Amino acid 72 of mouse and human GDF9 mature domain is responsible for altered homodimer bioactivities but has subtle effects on GDF9:BMP15 heterodimer activities.
Growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) are oocyte-secreted paralogs of the transforming growth factor beta (TGFbeta) superfamily. In mammals, these two growth factors play critical roles in folliculogenesis. As previously reported, an arginine in the pre-helix loop of GDF5 defines the high binding specificity to its type 1 receptor. Interestingly, bioactive mouse GDF9 and human BMP15 share the conserved arginine in the pre-helix loop, but their low-activity counterparts (mouse BMP15 and human GDF9) have a glycine or a proline instead. To address the question of whether the arginine residue defines the different activities of GDF9 and BMP15 homodimers and their heterodimers in human and mouse, we used site-directed mutagenesis to change the species-specific residues in human and mouse proteins, and examined their activities in our in vitro assays. Although amino acid 72 of mature GDF9 is responsible for altered homodimer bioactivities, neither the corresponding BMP15 amino acid 62 nor the intact pre-helix loop is indispensable for BMP15 homodimer activity. However, amino acid 72 in GDF9 only has only subtle effects on GDF9:BMP15 heterodimer activity. Based on previous studies and our recent findings, we provide hypothetical models to understand the molecular mechanism to define activities of the homodimeric and heterodimeric ligands. The arginine residue in the pre-helix loop of GDF9 homodimer may prevent the inhibition from its pro-domain or directly alter receptor binding, but this residue in GDF9 does not significantly affect the heterodimer activity, because of suggested conformational changes during heterodimer formation.


[Electrophoretic mobility of T and B lymphocytes in bone tumor patients].
Electrophoretic mobility of peripheral blood T and B lymphocytes in bone tumor patients (malignant 24 and benign 16) was studied. Its variations associated with cellular immunity were evaluated in this paper. The electrophoretic mobility of cells was measured at 25 +/- 0.5 degrees C with a cytopherometer. The results showed that electrophoretic mobility of T lymphocytes and E-rosettes in malignant tumor patients were significantly low (P less than 0.05) as compared with that in the normal subjects, while those in the benign tumor patients were not significantly different (P greater than 0.05).


Curcumin-reduced gold nanoparticles facilitate IL-12 delivery to a cervical cancer in vitro cell model.
Aim: To synthesize curcumin-reduced gold nanoparticles (AuNPs) for the efficient delivery to and expression of the IL-12 gene in cervical cancer (HeLa) cells in vitro. Methods: Curcumin-reduced AuNPs were synthesized, stabilized with poly-L-lysine and PEG, conjugated to IL-12 DNA and physicochemically characterized. Cytotoxicity and IL-12 expression were accessed in vitro. Results & discussion: Stable, spherical AuNPs effectively compacted and protected the IL-12 DNA and tolerated well in vitro. Real-time quantitative PCR and ELISA confirmed the successful delivery and expression of the IL-12 gene in HeLa cells. Conclusion: The favorable attributes of this AuNP-delivery system and the significant IL-12 expression obtained augur well for cytokine-based therapy or immunotherapy in cervical cancer.


Deep proteome profiling of Trichoplax adhaerens reveals remarkable features at the origin of metazoan multicellularity.
Genome sequencing of arguably the simplest known animal, Trichoplax adhaerens, uncovered a rich array of transcription factor and signalling pathway genes. Although the existence of such genes allows speculation about the presence of complex regulatory events, it does not reveal the level of actual protein expression and functionalization through posttranslational modifications. Using high-resolution mass spectrometry, we here semi-quantify 6,516 predicted proteins, revealing evidence of horizontal gene transfer and the presence at the protein level of nodes important in animal signalling pathways. Moreover, our data demonstrate a remarkably high activity of tyrosine phosphorylation, in line with the hypothesized burst of tyrosine-regulated signalling at the instance of animal multicellularity. Together, this Trichoplax proteomics data set offers significant new insight into the mechanisms underlying the emergence of metazoan multicellularity and provides a resource for interested researchers.


Spontaneous pneumothorax as manifestation of Marfan syndrome.
The authors describe a 16-year-old boy, previously healthy, who was admitted to our hospital for left-sided spontaneous pneumothorax. On physical examination he presented with marfanoid habitus. Pneumothorax was managed conservatively with resolution. Four months later he had a recurrence of left-sided pneumothorax and 1 week after that he presented with contralateral pneumothorax. He underwent video-assisted thoracoscopic surgery twice for bullectomy and pleurodesis. No further recurrence was stated. Additional investigation showed a prolapsed cardiac mitral valve and Marfan syndrome was confirmed genetically.


Environmental and school influences on physical activity in South Asian children from low socio-economic backgrounds: A qualitative study.
South Asian (SA) children are less active but have enhanced metabolic risk factors. Physical activity (PA) is a modifiable risk factor for metabolic disease. Evidence suggests that environmental factors and socio-economic status influence PA behaviour. The purpose of this study was to understand PA environments, barriers and facilitators of PA in deprived environments for children from SA backgrounds. Focus groups were conducted with 5 groups of children aged 7-9 years (n = 33; male = 16, female = 17; SA = 17, White = 8 and Black = 8) from two schools in deprived wards of Coventry, England. Thematic analysis was used to identify key themes and subthemes across all transcripts. From the results, emergent themes included school and home environment, outdoor activity, equipment, weather, parental constraints and safety. Ethnic differences were apparent for sources of beliefs and knowledge and religious practice as constraints for PA. The findings suggest that school provides a good foundation for PA attitude, knowledge and behaviour, especially for SA children. To increase PA, multi-component interventions are needed, which focus on changing the home environment (i.e. junk food and media time), encouraging outdoors activity, changing perceptions of safety and weather conditions, which provide parental constraints for children. Interventions also need to be considerate to religious practices that might constrain time.


Asymmetric Oxidation of Enol Derivatives to α-Alkoxy Carbonyls Using Iminium Salt Catalysts: A Synthetic and Computational Study.
We report herein the first examples of asymmetric oxidation of enol ether and ester substrates using iminium salt organocatalysis, affording moderate to excellent enantioselectivities of up to 98% ee for tetralone-derived substrates in the α-hydroxyketone products. A comprehensive density functional theory study was undertaken to interpret the competing diastereoisomeric transition states in this example in order to identify the origins of enantioselectivity. The calculations, performed at the B3LYP/6-31G(D) level of theory, gave good agreement with the experimental results, in terms of the magnitude of the effects under the specified reaction conditions, and in terms of the preferential formation of the ( R)-enantiomer. Just one of the 30 characterized transition states dominates the enantioselectivity, which is attributed to the adoption of an orientation relative to stereochemical features of the chiral controlling element that combines a CH-π interaction between a CH2 group in the substrate and one of the aromatic rings of the biaryl section of the chiral auxiliary with a good alignment of the acetoxy group with the other biaryl ring, and places the smallest substituent on the alkene (a hydrogen atom) in the most sterically hindered position.


Cardiac vagal control and theoretical models of co-occurring depression and anxiety: a cross-sectional psychophysiological study of community elderly.
In order to elucidate the complex relationship between co-occurring depression and anxiety with cardiac autonomic function in the elderly, this study examined the correlation between cardiac vagal control (CVC) and pre-defined, theoretical factors from the Hospital Anxiety and Depression Scale (HADS). Three hundred fifty-four randomly selected Chinese male subjects aged ≥ 65 years and living in the community were enrolled. CVC was measured using a frequency-domain index of heart rate variability. Confirmatory factor analysis showed that the flat tripartite model of HADS provided a modest advantage in model fit when compared with other theoretical factor solutions. In the flat tripartite model, there was a significant negative association between anhedonic depression and CVC. In contrast, autonomic anxiety showed a significant positive correlation with CVC. In the hierarchical tripartite model, negative affectivity was not directly associated with CVC; instead, it had positive and negative indirect effects on CVC via autonomic anxiety and anhedonic depression, respectively. As scores for negative affectivity increased, these specific indirect effects diminished. Among competing models of co-occurring depression and anxiety, constructs from tripartite models demonstrate fair conformity with the data but unique and distinct correlations with CVC. Negative affectivity may determine the relationship of anhedonic depression and autonomic anxiety with CVC. Separating affective symptoms under the constructs of the tripartite models helps disentangle complex associations between co-occurring depression and anxiety with CVC.


The fidgety yeast: focus on high-resolution live yeast cell microscopy.
Despite its small size of 5-8 mum - only one order of magnitude above the wavelength of visible light - yeast has developed into an attractive system for light microscopic analysis. First, the ease of genetic manipulation and integrative transformation have opened numerous experimental strategies for genome-wide tagging approaches, e.g., with fluorescent proteins (as discussed in several chapters of this issue). Second, the large number of cells that can be simultaneously visualized provides an excellent basis for statistical image analysis, resulting in reliable morphological or localization information. Third, the flexibility of yeast cultivation in terms of biochemical manipulation, rapid cellular growth, mutant isolation or drug susceptibility offers an unprecedented spectrum of possibilities for in vivo functional studies, and analysis of cellular dynamics and organelle inheritance. Although yeast in itself is an interesting cellular system, its "prototype character" in understanding cellular metabolism, physiology, and signaling in eukaryotes accounts for its popular use in technology development and biomedical research.Here we discuss experimental strategies for live yeast cell imaging, geared towards imaging-based large-scale screens. Major emphasis is on the methods for immobilizing cells under "physiological" conditions, with minimum impact on yeast. We also point out potential pitfalls resulting from live cell imaging that once again stresses the necessity for extremely careful experimental design and interpretation of data resulting from imaging experiments. It goes without saying that these problems are not restricted to yeast and are also highly relevant to "large" cells. If an image tells more than a thousand (perhaps misleading?) words, the ease of obtaining "images" thus rather suggests analyzing many thousands of images, to come up with one relevant and biologically significant conclusion.


Targeted linearization of DNA in vivo.
In the past decade, site-specific chromosomal DNA cleavage mediated by DNA endonucleases has been used to examine diverse aspects of chromosome structure and function in eukaryotes, such as DNA topology, replication, transcription, recombination, and repair. Here we describe a method with which chromosomes can be linearized at any predefined position in vivo. Yeast homothallic switching endonuclease (HO endo), a sequence-specific double-strand nuclease involved in mating-type switching, is employed for targeting DNA cleavage. HO endo contains discrete functional domains: a N-terminal nuclease and a C-terminal DNA-binding domain, thereby allowing construction of a chimeric nuclease with the cutting site distinct from the original HO recognition sequence. The expression of the nuclease is engineered to be controlled by a tightly regulated, inducible promoter. The cut sites recognized by HO endo or its derivatives are introduced specifically at desired positions in the yeast genome by homologous recombination. Here we present experimental procedures and review some applications based on this approach in yeast and other biological systems.


Interinstrument reliability of the Jamar electronic dynamometer and pinch gauge compared with the Jamar hydraulic dynamometer and B&L Engineering mechanical pinch gauge.
OBJECTIVE. This study sought to determine interinstrument reliability of the Jamar electronic dynamometer and pinch gauge compared with the commonly used Jamar hydraulic dynamometer and B&L Engineering mechanical pinch gauge. METHOD. Twenty men and 20 women were tested for grip strength with the two different dynamometers, and 17 men and 25 women were tested for lateral pinch strength with the two different pinch gauges. RESULTS. Grip strength measurements were approximately 10% higher with the hydraulic dynamometer, and lateral pinch strength measurements were approximately 18% higher with the mechanical pinch gauge. Paired t tests and intraclass correlation coefficients (ICCs) were used for statistical analyses. The two-tailed p value was <.0001, and the ICC indicated poor to moderate reliability. CONCLUSION. When retesting patients, it is recommended that occupational therapists use the same instrument to measure hand strength because interinstrument reliability may be lacking.


Different change patterns of the isozymes of cytochrome P450 and glutathione S-transferases in chemically induced liver damage in rat.
In this experiment, we studied the different changes in activities and protein levels of each subform of hepatic cytochrome P450 and glutathione S-transferase (GST), in chemical-induced liver injury in rats. Rats were administered 1,1-dichloroethylene (DCE), allyl alcohol (AA), bromobenzene (BB) and N,N-dimethylformamide (DMF) p.o. once every two days for 7 times, and decapitated 18 hr after the last administration. DCE and AA showed stronger hepatic toxicity than BB and DMF, as serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were higher in DCE and AA treated rats than in BB and DMF groups. Anti-cytochrome P450 inhibitable activity of toluene metabolism and/or immunoblot analysis showed that CYP2E1 and CYP2B1/2 were induced by BB and DMF, but not by the other two chemicals; CYP2C11 was greatly decreased by all of the four toxicants; and CYP1A1/2 was slightly reduced by the four treatments. These changes were reflected in testosterone metabolism. Formation of 6 beta- and 7 alpha-hydroxytestosterone from testosterone was enhanced only in DMF-treated rats, whereas that of 2 alpha- and 16 alpha-hydroxytestosterone was reduced by all of the four chemicals. Serum GST activity was increased only in BB and DMF treated rats, but liver cytosolic GST activity was enhanced by all of the four hepatotoxicants, with higher values in BB and DMF groups than in DCE and AA groups. Immunoblot analysis demonstrated that GST Yp was induced by BB and DMF treatments, and Ya and Yc were increased only by BB. GST Yk and Yb1 were not affected by the treatments. The different change patterns of enzymes by a specific toxin and the similar modifying effect on a specific enzyme by different toxins were discussed in relation to the liver damage and to the heterogeneous distribution of enzymes in liver.


[Tuberculous meningitis: diagnosis and therapeutic difficulties].
Tuberculosis remains a public-health problem in 2010 with 9 millions cases and 1,7 million deaths worldwide each year. Tuberculosis meningitis is rare (0.5 to 1%) but is associated with high mortality and disability among survivors. An early starting of treatment is crucial. Despite molecular biology methods, microbiological diagnosis remains a challenge for the biologist. We report here 2 cases of tuberculous meningitis with different clinical and biological presentations, which underline diagnosis and therapeutic difficulties encountered in the management of this disease. The first one occurred in an HIV infected patient and the second one was caused by a multidrug-resistant strain. Clinical issues were severe with important neurological residual disability and death. Biological methods available for tuberculous meningitis diagnosis are exposed.


Investigation of the fluorescence of myosin and its fragments, heavy and light meromyosins, in concentrated solutions of neutral salts.
It is shown that when myosin and heavy (HMM) and light (LMM) meromyosins are treated with concentrated solutions of neutral salts there is change in a number of parameters of the fluorescence spectra of these proteins. For myosin and HMM this change takes place in the region of the same concentrations of LiCl, MgDl2, KSCN where there occurs dissociation of the light chains of the myosin molecule. Change in the fluorescence characteristics of myosin and HMM in the presence of these salts may be caused by two effects: change in the native conformation of the myosin molecule and dissociation of its light chains. The effect of salts on LMM fluorescence is in good agreement with general theory of the influence of concentrated salts on macromolecules.


Evolution of drug resistance: insight on TEM β-lactamases structure and activity and β-lactam antibiotics.
Since the discovery of the first penicillin bacterial resistance to β-lactam antibiotics has spread and evolved promoting new resistances to pathogens. The most common mechanism of resistance is the production of β-lactamases that have spread thorough nature and evolve to complex phenotypes like CMT type enzymes. New antibiotics have been introduced in clinical practice, and therefore it becomes necessary a concise summary about their molecular targets, specific use and other properties. β-lactamases are still a major medical concern and they have been extensively studied and described in the scientific literature. Several authors agree that Glu166 should be the general base and Ser70 should perform the nucleophilic attack to the carbon of the carbonyl group of the β-lactam ring. Nevertheless there still is controversy on their catalytic mechanism. TEMs evolve at incredible pace presenting more complex phenotypes due to their tolerance to mutations. These mutations lead to an increasing need of novel, stronger and more specific and stable antibiotics. The present review summarizes key structural, molecular and functional aspects of ESBL, IRT and CMT TEM β-lactamases properties and up to date diagrams of the TEM variants with defined phenotype. The activity and structural characteristics of several available TEMs in the NCBI-PDB are presented, as well as the relation of the various mutated residues and their specific properties and some previously proposed catalytic mechanisms.


Oxidative reactions of normal and abnormal hemoglobins in the presence of phosphatidylserine vesicles.
We have monitored a sequence of reactions involving hemoglobin (Hb) oxidation in the presence of phospholipid bilayers in carefully characterized buffer systems in order to delineate molecular events that may have cytotoxic effects, some of which may even lead to cellular death of erythrocytes. We followed reactions of subunit cross-linking, heme iron oxidation, heme destruction and iron release in normal (Hb A) and sickle [Hb S or beta6(A3)Glu-->Val] Hbs under different experimental conditions. Our results show that, in the presence of lipid surfaces, the reaction rates for heme Fe+2 to Fe+3 oxidation in Hb A and Hb S molecules were both enhanced by lipid surfaces. However, the extent of the enhancement in Hb A and Hb S was quite different in T5K6.5, a Tris buffer with low ionic strength and low pH. In T5K6.5, the rate constants were 0.8 h(-1) for Hb A and 4.3 h(-1) for Hb S, a 5-fold difference. This finding supports a published suggestion that sickle Hb exhibits abnormal heme oxidation in erythrocytes of sickle cell disease patients. We found that the rates were quite similar in P110N7.4, a phosphate buffer near physiological conditions. Even in T5K6.5, the lipid surface did not enhance Hb S reactions involving subunit cross-linking, heme destruction, or iron release. These findings suggest that heme oxidation and related reactions may not be responsible for detrimental cellular events in sickle erythrocytes under physiological conditions, as suggested in the literature.


Biochemical requirements for inhibition of Connexin26-containing channels by natural and synthetic taurine analogs.
Previous work has shown that protonated taurine and aminosulfonate pH buffers, including HEPES, can directly and reversibly inhibit connexin channels that contain connexin26 (Cx26) (Bevans, C. G., and Harris, A. L. (1999) J. Biol. Chem. 274, 3711-3719). The structural requirements for this inhibition were explored by studies of the effects of structural analogs of taurine on the activity of Cx26-containing reconstituted hemichannels from native tissue. Several analogs inhibited the channels, with a range of relative affinities and efficacies. Each active compound contains a protonated amine separated from an ionized sulfonate or sulfinate moiety by several methylene groups. The inhibition is eliminated if the sulfonate/sulfinate moiety or the amine is not present. Compounds that contain a protonated amine but lack a sulfonate/sulfinate moiety do not inhibit but do competitively block the effect of the active compounds. Compounds that lack the protonated amine do not significantly inhibit or antagonize inhibition. The results suggest involvement of the protonated amine in binding and of the ionized sulfur-containing moiety in effecting the inhibition. The maximal effect of the inhibitory compounds is enhanced when a carboxyl group is linked to the alpha-carbon. Inhibition but not binding is stereospecific, with l-isomers being inhibitory and the corresponding d-isomers being inactive but able to antagonize inhibition by the l-isomers. Whereas not all connexins are sensitive to aminosulfonates, the well defined structural requirements described here argue strongly for a highly specific regulatory interaction with some connexins. The finding that cytoplasmic aminosulfonates inhibit connexin channels whereas other cytoplasmic compounds antagonize the inhibition suggests that gap junction channels are regulated by a complex interplay of cytoplasmic ligands.


Downregulation of SPARC expression inhibits cell migration and invasion in malignant gliomas.
The secreted protein acidic and rich in cysteine (SPARC) is a secreted glycoprotein that plays an essential role in promoting the motility of invasive tumor cells. In the present study, we investigated the role of SPARC in the motile and invasive activities of human glioma cells by silencing the SPARC gene. Introduction of SPARC-targeted small interfering RNA (siRNA) into glioma cell lines resulted in downregulation of SPARC expression, and significantly suppressed glioma cell migration in vitro. Furthermore, invasiveness was significantly reduced in the cells transfected with SPARC siRNA compared with those transfected with control siRNA. In an organotypic brain slice model, co-culture of glioma spheroids and rat brain slices showed that SPARC siRNA-transfected glioma cells failed to invade the surrounding normal brain tissue. In addition, intracerebral injection of glioma cells transfected with SPARC siRNA in nude mice resulted in the formation of a non-invasive tumor, whereas injection of cells transfected with control siRNA resulted in diffuse invasive tumors. Since SPARC was exclusively expressed in the invasive zone of the tumor margin and the area surrounding tumor necrosis, we investigated the relationship between SPARC expression and hypoxic stress. SPARC expression was upregulated under hypoxic stress of 1% oxygen concentration in glioma cells. Silencing hypoxia-inducible factor-1alpha with siRNA reduced the overexpression of SPARC induced under hypoxic conditions. These results suggest that SPARC plays an essential role in the invasive activity of human glioma cells, under hypoxic conditions. Downregulation of SPARC may be a novel anti-invasion therapeutic strategy for malignant gliomas.


Circular RNA expression profiles in extracellular vesicles from the plasma of patients with pancreatic ductal adenocarcinoma.
Tumor-derived extracellular vesicles (EVs) and their contents are involved in the development of human malignancies. Circular RNAs (circRNAs), enriched in EVs, can regulate diverse cellular processes by acting as microRNA (miRNA) sponges or through other mechanisms. In the present study, we explored the potential roles of circRNAs in EVs in the development of pancreatic ductal adenocarcinoma (PDAC). First, plasma was obtained from patients with PDAC (n = 8) and healthy volunteers (n = 8), and EVs were isolated by the ultracentrifugation method. Nanoparticle tracking analysis and transmission electron microscopy confirmed the size and form of the isolated EVs. The circRNA expression profiles of EVs were investigated by high-throughput whole transcriptome sequencing. We then further validated the accuracy of the circRNA sequencing data by quantitative real-time PCR analysis using plasma samples and PC cell lines, and subsequently performed bioinformatics analysis to reveal the potential functional roles of the differentially expressed circRNAs and to construct a circRNA-miRNA interaction network to predict the target miRNAs of these circRNAs. Our work provides novel targets for further studies concerning the pathogenesis of PDAC.


Influence of interleukin-28B single-nucleotide polymorphisms on progression to liver cirrhosis in human immunodeficiency virus-hepatitis C virus-coinfected patients receiving antiretroviral therapy.
Single-nucleotide polymorphisms (SNPs) near the IL28B gene have recently been associated with spontaneous hepatitis C virus (HCV) clearance and response to interferon-based therapies in patients with chronic hepatitis C. Because human immunodeficiency virus (HIV) coinfection appears to accelerate HCV-related liver fibrosis progression, any influence of IL28B SNP on the risk of developing cirrhosis might be more easily recognized in the coinfected population. All HIV-HCV-coinfected patients who underwent hepatic elastography before initiating a course of pegylated interferon plus ribavirin therapy at 2 Spanish clinics were retrospectively identified. Liver cirrhosis was defined as >14.5 kPa by transient elastography. The IL28B rs12979860 SNP was examined in a blinded fashion. A total of 304 HIV-HCV-coinfected individuals were analyzed (mean age, 43 years; 80% were male; and 85% were receiving antiretroviral therapy), of whom 18% had cirrhosis. IL28B genotype distribution was as follows: CC, 46%; CT, 43%; and TT, 11%. Cirrhosis was more frequent in CC than CT/TT carriers (24% vs 13%; P = .01). Logistic regression analysis revealed that older age (odds ratio [OR], 1.05; 95% confidence interval [CI], 0.99-1.12]; P = .08), past alcohol abuse (OR, 1.97; 95% CI, 0.95-4.06; P = .07), and CC IL28B genotype (OR, 2.32; 95% CI, 1.22-4.41; P = .01) were predictors of cirrhosis. Interestingly, mean (SD) alanine aminotransferase (ALT) levels were greater (90 ± 53 vs 71 ± 33 IU/L;, P = .01) in IL28B CC than CT/TT carriers during the prior 4.8 ± 3.8 years. The IL28B rs12979860 CC genotype is associated with a higher prevalence of cirrhosis in HIV-HCV-coinfected patients than CT/TT genotypes, suggesting that IL28B CC carriers may experience a more rapid progression of HCV-related liver fibrosis, perhaps as result of increased liver inflammation. Thus, access to HCV treatment is of utmost importance in IL28B CC carriers, in whom treatment response is better and in whom progression to cirrhosis might occur more rapidly.


Molecular Diagnosis of Brettanomyces bruxellensis' Sulfur Dioxide Sensitivity Through Genotype Specific Method.
The yeast species Brettanomyces bruxellensis is associated with important economic losses due to red wine spoilage. The most common method to prevent and/or control B. bruxellensis spoilage in winemaking is the addition of sulfur dioxide into must and wine. However, recently, it was reported that some B. bruxellensis strains could be tolerant to commonly used doses of SO2. In this work, B. bruxellensis response to SO2 was assessed in order to explore the relationship between SO2 tolerance and genotype. We selected 145 isolates representative of the genetic diversity of the species, and from different fermentation niches (roughly 70% from grape wine fermentation environment, and 30% from beer, ethanol, tequila, kombucha, etc.). These isolates were grown in media harboring increasing sulfite concentrations, from 0 to 0.6 mg.L-1 of molecular SO2. Three behaviors were defined: sensitive strains showed longer lag phase and slower growth rate and/or lower maximum population size in presence of increasing concentrations of SO2. Tolerant strains displayed increased lag phase, but maximal growth rate and maximal population size remained unchanged. Finally, resistant strains showed no growth variation whatever the SO2 concentrations. 36% (52/145) of B. bruxellensis isolates were resistant or tolerant to sulfite, and up to 43% (46/107) when considering only wine isolates. Moreover, most of the resistant/tolerant strains belonged to two specific genetic groups, allowing the use of microsatellite genotyping to predict the risk of sulfur dioxide resistance/tolerance with high reliability (>90%). Such molecular diagnosis could help the winemakers to adjust antimicrobial techniques and efficient spoilage prevention with minimal intervention.


A Phase I safety and immunogenicity trial of UBI microparticulate monovalent HIV-1 MN oral peptide immunogen with parenteral boost in HIV-1 seronegative human subjects.
Thirty-three HIV-seronegative adults were recruited into a Phase I safety and immunogenicity HIV-1 vaccine trial. The immunogens were as follows: a synthetic, monovalent, octameric HIV-1 MN V3 peptide in aluminum hydroxide (alum) adjuvant administered by intramuscular delivery; and a similar product encapsulated in biodegradable micro-spheres composed of co-polymers of lactic and glycolic acids, administered by the oral route. These were administered in three sequential oral doses, followed by a parenteral boost. No serious adverse experiences were observed. Oral administration of this vaccine, alone or in combination with parenteral boosting, resulted in no significant humoral, cellular, or mucosal immune responses.


Mimicking the fibrinolytic system on material surfaces.
Clotting and thrombosis remain the most serious problems in the development of blood contacting devices such as heart valves, vascular stents, grafts and catheters. No material exists that does not provoke these phenomena and coagulation appears to be inevitable when a foreign (i.e. non-endothelial) surface is in contact with blood. As an alternative to a surface that prevents coagulation, the concept of a clot-lysing or fibrinolytic surface is attractive. By designing the surface effectively to mimic the fibrinolytic system in the vasculature, it may be possible for clots to be lysed (effectively dissolved) as they form. In this review we elaborate on this concept and discuss ways in which such a surface could be realized. Developments in this area to the present time are reviewed, and some perspectives for future research are presented.


Association of Cortical Lesion Burden on 7-T Magnetic Resonance Imaging With Cognition and Disability in Multiple Sclerosis.
Cortical lesions (CLs) contribute to physical and cognitive disability in multiple sclerosis (MS). Accurate methods for visualization of CLs are necessary for future clinical studies and therapeutic trials in MS. To evaluate the clinical relevance of measures of CL burden derived from high-field magnetic resonance imaging (MRI) in MS. An observational clinical imaging study was conducted at an academic MS center. Participants included 36 individuals with MS (30 relapsing-remitting, 6 secondary or primary progressive) and 15 healthy individuals serving as controls. The study was conducted from March 10, 2010, to November 23, 2012, and analysis was performed from June 1, 2011, to September 30, 2014. Seven-Tesla MRI of the brain was performed with 0.5-mm isotropic resolution magnetization-prepared rapid acquisition gradient echo (MPRAGE) and whole-brain, 3-dimensional, 1.0-mm isotropic resolution magnetization-prepared, fluid-attenuated inversion recovery (MPFLAIR). Cortical lesions, seen as hypointensities on MPRAGE, were manually segmented. Lesions were classified as leukocortical, intracortical, or subpial. Images were segmented using the Lesion-TOADS (Topology-Preserving Anatomical Segmentation) algorithm, and brain structure volumes and white matter (WM) lesion volume were reported. Volumes were normalized to intracranial volume. Physical disability was measured by the Expanded Disability Status Scale (EDSS). Cognitive disability was measured with the Minimal Assessment of Cognitive Function in MS battery. Cortical lesions were noted in 35 of 36 participants (97%), with a median of 16 lesions per participant (range, 0-99). Leukocortical lesion volume correlated with WM lesion volume (ρ = 0.50; P = .003) but not with cortical volume; subpial lesion volume inversely correlated with cortical volume (ρ = -0.36; P = .04) but not with WM lesion volume. Total CL count and volume, measured as median (range), were significantly increased in participants with EDSS scores of 5.0 or more vs those with scores less than 5.0 (count: 29 [11-99] vs 13 [0-51]; volume: 2.81 × 10-4 [1.30 × 10-4 to 7.90 × 10-4] vs 1.50 × 10-4 [0 to 1.01 × 10-3]) and in cognitively impaired vs unimpaired individuals (count: 21 [0-99] vs 13 [1-54]; volume: 3.51 × 10-4 [0 to 1.01 × 10-4] vs 1.19 × 10-4 [0 to 7.17 × 10-4]). Cortical lesion volume correlated with EDSS scores more robustly than did WM lesion volume (ρ = 0.59 vs 0.36). Increasing log[CL volume] conferred a 3-fold increase in the odds of cognitive impairment (odds ratio [OR], 3.36; 95% CI, 1.07-10.59; P = .04) after adjustment for age and sex and a 14-fold increase in odds after adjustment for WM lesion volume and atrophy (OR, 14.26; 95% CI, 1.06-192.37; P = .045). Leukocortical lesions had the greatest effect on cognition (OR for log [leukocortical lesion volume], 9.65; 95% CI, 1.70-54.59, P = .01). This study provides in vivo evidence that CLs are associated with cognitive and physical disability in MS and that leukocortical and subpial lesion subtypes have differing clinical relevance. Quantitative assessments of CL burden on high-field MRI may further our understanding of the development of disability and progression in MS and lead to more effective treatments.


EBV-positive diffuse large B-cell lymphoma, not otherwise specified: 2022 update on diagnosis, risk-stratification, and management.
Epstein Barr virus-positive (EBV+) diffuse large B-cell lymphoma (DLBCL), not otherwise specified (NOS) is an entity included in the WHO classification of lymphoid neoplasms since 2016. EBV+ DLBCL, NOS, is an aggressive B-cell lymphoma associated with EBV infection, and a poor prognosis with standard chemotherapeutic approaches. The diagnosis is made through a careful pathological evaluation. Detection of EBV-encoded RNA (EBER) is considered standard for diagnosis; however, a clear cutoff for percentage of positive cells has not been defined. The differential diagnosis includes plasmablastic lymphoma (PBL), DLBCL associated with chronic inflammation, primary effusion lymphoma (PEL), among others. The International Prognostic Index (IPI) and the Oyama score can be used for risk-stratification. The Oyama score includes age >70 years and presence of B symptoms. The expression of CD30 and PD-1/PD-L1 are emerging as potential adverse but targetable biomarkers. Patients with EBV+ DLBCL, NOS, should be staged and managed following similar guidelines than patients with EBV-negative DLBCL. EBV+ DLBCL, NOS, however, might have a worse prognosis than EBV-negative DLBCL in the era of chemoimmunotherapy. Therefore, the inclusion of patients in clinical trials when available is recommended. There is an opportunity to study and develop targeted therapy in the management of patients with EBV+ DLBCL, NOS.


Hepatitis B virus (HBV) particles are produced in a cell culture system by transient expression of transfected HBV DNA.
An in vitro system for the production of hepatitis B virus (HBV) particles was established by the transient expression of transfected HBV DNA using a human hepatocellular carcinoma cell, HuH-7, as a recipient. The 3.6- and 2.2-kilobase transcripts observed were similar to those in virus-infected liver cells. Both transcripts revealed the microheterogeneity of their 5' ends. The formation of virus-related particles subsequent to the RNA transcription was demonstrated. The core particles observed in the cytoplasm and the virus particles secreted in the culture medium contained the replicative intermediates of HBV DNA and banded at densities of 1.35-1.36 g/cm3 and 1.22-1.24 g/cm3, respectively. Furthermore, the in vitro mutagenesis of the template HBV DNA demonstrated that the P gene as well as the C gene products were essential for the production of HBV particles.


What is your diagnosis? Multiple sclerosis.
Multiple sclerosis is a demyelinating disease of the central nervous system that commonly presents with ocular manifestations. These ocular manifestations include vision loss and optomotor deficit. Treatment modalities should include treatment of the systemic disease as well as the ocular disease. New treatment protocol suggests that the best way to treat optic neuritis is with IV methylprednisolone, unless there is some serious contraindication for the patient. The use of interferon and cyclosporin as well as other anti-inflammatory agents may be useful in the future.


High-density lipoprotein cholesterol changes after continuous egg consumption in healthy adults.
To determine the relationship between continuous egg consumption with Thai life-style dietary and serum lipids of healthy young people. Fifty-six participants with an average age of 35 were enrolled. In an experimental method of cholesterol intake, all participants were fed an additional egg per day to their basic diet. This project ran for 12 weeks. The 12-week egg consumption significantly increased serum total cholesterol by 0.27 +/- 0.15 mmol/L (10.43 +/- 5.80 mg/dL) (p < 0.05). The HDL-cholesterol (HDL-c) increased significant by 0.55 +/- 0.06 mmol/L (21.80 +/- 2.25 mg/dL) (p < 0.001) while the total cholesterol (TC) decreased as the HDL-c ratio was 0.94 +/- 1.1 (p < 0.001). No significant changes were found in LDL-cholesterol (LDL-c) and triglyceride levels. The present study showed that small serum LDL-c changed in response to change of egg consumption. Additionally, 12-week egg consumption also resulted in an increasing HDL-c level. In the majority of healthy adults, an addition of one egg per day to a normal fat diet could raise HDL-c levels and decreased the ratio of TC toHDL-c. Therefore, egg consumption might benefit blood cholesterol.


Gastric capacity, test meal intake, and appetitive hormones in binge eating disorder.
Binge eating disorder (BED), characterized by ingestion of very large meals without purging afterwards, is found in a subset of obese individuals. We showed previously that stomach capacity is greater in obese than in lean subjects, and in this study, we investigated capacity in obese individuals with BED. We also determined ad-libitum intake of a test meal until extremely full. Furthermore, we measured various appetitive hormones (insulin, leptin, glucagon, CCK, ghrelin) and glucose before a fixed meal and for 120 min afterwards. An acetaminophen tracer was used to assess gastric emptying rate. We compared three groups of overweight women: 11 BED, 13 BE (subthreshold BED), and 13 non-binge-eating normals. The BED individuals had the largest stomach capacity as assessed by either maximum volume tolerated (P=.05) or by gastric compliance to pressure (P=.02) using an intragastric balloon. Although test meal intake did not differ between groups, it correlated (P=.03) with gastric capacity. The BED group showed a tendency (P=.06) to have greater area under the curve (AUC) and had higher values at 5 and 60 min (P<.05) for insulin compared to normals. Moreover, the BED subjects had lower ghrelin baselines premeal, and lower AUC for ghrelin, which then declined less postmeal than for the normals (P<.05). None of the other blood values differed, including glucose, leptin glucagon, and CCK, as well as acetaminophen, reflecting gastric emptying. The lower ghrelin in BED, although contrary to what was expected, is consistent with lower ghrelin in obesity, and suggests down-regulation of ghrelin by overeating. The lack of differences in CCK is consistent with the lack of differences in gastric emptying rate, given that CCK is released when nutrients reach the intestine. The results show that BED subjects have a large gastric capacity as well as abnormalities in meal-related ghrelin and insulin patterns that may be factors in binge eating.


DNA methylation is the primary silencing mechanism for a set of germ line- and tumor-specific genes with a CpG-rich promoter.
A subset of male germ line-specific genes, the MAGE-type genes, are activated in many human tumors, where they produce tumor-specific antigens recognized by cytolytic T lymphocytes. Previous studies on gene MAGE-A1 indicated that transcription factors regulating its expression are present in all tumor cell lines whether or not they express the gene. The analysis of two CpG sites located in the promoter showed a strong correlation between expression and demethylation. It was also shown that MAGE-A1 transcription was induced in cell cultures treated with demethylating agent 5'-aza-2'-deoxycytidine. We have now analyzed all of the CpG sites within the 5' region of MAGE-A1 and show that for all of them, demethylation correlates with the transcription of the gene. We also show that the induction of MAGE-A1 with 5'-aza-2'-deoxycytidine is stable and that in all the cell clones it correlates with demethylation, indicating that demethylation is necessary and sufficient to produce expression. Conversely, transfection experiments with in vitro-methylated MAGE-A1 sequences indicated that heavy methylation suffices to stably repress the gene in cells containing the transcription factors required for expression. Most MAGE-type genes were found to have promoters with a high CpG content. Remarkably, although CpG-rich promoters are classically unmethylated in all normal tissues, those of MAGE-A1 and LAGE-1 were highly methylated in somatic tissues. In contrast, they were largely unmethylated in male germ cells. We conclude that MAGE-type genes belong to a unique subset of germ line-specific genes that use DNA methylation as a primary silencing mechanism.


Nonsurgical reduction of prolapsed colocolic intussusception in 2 puppies.
To describe the nonsurgical management of prolapsed colocolic intussusception in 2 puppies. Two 3-month-old male intact puppies (Pug and Labrador Retriever) developed intussusceptions that had prolapsed from the anus after they had suffered from tenesmus with soft feces or diarrhea. Reduction of the prolapsed tissue was performed and colocolic intussusception was diagnosed ultrasonographically in 1 dog. Rectal manipulation and saline enema were performed in both dogs, and reduction of the intussusception was confirmed endoscopically or ultrasonographically. Bupivacaine and prednisolone foam enemas were administered to reduce tenesmus. Abdominal ultrasonography the following day in 1 dog confirmed a normal appearing colon. No further tenesmus was noted in 1 dog; the other had a brief rectal prolapse after an episode of tenesmus that was manually reduced. Intussusception had not recurred 4 months and 1 year following manual reduction procedures. This is the first report of successful nonsurgical management of colocolic intussusception in a companion animal. Nonsurgical management is a first-line treatment of intussusception in children and could be considered in dogs with colocolic intussusception. Further investigation is required to define specific aspects, including rectal manipulation, saline enema, or anesthesia, that aid in resolution of this type of intussusception.


[Plastic properties of neuronal synapses in hippocampal slices].
Long-lasting potentiation was examined in the dentate gyrus -- area CA3 pathway by extra- and intracellular recording from slices of the mouse hippocampus. Tetanization of the dentate gyrus led to a shortening of the latency of action potentials (APs) and EPSPs recorded from the area CA3, to an increase in the amplitude of EPSPs and in the steepness of their ascending slope and to an augmented probability of APs. These changes persisted for a period of several seconds to 30 min after tetanization. The results suggest that monosynaptic EPSPs also can undergo long-lasting (up to 30 min) post-potentiation.


Household costs and time to seek care for pregnancy related complications: The role of results-based financing.
Results-based financing (RBF) schemes-including performance based financing (PBF) and conditional cash transfers (CCT)-are increasingly being used to encourage use and improve quality of institutional health care for pregnant women in order to reduce maternal and neonatal mortality in low-income countries. While there is emerging evidence that RBF can increase service use and quality, little is known on the impact of RBF on costs and time to seek care for obstetric complications, although the two represent important dimensions of access. We conducted this study to fill the existing gap in knowledge by investigating the impact of RBF (PBF+CCT) on household costs and time to seek care for obstetric complications in four districts in Malawi. The analysis included data on 2,219 women with obstetric complications from three waves of a population-based survey conducted at baseline in 2013 and repeated in 2014(midline) and 2015(endline). Using a before and after approach with controls, we applied generalized linear models to study the association between RBF and household costs and time to seek care. Results indicated that receipt of RBF was associated with a significant reduction in the expected mean time to seek care for women experiencing an obstetric complication. Relative to non-RBF, time to seek care in RBF areas decreased by 27.3% (95%CI: 28.4-25.9) at midline and 34.2% (95%CI: 37.8-30.4) at endline. No substantial change in household costs was observed. We conclude that the reduced time to seek care is a manifestation of RBF induced quality improvements, prompting faster decisions on care seeking at household level. Our results suggest RBF may contribute to timely emergency care seeking and thus ultimately reduce maternal and neonatal mortality in beneficiary populations.


Buckling-Fracture Transition and the Geometrical Charge of a Crack.
We present a unifying approach that describes both surface bending and fracture in the same geometrical framework. An immediate outcome of this view is a prediction for a new mechanical transition: the buckling-fracture transition. Using responsive gel strips that are subjected to nonuniform osmotic stress, we show the existence of the transition: Thin plates do not fracture. Instead, they release energy via buckling, even at strains that can be orders of magnitude larger than the Griffith fracture criterion. The analysis of the system reveals the dependence of the transition on system's parameters and agrees well with experimental results. Finally, we suggest a new description of a mode I crack as a line distribution of Gaussian curvature. It is thus exchangeable with extrinsic generation of curvature via buckling. The work opens the way for the study of mechanical problems within a single nonlinear framework. It suggests that fracture driven by internal stresses can be completely avoided by a proper geometrical design.


Gray platelet syndrome. Demonstration of alpha granule membranes that can fuse with the cell surface.
Platelets from patients with the gray platelet syndrome have decreased recognizable alpha granules and are markedly deficient in some alpha-granule secretory proteins. Using immunocytochemical techniques with antibodies to an alpha-granule membrane protein, GMP-140, we identified the membranes of intracellular vesicles in gray platelets as alpha-granule membranes. Gray platelets contained normal amounts of GMP-140 as measured by electroimmunoassay. The activation of gray platelets with thrombin caused GMP-140 to be redistributed to the plasma membrane surface, as in normal platelets. In agreement with previous studies, an endogenously synthesized secretory protein, platelet factor 4, was undetectable in gray platelets. However, the alpha-granule proteins albumin and IgG, which are thought to be derived from endocytosis of plasma proteins into megakaryocytes, were present in substantial quantities and were secreted efficiently from gray platelets. Therefore, the fundamental defect in the gray platelet syndrome may be in the targeting of endogenously synthesized secretory proteins to developing alpha granules in megakaryocytes.


Identification of a cancer associated fibroblasts-related index to predict prognosis and immune landscape in ovarian cancer.
Cancer-associated fibroblasts (CAFs) play a role in ovarian cancer (OV) evolution, immunosuppression and promotion of drug resistance. Exploring the value of CAFs-related biomarker in OV is of great importance. In the present work, we developed a CAFs-related index (CAFRI) based on an integrated analysis of single-cell and bulk RNA-sequencing and highlighted the value of CAFRI in predicting clinical outcomes in individuals with OV, tumour immune microenvironment (TIME) and response to immune checkpoint inhibitors (ICIs). The GSE151214 cohort was used for cell subpopulation localization and analysis, the TCGA-OV patients as a training set. Moreover, the ICGC-OV, GSE26193, GSE26712 and GSE19829 cohorts were used for the validation of CAFRI. The TIMER 2.0, CIBERSORT and ssGSEA algorithms were used for analysis of TIME characteristics based on the CAFRI. The GSVA, GSEA, GO, KEGG and tumour mutation burden (TMB) analyses were used for mechanistic exploration. Additionally, the IMvigor210 cohort was conducted to validate the predictive value of CAFRI on the efficacy of ICIs. Finally, CAFRI-based antitumour drug sensitivity was analysed. The findings demonstrate that the CAFRI can served as an excellent predictor of prognosis for individuals with OV, as well as identifying patients with different TIME characteristics, differentiating between immune 'hot' and 'cold' tumour populations, and providing new insights into the selection of ICIs and personalised treatment regimens. CAFRI provides new perspectives for the development of novel prognostic and immunotherapy efficacy predictive biomarkers for OV.


A computationally efficient superresolution image reconstruction algorithm.
Superresolution reconstruction produces a high-resolution image from a set of low-resolution images. Previous iterative methods for superresolution had not adequately addressed the computational and numerical issues for this ill-conditioned and typically underdetermined large scale problem. We propose efficient block circulant preconditioners for solving the Tikhonov-regularized superresolution problem by the conjugate gradient method. We also extend to underdetermined systems the derivation of the generalized cross-validation method for automatic calculation of regularization parameters. The effectiveness of our preconditioners and regularization techniques is demonstrated with superresolution results for a simulated sequence and a forward looking infrared (FLIR) camera image sequence.


Vectorcardiographic diagnosis and electrocardiographic correlation in left ventricular asynergy due to coronary artery disease. I. Severe asynergy of the anterior and apical segments.
The transverse plane QRS loop vectorcardiogram (VCG) was analyzed in 102 consecutive patients, angiographically proven to have coronary artery disease (CAD), whose right anterior oblique left ventriculogram showed severe asynergy (akinesis or marked hypokinesis) of the anterior and apical segments. Eighty-four of the 102 loops were considered to be diagnostic of severe asynergy because similar loops were not found in patients with CAD with less severe asynergy of either or both segments or in normal control subjects. Eighty-two of the 84 showed a uniformly abnormal pattern. Posterior vectors exhibited partial or complete clockwise "rotation" and were much more prominent than early anterior forces. In fact, the latter were often absent. In the other two VCG's at least the early half of the loop displayed prominent anterior vectors. The ECGs showed marked variability of the QRS in leads V-2-V-6. Abnormal Q waves were present in only 63 of the 102. The numbers of these leads showing abnormal Q waves varied from one to all of these positions.


Hepatitis E virus (HEV)-1 harbouring HEV-4 non-structural protein (ORF1) replicates in transfected porcine kidney cells.
Hepatitis E virus (HEV) is a causative agent of acute hepatitis and a major public health problem in India. There are four mammalian HEV genotypes worldwide. In India, genotype 1 (HEV-1) is restricted to humans whereas genotype 4 (HEV-4) circulates in pigs. Studies from our laboratory have shown that HEV-4 (swine) virus can establish experimental infection in rhesus monkeys; however, HEV-1 (human) virus cannot infect pigs. Viral and/or cellular factors responsible for this host specificity are not yet known. We developed 12 different genotype 1-4 chimeric full genome clones with pSK-HEV2 as the backbone and by replacing structural (ORF2 and ORF3), non-structural (ORF1) and non-coding regions (NCR) with corresponding segments from the HEV-4 clone. S10-3 (human hepatoma) and PK-15 (pig kidney) cells were transfected with transcripts generated from the above clones to test their replication competence. Transfected cells were monitored for successful virus replication by detecting replicative intermediate RNA and capsid protein (immunofluorescence assay). All the chimeric constructs were able to replicate in S10-3 cells. However, only two chimeric clones, HEV-1 (HEV-4 5'NCR-ORF1) and HEV-1 (HEV-4 ORF1), containing 5'NCR-ORF1 and ORF1 regions from the HEV-4 clone, respectively, were able to replicate in PK-15 cells. We demonstrate for the first time the crucial role of ORF1 polyprotein in crossing the species barrier at the cellular level. These results indicate the importance of interactions between ORF1 protein domains and host cell specific factors during HEV replication and the critical role of cellular factors as post-entry barrier/s in virus establishment.


Non-canonical AR activity facilitates endocrine resistance in breast cancer.
The role of androgen receptor (AR) in endocrine-resistant breast cancer is controversial and clinical trials targeting AR with an AR antagonist (e.g., enzalutamide) have been initiated. Here, we investigated the consequence of AR antagonism using in vitro and in vivo models of endocrine resistance. AR antagonism in MCF7-derived tamoxifen-resistant (TamR) and long-term estrogen-deprived breast cancer cell lines were achieved using siRNA-mediated knockdown or pharmacological inhibition with enzalutamide. The efficacy of enzalutamide was further assessed in vivo in an estrogen-independent endocrine-resistant patient-derived xenograft (PDX) model. Knockdown of AR inhibited the growth of the endocrine-resistant cell line models. Microarray gene expression profiling of the TamR cells following AR knockdown revealed perturbations in proliferative signaling pathways upregulated in endocrine resistance. AR loss also increased some canonical ER signaling events and restored sensitivity of TamR cells to tamoxifen. In contrast, enzalutamide did not recapitulate the effect of AR knockdown in vitro, even though it inhibited canonical AR signaling, which suggests that it is the non-canonical AR activity that facilitated endocrine resistance. Enzalutamide had demonstrable efficacy in inhibiting AR activity in vivo but did not affect the growth of the endocrine-resistant PDX model. Our findings implicate non-canonical AR activity in facilitating an endocrine-resistant phenotype in breast cancer. Unlike canonical AR signaling which is inhibited by enzalutamide, non-canonical AR activity is not effectively antagonized by enzalutamide, and this has important implications in the design of future AR-targeted clinical trials in endocrine-resistant breast cancer.


Acinetobacter baumannii and Cefiderocol, between Cidality and Adaptability.
Among the bacterial species included in the ESKAPE group, Acinetobacter baumannii is of great interest due to its intrinsic and acquired resistance to many antibiotics and its ability to infect different body regions. Cefiderocol (FDC) is a novel cephalosporin that is active against Gram-negative bacteria, with promising efficacy for A. baumannii infections, but some studies have reported therapeutic failures even in the presence of susceptible strains. This study aims to investigate the interactions between FDC and 10 A. baumannii strains with different susceptibilities to this drug. We confirmed diverse susceptibility profiles, with resistance values close to the EUCAST-proposed breakpoints. The minimal bactericidal concentration (MBC)/MIC ratios demonstrated bactericidal activity of the drug, with ratio values of ≤4 for all of the strains except ATCC 19606; however, bacterial regrowth was evident after exposure to FDC, as were changes in the shapes of colonies and bacterial cells. A switch to a nonsusceptible phenotype in the presence of high FDC concentrations was found in 1 strain as an adaptation mechanism implemented to overcome the cidal activity of this antibiotic, which was confirmed by the presence of heteroresistant, unstable subpopulations in 8/10 samples. Genomic analyses revealed the presence of mutations in penicillin-binding protein 3 (PBP3) and TonB3 that were shared by all of the strains regardless of their resistance phenotype. Because our isolates harbored β-lactamase genes, β-lactamase inhibitors showed the ability to restore the antimicrobial activity of FDC despite the different nonsusceptibility levels of the tested strains. These in vitro results support the concept of using combination therapy to eliminate drug-adapted subpopulations and regain full FDC activity in this difficult-to-treat species. IMPORTANCE This work demonstrates the underrated presence of Acinetobacter baumannii heteroresistant subpopulations after exposure of A. baumannii strains to FDC and its instability. Both A. baumannii and FDC are of great interest for the scientific community, as well as for clinicians; the former represents a major threat to public health due to its resistance to antibiotics, with related costs of prolonged hospitalization, and the latter is a novel, promising cephalosporin currently under the magnifying glass.


A comparative pilot study on ultraviolet-induced skin changes assessed by noninvasive imaging techniques in vivo.
The effects of acute and chronic ultraviolet (UV) on the morphology of human skin have been extensively studied ex vivo by means of histological investigations. However, innovative skin imaging techniques enable visualization of micromorphological structures in vivo. We aimed to perform a correlation study evaluating in vivo dose and time dependent skin changes following solar-simulated irradiation using noninvasive techniques such as optical coherence tomography (OCT) and confocal laser scanning microscopy (CLSM). The forearms of 10 healthy subjects were exposed to 1 minimal erythema dose (MED) and 3 MED of solar-simulated radiation. Noninvasive measurements were performed before and 24 h and 72 h after UV exposures. We demonstrate definite OCT and CLSM findings obtained from UV-exposed skin, including an increase in epidermal thickness (hyperproliferation, acanthosis), a reduction in dermal reflectivity (dermal edema), an increase in brightness of the basal layer (pigmentation), and an increase in vessel diameter within the dermal papillae (vasodilatation). A moderate to strong linear association between the methods employed was observed. In conclusion, noninvasive high-resolution imaging techniques such as OCT and CLSM may be promising tools for photobiological studies aimed at assessing photoadaptive and/or phototoxic processes in vivo. However, larger studies are needed to demonstrate the applicability of the findings presented in this pilot study.


Relevance of routine blood pressure assessment among dental patients in Lagos, Nigeria.
To determine the relevance of routine blood pressure (BP) measurement in patients attending the dental clinic. This was a retrospective survey of patients who attended the dental clinic in the Preventive Dentistry Department of a tertiary Hospital in Lagos, Nigeria over an eighteen month period. Data retrieved from the patient's hospital records included age, sex, BP and history of hypertension. Data was analyzed using Epi info 2011 package. p-value <0.05 was considered statistically significant. A total of 1910 patients' records were assessed. Females represented 53.4% (n = 1020) of the study population. The mean age was 37.1 ± 15.0 years (range 18-88 years). The prevalence of hypertension was 39.9% (n = 763). Hypertension was stage 1 in 25.5%, stage 2 in 9.8% and severe in 4.6% of the dental patients. Past medical history of hypertension was obtained for 952 patients (49.8%). Of the 763 patients with high blood pressure, 14.8% had a known history of hypertension, while 42.1% were previously undiagnosed (p < 0.0001). Systolic and diastolic BP increased with increasing age (p < 0.05) and was significantly higher in males than females (p < 0.05). The high prevalence of hypertension noted among the study population with its potential consequence during dental procedures makes the measurement of blood pressure a valuable assessment in a dental clinic. The high prevalence of hypertension particularly the previously undiagnosed cases among the dental patients highlights the relevance of routine blood pressure assessment prior to dental procedures in contemporary dental practice, so as to minimize the potential complications that could occur.


Growth factors enhance endothelial progenitor cell proliferation under high-glucose conditions.
The purpose of this study was to investigate the impact of growth regulators, including growth hormone (GH), insulin-like growth factor 1 (IGF-1), and mechano growth factor (MGF), on endothelial progenitor cell (EPC) proliferation at different glucose concentrations. EPCs were isolated and cultured from peripheral blood samples of healthy volunteers and immunocytochemically characterized after 7 days. The effects of glucose and growth regulators on EPC proliferation were determined with the Alamar Blue and Trypan Blue assays. The effect of glucose supplementation at 2.5, 11.1, and 25.0 mM was examined using cells seeded at densities of 15000, 30000, and 45000 cells/ml. For the GH-treated cells, enhancement of EPC proliferation was detected in the samples supplemented with 11.1 and 25.0 mM glucose. A slight elevation in EPC proliferation was only observed in the IGF-1-treated cells supplemented with 25.0 mM glucose. Significant enhancement of EPC proliferation was observed in MGF-treated cells supplemented with 11.1 and 25.0 mM glucose. All three growth factors demonstrated enhancement of cellular proliferation when the cells were supplemented with 25.0 mM glucose. No enhancement of EPC proliferation by the growth factors was detected in any of the cells supplemented with 2.5 mM glucose. GH, IGF-1, and MGF enhance EPC proliferation under 25.0 mM glucose conditions. The presence of these growth regulators in EPC culture may contribute to protecting EPCs from high-glucose conditions. This action may be of therapeutic relevance contributing to beneficial cardiovascular effects for diabetic patient.


Transferrin Receptor-Targeted Nonspherical Microbubbles for Blood-Brain Barrier Sonopermeation.
Microbubbles (MB) are widely used for ultrasound (US) imaging and drug delivery. MB are typically spherically shaped, due to surface tension. When heated above their glass transition temperature, polymer-based MB can be mechanically stretched to obtain an anisotropic shape, endowing them with unique features for US-mediated blood-brain barrier (BBB) permeation. It is here shown that nonspherical MB can be surface-modified with BBB-specific targeting ligands, thereby promoting binding to and sonopermeation of blood vessels in the brain. Actively targeted rod-shaped MB are generated via 1D stretching of spherical poly(butyl cyanoacrylate) MB and via subsequently functionalizing their shell with antitransferrin receptor (TfR) antibodies. Using US and optical imaging, it is demonstrated that nonspherical anti-TfR-MB bind more efficiently to BBB endothelium than spherical anti-TfR-MB, both in vitro and in vivo. BBB-associated anisotropic MB produce stronger cavitation signals and markedly enhance BBB permeation and delivery of a model drug as compared to spherical BBB-targeted MB. These findings exemplify the potential of antibody-modified nonspherical MB for targeted and triggered drug delivery to the brain.


Variola minor in Braganca Paulista County, 1956: household aggregation of the disease and the influence of household size on the attack rate.
Household aggregation of cases, one possible characteristic of person-to-person transmitted disease, was formally tested in one epidemic of variola minor by using a pair statistic. A significant result was found for all households as well as for households grouped by the type of environment, or by the phase of the epidemic growth in time. Secondary attack rates, when related to household size (number of susceptibles) showed only a marginal trend in rural households but no trend in urban or semi-rural households.


Socio-economic and demographic variations in school lunch participation of French children aged 3-17 years.
To assess school canteen attendance in a French nationally representative sample of children and to analyse its association with the socio-economic and demographic characteristics of the children and their families. Data from the second French national cross-sectional food consumption survey (INCA2), performed in 2006-2007, were used. Information on usual weekly school canteen attendance was collected through a self-reported questionnaire, and demographic and socio-economic variables through a face-to-face questionnaire. The associations between school canteen attendance and the socio-economic and demographic variables were investigated by multivariate logistic regression analyses. The INCA2 sample was representative of the children aged 3-17 years in France. Analysis was performed on 1413 schoolchildren who completed the school canteen attendance questions. Some 65·6 % of schoolchildren aged 3-17 years had school lunch at least once weekly. This rate of attendance was positively correlated with age. Whatever the school level, school canteen attendance was positively associated with the educational level of the caregiver/parent. In pre- and elementary-school children, enrolment at the school canteen was also higher when the caregiver/parent worked, or in single-parent families. In secondary-school children, school lunch participation decreased with children living in more densely populated areas and increased with the level of the household's living standards. School canteen attendance was positively associated with children's socio-economic background. This could reduce the effectiveness of the forthcoming school meal composition regulations designed to improve the diet of children from deprived backgrounds, who are more likely to have unhealthy food habits.


[Occupational safety and health promotion: competition or cooperation?].
Between 1988-1998 in Germany Occupational Safety and Health (OSH) rivalled with Workplace Health Promotion (WHP). Now that European legislation has influenced modernisation of the German OSH, both can embark on useful cooperation. Safety Services and personnel are required to evaluate risk assessment accurately; the results can be helpful for WHP. Safety communication and workers' participation will explore and ensure new avenues in WHP and--in consequence--scientific knowledge concerning work. This knowledge, in turn, can now support modern OSH.


Sepsis and pulmonary edema.
Sepsis is the most common cause of adult respiratory distress syndrome and is associated with the highest mortality. This article describes the pathophysiology of septic pulmonary edema, which is the culmination over time of many complex responses related to sepsis.


Effects of antiepileptic drugs on cortical excitability in humans: A TMS-EMG and TMS-EEG study.
Brain responses to transcranial magnetic stimulation (TMS) recorded by electroencephalography (EEG) are emergent noninvasive markers of neuronal excitability and effective connectivity in humans. However, the underlying physiology of these TMS-evoked EEG potentials (TEPs) is still heavily underexplored, impeding a broad application of TEPs to study pathology in neuropsychiatric disorders. Here we tested the effects of a single oral dose of three antiepileptic drugs with specific modes of action (carbamazepine, a voltage-gated sodium channel (VGSC) blocker; brivaracetam, a ligand to the presynaptic vesicle protein VSA2; tiagabine, a gamma-aminobutyric acid (GABA) reuptake inhibitor) on TEP amplitudes in 15 healthy adults in a double-blinded randomized placebo-controlled crossover design. We found that carbamazepine decreased the P25 and P180 TEP components, and brivaracetam the N100 amplitude in the nonstimulated hemisphere, while tiagabine had no effect. Findings corroborate the view that the P25 represents axonal excitability of the corticospinal system, the N100 in the nonstimulated hemisphere propagated activity suppressed by inhibition of presynaptic neurotransmitter release, and the P180 late activity particularly sensitive to VGSC blockade. Pharmaco-physiological characterization of TEPs will facilitate utilization of TMS-EEG in neuropsychiatric disorders with altered excitability and/or network connectivity.


New mechanisms driving muscle stem cell regenerative decline with aging.
Stem cells must preserve their function in order to sustain organ and tissue formation, homeostasis and repair. Adult stem cells, particularly those resident in tissues with little turnover, remain quiescent for most of their life, activating only in response to regenerative demands. Among the best studied long-lived quiescent stem cells are skeletal muscle stem cells, which are fully equipped to sustain repair in response to tissue trauma. Recent evidence indicates that the preservation of muscle stem-cell quiescence and regenerative capacity depends on intracellular networks linking metabolism and protein homeostasis. Here, we review recent research into how these networks control stem cell function and how their dysregulation contributes to aging, with a particular focus on senescence entry in extreme old age. We also discuss the implications of these new findings for anti-aging research in muscle stem-cell-based regenerative medicine.


Online Kernel Learning With Adaptive Bandwidth by Optimal Control Approach.
Online learning methods are designed to establish timely predictive models for machine learning problems. The methods for online learning of nonlinear systems are usually developed in the reproducing kernel Hilbert space (RKHS) associated with Gaussian kernel in which the kernel bandwidth is manually selected and remains steady during the entire modeling process in most cases. This setting may make the learning model rigid and inappropriate for complex data streams. Since the bandwidth appears in a nonlinear term of the kernel model, it raises substantial challenges in the development of learning methods with an adaptive bandwidth. In this article, we propose a novel approach to address this important open issue. By a carefully casted linearization scheme, the nonlinear learning problem is reasonably transformed into a state feedback control problem for a series of controllable systems. Then, by employing optimal control techniques, an effective algorithm is developed, and the parameters in the learning model including kernel bandwidth can be efficiently updated in a real-time manner. By taking advantage of the particular structure of the Gaussian kernel model, a theoretical analysis on the convergence and rationality of the proposed method is also provided. Compared with the kernel algorithms with a fixed bandwidth, our novel learning framework can not only achieve adaptive learning results with a better prediction accuracy but also show performance that is more robust with a faster convergence speed. Encouraging numerical results are provided to demonstrate the advantages of our new method.


Matrix-producing mammary carcinoma: a rare breast tumor.
Matrix-producing carcinoma of the breast is a unique subclass of metaplastic carcinoma which is characterized by the existence of a ductal carcinomatous component with direct transition to areas showing cartilagenous or osseous differentiation, lacking an interspersed spindle cell component. This article reports one such rare case in a 50-year-old woman who had a right breast mass. f0 ine needle aspiration (FNA) smears showed abundant chondromyxoid extracellular matrix to which were variably admixed carcinomatous cells. Histological examination revealed a neoplasm composed of invave ductal carcinoma with a direct transition to chondrosarcomatous areas. The case is reported not only for its peculiar microscopic characteristics but also to highlight its better prognostic features and hence, the need for its recognition.


Lensless coherent imaging by a deterministic phase retrieval method with an aperture-array filter.
Recently we have proposed a noniterative and analytic phase retrieval method using the filter of an aperture array to reconstruct a complex-valued object from a diffraction intensity pattern [Phys. Rev. Lett.98, 223901 (2007)], but this method suffers from the restriction of the far-field condition of two distances between the object and the filter and between the filter and the detector for the intensity measurement. An improved method, which extends the adaptable condition of those distances to the region of Fresnel diffraction, is proposed here. In addition, the procedure for reducing the influence of noises on the phase retrieval is presented, in which the phase information contained in multiple groups of sampling data of a single intensity distribution is utilized. The usefulness of this method is shown in computer-simulated examples of the object reconstructions, including an object with phase vortices.


Enhanced cyclooxygenase-2 gene expression in alcoholic liver disease in the rat.
Inflammatory stimuli and lipid peroxidation up-regulate cyclooxygenase (COX)-2. This study evaluated the relationship between inflammatory mediators, COX expression, and pathological changes in experimental alcoholic liver disease. Rats (5 per group) were fed ethanol and a diet containing saturated fat, corn oil, or fish oil by intragastric infusion. Dextrose isocalorically replaced ethanol in controls. In the first set of experiments, whole livers were analyzed. In the second set of experiments, Kupffer cells, endothelial cells, and hepatocytes were isolated from rats in each group. Pathological analyses and measurements of lipid peroxidation, tumor necrosis factor (TNF)-alpha, COX-1 and COX-2 messenger RNA (mRNA), endotoxin, and liver and plasma thromboxane were performed. Increased expression of COX-2 mRNA was detected in the livers of rats showing necroinflammatory changes. The Kupffer cell was the cell primarily responsible for the increase in COX-2 mRNA level. Increased expression of COX-2 was associated with increased levels of endotoxin, TNF-alpha mRNA, lipid peroxidation, and synthesis of thromboxane. COX-1 mRNA was decreased in Kupffer cells in rats with the most severe liver injury. Up-regulation of COX-2 in alcoholic liver injury occurred in the presence of proinflammatory stimuli and resulted in increased synthesis of inflammatory and vasoactive eicosanoids. Down-regulation of COX-1 may result in decreased synthesis of cytoprotective eicosanoids and additionally exacerbate liver injury.


Contribution of dermal macrophage trafficking in the sensitization phase of contact hypersensitivity.
We investigated cellular trafficking of dermal macrophages that express a macrophage calcium-type lectin (MMGL) during the sensitization of delayed-type hypersensitivity. In skin, dermal macrophages, but not epidermal Langerhans cells, have been shown to express MMGL. Epicutaneous sensitization by FITC produced a transient increase in MMGL-positive cells in regional lymph nodes. To directly investigate whether the increase was due to cell migration from dermis, MMGL-positive cells purified from skin were intradermally injected into syngeneic mice after labeling with a fluorescent cell tracer, followed by epicutaneous sensitization over the site of injection. MMGL-positive cells containing the tracer were found in the regional lymph nodes after sensitization. The majority of the MMGL-positive cell migrants were negative for FITC fluorescence despite the presence of FITC-labeled cells that included Langerhans cell migrants. Because the extent of MMGL-positive cell migration was greatly influenced by the selection of vehicles to dissolve FITC, the efficiency of sensitization was compared using the ear swelling test. Migration of both Langerhans cells (FITC-labeled cells) and MMGL-positive cells contributed positively to the efficiency of sensitization. Interestingly, MMGL-positive cell migration was induced by vehicle alone, even in the absence of FITC. These results suggest that migration of dermal MMGL-positive cells accounts for the adjuvant effects of vehicles at least in part.


Soluble LDL-receptor is induced by TNF-α and inhibits hepatocytic clearance of LDL-cholesterol.
Defective LDL-C clearance and hence its elevation in the circulation is an established risk factor for cardiovascular diseases (CVDs) such as myocardial infarction (MI). A soluble LDL-receptor (sLDL-R) has been detected in human plasma which correlates strongly with circulating LDL-C and classical conditions that promote chronic inflammation. However, the mechanistic interplay between sLDL-R, inflammation, and CVDs remains to be investigated. Here, we report that stimulation of HepG2 cells with TNF-α induces the release of sLDL-R into culture supernatants. In addition, TNF-α induces gene expression of peptidases ADAM-17 and MMP-14 in HepG2 cells, and inhibiting these peptidases using TMI 1 significantly reduces the TNF-α induced sLDL-R release. We found that a soluble form of recombinant LDL-R (100 nM) can strongly bind to LDL-C and form a stable complex (KD = E-12). Moreover, incubation of HepG2 cells with this recombinant LDL-R resulted in reduced LDL-C uptake in a dose-dependent manner. In a nested case-control study, we found that baseline sLDL-R in plasma is positively correlated with plasma total cholesterol level. Furthermore, a twofold increase in plasma sLDL-R was associated with a 55% increase in the risk of future MI [AOR = 1.55 (95% CI = 1.10-2.18)]. Nevertheless, mediation analyses revealed that a significant proportion of the association is mediated by elevation in plasma cholesterol level (indirect effect β = 0.21 (95% CI = 0.07-0.38). Collectively, our study shows that sLDL-R is induced by a pro-inflammatory cytokine TNF-α via membrane shedding. Furthermore, an increase in sLDL-R could inhibit hepatic clearance of LDL-C increasing its half-life in the circulation and contributing to the pathogenesis of MI. KEY MESSAGES: TNF-α causes shedding of hepatocytic LDL-R through induction of ADAM-17 and MMP-14. sLDL-R binds strongly to LDL-C and inhibits its uptake by hepatocytic cells. Plasma sLDL-R is positively correlated with TNF-α and cholesterol. Plasma sLDL-R is an independent predictor of myocardial infarction (MI). Plasma cholesterol mediates the association between sLDL-R and MI.


Lamivudine monotherapy in HIV-1-infected patients harbouring a lamivudine-resistant virus: a randomized pilot study (E-184V study).
We compared the immunological and clinical outcomes of lamivudine monotherapy and complete therapy interruption in the treatment of HIV-1-infected patients harbouring lamivudine-resistant virus. This 48-week, open-label pilot study randomly assigned HIV-infected patients receiving lamivudine-containing HAART and harbouring the M184V mutation to monotherapy with lamivudine 300 mg once daily (lamivudine group) or the discontinuation of all antiretroviral drugs (TI group). The primary endpoint was the occurrence of immunological or clinical failure; immunological failure was defined as the first report of a CD4 T-cell count less than 350 cells/microl, and clinical failure as the occurrence of a Centers for Disease Control and Prevention grade B or C event. The data were analysed on the basis of the intention-to-treat principle. By week 48, 20 of 29 patients in the TI group (69%; 95% CI 51-83%) and 12 of 29 in the lamivudine group (41%; 95% CI 26-59%) had discontinued the study because of immunological or clinical failure, which was significantly delayed in the lamivudine group (P = 0.018). Only patients in the TI group (6/29, 20.7%) experienced grade 3-4 clinical adverse events at least possibly related to HIV-1 (P = 0.02). The mean decline in CD4 cell percentage, viral rebound and recovery of HIV-1 replication capacity were significantly lower in the lamivudine group. The 24-week virological and immunological response after therapy resumption in patients who prematurely discontinued the study was similar in the two groups. In HIV-1-infected patients harbouring a lamivudine-resistant virus, lamivudine monotherapy may lead to a better immunological and clinical outcome than complete therapy interruption.


Pyridoxine deficiency: new approaches in immunosuppression and chemotherapy.
Pyridoxine deficiency leads to impairment of immune responses. It appears that the basic derangement is the decreased rate of production of one-carbon units necessary for the synthesis of nucleic acids. The key factor is a pyridoxine enzyme, serine hydroxymethyltransferase. This enzyme is very low in resting lymphocytes but increases significantly under the influence of antigenic or mitogenic stimuli, thus supplying the increased demand for nucleic acid synthesis during an immune response. Serine hydroxymethyltransferase activity is depressed by deoxypyridoxine, a potent antagonist of pyridoxal phosphate, and also by known immunosuppressive or antiproliferative agents. The combination of these agents is additive. Our results lead us to suggest the following medical applications: (a) combination of deoxypyridoxine with immunosuppressive or chemotherapeutic drugs may be effective in cases of immunosuppressive therapy or organ transplantation, (b) the development of special agents directed against the serine hydroxymethyltransferase apoprotein may prove to be a valuable medical tool, since this enzyme presents an excellent target for chemotherapy, (c) lymphocytes of individual patients could be used to design tailor-made specific immunosuppressive or chemotherapeutic treatment, and (d) the serine hydroxymethyltransferase activity of lymphocyte culture presents an excellent indicator for the evaluation of potency of immunosuppressive, chemotherapeutic or genotoxic compounds in a simple and rapid test.


The sleep log and actigraphy: congruency of measurement results for heart failure patients.
: Quality of life in patients with heart failure (HF) can be significantly impacted by poor sleep and its daytime consequences. As more attention is being paid to the sleep problems of HF patients, it is important to evaluate the degree of congruence between subjective and objective sleep measurements in this patient group. : This study was developed to evaluate the congruence between sleep parameters as measured using a wrist-worn ActiGraph and a daily sleep log in patients with stable HF. : Forty-three HF patients aged 40-92 years served as subjects. Sleep parameters were derived from actigraphy and a daily sleep log by averaging scores for 7 nights. : There were significant differences in wake time after sleep onset (WASO) and total sleep time between the sleep log and the ActiGraph (both ps < .001). Neither WASO nor sleep onset latency, both derived from the sleep log, correlated significantly with actigraphy variables. The mean bias for WASO and total sleep time between methods was 54.1 min (SD = 47.5 min) and 109.3 min (SD = 91.68 min) as assessed using a Bland-Altman analysis. A majority (83.7%) of participants experienced sleep disturbances as assessed by actigraphy. However, fewer (53.5%) had sleep disturbances as assessed using the sleep log. : A considerable degree of incongruence between actigraphy- and sleep log-derived measures of sleep exists in patients with stable HF.


Pregnancy and Contraception Experiences in Women With Narcolepsy: A Narcolepsy Network Survey.
To explore knowledge and experiences of women with narcolepsy on pregnancy and contraception issues and their relationships with narcolepsy pharmacotherapy. An 18-item survey was administered through the Narcolepsy Network website for 8 weeks during the fall of 2012. The survey ascertained demographic information; prescription narcolepsy medication use and discontinuation during pregnancy; physician counseling regarding pregnancy, contraception, and medication usage; and pregnancy history and outcomes. Frequencies of responses were analyzed and compared between pharmacotherapy groups. Surveys from 182 women (age 41.5 ± 15.2 years) with narcolepsy were analyzed. Most of the respondents (78.7%) who reported a history of pregnancy did not use pharmacotherapy during pregnancy. Most of them discontinued narcolepsy pharmacotherapy during pregnancy because of their own fear of harming the fetus (82.9%), and 58.5% noted advice of discontinuation from their narcolepsy physician as a factor in their decision. As an alternative to pharmacotherapy, 72.1% of women extended their sleep time, 32.6% discontinued working, and 27.9% discontinued driving. Similar pregnancy and fetal outcomes were reported between women using monotherapy, polytherapy, or no therapy during pregnancy, but some outcomes were worse than national averages. In general, women with narcolepsy were dissatisfied with the amount and type of counseling that they received regarding pregnancy and contraception. Improved health education counseling and symptom management options are needed for women with narcolepsy to improve pregnancy management and outcomes in this population.


Human immunodeficiency virus-associated eosinophilic folliculitis. A unique dermatosis associated with advanced human immunodeficiency virus infection.
We studied 13 patients with human immunodeficiency virus (HIV) infection and a chronic pruritic folliculitis that was unresponsive to systemic treatment with bactericidal anti-staphylococcal antibiotics. The skin eruption was characterized by multiple urticarial follicular papules scattered on the trunk (100%), the head and neck (85%), and the proximal aspect of the extremities (62%). Absolute peripheral eosinophil counts were increased in six of 13 patients; a relative peripheral eosinophilia was present in 10 of 13 patients. Serum IgE levels were elevated in all seven patients tested (range, 88 to 9050 IU). Histopathologic features included a folliculitis with eosinophils. Pathogenic bacteria were not consistently found by routine bacterial skin cultures, cultures of skin biopsy specimens, or histopathologic evaluation. CD4 counts were decreased in all of the 12 patients tested (less than 300 cells per cubic millimeter) and were below 250 cells per cubic millimeter in 10 patients. A clinical response was noted to astemizole, to ultraviolet light in the B range, and to topical clobetasol propionate. These observations demonstrate that HIV-associated eosinophilic folliculitis is a unique HIV-related cutaneous disorder that is characterized by a culture-negative, chronic, pruritic folliculitis and a characteristic histopathologic picture. Of special importance, because it is associated with CD4 counts of less than 250 to 300 cells per cubic millimeter, eosinophilic folliculitis appears to be an important clinical marker of HIV infection and, particularly, of patients at increased risk of developing opportunistic infections. We suggest that the term eosinophilic pustular folliculitis (Ofuji's disease), previously used to describe this dermatosis in HIV-infected patients, should be discarded.


Application of enzyme-linked immunosorbent assay analysis for determination of monensin in environmental samples.
There is growing concern that antibiotic use in livestock production is contributing to contamination of soil and aquatic environments. Monensin, a polyether ionophore antibiotic, accounts for approximately 13% of total subtherapeutic livestock antibiotic usage in the USA and has been widely detected in aquatic environments. Due to insufficient ultraviolet absorbance, liquid chromatography analysis of monensin in environmental samples is limited to equipment with mass spectrometry (e.g., liquid chomatography-mass spectrometry [LC-MS]). However, LC-MS equipment is costly, and extensive sample preparation and clean-up is often required. Rapid, low-cost analytical techniques are needed to monitor for monensin residues in the environment. In this study, a commercially available enzyme-linked immunosorbent assay (ELISA) for detecting monensin in animal feed extracts was evaluated for determination of monensin in water, soil, and manure. The monensin ELISA was highly sensitive, with limits of detection and quantification at 1.5 and 3.0 microg L(-1), respectively. Recovery of monensin in spiked water samples was approximately 100%. Cross-reactivity was not observed with similar polyether ionophores, tetracyclines, macrolides, or sulfonamides. Concentrations of monensin using ELISA were greater than concentrations measured with LC-MS. This is attributed to cross-reactivity of the monensin ELISA toward structurally similar monensin compounds, such as factors (slight naturally produced structural variants) and metabolites. Results from this study showed that ELISA can be a reliable, rapid, and low-cost alternative to LC-MS analysis of environmental samples.


Liquid crystal hydroglass formed via phase separation of nanocellulose colloidal rods.
A new anisotropic soft material - a liquid crystal 'hydroglass' (LCH) - is created from aqueous suspensions of nanocrystalline cellulose (NCC) colloidal rods. Under specific conditions, the NCC suspension separates into a colloid-rich attractive glass matrix phase and a coexisting liquid crystal phase. LCH provides similar viscoelastic properties to polymer and colloidal gels, but permits reversibly-orientating the colloidal rods through shear forces.


Neutral endopeptidase-resistant C-type natriuretic peptide variant represents a new therapeutic approach for treatment of fibroblast growth factor receptor 3-related dwarfism.
Achondroplasia (ACH), the most common form of human dwarfism, is caused by an activating autosomal dominant mutation in the fibroblast growth factor receptor-3 gene. Genetic overexpression of C-type natriuretic peptide (CNP), a positive regulator of endochondral bone growth, prevents dwarfism in mouse models of ACH. However, administration of exogenous CNP is compromised by its rapid clearance in vivo through receptor-mediated and proteolytic pathways. Using in vitro approaches, we developed modified variants of human CNP, resistant to proteolytic degradation by neutral endopeptidase, that retain the ability to stimulate signaling downstream of the CNP receptor, natriuretic peptide receptor B. The variants tested in vivo demonstrated significantly longer serum half-lives than native CNP. Subcutaneous administration of one of these CNP variants (BMN 111) resulted in correction of the dwarfism phenotype in a mouse model of ACH and overgrowth of the axial and appendicular skeletons in wild-type mice without observable changes in trabecular and cortical bone architecture. Moreover, significant growth plate widening that translated into accelerated bone growth, at hemodynamically tolerable doses, was observed in juvenile cynomolgus monkeys that had received daily subcutaneous administrations of BMN 111. BMN 111 was well tolerated and represents a promising new approach for treatment of patients with ACH.


Creaming and Parking in Quasi-Marketised Welfare-to-Work Schemes: Designed Out Of or Designed In to the UK Work Programme?
'Creaming' and 'parking' are endemic concerns within quasi-marketised welfare-to-work (WTW) systems internationally, and the UK's flagship Work Programme for the long-term unemployed is something of an international pioneer of WTW delivery, based on outsourcing, payment by results and provider flexibility. In the Work Programme design, providers' incentives to 'cream' and 'park' differently positioned claimants are intended to be mitigated through the existence of nine payment groups (based on claimants' prior benefit type) into which different claimants are allocated and across which job outcome payments for providers differ. Evaluation evidence suggests however that 'creaming' and 'parking' practices remain common. This paper offers original quantitative insights into the extent of claimant variation within these payment groups, which, contrary to the government's intention, seem more likely to design in rather than design out 'creaming' and 'parking'. In response, a statistical approach to differential payment setting is explored and is shown to be a viable and more effective way to design a set of alternative and empirically grounded payment groups, offering greater predictive power and value-for-money than is the case in the current Work Programme design.


Computational modeling of the olfactory receptor Olfr73 suggests a molecular basis for low potency of olfactory receptor-activating compounds.
The mammalian olfactory system uses hundreds of specialized G-protein-coupled olfactory receptors (ORs) to discriminate a nearly unlimited number of odorants. Cognate agonists of most ORs have not yet been identified and potential non-olfactory processes mediated by ORs are unknown. Here, we used molecular modeling, fingerprint interaction analysis and molecular dynamics simulations to show that the binding pocket of the prototypical olfactory receptor Olfr73 is smaller, but more flexible, than binding pockets of typical non-olfactory G-protein-coupled receptors. We extended our modeling to virtual screening of a library of 1.6 million compounds against Olfr73. Our screen predicted 25 Olfr73 agonists beyond traditional odorants, of which 17 compounds, some with therapeutic potential, were validated in cell-based assays. Our modeling suggests a molecular basis for reduced interaction contacts between an odorant and its OR and thus the typical low potency of OR-activating compounds. These results provide a proof-of-principle for identifying novel therapeutic OR agonists.


Application of peripherally inserted central catheters in critically ill newborns experience from a neonatal intensive care unit.
Peripherally inserted central catheters (PICCs) can provide nutritional and medical support for very low birth weight or critically ill newborns. The aim of this study was to retrospectively analyze the use of PICCs in our clinic for critically ill newborns to evaluate the relationship between catheter related factors and the occurrence of complications.Retrospective analysis was conducted for all newborns consecutively admitted at the Neonatal Intensive Care Unit (NICU), Chongqing Health Center for Women and Children, who underwent PICC insertion between May 2011 and March 2018. Data collected included total puncture success rate, one puncture success rate, infection rate, complication rate, unplanned catheter withdrawal rate, device days, and catheter indwelling time.Five-hundred eighty-eight infants (304 males and 284 females) aged 3.4 ± 3.9 days, mean gestational age of 30.9 ± 2.7 weeks and a mean body mass of 1.38 ± 0.47 kg at insertion were included. Total puncture success rate was 99.65%, one puncture success rate was 77.77%. The mean catheter retention was 13.6 ± 6.7 days: more than 30 days in 15 (2.61%) cases, 20 to 30 days in 60 (10.43%) cases, 10 to 19 days in 372 (64.70%) cases, and 62 days in 1 case. Complications occurred in 63 (10.71%) cases: with PICC insertion within 24 hours after birth in 29 (15.43%), within 48 hours in 13 (6.63%), and after 48 hours in 21 (10.99%) cases. Catheter tip culture was positive in 3 cases and there was 1 case of catheter-related bloodstream infection.Nursing measures of the maintenance of body temperature and the evaluation of blood vessels were important conditions for improving the success rate of one puncture in critically ill neonates. PICC catheterization as early as 48 hours will not increase the difficulty of PICC puncture. Nor did it increase the incidence of PICC complications.