Māori and Pacific children, a population with strikingly high rates of S. aureus infection. A cross-sectional study of predominantly Māori and Pacific school-aged children in Auckland, New Zealand was performed in October 2013, and swabs were taken from the nares and oropharynx. Sampling was repeated from the same schools in October 2014. All S. aureus isolates underwent antimicrobial susceptibility testing and spa typing. Overall, 506/893 (56.7%) children were colonized with S. aureus, and the colonization prevalence was significantly higher in the oropharynx than nares (41.1% vs. 31.5%; P < 0.001). Longitudinal colonization was significantly higher in the oropharynx than the nares, and children with longitudinal oropharyngeal colonization were more likely to be colonized with the same spa type than those colonized in the nares (67.6% vs. 37.0%; P = 0.01). Approximately 40% of children had discordant spa types at the nares and oropharynx. Oropharyngeal S. aureus colonization represents a significant reservoir of S. aureus and it is possible that the oropharynx may represent a protected anatomical niche, enabling persistent colonization with the same S. aureus strain. Future study should attempt to better understand the determinants of oropharyngeal carriage.


Altered Global 5-Hydroxymethylation Status in Hidradenitis Suppurativa: Support for an Epigenetic Background.
The pathogenesis of hidradenitis suppurativa (HS), with its complex inflammatory network, is still elusive. Imbalances in DNA methylation can lead to genome destabilization and have been assumed to play a role in inflammatory diseases. Global DNA methylation and hydroxymethylation have not been studied in HS yet. We conducted this study to investigate the global DNA methylation and hydroxymethylation status in lesional and perilesional HS skin compared to healthy controls. Immunohistochemical analysis was performed for 5-methylcytosine (5-mC) and 5-hydroxymethylcytosine (5-hmC) in 30 lesional and 30 corresponding healthy-appearing perilesional HS tissue samples. We included 30 healthy subjects as an interindividual control group. 5-hmC levels were significantly lower in healthy-appearing perilesional (p < 0.0001) and lesional HS skin (p < 0.0001) when compared to healthy controls. There was no significant difference between lesional HS skin and perilesional HS skin regarding 5-hmC levels (p = 0.6654). In contrast to 5-hmC, 5-mC staining showed no significant changes between the 3 groups. Univariate analysis revealed no significant association between patients' characteristics, disease severity, and the levels of 5-mC and 5-hmC. Our findings indicate that imbalances in DNA hydroxymethylation may play a role in the pathogenesis of HS rather than DNA methylation. Further studies are warranted to investigate the significance of DNA hydroxymethylation and the regulating enzymes in HS in order to advance our knowledge of the inflammatory network in this disease.


Complications of eyelid reconstruction using a semicircular flap.
Complications of the semicircular flap technique of eyelid reconstruction include lateral canthal webbing, ectropion, lid notching, symblepharon, and tissue fullness of the lateral lid. These difficulties occur infrequently and rarely cause significant problems. Attention to surgical details, however, can minimize unfavorable results. It should be emphasized that the semicircular flap remains a superb technique for reconstruction of defects involving up to 80% of the lower eyelid.


Over-the-Counter Tinnitus "Cures": Marketers' Promises Do Not Ring True.
The Clinical Practice Guideline of the American Academy of Otolaryngology-Head & Neck Surgery (2014) stated that clinicians should not recommend dietary supplements for the treatment of tinnitus. The aim of this study is to characterize over-the-counter tinnitus remedies (OTCTR) on the U.S. market, describe the ingredients and prices, and characterize the methods of promoting these products. OTCTR were identified via Web search and visits to retail establishments. Information was collected regarding OTCTR chemical composition, product labeling, advertisements and marketing, price, and customers reviews. A wide array of unproven OTCTR exist on today's market. All make unfounded claims of relief from ear ringing. Most of the products considered in this study consist of mixtures of inexpensive and common vitamins, minerals, and/or herbs sold at a premium compared to similar preparations not expressly advertised for tinnitus. Certain brands, most notably Arches Tinnitus Formula (Arches Natural Products Inc., Salt Lake City, UT) and Lipo-Flavonoid (Clarion Brands Inc., Solon, OH), target otolaryngologists by advertising in specialty journals and prominently featuring supposed endorsement by "Ear-Nose-and-Throat Doctors" in their marketing. It is important for otolaryngologists who are caring for tinnitus sufferers to be aware that a robust and diverse market exists for unproven OTC tinnitus remedies. It is troubling that heavily advertised brands profess support by otolaryngologists. Responsible specialty organizations in the field should consider opposing such commercially motivated representations. Otolaryngology journals may wish to adopt a policy along the lines of The Journal of the American Medical Association publications to decline advertisements of dietary supplements that make unproven therapeutic claims. 5 Laryngoscope, 129:1898-1906, 2019.


Amphiphilic and nonamphiphilic forms of bovine and human dopamine beta-hydroxylase.
We show that human and bovine dopamine beta-hydroxylases (DBH) exist under three main molecular forms: a soluble nonamphiphilic form and two amphiphilic forms. Sedimentation in sucrose gradients and electrophoresis under nondenaturing conditions, by comparison with acetylcholinesterase (AChE), suggest that the three forms are tetramers of the DBH catalytic subunit and bind either no detergent, one detergent micelle, or two detergent micelles. By analogy with the Gna4 and Ga4 AChE forms, we propose to call the nonamphiphilic tetramer Dna4 and the amphiphilic tetramers Da4I and Da4II. In addition to the major tetrameric forms, DBH dimers occur as very minor species, both amphiphilic and nonamphiphilic. Reduction under nondenaturing conditions leads to a partial dissociation of tetramers into dimers, retaining their amphiphilic character. This suggests that the hydrophobic domain is not linked to the subunits through disulfide bonds. The two amphiphilic tetramers are insensitive to phosphatidylinositol phospholipase C, but may be converted into soluble DBH by proteolysis in a stepwise manner; Da4II----Da4I----Dna4. Incubation of soluble DBH with various phospholipids did not produce any amphiphilic form. Several bands corresponding to the catalytic subunits of bovine DBH were observed in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, but this multiplicity was not simply correlated with the amphiphilic character of the enzyme. In the case of human DBH, we observed two bands of 78 and 84 kDa. As previously reported by others, the presence of the heavy subunit characterizes the amphiphilic forms of the enzyme. We discuss the nature of the hydrophobic domain, which could be an uncleaved signal peptide, and the organization of the different amphiphilic and nonamphiphilic DBH forms. We present two models in which dimers may possess either one hydrophobic domain or two domains belonging to each subunit; in both cases, a single detergent micelle would be bound per dimer.


Estradiol: a protective factor in the adult brain.
Clinical studies suggest that estradiol acts as a protective factor in the adult brain. Postmenopausal women suffer from an increased risk of brain injury associated with neurodegenerative diseases such as Alzheimer's disease, and estrogen replacement therapy appears to decrease the risk and severity of this neurodegenerative condition. Studies using animal models show that estradiol exerts similar effects in rodents and can enhance cell survival. Therefore, we designed experiments to determine whether estradiol treatment can decrease brain injury induced by an experimental model of ischemia. Our experiments used a permanent middle cerebral artery occlusion model and physiological levels of estradiol replacement therapy. The results demonstrate that estradiol exerts profound protective effects against ischemic brain injury induced by cerebral artery occlusion and that this protective action correlates with changes in the level of gene expression of estradiol receptors and members of the Bcl-2 family. These data suggest that estrogen replacement therapy may provide important protection against age- and disease-related degenerative processes in the brain.


[Rights and responsibilities of the radiologist in determining indications and applying radiologic procedures].
A radiological examination or an interventional procedure is a medical invasion of the patient's freedom and does not constitute an offense of causing bodily harm there is a medical indication, when the patient has given consent after being adequately informed, and the intervention is carried out in accord with the valid rules of medical conduct. During the assessment of possible contraindications, the radiologist must also evaluate the indicationy, i.e., he or she must decide if the medical question can be answered by the requested radiological examination. The indication must be viewed the more critically the greater the risk for a complication is. A radiological diagnosis with the use of X-rays may only be employed when it will furnish information with therapeutic consequences. The more urgent the indication, the less strict are the requirements for informed consent and agreement, and vice versa. Thus, the information need for a purely diagnostic procedure without any direct therapeutic value is particularly high. The consent of the patient is only legally binding when he has been informed in advance of the objectives, extent, necessity and urgency, type, procedures, and alternatives of a medical investigation or therapeutic intervention and also of the accompanying risks. All risks on the basis of which a responsible patient can make the decision to consent or to reject a specific medical intervention are to be considered as relevant and must be explained appropriately.


Effect of Streptococcus uberis on Gamma Delta T Cell Phenotype in Bovine Mammary Gland.
In this study, we focused analyzing γδ T cells during bovine mammary gland inflammation induced by Streptococcus uberis. A mammary gland cell suspension was obtained using lavage 24, 48, 72, and 168 h after intramammary-induced infection. The proportion of lymphocytes increased during the entire week in which inflammation was present. The γδ T cells were also elevated during inflammation, reaching their peak at 72 h following induced inflammation. The percentage of apoptotic lymphocytes continually increased, with the highest proportion occurring 168 h after S. uberis infection. The results show that γδ T cells may be involved in the resolution of inflammation in bovine mammary glands, with the apoptosis of those cells potentially playing an important role.