[Reading behavior and preferences regarding subscriptions to scientific journals : Results of a survey of members of the German Society for General and Visceral Surgery].
The purpose of surgical literature is to publish the latest study results and to provide continuing medical education to readers. For optimal allocation of resources, institutional subscribers, professional societies and scientific publishers require structured data on reading and subscription preferences of potential readers of surgical literature. To obtain representative data on the preferences of German general and visceral surgeons regarding reading of and subscription to scientific journals. All members of the German Society for General and Visceral Surgery (DGAV) were invited to participate in a web-based survey. Questions were asked on the affiliation and position of the member, individual journal subscriptions, institutional access to scientific journals, preferences regarding electronic or print articles and special subscriptions for society members. Answers were descriptively analyzed. A total of 630 out of 4091 (15 %) members participated in the survey and 73 % of the respondents had at least 1 individual subscription to a scientific journal. The most frequently subscribed journal was Der Chirurg (47 % of respondents). The institutional access to journals was deemed insufficient by 48 % of respondents, predominantly in primary care hospitals and outpatient clinics. Almost half of the respondents gave sufficient importance to reading printed versions of articles for which they would pay extra fees. A group subscription for society members was perceived as advantageous as long as no relevant extra costs were incurred. This structured survey among members of the DGAV provides data on preferences regarding reading of and subscription to scientific journals. Individual subscriptions to journals are still common, possibly due to suboptimal institutional access particularly at smaller non-academic institutions. In an age of online publications it seems surprising that many respondents place a high value on printed versions. The results are relevant for potential institutional subscribers, professional societies and scientific publishers.


The identification of alpha-adrenergic receptors in adipose tissue by [3H]dihydroergocryptine binding.
The 3H-labelled alpha-adrenergic antagonist dihydroergocryptine has been found to bind specifically to hamster white adipocyte membranes. Binding is rapid and is inhibited by alpha but not beta antagonists. In addition, adrenergic agonists compete for this binding site in order of affinities equal to their activity as alpha-adrenergic agonists. The (-) stereoisomer of adrenaline is 10 times more potent than the (+) stereoisomer in inhibiting binding.


Supramolecular Hydrogels Fabricated from Supramonomers: A Novel Wound Dressing Material.
Daily repeated wound dressing changes will lead to additional trauma to newly formed tissue and prolonging the healing process. In this letter, we designed and fabricated an easily removable wound dressing material. To accomplish this, we first generated cross-linkable supramonomers through host-guest noncovalent interaction, followed by radical copolymerization of acrylamide with supramonomer as cross-linker to fabricate supramolecular hydrogels. Benefiting from the dynamic nature of the supramonomer, the supramolecular hydrogel is able to dissolve upon exposure to memantine, an FDA approved drug, making it easily removed from a wound, representing a promising candidate for the new generation of wound dressing.


Thermodynamic Differentiation of the Two Sides of Azabuckybowl through Complexation with Square Planar Platinum(II).
The precise control of the two faces, concave/convex faces, is an attractive challenge to realizing novel dynamic molecular systems. Herein, we report the synthesis, X-ray crystal structure, and bowl-to-bowl inversion behavior of a platinum complex with azabuckybowl as a monodentate ligand. X-ray crystallography revealed that the azabuckybowl is orthogonally coordinated to the plane containing the Pt center and other ligands. One and two-dimensional NMR studies have also confirmed that this complex was observed as mixtures of two isomers, although the isomeric ratio was highly biased. Theoretical calculations indicate that the difference in thermodynamic stability of these isomers is due to the direction of the concave/convex face of an azabuckybowl ligand.


A novel shape-locking guide for prevention of sigmoid looping during colonoscopy.
Looping of the endoscope in the sigmoid colon and other colonic segments often represents a significant challenge to the performance of comfortable, complete, and swift colonoscopy. This report describes the design and operation of a new device that addresses this problem, together with preliminary preclinical experience with this use of this shape-locking guide (SG-1). The shape-locking guide is an overtube that can be converted from a flexible to a rigid configuration on demand. When in the rigid configuration, the shape-locking guide is designed to protect the colon wall from lateral forces exerted by the colonoscope. The shape-locking guide was evaluated in vitro by using an artificial colon model to learn how to operate it, and to assess feasibility for prevention of colon looping. In addition, safety was assessed in vivo in a pig model. In vitro, the shape-locking guide prevented colonic looping and, thereby, aided completion of "colonoscopy" in the artificial colon model. Subsequent in vivo studies demonstrated that use of the shape-locking guide is safe and feasible; it performed well with respect to ease of insertion and avoidance of sigmoid looping. There was no evidence of significant injury to the colon or adjacent abdominal viscera. This preliminary study shows that use of the shape-locking guide is safe and that it has performance characteristics that may assist the performance of colonoscopy. Human trials are being undertaken.


[Measles: immunity and vaccination status in Greater Frankfurt on the Main].
The elimination of measles is a goal of the WHO which has not been achieved in Europe yet. As measles is a vaccine-preventable disease and can be eliminated by immunization the Standing Committee on Vaccination (STIKO) has updated the recommendation of measles vaccination. The goal of this study was to evaluate the immune status by serological testing and by raising immunisation status. Antibody-testing from the routine diagnostic of the Institute of Medical Virology (Frankfurt/Main) for patients, health care workers and medical students was investigated retrospectively. Overall all significant increase in seropositivity was shown from 64.8 to 86.2 %. Nevertheless immunity gaps still exist in population, especially in persons under 19 years of age. In 1999 80.7 % of the 5-9 year age group and 75.3 % of the 10-19 year age group were seropositive. In 2009 the percentage in these age groups has decreased to 68.2 % and 71.1 % respectively. By obtaining the immunity status of 230 travellers at our vaccination centre immunity gaps have been recognised as well. Only 29.6 % were completely immunized (at least two vaccination). 54.4 % have been vaccinated incomplete and in 16.1 % vaccination status was unknown. Our results indicated that measles immunization status and seroprevalence are still not sufficient to achieve the elimination of measles.


Synchronization stability of memristor-based complex-valued neural networks with time delays.
This paper focuses on the dynamical property of a class of memristor-based complex-valued neural networks (MCVNNs) with time delays. By constructing the appropriate Lyapunov functional and utilizing the inequality technique, sufficient conditions are proposed to guarantee exponential synchronization of the coupled systems based on drive-response concept. The proposed results are very easy to verify, and they also extend some previous related works on memristor-based real-valued neural networks. Meanwhile, the obtained sufficient conditions of this paper may be conducive to qualitative analysis of some complex-valued nonlinear delayed systems. A numerical example is given to demonstrate the effectiveness of our theoretical results.


Prognostic Impact of Discordance in Hormone Receptor Status Between Primary and Recurrent Sites in Patients With Recurrent Breast Cancer.
Recent retrospective studies have reported discordance rate of hormone receptor (HR) and human epidermal growth factor receptor 2 (HER2) statuses between primary and recurrent tumors and prognostic values of discordance. However, the results of these reports may possibly include analytical error. We analyzed 153 patients from whom pathological specimens of tumor tissues were available from both primary and recurrent sites. For all specimens, immunohistochemistry was performed for these statuses with a standardized method. Two experienced pathologists evaluated these specimens in a blinded fashion. The discordance rates for estrogen receptor, progesterone receptor, and HER2 were 18%, 26%, and 7%, respectively. Subtype changes based on HR and HER2 status occurred in 21% of patients. Clinical outcome was significantly worse in the patients with the tumors that were primarily HR-positive (HR(+)) converted to HR-negative (HR(-)) at recurrent sites than in the patients with the tumors in which HR status did not change or converted from HR(-) to HR(+) (P = .001). Clinical outcome was also significantly worse in the patients with the primarily HR(+) tumor that converted to triple negative in the recurrence sites than in the patients with a constantly HR(+) tumor (P < .001). By the Cox multivariate analyses, loss of HR expression and conversion to triple negative at the recurrence sites were independent indicators of worse clinical outcome. Discordance in HR and HER2 status often occurred between primary and recurrent breast cancer and had independent prognostic impact in the patients with recurrent breast cancer.


Glycosyltransferases in SWISS-PROT.
SWISS-PROT is a curated protein sequence database with a high level of annotation (such as description of the function of a protein, its domain structure, post-translational modification, variants, etc), a minimal level of redundancy and a high level of integration with other databases. An ongoing project is to maintain the glycosyltransferase family of enzymes with comprehensive annotation and documentation in the SWISS-PROT database and to represent the most recent research developments.


Effects of transport and racing on ionic changes in thoroughbred race horses.
1. Packed cell volume (PCV), blood glucose, total plasma proteins (TPP) and plasma electrolytes, osmolality, cortisol and aldosterone alterations produced by transport and racing, were investigated in race horses. 2. Plasma cortisol, sodium and blood glucose, found after transport, were higher, while aldosterone was lower than control levels. 3. After racing, PCV, blood glucose, TPP and plasma cortisol, sodium and osmolality were higher than control, while chloride diminished and aldosterone returned to control values. 4. These results demonstrate that transport and racing are different kinds of stressors, suggesting that the sympathetic system and hypophysis-suprarenal cortex axis have a dissimilar contribution to the physiological response.


Different sites of action of electrical and magnetic stimulation of the human brain.
The latency of the surface recorded electromyographic response to either an electrical or magnetic stimulus applied to the scalp has been measured in the first dorsal interosseous and abductor pollicis brevis muscles of 3 subjects. In the contracting muscle the response latency to the magnetic stimulus was longer by 1.4-2.7 ms compared to the electrical stimulus. Poststimulus time histograms of the firing of single motor units of first dorsal interosseous muscle were studied in 4 subjects. The first period of increased probability of firing of the single motor units showed a similar latency difference (mean 2.8 ms) to the two modes of stimulation. It is concluded that the extra delay to magnetic stimulation is consumed in central motor pathways. This implies that the two modes of stimulation activate the brain at different sites. It is suggested that the magnetic stimulus excites the corticospinal neurones transynaptically, whereas the electrical stimulus excites these neurones directly.


Is adipose tissue metabolic activity a predictor of pathological responses to neoadjuvant treatment in breast cancer.
Prediction of the pathologic response to neoadjuvant chemotherapy (NAC) in patients with locally advanced breast cancer is essential for optimal treatment strategy. The current approach of adjuvant or neoadjuvant treatment is based on the molecular subtype. Obesity may have affected chemotherapy response. This study aims to evaluate the relationship between metabolic activity of adipose tissue (AT) and pathological responses to NAC. And to define the association with body mass index (BMI) and metabolic parameters of standardized uptake value (SUV) of adipose tissue measured by positron emission computed tomography (PET/CT). One-hundred and sixteen consecutive patients with stage II and III breast cancer who underwent PET/CT before receiving NAC, were evaluated in the study. Metabolic parameters of visceral adipose tissue (VAT-SUV), subcutaneous adipose tissue (SAT-SUV), and calculated SUV of visceral-to-subcutaneous ratio (V/S-ratio) were regarded. The relationship between SUV of AT and pathologic response was evaluated from medical records retrospectively. Univariate-analysis revealed that good pathological response was significantly associated with clinical stage (P<.001), HER-2 positivity (P<.001), VAT-SUV (P=.037), VAT-density (P=.043) and V/S-ratio (P=.003). In multivariate-analysis clinical stage, HER-2 positivity and V/S-ratio were found to have statistically effect on pathological response. VAT-volume (P<.001), VAT-SUV (P=.016), SAT-volume (P<.001) and SAT-SUV (P<.001) has positive correlation with BMI value. On the other hand, V/S-ratio (P=.039) and SAT-density (P=.003) has negative correlation with BMI. Metabolic activity of AT is associated with BMI and effected chemotherapy responses. LowV/S ratio was associated with high BMI and poor pathological response to NAC. V/S ratio may be a useful marker for the prediction of NAC responses.


Cathepsin L--a latent proteinase in guinea pig sperm.
Guinea pig spermatozoa were found to contain a fully-latent cysteine proteinase that could be unmasked by incubating epididymal sperm for 2 hr at pH 3.5 and 37 degrees C. The proteinase was identified as cathepsin L (EC 22.214.171.124) on the basis of its optimal hydrolysis of benzyloxycarbonyl-Phe-Arg-7-(4-methyl)coumarylamide (Z-Phe-Arg-NMec) at pH 5.5; lack of action on Z-Arg-Arg-NMec and Arg-NMec; urea-enhanced digestion of azocasein; marked sensitivity to thiol reagents, leupeptin, Z-Phe-Phe-CHN2, and L-trans-epoxy-succinylleucylamido(3-methyl)butane (Ep-475 or E-64-c); and insensitivity to pepstatin and serine proteinase inhibitors. Gossypol, a male antifertility agent, was inhibitory. The unmasking phenomenon was reversibly inhibited by HgCl2 and mersalyl acid, and prevented by leupeptin and Ep-475, but not by pepstatin.


A numerical solution of the mechanical bidomain model.
The mechanical bidomain model predicts forces on integrin proteins in the membrane. It has been solved analytically for idealized examples, but a numerical algorithm is needed to address realistic problems. The bidomain equations are approximated using finite differences. An ischemic region is modeled as a circular area having no active tension, surrounded by normal tissue. The membrane force is large in the ischemic border zone, but is small elsewhere. Strain is distributed widely throughout the ischemic region and surrounding tissue. This calculation provides a testable prediction for the mechanism of mechanotransduction and remodeling in cardiac tissue.


Effects of soil drought and nitrogen deposition on BVOC emissions and their O3 and SOA formation for Pinus thunbergii.
Soil drought and nitrogen (N) deposition can influence the biogenic volatile organic compound (BVOC) emissions and thereby their ozone (O3) and secondary organic aerosol (SOA) formation. This study addressed their single and combined effects on BVOC emissions of Pinus thunbergii by laboratory simulation experiments. The results showed that light drought (LD, 50% soil volumetric water content (VWC)) stimulated isoprene, monoterpene, sesquiterpene, and total BVOC emissions, while moderate drought (MD, 30% and 40% VWC) and severe drought (SD, 10% and 20% VWC) inhibited their emissions (except for sesquiterpene in 20% VWC). N deposition decreased other VOC emissions and increased isoprene and sesquiterpene emissions. Total BVOCs and monoterpene were stimulated in low N deposition (LN, 2 g N/(m2·yr)) and inhibited in moderate (MN, 5 g N/(m2·yr)) and high N deposition (HN, 10 g N/(m2·yr)). Under combined treatment of soil drought and N deposition, total BVOC, monoterpene, and other VOC emissions were inhibited, sesquiterpene had no significant change, and isoprene emission was inhibited in MD combined treatment but promoted in SD. The O3 formation potential (OFP) and SOA formation potential (SOAP) from the changed BVOC emissions were calculated, OFP and SOAP of BVOC emissions and their compositions varied significantly among the treatments. Our study provided theoretical basis for assessing the impact of climate change and atmospheric pollution on BVOC emissions and their contribution to the formation of secondary atmospheric pollution.


Abstract art paintings, global image properties, and verbal descriptions: An empirical and computational investigation.
While global image properties (GIPs) relate to preference ratings in many categories of visual stimuli, this relationship is typically not seen for abstract art paintings. Using computational network science and empirical methods, we further investigated GIPs and subjective preferences. First, we replicated the earlier observation that GIPs do not relate to preferences for abstract art. Next, we estimated the network structure of abstract art paintings using two approaches: the first was based on verbal descriptions and the second on GIPs. We examined the extent to which network measures computed from these two networks (1) related to preference for abstract art paintings and (2) determined affiliation of images to specific art styles. Only semantic-based network predicted the subjective preference ratings and art style. Finally, preference and GIPs differed for sub-groups of abstract art paintings. Our results demonstrate the importance of verbal descriptors in evaluating abstract art, and that it is not useful in empirical aesthetics to treat abstract art paintings as a single category.


Biochemical characterisation and localization in brain of a human brain-leucocyte membrane glycoprotein recognised by a monoclonal antibody.
The F10-44-2 monoclonal antibody was originally shown to interact with a determinant found predominantly in human brain and leucocytes. In this study we demonstrate by quantitative absorption analysis with homogenates of the head of the caudate nucleus, putamen, thalamus, cerebral grey matter, cerebral white matter, corpus callosum and cerebellar folia that the determinant is restricted to the white matter of the CNS. Immunofluorescence studies on frozen sections of the above brain subregions confirm the absorption analyses, showing staining only of white matter. In addition, and unexpectedly, we found very bright staining around blood vessels, particularly in the cerebellum. Biochemical studies established that the molecule in white matter bearing the F10-44-2 determinants is a sialylated membrane glycoprotein with an apparent molecular weight of 90,000, which is similar to but slightly smaller than the T lymphocyte form of the antigen. Developmental studies comparing 16-week foetal and adult cerebrum showed a fivefold increase in F10-44-2 antigen content. Thus, in the human CNS, the F10-44-2 antigen is a medium-sized glycoprotein which is restricted to white matter and shows a marked increase in concentration during development. No such molecule has been described previously.


Association between CARD15/NOD2 gene polymorphisms and paratuberculosis infection in cattle.
Paratuberculosis represents a major problem in farmed ruminants and at the present is considered a potential zoonosis. The disease is caused by Mycobacterium avium subsp. paratuberculosis, and susceptibility to infection is suspected to have a genetic component. Caspase recruitment domain 15 (CARD15) gene encodes for a cytosolic protein implicated in bacterial recognition during innate immunity. Crohn's disease (CD) is an idiopathic inflammatory bowel disease in humans comparable in many features to bovine paratuberculosis involving an abnormal mucosal immune response. The association between mutations in the CARD15 gene and increased risk of Crohn's disease has been described. The objective of this candidate gene case-control study was to characterize the distribution of three polymorphisms in the bovine CARD15 gene and test their association with paratuberculosis infection in cattle. Three previously reported single nucleotide polymorphisms (E2[-32] intron 1; 2197/C733R and 3020/Q1007L) were screened for the study population (431 adult cows). The statistical analysis resulted in significant differences in allelic frequencies between cases and controls for SNP2197/C733R (P<0.001), indicating a significant association between infection and variant allele. In the analysis of genotypes, a significant association was also found between SNP2197/C733R and infection status (P<0.0001); cows with the heterozygous genotype were 3.35 times more likely to be infected than cows with the reference genotype (P=0.01). Results suggest a role for CARD15 gene in the susceptibility of cattle to paratuberculosis infection. These data contribute to the understanding of paratuberculosis, suggest new similarities with Crohn's disease and provide new information for the control of bovine paratuberculosis.


Transcriptional regulation of MHC class II genes.
MHC class II molecules play a fundamental role in the homeostasis of the immune response, functioning as receptors for antigenic peptides to be presented to regulatory T cells. Both quantitative and qualitative alterations in the expression of these molecules on the cell surface dramatically affect the onset of the immune response, and may be the basis of a wide variety of disease states, such as autoimmunity, immunodeficiencies, and cancer. Most regulation of MHC class II molecule expression is under the control of transcription mechanisms which are both cell type and development specific. In the last few years classical genetics together with molecular biology have greatly contributed to the widening of our knowledge on the regulatory mechanisms operating in the control of class II gene expression. This review deals with the latest developments in this fundamental area of immunology.


Role of renal biopsy in end stage renal failure.
The role of renal biopsy in 46 patients with end stage renal failure was assessed. Renal tissue was obtained by open renal biopsy in 24 patients, by needle biopsy in 5, and at bilateral nephrectomy in 17. In 4 patients the renal biopsy specimens showed advanced changes that could only be diagnosed as "end stage kidney". In 42 patients (91 per cent), a precise renal diagnosis was made. In 20 patients (43 per cent) the prebiopsy clinical impression differed from the histologic diagnosis. In the 29 patients undergoing renal biopsy, minor postoperative morbidity occurred in 5. We conclude that a renal biopsy in patients with end stage renal failure will result in a change in clinical diagnosis in a significant number of patients and can be performed with low morbidity.


Five oximes (K-27, K-33, K-48, BI-6 and methoxime) in comparison with pralidoxime: in vitro reactivation of red blood cell acetylcholinesterase inhibited by paraoxon.
Oximes are cholinesterase reactivators of use in poisoning with organophosphorus compounds. Pralidoxime (PRX) is used clinically as an adjunct to atropine in such exposure. Clinical experience with PRX (and other oximes) is, however, disappointing and routine use has been questioned. In addition it is known that oximes are not equally effective against all existing organophosphorus compounds. There is a clear demand for 'broad spectrum' cholinesterase reactivators with a higher efficacy than PRX. Over the years new reactivators of cholinesterase of potential clinical utility have been developed. Their chemical structures were derived from those of existing esterase reactivators, especially pralidoxime, obidoxime and HI-6. The purpose of the study was to quantify in vitro the extent of oxime (pralidoxime, K-27, K-33, K-48, methoxime and BI-6) conferred protection, using paraoxon as an inhibitor. Paraoxon (POX), the active metabolite of parathion (O,O-diethyl-O-p-nitro-phenyl phosphorothioate) is a non-neuropathic organophosphate. Red blood cell (RBC) acetylcholinesterase (AChE) activities in whole blood were measured photometrically in the presence of different POX concentrations and the IC50 was calculated. Determinations were repeated in the presence of increasing oxime concentrations. The IC50 of POX increases with the oxime concentration in a linear manner. The calculated IC50 values were plotted against the oxime concentrations to obtain an IC50 shift curve. The slope of the shift curve (tg alpha) was used to quantify the magnitude of the protective effect (nm IC50 increase per microm reactivator). Based on our determinations the new K series of reactivators is far superior to pralidoxime, methoxime and BI-6, K-27 being the outstanding compound with a tg alpha value of 3.7 (nm IC50 increase per microm reactivator) which is approximately 13 times the reactivator ability of PRX. In general there is an (expected) inverse relationship between the binding constant K and the slope of the IC50 shift curve (tg alpha) for all oximes examined. K-27 (the most protective substance judging by the tg alpha) has the lowest K value (highest affinity). In vivo testing of the new oximes as an organophosphate protective agent is necessary.


Outward- and inward-facing structures of a putative bacterial transition-metal transporter with homology to ferroportin.
In vertebrates, the iron exporter ferroportin releases Fe(2+) from cells into plasma, thereby maintaining iron homeostasis. The transport activity of ferroportin is suppressed by the peptide hormone hepcidin, which exhibits upregulated expression in chronic inflammation, causing iron-restrictive anaemia. However, due to the lack of structural information about ferroportin, the mechanisms of its iron transport and hepcidin-mediated regulation remain largely elusive. Here we report the crystal structures of a putative bacterial homologue of ferroportin, BbFPN, in both the outward- and inward-facing states. Despite undetectable sequence similarity, BbFPN adopts the major facilitator superfamily fold. A comparison of the two structures reveals that BbFPN undergoes an intra-domain conformational rearrangement during the transport cycle. We identify a substrate metal-binding site, based on structural and mutational analyses. Furthermore, the BbFPN structures suggest that a predicted hepcidin-binding site of ferroportin is located within its central cavity. Thus, BbFPN may be a valuable structural model for iron homeostasis regulation by ferroportin.


Detection and purification of glucose 6-phosphate dehydrogenase, malic enzyme, and NADP-dependent isocitrate dehydrogenase by blue native polyacrylamide gel electrophoresis.
We describe a blue native polyacrylamide gel electrophoretic technique that allows the facile detection, quantitation and purification of three NADPH-producing enzymes. Glucose 6-phosphate dehydrogenase, malic enzyme and NADP-dependent isocitrate dehydrogenase were detected simultaneously. Activity staining based on the formation of NADPH from the respective substrates and the subsequent precipitation of formazan enabled the relative quantitation of enzymatic activities, while Coomassie staining on one-dimensional or two-dimensional gels helped monitor the amount of protein associated with these enzymatic activities. This technique provides a simple and effective route to obtain homogeneous protein for further analyses and also enables the screening of these NADPH-producing enzymes in various cellular systems.


Morphological transformation of early passage golden Syrian hamster embryo cells derived from cryopreserved primary cultures as a reliable in vitro bioassay for identifying diverse carcinogens.
Cryopreserved primary cultures of golden Syrian hamster embryo cells were used as the source of target and feeder cells for establishing an in vitro carcinogenesis bioassay. The primary culture giving the best overall response in a pretest before freezing gave positive results in 20 consecutive experiments when retested with 3-methylcholanthrene after cryopreservation, indicating that pretested cryopreserved cultures can serve as a source of susceptible target cells in an in vitro carcinogenesis bioassay. Similarly prepared and cryopreserved cultures served satisfactorily as feeder cells. Susceptible positive cultures were used to test a large number of carcinogenic and non-carcinogenic chemicals in this system. The results showed a very high positive correlation (90.8%) between morphological transformation and the reported carcinogenic activity of the chemicals. Transformation was not observed when cells were tested with a few carcinogens that may not be metabolized to their active forms by early passage hamster embryo cells. N-2-acetylaminofluorene transformed cells only when tested in the presence of hamster liver microsomes. No false positive results were obtained when non-carcinogens were bioassayed, nor was spontaneous transformation observed in control cultures treated with medium alone, 0.2% dimethylsulfoxide or other solvents. Cultures derived from morphologically transformed colonies arising after treatment of cells with several known carcinogens were tumorigenic in vivo, confirming the correlation of morphological transformation with tumorigenicity and the validity of altered morphology as an in vitro criterion for carcinogenicity in vivo.


A Network of Phosphate Starvation and Immune-Related Signaling and Metabolic Pathways Controls the Interaction between Arabidopsis thaliana and the Beneficial Fungus Colletotrichum tofieldiae.
The beneficial root-colonizing fungus Colletotrichum tofieldiae mediates plant growth promotion (PGP) upon phosphate (Pi) starvation in Arabidopsis thaliana. This activity is dependent on the Trp metabolism of the host, including indole glucosinolate (IG) hydrolysis. Here, we show that C. tofieldiae resolves several Pi starvation-induced molecular processes in the host, one of which is the downregulation of auxin signaling in germ-free plants, which is restored in the presence of the fungus. Using CRISPR/Cas9 genome editing, we generated an Arabidopsis triple mutant lacking three homologous nitrilases (NIT1 to NIT3) that are thought to link IG-hydrolysis products with auxin biosynthesis. Retained C. tofieldiae-induced PGP in nit1/2/3 mutant plants demonstrated that this metabolic connection is dispensable for the beneficial activity of the fungus. This suggests that either there is an alternative metabolic link between IG-hydrolysis products and auxin biosynthesis, or C. tofieldiae restores auxin signaling independently of IG metabolism. We show that C. tofieldiae, similar to pathogenic microorganisms, triggers Arabidopsis immune pathways that rely on IG metabolism as well as salicylic acid and ethylene signaling. Analysis of IG-deficient myb mutants revealed that these metabolites are, indeed, important for control of in planta C. tofieldiae growth: however, enhanced C. tofieldiae biomass does not necessarily negatively correlate with PGP. We show that Pi deficiency enables more efficient colonization of Arabidopsis by C. tofieldiae, possibly due to the MYC2-mediated repression of ethylene signaling and changes in the constitutive IG composition in roots.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.


Super-oxidized "activated graphene" as 3D analogue of defect graphene oxide: Oxidation degree vs U(VI) sorption.
Porous carbons are not favorable for sorption of heavy metals and radionuclides due to absence of suitable binding sites. In this study we explored the limits for surface oxidation of "activated graphene" (AG), porous carbon material with the specific surface area of ∼2700 m2/g produced by activation of reduced graphene oxide (GO). Set of "Super-Oxidized Activated Graphene" (SOAG) materials with high abundance of carboxylic groups on the surface were produced using "soft" oxidation. High degree of oxidation comparable to standard GO (C/O=2.3) was achieved while keeping 3D porous structure with specific surface area of ∼700-800 m2/. The decrease in surface area is related to the oxidation-driven collapse of mesopores while micropores showed higher stability. The increase in the oxidation degree of SOAG is found to result in progressively higher sorption of U(VI), mostly related to the increase in abundance of carboxylic groups. The SOAG demonstrated extraordinarily high sorption of U(VI) with the maximal capacity up to 5400 μmol/g, that is 8.4 - fold increase compared to non-oxidized precursor AG, ∼50 -fold increase compared to standard graphene oxide and twice higher than extremely defect-rich graphene oxide. The trends revealed here show a way to further increase sorption if similar oxidation degree is achieved with smaller sacrifice of surface area.


Prenatal diagnosis of partial trisomy 3q resulting from t(3;14) in a fetus with multiple anomalies including vermian hypoplasia.
While genetic origin of Dandy-Walker complex has not yet fully elucidated, the complex has been known to be associated with structural and chromosomal abnormalities. A partial trisomy 3q was also identified in patients with DWC. 3q duplication syndrome is defined as duplications of large parts of 3q, especially 3q21-qter. Most cases with 3q duplication are diagnosed postnatally and the patients show typical features including various facial dysmorphisms, congenital heart defects, genitourinary malformations, and mental and growth retardation. Here we report a 28 year old nulliparous woman who was referred from the infertility clinic at 21 gestational weeks. Fetal ultrasonographic examination showed various abnormal findings including a ventricular septal defect, hydrocephalus, and hypoplasia of the cerebellar vermis. Fetal chromosome analysis was initially reported as 46,XY,der(14)(?::p11.2→qter). Array CGH followed by FISH allowed precise characterization of the der(14) chromosome and the initial karyotype of the fetus had been changed to 46,XY,add(14)(p11).ish der(14)t(3;14)(q26.1;p11)(tel3q+).arr 3q26.1q29(166249469-199288361)x3. Though further studies are required, gene clusters rather than a single gene might be responsible for the clinical features of the Dandy-Walker complex.


Hereditary hemorrhagic macular dystrophy.
Four members of a family with dominantly inherited macular dystrophy demonstrated the essential features of hereditary hemorrhagic macular dystrophy. They reported decreased visual acuity in the third decade and eventually developed bilateral disease. Fundus evaluations revealed hemorrhagic and exudative maculopathy associated with pigment atrophy, pigment clumping, and eventual glial scar formation. Early stages of the disease manifested a macular subretinal neovascular network. Photocoagulation treatment of subretinal neovascularization was successful in Case 3, unsuccessful in Case 1. Follow-up of these four brothers lasted over a period of two to 12 years. After they reached the cicatricial phase, the size of the lesion and visual symptoms remained stable. Visual function tests in the early and late stages indicated a local or geographic disease. We differentiated this dystrophy from other hereditary causes for subretinal neovascularization. We recommend early obliteration of the subretinal neovascular network with intense photocoagulation because the outcome of untreated hereditary hemorrhagic macular dystrophy is legal blindness.


Supplemental Nutrition Assistance Program Education reductions during COVID-19 may have exacerbated health inequities.
Describe, and assess disparities in, the changes in Supplemental Nutrition Assistance Program Education (SNAP-Ed) that occurred the year before vs. the year when COVID-19 restrictions were implemented. Observational study comparing reach, intensity, and dose of California Local Health Department (LHD) SNAP-Ed interventions in Federal Fiscal years 2019 and 2020 (FFY19, FFY20). Student t-tests determined significance of differences in the number of Direct Education (DE) programs, Policy, Systems and Environmental change (PSE) sites, people reached, and intervention intensity and dose between FFY19 and FFY20 using data reported online by LHDs. Linear regression assessed associations between census tract-level characteristics (urbanicity; percentages of population with income <185% of federal poverty level, under 18 years of age, and belonging to various racial/ethnic groups; and California Healthy Places Index) and changes in number of DE programs, PSE sites, people reached, and intervention dose between FFY19 and FFY20. From FFY19 to FFY20, the number of DE programs, PSE sites, people reached, and census tract-level intervention intensity and dose decreased. Higher census tract poverty, higher proportions of Black and Latino residents, and less healthy neighborhood conditions were associated with greater decreases in some intervention characteristics including PSE sites, PSE reach, DE programs, and DE dose. These reductions in LHD SNAP-Ed interventions indicate reduced access to education and environments that support healthy eating and obesity prevention during a time when this support was especially needed to reduce risk of COVID-19 infection and complications. Disproportionately reduced access, may have worsened health disparities in already-disadvantaged communities. Assuring maintenance of SNAP-Ed interventions, especially in disadvantaged communities, should be a priority during public health emergencies.


Diagnostic yield of uroendoscopy compared to ultrasonography for evaluating lower urinary tract disorders in dogs.
Cystourethroscopy and vaginoscopy (uroendoscopy) is often used in the diagnostic evaluation of dogs with lower urinary tract disorders (LUTD). To evaluate if uroendoscopy is warranted in dogs with various LUTD, the agreement between uroendoscopic and ultrasonographic diagnoses were compared. Dogs with recurrent urinary tract infections (rUTI) will have the highest diagnostic agreement between uroendoscopy and ultrasonography (US) compared to dogs presenting for other LUTD. Two hundred thirty-seven dogs presenting between 2014 and 2019 with lower urinary tract signs (LUTS) that had US within 60 days preceding uroendoscopy. Retrospective study. Dogs were categorized by primary indication for ultrasound. Pertinent uroendoscopic findings were recorded and agreements (κ analysis) between the final uroendoscopic diagnosis were compared with the final ultrasonographic diagnosis. Pertinent uroendoscopic findings were recorded for 69/237 (29%) cases. For dogs presenting primarily for urinary incontinence (UI), agreement between uroendoscopy and US was 71% (46/65; κ = 0.47, 95% CI 0.28-0.66), for dogs with stranguria, 58% (29/50; κ = 0.47, 95% CI 0.31-0.62) and for dogs with rUTI the agreement was substantial at 87% (26/30; κ = 0.70, 95% CI 0.43-0.98). Urethral strictures were the majority (14/21; 67%) of pertinent uroendoscopic findings for dogs with stranguria, of which 12 were male dogs. Agreement between uroendoscopy and US was moderate for all dogs. Based on these data, recommendation for uroendoscopy should be tailored to individual clinical presentation and signalment; transabdominal US is not the preferred modality for urethral lesions.


Enzymatic hydrolysis optimization of microwave alkali pretreated wheat straw and ethanol production by yeast.
Microwave alkali pretreated wheat straw was used for in-house enzyme production by Aspergillusflavus and Trichodermareesei. Produced enzymes were concentrated, pooled and assessed for the hydrolysis of pretreated wheat straw. Factors affecting hydrolysis were screened out by Placket-Burman design (PBD) and most significant factors were further optimized by Box-Behnken design (BBD). Under optimum conditions, 82% efficiency in hydrolysis yield was observed. After the optimization by response surface methodology (RSM), a model was proposed to predict the optimum value confirmed by the experimental results. The concentrated enzymatic hydrolyzate was fermented for ethanol production by Saccharomyces cerevisiae, Pichia stipitis and co-culture of both. The yield of ethanol was found to be 0.48 g(p)/g(s), 0.43 g(p)/g(s) and 0.40 g(p)/g(s) by S. cerevisiae, P. stipitis and by co-culture, respectively, using concentrated enzymatic hydrolyzate. During anaerobic fermentation 42.31 μmol/mL, 36.69 μmol/mL, 43.35 μmol/mL CO(2) was released by S. cerevisiae, P. stipitis and by co-culture, respectively.


Smoking asthma phenotype: diagnostic and management challenges.
The prevalence of active smoking in adults with asthma is similar to the general population. Smoking asthma is associated with poorer disease control, impaired response to corticosteroid therapy, accelerated decline in lung function, and increased rate of healthcare utilization. Current asthma guidelines do not provide specific treatment advice for smoking asthmatic patients. There is an urgent need for better understanding of the underlying mechanisms and effective treatment for smoking asthmatic patients. An association between both active and passive smoking and adult-onset asthma is supported by many studies.The asthma-COPD overlap syndrome (ACOS) has recently gained particular interest and smoking asthmatic patients should be evaluated for ACOS.Treatment regimens for smoking asthma include higher doses of inhaled corticosteroids (ICS), extrafine particle ICS formulations, antileukotrienes, and combinations of these options.Asthma is associated with increased risk of cardiovascular comorbidities whereas smoking is an additional strong independent risk factor for pulmonary and cardiovascular diseases. Tobacco smoking and not asthma per se seems to be the reason of poor prognosis, especially with regard to lung cancer, cardiovascular diseases, and mortality in asthmatic patients. Smoking asthma represents a common challenge to the clinician both in terms of diagnosis and management. These aspects have not been thoroughly evaluated and deserve further investigation.


Transduction of full-length dystrophin to multiple skeletal muscles improves motor performance and life span in utrophin/dystrophin double knockout mice.
Duchenne muscular dystrophy (DMD) is a fatal, progressive, muscle-wasting disease caused by defects in the dystrophin. No viral vector except the helper-dependent adenovirus vector (HDAdv) can package 14-kilobase (kb) full-length dystrophin complementary DNA (cDNA), and HDAdv is considerably safer than old-generation adenovirus vectors because of the large-size deletion in its genome. We have generated HDAdv that carries myc-tagged murine full-length dystrophin cDNA (HDAdv-myc-mFLdys). We injected it into multiple proximal muscles of 7-day-old utrophin/dystrophin double knockout mice (dko mice) (which typically show symptoms quite similar to human DMD) because the proximal muscles are affected in DMD patients. Eight weeks after the injections, the transduced dystrophin was widely expressed, and we found a significant reduction in centrally nucleated myofibers and the restoration of the dystrophin-associated proteins, beta-dystroglycan (beta-DG) and alpha-sarcoglycan (alpha-SG), as well as neuronal nitric oxide synthase (nNOS). The injected dko mice also showed an increase in body weight, an improvement in motor performance, and a prolongation of life span. Using HDAdv, we could treat DMD model mice even by transferring the therapeutic gene into multiple skeletal muscles. Our results suggest that multiple intramuscular administrations of HDAdv carrying full-length dystrophin cDNA may reduce symptoms and compensate for lost functions in DMD patients.


Subcutaneous erythropoietin therapy: efficacy and economic implications.
Recent studies have suggested that lower doses of erythropoietin (rHuEPO) may be required to achieve a target hematocrit when the hormone is administered subcutaneously compared with intravenously. In this article, previous studies are reviewed with a focus on the recently completed Department of Veterans Affairs multicenter trial comparing the two routes of administration. In this randomized, parallel group study of 208 patients, the average rHuEPO dose required to maintain a target hematocrit of 30% to 33% for 26 weeks was 95.1 +/- 75.0 U/kg/wk in the subcutaneous-therapy group and 140.3 +/- 88.5 U/kg/wk in the intravenous-therapy group (P < 0.0001). The specific mechanisms that result in the greater efficiency of the subcutaneous route are unknown but are probably related to the prolonged half-life of the hormone with subcutaneous administration. Possible mechanisms resulting in greater efficiency with subcutaneous therapy include sustained stimulation of the erythroid progenitor cells, diminished inhibition of erythropoiesis by proinflammatory cytokines, and prevention of neocytolysis, the hemolysis of newly formed red blood cells. Because most hemodialysis patients in the United States are receiving rHuEPO by the intravenous route, switching to the subcutaneous route may result in significant cost savings for the health care system.


Transfer and expression of a murine UDP-Gal:beta-D-Gal-alpha 1,3-galactosyltransferase gene in transfected Chinese hamster ovary cells. Competition reactions between the alpha 1,3-galactosyltransferase and the endogenous alpha 2,3-sialyltransferase.
The cDNA encoding a murine UDP-Gal:beta-D-Gal-alpha 1,3-galactosyltransferase has recently been cloned and sequenced using a transient expression method (Larsen, R.D., Rajan, V.P., Ruff, M.M., Kukowska-Latallo, J., Cummings, R.D., and Lowe, J.B. (1989) Proc. Natl. Acad. Sci. U.S.A. 86, 8227-8231). This report describes the construction and analysis of a Chinese hamster ovary (CHO) cell line in which in vitro expression alpha 1,3-galactosyltransferase activity has been achieved via transfer and expression of the murine alpha 1,3-galactosyltransferase gene. A primary aim of this research was to explore the role of the alpha 1,3-galactosyltransferase in regulating glycoprotein and glycolipid biosynthesis. CHO cells were cotransfected with murine genomic DNA fragments from F9 cells and plasmid DNA containing a resistance gene to the antibiotic G418. Cells resistant to G418 were then selected for expression of surface glycoconjugates containing terminal alpha 1,3-galactosyl residues by isolating cells bound to immobilized Griffonia simplicifolia-I-B4, a lectin which binds to alpha 1,3-galactosyl residues. A positive, stable transfectant clone, designated Clone 3, was obtained and analyzed for expression of the murine of alpha 1,3-galactosyltransferase. Fluorescence-activated cell sorting demonstrated that Clone 3, but not parental, CHO cells bound significant amounts of fluorescein isothiocyanate-labeled G. simplicifolia-I-B4. Southern and Northern blot analyses using the murine alpha 1,3-galactosyltransferase cDNA demonstrated that clone 3, but not parental, CHO cells contain murine alpha 1,3-galactosyltransferase genomic DNA sequences, and express a homologous transcript that comigrates with the authentic 3.6 kilobase alpha 1,3-galactosyltransferase murine mRNA. Enzyme assays confirmed that clone 3, but not parental CHO cells, contained the alpha 1,3-galactosyltransferase activity and that the level of activity is comparable to that found in F9 cells. [3H]Galactose-labeled glycopeptides and glycolipids were obtained from metabolically radiolabeled parental and Clone 3 cells and were analyzed for the presence of terminal alpha 1,3-galactosyl residues. Complex-type, Asn-linked oligosaccharides from both parental and Clone 3 cells contain the repeating disaccharide [3Gal beta 1, 4GlcNAc beta 1]n or poly-N-acetyllactosamine sequences, but only the poly-N-acetyllactosamine chains from clone 3 cells contained the terminal sequence Gal alpha 1,3Gal beta 1,4GlcNAc beta 1-R.(ABSTRACT TRUNCATED AT 400 WORDS)


Sphingomyelin and progression of renal and coronary heart disease in individuals with type 1 diabetes.
Lipid abnormalities are associated with diabetic kidney disease and CHD, although their exact role has not yet been fully explained. Sphingomyelin, the predominant sphingolipid in humans, is crucial for intact glomerular and endothelial function. Therefore, the objective of our study was to investigate whether sphingomyelin impacts kidney disease and CHD progression in individuals with type 1 diabetes. Individuals (n = 1087) from the Finnish Diabetic Nephropathy (FinnDiane) prospective cohort study with serum sphingomyelin measured using a proton NMR metabolomics platform were included. Kidney disease progression was defined as change in eGFR or albuminuria stratum. Data on incident end-stage renal disease (ESRD) and CHD were retrieved from national registries. HRs from Cox regression models and regression coefficients from the logistic or linear regression analyses were reported per 1 SD increase in sphingomyelin level. In addition, receiver operating curves were used to assess whether sphingomyelin improves eGFR decline prediction compared with albuminuria. During a median (IQR) 10.7 (6.4, 13.5) years of follow-up, sphingomyelin was independently associated with the fastest eGFR decline (lowest 25%; median [IQR] for eGFR change: <-4.4 [-6.8, -3.1] ml min-1 [1.73 m-2] year-1), even after adjustment for classical lipid variables such as HDL-cholesterol and triacylglycerols (OR [95% CI]: 1.36 [1.15, 1.61], p < 0.001). Similarly, sphingomyelin increased the risk of progression to ESRD (HR [95% CI]: 1.53 [1.19, 1.97], p = 0.001). Moreover, sphingomyelin increased the risk of CHD (HR [95% CI]: 1.24 [1.01, 1.52], p = 0.038). However, sphingomyelin did not perform better than albuminuria in the prediction of eGFR decline. This study demonstrates for the first time in a prospective setting that sphingomyelin is associated with the fastest eGFR decline and progression to ESRD in type 1 diabetes. In addition, sphingomyelin is a risk factor for CHD. These data suggest that high sphingomyelin level, independently of classical lipid risk factors, may contribute not only to the initiation and progression of kidney disease but also to CHD. Graphical abstract.


A comparison of the Food and Drug Administration's and Health Canada's regulatory decisions about failed confirmatory trials for oncology drugs: an observational study.
Oncology drugs are frequently approved on the basis of surrogate outcomes that require further trials to confirm the benefits, but at times these trials fail and regulators need to decide whether to withdraw approval for the indication and/or to remove the drug from the market. This study compares decisions by the Food and Drug Administration and Health Canada about oncology drugs that were approved using either Accelerated Approval (FDA) or Notice of Compliance with conditions (NOC/c, Health Canada) and that failed confirmatory trials. Drug/indications approved by the FDA through its Accelerated Approval Pathway and that later failed confirmatory studies were identified from a published study and additional information on these drugs was collected from Drugs@FDA. Health Canada websites were searched on September 11, 2021 for the same group of drugs to determine if they were approved in Canada under the NOC/c pathway for the same indication as in the US. Information from both the FDA and Health Canada about these products was entered into an Excel spreadsheet. Decisions about whether to withdraw the drugs or remove the failed indication for the drug and requirements for confirmatory studies were compared. In addition, the dates of decisions by the two agencies were compared. Ten drug/indications were available for comparison. Regulatory decisions were similar in 4 cases, different in 1 case and could not be determined in the remaining 5, in 1 case because decisions were pending in both countries and in the other 4, because the NOC/c had not been fulfilled in Canada. The requirements for the confirmatory studies were similar in both countries. Decisions were made earlier in the United States. This study shows that decisions made by Health Canada and the FDA about whether to withdraw a drug or remove a failed indication when drug/indications fail a confirmatory trial are usually similar, although the sample size on which this conclusion is made is small. The clinical implications of these similarities and differences should be explored.


Ion-exchange displacement chromatography of proteins, using narrow-range carboxymethyldextrans and a new index of affinity.
A new method of characterizing carboxymethyldextrans with respect to their relative affinities for an anion exchanger can be applied not only to pure preparations but also to fractions of a displacement chromatogram in which these polyanions have been used as spacers. This, along with the development of a means of preparing carboxymethyldextrans with narrow ranges of affinity, as contrasted with the very heterogeneous preparations previously used, has greatly facilitated the focusing of resolving power on critical regions of the chromatogram. The effectiveness of this approach is illustrated by the separation of proteins that differ only slightly in affinity, using the genetic variants of beta-lactoglobulin as one model and the numerous components of purified ovalbumin as another.


Peripheral Monocyte Percentage as a Potential Indicator of Prognosis in Patients with Chronic Subdural Hematoma Receiving Conservative Therapy.
Studies have confirmed active and abnormal inflammation in the hematoma cavity of chronic subdural hematoma (CSDH). However, a relationship between the peripheral blood status and the prognosis of CSDH patients has not been demonstrated. We retrospectively analyzed 245 CSDH patients who received conservative therapy (67 under close follow-up observation, 103 treated with atorvastatin, and 75 treated with atorvastatin combined with dexamethasone) from 2014 to 2021 to evaluate the role of major inflammation-associated cells in the prognostic assessment of patients. Univariate and multivariate analyses were performed to assess the potential factors that could indicate the prognosis among the 103 patients who underwent observation only or atorvastatin therapy. Changes in peripheral blood inflammation-associated cells at different time points were compared between patients with good and poor outcomes. Furthermore, the changes in inflammatory cells in 75 patients who received atorvastatin combined with dexamethasone were analyzed. The monocyte percentage was the only independent influencing factor in subsequent follow-up assessments. Patients with good outcomes had obviously lower circulating monocyte percentages in their peripheral blood counts throughout the treatment period. The monocyte percentage was also significantly decreased in the patients who responded well to atorvastatin combined with dexamethasone. The peripheral monocyte percentage was significantly higher in patients who transitioned to surgery because of a poor response to pharmacotherapy. The peripheral monocyte percentage may be a convenient and effective indicator for predicting the outcome of CSDH for patients receiving conservative treatment. A higher percentage of monocytes could be a risk factor for a poor response.


Novel terephthaloyl thiourea cross-linked chitosan hydrogels as antibacterial and antifungal agents.
Four novel terephthaloyl thiourea chitosan (TTU-chitosan) hydrogels were synthesized via a cross-linking reaction of chitosan with different concentrations of terephthaloyl diisothiocyanate. Their structures were investigated by elemental analyses, FTIR, SEM and X-ray diffraction. The antimicrobial activities of the hydrogels against three species of bacteria (Bacillis subtilis, Staphylococcus aureus and Escherichia coli) and three crop-threatening pathogenic fungi (Aspergillus fumigatus, Geotrichum candidum and Candida albicans) are much higher than that of the parent chitosan. The hydrogels were more potent in case of Gram-positive bacteria than Gram-negative bacteria. Increasing the degree of cross-linking in the hydrogels resulted in a stronger antimicrobial activity.


Phentolamine and structurally related compounds selectively antagonize the vascular actions of the K+ channel opener, cromromakalim.
1. The effects of cromakalim, a novel vasodilator agent believed to open K+ channels, were studied in a range of large and small arteries in vitro. In dog isolated coronary artery, precontracted with U46619 (a thromboxane A2-mimetic), cromakalim caused concentration-dependent relaxation which could be inhibited by phentolamine (10-100 microM). 2. The ability of phentolamine to antagonize cromakalim was selective since it did not affect responses to a number of other vasodilators including isoprenaline, nitroprusside or nicorandil. 3. The effect of phentolamine was not related to its alpha-adrenoceptor blocking actions since other alpha-adrenoceptor antagonists (prazosin 10 microM, rauwolscine 10 microM and phenoxybenzamine 1 microM) failed to influence the action of cromakalim. 4. A number of compounds structurally related to phentolamine were also able to block the vaso-relaxant response to cromakalim in the dog isolated coronary artery. The rank order of potency was alinidine = phentolamine = ST91 greater than tramazoline = naphazoline. Clonidine and tolazoline were inactive. The most potent compounds (alinidine and phentolamine) were effective only at concentrations above 1 microM. 5. Electrophysiological studies, in which resting membrane potential and tension were measured simultaneously, were carried out on rat isolated femoral artery. Phentolamine (30 microM) antagonized both the vasorelaxation and hyperpolarization caused by cromakalim. 6. These results suggest that phentolamine and some structurally related compounds, may inhibit K+ channel opening, an action which would account for their ability to antagonize the actions of cromakalim. Such compounds may prove useful in determining the role of K+ channels in regulating vascular smooth muscle tone in vivo and in vitro.


Novel Tretinoin 0.05% Lotion for the Once-daily Treatment of Moderate-to-Severe Acne Vulgaris in an Adult and Adolescent Female Population
Background: Acne vulgaris (acne) is a common dermatological condition typically associated with adolescents, affecting about 85% of young people. However, it is also prevalent and persistent into adulthood, particularly in females. The efficacy of tretinoin in acne is well documented with large pivotal studies. The first lotion formulation of tretinoin was developed to provide an important alternative option to treat acne patients who may be sensitive to the irritant effects of other tretinoin formulations. Objective: To determine whether efficacy and safety of tretinoin 0.05% lotion was similar in adolescent (<18 years) and adult (>=18 years) women with moderate-to-severe acne. Methods: Post hoc analysis of two multicenter, randomized, double-blind, vehicle-controlled Phase 3 studies in moderate or severe acne. Female subjects (aged 9 to 58 years, N=909) randomized (1:1) to receive tretinoin 0.05% lotion or vehicle, once-daily for 12 weeks. Efficacy assessments included changes in baseline inflammatory and noninflammatory lesions and treatment success (at least 2-grade reduction in Evaluator's Global Severity Score [EGSS] and clear/almost clear). Safety, adverse events (AEs), and cutaneous tolerability were evaluated throughout. Results: At week 12, mean percent reduction in inflammatory and noninflammatory lesion counts in female subjects were 56.9% and 51.7%, respectively, compared with 47.1% and 34.9% with vehicle (P=<0.001). Similar results were seen in adult and adolescent females in terms of reduction in inflammatory lesion counts with tretinoin 0.05% lotion; reduction in noninflammatory lesions was significantly greater in adult females (P=0.002). Treatment success was achieved by 23.6% of female subjects by week 12, compared with 13.5% on vehicle (P<0.001). Although treatment success was somewhat greater in adult females (24.6% versus 21.6%), the difference was not significant. The majority of AEs were mild and transient. There were five serious AEs (SAEs) reported (4/1, adult/adolescent, respectively). The most frequently reported treatment related AEs with tretinoin 0.05% lotion were application site pain (3.0%/5.7%), and application site dryness (4.9%/6.4%). Local cutaneous safety and tolerability assessments were generally mild-to-moderate and improved by week 12. Slight increases in mean scores were observed for scaling, burning and stinging within the first four weeks and appeared to be transient. Conclusions: Tretinoin 0.05% lotion was significantly more effective than its vehicle in achieving treatment success and reducing inflammatory and noninflammatory lesions in female acne. Noninflammatory lesion count reduction was significantly greater in adult females compared with adolescent females. The new lotion formulation was well-tolerated. J Drugs Dermatol. 2019;18(2):178-188.


The mechanism of Ca2+ stimulation of citrulline and N-acetylglutamate synthesis by mitochondria.
ATP-driven citrulline synthesis by mitochondria treated with oligomycin, uncoupler and Ca2+ ionophore is stimulated more than 2-fold by an increase in extramitochondrial free [Ca2+] in the range 1-5 microM. Stimulation increases with the length of preincubation of mitochondria with Ca2+. EGTA prevents stimulation if added at the start of the preincubation period, but is without effect if added at the end, suggesting that Ca2+ acts indirectly on citrulline synthesis via the accumulation of an intermediate. Neither carbamoyl-phosphate synthase (ammonia) nor ornithine carbamoyltransferase are stimulated by up to 50 microM free Ca2+ in mitochondrial extracts, but N-acetylglutamate synthase is stimulated about 30% by 10 microM free Ca2+. We propose that an increase in the activity of N-acetylglutamate synthase in response to an increase in free [Ca2+] in the mitochondrial matrix may contribute to hormonal stimulation of the urea cycle.


Characterization of yellow-pigmented and motile enterococci isolated from intestines of the garden snail Helix aspersa.
Enterococci associated with garden snails (Helix aspersa) were studied in order to obtain reliable species identification and characterization. Twelve yellow-pigmented and motile enterococci, isolated from the intestines of garden snails, were phenotypically close to Enterococcus casseliflavus, but they showed certain unusual biochemical characteristics. tRNA intergenic length polymorphism analysis (tDNA-PCR) divided all strains studied into two groups, in full agreement with biochemical test results. 16S rDNA sequencing, DNA base composition analysis and DNA-DNA hybridization results showed unambiguously that the enterococci studied belonged to the species Ent. casseliflavus. The representative strains of described ecovars were deposited in the Czech Collection of Microorganisms (CCM) as Ent. casseliflavus CCM 4868, 4869, 4870 and 4871. Enterococcus casseliflavus associated with garden snails can be subdivided into groups. Enterococcus casseliflavus differs from other enterococcal species in that it is typically associated with plants, soil, water and invertebrate animals. The different groups that can be found in these widely occurring bacteria are possibly source-specific ecovars, as exemplified by the Ent. casseliflavus inhabiting the intestines of snails.


Differentiation of the epiblastic part of chick Hensen's node in coelomic cavity.
When implanted into the coelomic cavity the epiblastic, non-invaginated part of Hensen's node of chick blastoderm at Hamburger--Hamilton stage 3 to 3+ can differentiate into various tissues. These include neural, mesodermal, and endodermal ones, which are not essentially different from those derived from a complete node in similar conditions.


The Rising Problem of Hip Fractures in Geriatric Patients-Analysis of Surgical Influences on the Outcome.
Hip fractures in geriatric patients often have a poor outcome in terms of mortality, mobility as well as independence. Different surgical influence factors are known that improve the outcome. In this observational cohort study, 281 patients of a geriatric trauma unit were analyzed prospectively. Demographic factors, as well as data regarding the trauma mechanism and perioperative treatment, were recorded. The nutritional status was also analyzed. The follow-up was set to 120 days. The key conclusion of this study is that a high ASA classification, the use of anticoagulatory medicine and malnutrition are significantly associated with higher mortality together with worse independence (p < 0.05). There is no significant difference in outcome concerning the time to surgery within the first 24 h. Malnutrition seems to be an important risk factor for an adverse outcome of geriatric patients and therefore warrants a focus in multidisciplinary treatment. The risk factor ASA cannot be improved during the pre-surgery phase, but requires intensified care by a multidisciplinary team specialized in orthogeriatrics.


Daily exposure to toxic metals through urban road dust from industrial, commercial, heavy traffic, and residential areas in Petaling Jaya, Malaysia: a health risk assessment.
Human health is threatened by significant emissions of heavy metals into the urban environment due to various activities. Various studies describing health risk analyses on soil and dust have been conducted previously. However, there are limited studies that have been carried out regarding the potential health risk assessment of heavy metals in urban road dust of < 63-μm diameter, via incidental ingestion, dermal contact, and inhalation exposure routes by children and adults in developing countries. Therefore, this study evaluated the health risks of heavy metal exposure via ingestion, dermal contact, and inhalation of urban dust particles in Petaling Jaya, Malaysia. Heavy metals such as lead (Pb), chromium (Cr), zinc (Zn), copper (Cu), and manganese (Mn) were measured using dust samples obtained from industrial, high-traffic, commercial, and residential areas by using inductively coupled plasma mass spectrometry (ICP-MS). The principal component and hierarchical cluster analysis showed the dominance of these metal concentrations at sites associated with anthropogenic activities. This was suggestive of industrial, traffic emissions, atmospheric depositions, and wind as the significant contributors towards urban dust contamination in the study sites. Further exploratory analysis underlined Cr, Pb, Cu, and Zn as the most representative metals in the dust samples. In accommodating the uncertainties associated with health risk calculations and simulating the reasonable maximum exposure of these metals, the related health risks were estimated at the 75th and 95th percentiles. Furthermore, assessing the exposure to carcinogenic and non-carcinogenic metals in the dust revealed that ingestion was the primary route of consumption. Children who ingested dust particles in Petaling Jaya could be more vulnerable to carcinogenic and non-carcinogenic risks, but the exposure for both children and adults showed no potential health effects. Therefore, this study serves as an important premise for a review and reformation of the existing environmental quality standards for human health safety.


Research on aerobic oxidation of methane bacteria and its influencing factors in Chongqing central city section of the Yangtze River, China.
Bacterial communities play an important role in the carbon cycle of freshwater ecosystems. In order to understand the influencing factors of bacterial community in the process of carbon cycle and search for measures to reduce carbon emissions, Chongqing central city section of the Yangtze River and its tributaries were selected to be the study area in this research. High-throughput sequencing was applied to study aerobic oxidation of methane bacteria (MOB) in sampling area. The results showed that there were spatial differences in the community diversity of aerobic MOB in the Yangtze River in central Chongqing. The Shannon index in the sediment (2.389-2.728) was higher than that in the water (1.820-2.458), and the community diversity in the middle reaches of the main river was higher than that in the upstream and the downstream. The aerobic MOB community was mainly dominated by Type II (Methylocystis). Most of operational taxonomic units (OTUs) in the top ten had high homology with MOB from river and lake sediments, and a few OTUs had high homology with MOB from paddy fields, forests and wetland soils. The main environmental factors affecting the community structure of aerobic MOB were NH4+-N, dissolved oxygen (DO), temperature (T, p ≤ 0.001), pH (p ≤ 0.05), methane (CH4) and carbon dioxide (CO2).


Immunologic reconstitution during PEG-ADA therapy in an unusual mosaic ADA deficient patient.
We report detailed genetic and immunologic studies in a patient diagnosed with adenosine deaminase (ADA) deficiency and combined immune deficiency at age 5 years. At the time of diagnosis, although all other lymphocyte subsets were depleted, circulating CD8(+) T cells with a terminally differentiated phenotype were abundant and expressed normal ADA activity due to a reversion mutation in a CD8(+) T cell or precursor. Over the first 9 months of replacement therapy with PEG-ADA, the patient steadily accumulated mature naïve CD4(+) and CD8(+) T cells, as well as CD4(+)/FOXP3(+) regulatory T cells, consistent with restoration of a functional cellular immune system. While CD19(+) naïve B cells also accumulated in response to PEG-ADA therapy, a high proportion of these B cells exhibited an immature surface marker phenotype even after 9 months, and immunization with neoantigen bacteriophage varphiX174 demonstrated a markedly subnormal humoral immune response. Our observations in this single patient have important implications for gene therapy of human ADA deficiency, as they indicate that ADA expression within even a large circulating lymphocyte population may not be sufficient to support adequate immune reconstitution. They also suggest that an immature surface marker phenotype of the peripheral B cell compartment may be a useful surrogate marker for incomplete humoral immune reconstitution during enzyme replacement, and possibly other forms of hematopoietic cell therapies.


Effect of timing of initiation of adjuvant chemotherapy on disease-free survival in breast cancer.
Four hundred and sixty patients with stage II or III breast cancer following regional therapy were treated with an adjuvant combination chemotherapy consisting of fluorouracil, doxorubicin, and cyclophosphamide (FAC). The relationship between the length of disease-free survival and length of delays in initiation of chemotherapy after surgery was evaluated. Patients were divided into four subgroups according to the length of delay in initiation of chemotherapy (less than 10 weeks, 10-13, 14-17, and greater than or equal to 18 weeks). Overall four year disease-free survival was 64%, 68%, 60%, and 63% for patient groups with delays of less than 10 weeks, 10-13, 14-17, or greater than or equal to 18 weeks respectively (p = 0.39). There was no trend for longer delay in treatment to be associated with shorter disease-free survival, except in poor prognosis patients.


Protective immunity and lack of histopathological damage two years after DNA vaccination against infectious hematopoietic necrosis virus in trout.
The DNA vaccine pIHNw-G encodes the glycoprotein of the fish rhabdovirus infectious hematopoietic necrosis virus (IHNV). Vaccine performance in rainbow trout was measured 3, 6, 13, 24, and 25 months after vaccination. At three months all fish vaccinated with 0.1 microg pIHNw-G had detectable neutralizing antibody (NAb) and they were completely protected from lethal IHNV challenge with a relative percent survival (RPS) of 100% compared to control fish. Viral challenges at 6, 13, 24, and 25 months post-vaccination showed protection with RPS values of 47-69%, while NAb seroprevalence declined to undetectable levels. Passive transfer experiments with sera from fish after two years post-vaccination were inconsistent but significant protection was observed in some cases. The long-term duration of protection observed here defined a third temporal phase in the immune response to IHNV DNA vaccination, characterized by reduced but significant levels of protection, and decline or absence of detectable NAb titers. Examination of multiple tissues showed an absence of detectable long-term histopathological damage due to DNA vaccination.


Accelerated reactive dissolution model of drug release from long-acting injectable formulations.
Long-acting injectable formulations represent a rapidly emerging category of drug delivery systems that offer several advantages compared to orally administered medicines. Rather than having to frequently swallow tablets, the medication is administered to the patient by intramuscular or subcutaneous injection of a nanoparticle suspension that forms a local depot from which the drug is steadily released over a period of several weeks or months. The benefits of this approach include improved medication compliance, reduced fluctuations of drug plasma level, or the suppression of gastrointestinal tract irritation. The mechanism of drug release from injectable depot systems is complex, and there is a lack of models that would enable quantitative parametrisation of the process. In this work, an experimental and computational study of drug release from a long-acting injectable depot system is reported. A population balance model of prodrug dissolution from asuspension with specific particle size distribution has been coupled with the kinetics of prodrug hydrolysis to its parent drug and validated using in vitro experimental data obtained from an accelerated reactive dissolution test. Using the developed model, it is possible to predict the sensitivity of drug release profiles to the initial concentration and particle size distribution of the prodrug suspension, and subsequently simulate various drug dosing scenarios. Parametric analysis of the system has identified the boundaries of reaction- and dissolution-limited drug release regimes, and the conditions for the existence of a quasi-steady state. This knowledge is crucial for the rational design of drug formulations in terms of particle size distribution, concentration and intended duration of drug release.


Team function in obstetrics to reduce errors and improve outcomes.
Crew resource management (CRM), adapted from aviation for the practice of medicine, offers the potential of reducing medical errors, increasing employee retention, and improving patient satisfaction. CRM, however, requires a culture that promotes teamwork and acceptance of new concepts. Leadership is needed to transform the culture, as well as to train, coach, and sustain the behavior CRM demands. Culture change can be fostered through teamwork activities that, when made part of a daily routine, provides the basis for modeling teamwork skills and sets the stage for sustained culture change. New tools are available to measure processes as well as patient and staff satisfaction.


[The snapping scapula as a sympton of a tumor in the scapulothoracic region].
The snapping scapula syndrome is a grating sensation located in the scapulothoracic region that appears with movement.This sign is occasionally related to tumors. To show the high incidence of this relationship (clinical sign-tumor), and to be aware of it when performing a differential diagnosis. Retrospective study of the elastofibromas dorsi (ED) and scapular osteochondromas (SO), which may have presented with the sign under study in our center over the last 17 years (1993-2009). Thirty-seven ED and 6 SO were identified. The series was divided into group A (ED) and group B (SO). Mean follow-up was 7 years. The cohorts are made up of 23 women and 4 men with a mean age of 57 years (42-78) in group A, there were 2 women and 4 men with a mean age of 20 years (11-28) in group B. Action was taken to identify the initial medical sign at diagnosis, the treatment carried out, and the outcome. Around 21% of these tumors are reported to be associated with physical activity. The initial symptom was a painful mass in 81% of the patients, followed by a scapular snapping or clicking in 30 out of the 43 patients (70%). The treatment of choice was resection in both groups. A noticeable improvement in terms of pain was seen (VAS 7.5 preoperatively, 2.8 postoperatively). The presence of snapping scapula has a strong relationship to tumors of the scapulothoracic region. Therefore it is important to be aware of this.


Communications between the nucleotide- and actin-binding site of the myosin head in muscle fibers.
2'(or 3')-O-(2,4,6-trinitrophenyl) adenosine 5'-triphosphate (TNP-ATP), a fluorescent analog of ATP, and the derivatives were used to fluorescently label the myosin head in skinned skeletal muscle fibers. It has been known that a secondary site for TNP-ADP other than the active site exists in myosin S1. We found by fluorescence resonance energy transfer between mant ATP and TNP-AMP that the secondary site for TNP-nucleotides is located within 2 nm of the active site in skeletal muscle fibers. The changes in fluorescence intensity of muscle fibers in 20 microM TNP-AMP when nucleotides are bound may reflect changes of the structure of the active site of myosin heads. It was also shown that actin affected the extent of the fluorescence changes induced by ATP binding to the active site. Both ATP and caged ATP affected the fluorescence intensity, thus caged ATP interacts with the active site. When ATP was released from caged ATP by pulse photolysis in muscle fibers in TNP-AMP showed a transient increase in fluorescence intensity, and still greater fluorescence signal can be detected when the fiber actively contracted when Ca2+ was present.


Binding properties of cerebrospinal fluid IgG in multiple sclerosis and other neurological diseases.
A solid phase radioimmunoassay (RIA) was used to detect antibodies to myelin or myelin basic protein (MBP) in the cerebrospinal fluid (CSF) of patients with multiple sclerosis (MS) or other neurological diseases (OND). When measured at the same IgG concentration, MS samples had higher binding values than OND against myelin, but not against MBP. Using F(ab')2 fragments purified from pools of MS and OND CSF there was no difference in binding to myelin between MS and OND samples. These results indicate that anti-MBP antibodies are nt a feature of MS and binding of CSF IgG to myelin is not due to specific antibody, but is probably the result of non-specific binding to Fc receptors.


Procedure-Related Complications in Sham Surgeries for Parkinson's Clinical Trials: A Meta-analysis.
Double-blind, sham-controlled neurosurgical trials for neurodegenerative disorders are debated as an ethical dilemma, particularly regarding subjects randomized to the sham surgery group with general anesthesia. The objective of this study was to examine the safety of sham surgeries in Parkinson's disease (PD) clinical trials through complications related to the procedure. A systematic review and meta-analysis were performed following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Rates and odds ratios (OR) were compared using random effects analysis. Seven studies, all randomized, double-blind, sham surgery-controlled trials, with 309 patients with PD, were qualitatively and quantitatively analyzed: 141 patients in sham groups and 168 patients in the experimental arms of gene or cell therapy trials. Sham subjects had lower rates of gastrointestinal, positioning, incision-site, respiratory (hypoxic or hypercapnic respiratory failure), cardiovascular, thromboembolism, postoperative cognitive decline, skull fracture, and intracranial or spinal complications when compared with active treatment subjects. Sham subjects, however, had a higher rate of perioperative respiratory infections, such as pneumonia or sinusitis. Further, sham subjects were less likely to experience postoperative cognitive decline (OR, 0.23; 95% confidence interval [CI]: 0.11-0.47), intracranial or spinal complications (OR, 0.10; 95% CI: 0.01-0.75), total major morbidity (OR, 0.30; 95% CI: 0.19-0.47), or overall complications (OR, 0.59; 95% CI: 0.47-0.75) when compared with patients receiving experimental therapy. Patients with PD in the sham surgery control arm of cell transplantation or gene therapy clinical trials have a low risk of procedure-related adverse events overall and fewer complications than patients in the experimental groups. There were no reported deaths attributed to sham surgery-controlled PD clinical trials. © 2023 International Parkinson and Movement Disorder Society.


Pneumococcal vaccination for patients with spinal cord injury.
Pneumococcal vaccination is recommended for persons with increased risk for pneumococcal disease or its complications. Even though recommendations for pneumococcal vaccination of the spinal cord injury (SCI) population have not been made to date, many SCI patients may qualify for vaccination. The purpose of this study was to examine the antibody response of SCI patients to polyvalent pneumococcal vaccine. Immunoglobulin G antibodies to five commonly infecting pneumococcal capsular polysaccharides were determined in 40 SCI patients and in an age-matched control of 40 able-bodied persons before and 4 to 6 weeks after vaccination. There were no significant differences in the mean antibody levels and the percentage of persons who responded to the five capsular polysaccharides in the SCI versus control groups. Among SCI patients, the antibody response was generally not affected by age, time since injury, or level of lesion. The antibody response to pneumococcal vaccination in SCI patients seems adequate.


Sequence of thioredoxin reductase from Escherichia coli. Relationship to other flavoprotein disulfide oxidoreductases.
The DNA sequence of the Escherichia coli gene encoding thioredoxin reductase has been determined. The predicted protein sequence agrees with an earlier determination of the 17 amino-terminal amino acids and with a fragment of the protein containing the redox-active half-cystines. Similarity between E. coli thioredoxin reductase and other flavoprotein disulfide oxidoreductases is quite limited, but three short segments, two of which are probably involved in FAD and NADPH binding, are highly conserved between thioredoxin reductase, glutathione reductase, dihydrolipoamide dehydrogenase, and mercuric reductase.


Are there factors in the disease of peroneal necrotizing fasciitis which should be used when choosing the anesthetic technique?: Descriptive analysis of a cohort from two centers.
This study aims to examine the factors affecting the selection of anaesthesia method in Fournier's gangrene. A retrospective evaluation was made of 113 patients operated on because of Fournier's gangrene between January-May 2019. The operations were performed under spinal anaesthesia in 78 cases (Group S) and under general anaesthesia in 35 cases (Group G). The patients were evaluated regarding age, gender, the anaesthesia method used (spinal, general) anaesthetic agent applied, presence of sepsis, and biochemical, hematological and inflammatory parameters. When the patients were evaluated regarding the Fournier Gangrene Severity Index (FGSI), patients in Group S had lower scores (p=0.001). Examination of the tomography images revealed that in 13 (37.1%) patients, air values were seen in the right or left gluteal area, or both, extending to the subcutaneous tissue. In the evaluation of the factors affecting the selection of general anaesthesia, a positive correlation was determined between an increase in FGSI (r=0.482, p=0.001) and the presence of sepsis (r=0.485, p=0.001) and gluteal region involvement (r=0.628, p<0.001). The selection of anaesthesia method in Fournier gangrene patients is a complex process affected by factors, such as the patients' general condition, sepsis, and whether or not there is bleeding diathesis. The risk -benefit balance in the selection of anaesthesia method should be evaluated individually for patients.


[Comparative study of the psychotropic activity of tuftsin and its analogs].
Tuftsin and its Leu1 and D-Arg4 analogs displayed stimulating activity in experimental behavioral despair in mice. In rats with different types of emotional reactions and with destroyed catecholamine terminals (6-OHDA treatment), tuftsin increased exploratory activity, with fear manifestations being decreased and avoidance behavior improved. This was shown while testing the rats in the "open field" and according to the ability to accomplish an extrapolation task of avoiding critical stress-situation. Leu1-tuftsin increased the emotional stress and sharply hindered the avoidance reaction, while D-Arg4-tuftsin modulated the behavior of the animals with increased emotional reactivity and made the avoidance behavior prompter. Pentapeptide, an inhibitor of tuftsin stimulation of phagocytosis, had no significant effect on the behavior. Modifications in the structure of tuftsin resulted both in the changes in phagocytosis-stimulating activity and the appearance of other psychotropic effects.


Generation of a new human induced pluripotent stem cell (hiPSC) line from a South Asian Indian with a MYBPC3Δ25bp variant.
Myosin binding protein C3 (MYBPC3) is a thick filament contractile protein that interacts with myosin, titin and actin and regulates cardiac muscle contraction. Genetic variations in the MYBPC3 gene are known causal factors for cardiomyopathy and heart failure. Previously, we identified a recurrent MYBPC3 deletion (25 base pairs) among South Asians associated with cardiomyopathy and heart failure. Here, we generated an induced pluripotent stem cell (iPSC) line using peripheral blood mononuclear cells (PBMC) from an Indian harboring MYBPC3 deletion. This iPSC line displayed embryonic stem cell morphology, expressed pluripotency markers, differentiated into three germ layers and exhibited normal karyotype.


Studies on matrix vesicles isolated from chick epiphyseal cartilage. Association of pyrophosphatase and ATPase activities with alkaline phosphatase.
Fractions composed primarily of cells (Fraction I), membrane fragments (Fraction II) and matrix vesicles (Fraction III) were isolated from chick epiphyseal cartilage. The characteristics of the alkaline phosphatase (EC 184.108.40.206), pyrophosphatase (EC 184.108.40.206) and ATPase (EC 18.104.22.168) activities in the matrix vesicle fraction were studied in detail. Mg-2-+ was not absolutely essential to any of the activities, but at low levels was stimulatory in all cases. Higher concentrations inhibited both pyrophosphatase and ATPase activities. Both the stimulatory and inhibitory effects were pH-dependent. Ca-2-+ stimulated all activities weakly in the absence of Mg-2-+. However, when Mg-2-+ was present, Ca-2-+ was slightly inhibitory. Thus, none of the activities appear to have a requirement for Ca-2-+, and hence would not seem to be involved with active Ca-2-+ transport in the typical manner. The distribution of alkaline phosphatase, pyrophosphatase, and Mg-2-+ ATPase activities among the various cartilage fractions was identical, and concentrated primarily in the matrix vesicles. Conversely, the highest level of (Na-+ + K-+)-ATPase activity was found in the cell fraction. All activites showed nearly identical sensitivities to levamisole (4 - 10-3 M) which caused nearly complete inhibition of alkaline phosphatase and pyrophosphatase. About 10-15% of the ATPase activity was levamisole-insensitive. The data are consistent with the concept that the Mg-2-+-ATPase and pyrophosphatase activities of matrix vesicles stem from one enzyme, namely, alkaline phosphatase.


Early-onset brain alterations during postnatal development in a mouse model of CDKL5 deficiency disorder.
Mutations in the CDKL5 gene are the cause of CDKL5 deficiency disorder (CDD), a rare and severe neurodevelopmental condition characterized by early-onset epilepsy, motor impairment, intellectual disability, and autistic features. A mouse model of CDD, the Cdkl5 KO mouse, that recapitulates several aspects of CDD symptomology, has helped to highlight brain alterations leading to CDD neurological defects. Studies of brain morphogenesis in adult Cdkl5 KO mice showed defects in dendritic arborization of pyramidal neurons and in synaptic connectivity, a hypocellularity of the hippocampal dentate gyrus, and a generalized microglia over-activation. Nevertheless, no studies are available regarding the presence of these brain alterations in Cdkl5 KO pups, and their severity in early stages of life compared to adulthood. A deeper understanding of the CDKL5 deficient brain during an early phase of postnatal development would represent an important milestone for further validation of the CDD mouse model, and for the identification of the optimum time window for treatments that target defects in brain development. In sight of this, we comparatively evaluated the dendritic arborization and spines of cortical pyramidal neurons, cortical excitatory and inhibitory connectivity, microglia activation, and proliferation and survival of granule cells of the hippocampal dentate gyrus in hemizygous Cdkl5 KO male (-/Y) mice aged 7, 14, 21, and 60 days. We found that most of the structural alterations in Cdkl5 -/Y brains are already present in pups aged 7 days and do not worsen with age. In contrast, the difference in the density of excitatory and inhibitory terminals between Cdkl5 -/Y and wild-type mice changes with age, suggesting an age-dependent cortical excitatory/inhibitory synaptic imbalance. Confirming the precocious presence of brain defects, Cdkl5 -/Y pups are characterized by an impairment in neonatal sensory-motor reflexes.


The impact of organ dysfunction in cirrhosis: survival at a cost?
The incidence of cirrhosis and subsequent development of organ dysfunction (OD) requiring intensive care unit (ICU) support is rising. Historically, critically ill cirrhotics are perceived as having poor prognosis and substantial cost of care. The aim was to prospectively analyse resource utilisation and cost of a large cohort of patients (n=660) admitted to a Liver ICU from 2000 to 2007 with cirrhosis and OD. Child Pugh, MELD, SOFA, APACHE II, and organ support requirements were collected. The Therapeutic Intervention Scoring System (TISS) score, a validated tool for estimating cost in ICU, was calculated daily. Logistic regression was used to determine independent predictors of increased cost. Alcohol was the most common etiology (47%) and variceal bleeding (VB) the most common reason for admission (35%). Invasive ventilatory support was required in 74% of cases, vasopressors in 49%, and 50% required renal replacement therapy. Forty-nine per cent of non-transplanted patients survived to ICU discharge. Median TISS score and ICU cost per patient were 261 and €14,139, respectively. VB patients had the highest survival rates (53% vs. 24%; p<0.001) and lower associated cost. A combination of VB (OR 0.48), need for ventilation (OR 2.81), low PO(2)/FiO(2) on admission (OR 0.97), and lactate (OR 0.93) improved cost prediction on multivariate analysis (AUROC 0.7; p<0.001) but organ failure scores per se were poor predictors of cost. Patients with cirrhosis and OD result in considerable resource expenditure but have acceptable hospital survival. Further health economic assessment and outcome prediction tools are required to appropriately target resource utilisation.


Prediction of arterial pressure increase after fluid challenge.
Mean arterial pressure above 65 mmHg is recommended for critically ill hypotensive patients whereas they do not benefit from supranormal cardiac output values. In this study we investigated if the increase of mean arterial pressure after volume expansion could be predicted by cardiovascular and renal variables. This is a relevant topic because unnecessary positive fluid balance increases mortality, organ dysfunction and Intensive Care Unit length of stay. Thirty-six hypotensive patients (mean arterial pressure < 65 mmH) received a fluid challenge with hydroxyethyl starch. Patients were excluded if they had active bleeding and/or required changes in vasoactive agents infusion rate in the previous 30 minutes. Responders were defined by the increase of mean arterial pressure value to over 65 mmHg or by more than 20% with respect to the value recorded before fluid challenge. Measurements were performed before and at one hour after the end of fluid challenge. Twenty-two patients (61%) increased arterial pressure after volume expansion. Baseline heart rate, arterial pressure, central venous pressure, central venous saturation, central venous to arterial PCO2 difference, lactate, urinary output, fractional excretion of sodium and urinary sodium/potassium ratio were similar between responder and non-responder. Only 7 out of 36 patients had valuable dynamic indices and then we excluded them from analysis. When the variables were tested as predictors of responders, they showed values of areas under the ROC curve ranging between 0.502 and 0.604. Logistic regression did not reveal any association between variables and responder definition. Fluid challenge did not improve arterial pressure in about one third of hypotensive critically ill patients. Cardiovascular and renal variables did not enable us to predict the individual response to volume administration. ClinicalTrials.gov: NCT00721604.


The novel tumour suppressor gene ING1 is overexpressed in human melanoma cell lines.
Epidemiological evidence indicates that exposure to ultraviolet (UV) radiation is directly linked to the increase of both incidence and mortality of melanoma. However, the genetic changes caused by UV radiation that lead to melanoma formation remain poorly understood. Recently, a potential tumour suppressor gene ING1 (inhibitor of growth 1) was shown to inhibit cell growth and induce apoptosis in the presence of p53. We have demonstrated that the expression of ING1 is induced after UV irradiation and that ING1 enhances the repair of UV-damaged DNA. To investigate if ING1 plays a role in melanoma formation. We examined p33ING1 expression levels in 14 melanoma cell lines. We found that p33ING1 is overexpressed at both mRNA and protein levels in melanoma cell lines compared with normal melanocytes. Single-strand conformation polymorphism (SSCP) analysis showed band shifting in two melanoma cell lines. DNA sequencing confirmed that there were nucleotide alterations in the ING1 gene in Sk-mel-24 and Sk-mel-110 cell lines. Two silent nucleotide alterations in exon 1a were detected in Sk-mel-110. In Sk-mel-24, the A-->G nucleotide alteration at codon 260 resulted in an amino acid change from Asn to Ser, while seven other nucleotide alterations were silent. To determine if the silent nucleotide alterations in these two melanoma cell lines were due to polymorphism, SSCP analysis of ING1 gene was performed in 25 healthy volunteers. No band shift was observed in the SSCP analysis, suggesting that the nucleotide alterations in the melanoma cell lines are unlikely to be due to polymorphism. Taken together, our data demonstrate that ING1 is overexpressed, but infrequently mutated, in melanoma cell lines.


Medical Informatics in the Digital Personalized Health and Medicine Era: A SWOT Analysis and Actionable Strategies.
With the rapid proliferation of digital health technologies, the role of medical informatics in advancing personalized healthcare and wellbeing is emphasized. To examine the readiness of medical informatics for fostering digital health moving forward, a Strengths, Weaknesses, Opportunities, and Threats (SWOT) analysis was performed. As a result, actionable strategies have been identified to maximize the benefits of medical informatics and overcoming the faced challenges in the digital health era.


Surgical management of parapharyngeal lymph node metastases from thyroid carcinoma with transcervical approach.
Parapharyngeal lymph node metastasis is very rare in patients with thyroid carcinoma. In this study, we elucidated on the clinical features, diagnosis, surgical treatment, and outcomes for patients with parapharyngeal lymph node metastases from thyroid carcinoma. Patients with thyroid carcinoma who had been subjected to surgery through the transcervical approach at our institution between April 1999 and January 2019 were retrospectively enrolled in this study. Parapharyngeal lymph node metastases were successfully resected through the transcervical approach in all the 97 patients. There were 32 treatment-naïve patients, while 65 patients had a history of treatment before parapharyngeal lymph node metastases. All 97 cases of parapharyngeal lymph node metastases were detected by enhanced CT scans. Lateral neck lymph nodes metastases were confirmed in 96 patients. Unexpected postoperative neurological side effects were reported in 11 (11%) patients, including dysphagia in 5 (5%) patients, Horner's syndrome in 2 (2%) patients, glossal deviation in 3 (3%) patients, and asymmetrical mouth in 1 (1%) patient. During the follow-up period, 6 of the 97 patients exhibited parapharyngeal lymph node recurrence (recurrence rate 6.2%). The incidence of distant metastases was 44.3% (43 out of 97). The 5- and 10-year disease-specific survival was 93.4% and 88.9%, respectively. Parapharyngeal lymph node metastases should be considered in patients with widespread cervical lymph node metastases. Enhanced CT scan is one of the best methods for diagnosing such metastases. Surgical resection through the transcervical approach is an effective and safe option for patients with parapharyngeal lymph node metastases from thyroid carcinoma. Moreover, parapharyngeal lymph node metastasis is associated with distant metastases that may lead to poor prognosis.