Today´s medical self and the other: Challenges and evolving solutions for enhanced humanization and quality of care.
Recent scientific developments, along with growing awareness of cultural and social diversity, have led to a continuously growing range of available treatment options; however, such developments occasionally lead to an undesirable imbalance between science, technology and humanism in clinical practice. This study explores the understanding and practice of values and value clusters in real-life clinical settings, as well as their role in the humanization of medicine and its institutions. The research focuses on the values of clinical practice as a means of finding ways to enhance the pairing of Evidence-Based Medicine (EBM) with Values-based Medicine (VBM) in daily practice. The views and representations of clinical practice in 15 pre-CME and 15 post-CME interviews were obtained from a random sampling of active healthcare professionals. These views were then identified and qualitatively analyzed using a three-step hermeneutical approach. A clinical values space was identified in which ethical and epistemic values emerge, grow and develop within the biomedical, ethical, and socio-economic dimensions of everyday health care. Three main values-as well as the dynamic clusters and networks that they tend to form-were recognized: healthcare personnel-patient relationships, empathy, and respect. An examination of the interviews suggested that an adequate conceptualization of values leads to the formation of a wider axiological system. The role of clinician-as-consociate emerged as an ideal for achieving medical excellence. By showing the intricate clusters and networks into which values are interwoven, our analysis suggests methods for fine-tuning educational interventions so they can lead to demonstrable changes in attitudes and practices.


High-Quality Library Preparation for NGS-Based Immunoglobulin Germline Gene Inference and Repertoire Expression Analysis.
Next generation sequencing (NGS) of immunoglobulin (Ig) repertoires (Rep-seq) enables examination of the adaptive immune system at an unprecedented level. Applications include studies of expressed repertoires, gene usage, somatic hypermutation levels, Ig lineage tracing and identification of genetic variation within the Ig loci through inference methods. All these applications require starting libraries that allow the generation of sequence data with low error rate and optimal representation of the expressed repertoire. Here, we provide detailed protocols for the production of libraries suitable for human Ig germline gene inference and Ig repertoire studies. Various parameters used in the process were tested in order to demonstrate factors that are critical to obtain high quality libraries. We demonstrate an improved 5'RACE technique that reduces the length constraints of Illumina MiSeq based Rep-seq analysis but allows for the acquisition of sequences upstream of Ig V genes, useful for primer design. We then describe a 5' multiplex method for library preparation, which yields full length V(D)J sequences suitable for genotype identification and novel gene inference. We provide comprehensive sets of primers targeting IGHV, IGKV, and IGLV genes. Using the optimized protocol, we produced IgM, IgG, IgK, and IgL libraries and analyzed them using the germline inference tool IgDiscover to identify expressed germline V alleles. This process additionally uncovered three IGHV, one IGKV, and six IGLV novel alleles in a single individual, which are absent from the IMGT reference database, highlighting the need for further study of Ig genetic variation. The library generation protocols presented here enable a robust means of analyzing expressed Ig repertoires, identifying novel alleles and producing individualized germline gene databases from humans.


The Fungal Microbiome Is an Important Component of Vineyard Ecosystems and Correlates with Regional Distinctiveness of Wine.
The flavors of fermented plant foods and beverages are formed by microorganisms, and in the case of wine, the location and environmental features of the vineyard site also imprint the wine with distinctive aromas and flavors. Microbial growth and metabolism play an integral role in wine production, by influencing grapevine health, wine fermentation, and the flavor, aroma, and quality of finished wines. The contributions by which microbial distribution patterns drive wine metabolites are unclear, and while flavor has been correlated with fungal and bacterial composition for wine, bacterial activity provides fewer sensorially active biochemical conversions than fungi in wine fermentation. Here, we collected samples across six distinct wine-growing areas in southern Australia to investigate regional distribution patterns of fungi and bacteria and the association with wine chemical composition. Results show that both soil and must microbiota distinguish wine-growing regions. We found a relationship between microbial and wine metabolic profiles under different environmental conditions, in particular measures of soil properties and weather. Fungal communities are associated with wine regional distinctiveness. We found that the soil microbiome is a source of grape- and must-associated fungi and suggest that weather and soil could influence wine characteristics via the soil fungal community. Our report describes a comprehensive scenario of wine microbial biogeography where microbial diversity responds to the surrounding environment and correlates with wine composition and regional characteristics. These findings provide perspectives for thoughtful human practices to optimize food composition through understanding fungal activity and abundance.IMPORTANCE The composition of soil has long been thought to provide wine with characteristic regional flavors. Here, we show that for vineyards in southern Australia, the soil fungal communities are of primary importance for the aromas found in wines. We propose a mechanism by which fungi can move from the soil through the vine.


Supporting healthful lifestyles during pregnancy: a health coach intervention pilot study.
Excessive gestational weight gain (GWG) is associated with adverse health outcomes in both the mother and child. Many previous lifestyle interventions in women with excess weight during pregnancy encouraging appropriate GWG have been unsuccessful, and there remains no consensus about the content, format, or theoretical framework of GWG interventions. We assessed the feasibility and acceptability of a remote health coach intervention to promote healthful lifestyle behaviors and appropriate GWG among overweight pregnant women. At one northeastern US clinic, we enrolled 30 overweight (pre-pregnancy BMI ≥ 25 kg/m2) pregnant women at a median gestation of 12.5 weeks (IQR: 11-15) into a one-arm trial. We connected participants with a health coach to provide behavioral support to help participants adopt healthful lifestyles during pregnancy. Health coaches contacted participants by phone every 2-3 weeks to monitor goals, and sent emails and text messages between calls. Participants completed baseline (N = 30) and follow-up (N = 26) surveys at the end of the intervention (36 weeks gestation), as well as follow-up phone interviews (N = 18). Among 30 participants, median age was 32 years (IQR: 28-33), median self-reported pre-pregnancy BMI was 27.3 kg/m2 (IQR: 25.7-31.1), and 17/30 were white, 9/30 African-American, and 3/30 Asian. Three-quarters (22/29) of participants completed at least a college degree. Although 25/30 participants reported in baseline surveys that they worried about being able to lose the weight postpartum that they expected to gain during pregnancy, just 12/26 participants reported the same at follow-up (P < 0.001). In follow-up surveys, 21/26 participants reported that health coaches were helpful in keeping them motivated, and 22/26 thought the phone conversations helped them face problems and find solutions. Based on qualitative assessment, several themes emerged in follow-up interviews about the quality of the intervention including accountability and support from health coaches. Participants also expressed desire for more visual resources and integration with standard clinical care to improve the intervention. We demonstrated feasibility and high participant satisfaction with our remote health coach intervention during pregnancy. We identified areas in which we could refine the intervention for inclusion in a full-scale RCT, such as integration with clinical care and additional visual resources. Retrospectively registered at ClinicalTrials.gov ( NCT03080064 , 3/14/2017).


The Impact of Yoga Nidra and Seated Meditation on the Mental Health of College Professors.
World statistics for the prevalence of anxiety and mood disorders shows that a great number of individuals will experience some type of anxiety or mood disorder at some point in their lifetime. Mind-body interventions such as Hatha Yoga and seated meditation have been used as a form of self-help therapy and it is especially useful for challenging occupations such as teachers and professors. In this investigation, we aimed at observing the impact of Yoga Nidra and seated meditation on the anxiety and depression levels of college professors. Sixty college professors, men and women, aged between 30 and 55 years were randomly allocated in one of the three experimental groups: Yoga Nidra, seated meditation, and control group. Professors were evaluated two times throughout the 3-month study period. Psychological variables included anxiety, stress, and depression. Data analysis showed that the relaxation group presented better intragroup results in the anxiety levels. Meditation group presented better intragroup results only in the anxiety variable (physical component). Intergroup analysis showed that, except for the depression levels, both intervention groups presented better results than the control group in all other variables. Prepost results indicate that both interventions represent an effective therapeutic approach in reducing anxiety and stress levels. However, there was a tendency toward a greater effectiveness of the Yoga Nidra intervention regarding anxiety, which might represent an effective tool in reducing both cognitive and physiological symptoms of anxiety.


Calcium- and guanine-nucleotide-dependent exocytosis in permeabilized rat mast cells. Modulation by protein kinase C.
We have used a digitonin-permeabilized cell system to study the signal transduction pathways responsible for stimulus-secretion coupling in the rat peritoneal mast cell. Conditions were established for permeabilizing the mast cell plasma membrane without disrupting secretory vesicles. Exocytotic release of histamine from digitonin-permeabilized cells required a combination of micromolar concentrations of Ca2+ and the stable guanine nucleotide analogue guanosine 5'-[gamma-thio]triphosphate (GTP[S]), but was independent of exogenous ATP. In the presence of 40 microM-GTP[S], exocytosis was half-maximal at 1.3 microM-Ca2+ and maximal at 10 microM-Ca2+; GTP[S] alone (100 microM) had no effect on histamine release in the absence of added Ca2+. In the presence of 10 microM free Ca2+, 5 microM-GTP[S] was required for half-maximal exocytosis. To examine the possible role of protein kinase C (PKC) in exocytosis, we utilized 12-O-tetradecanoylphorbol 13-acetate (TPA) to activate PKC and studied its effect on histamine release from permeabilized mast cells. Cells that had been incubated with TPA (25 nM for 5 min) exhibited increased sensitivity to both GTP[S] and Ca2+. The PKC inhibitor staurosporine blocked the effect of TPA without inhibiting normal exocytosis in response to the combination of GTP[S] and Ca2+. In addition, down-regulation of mast-cell PKC by long-term TPA treatment (25 nM for 20 h) blocked the ability of the cells to respond to TPA and inhibited exocytosis in response to Ca2+ and GTP[S] by 40-50%. These results suggest that the sensitivity of the exocytotic machinery of the mast cell can be altered by PKC-catalysed phosphorylation events, but that activation of PKC is not required for exocytosis to occur.


[Effect of the antioxidant, 4-methyl-2,6 di-tert-butylphenol on the mitotic division of epithelial cells of the small-intestinal crypts of mice depending on the degree of differentiation].
Antioxidants increase the number of mitoses in epithelial cell populations being on differentiation, examined from various parts of small intestinal crypts of mice. 2 hours after antioxidant injection a considerable increase in mitotic index (MI) is found, with maximal value (by 3 times) seen for cells from higher crypt region where more differentiated cells occur. The increase of MI is observed also in later period, 14, 21--24 hours after antioxidant administration. The observed effect may be explained by the influence of antioxidants on the reserve pool of cells. However, it does not seem unlikely that the increase of MI after antioxidant injection may be due to the influence of antioxidants on the longevity of individual stages of mitotic cycle. Similar evidence was provided earlier by the authors for the effect of antioxidants on the mitotic division of liver cells of normal mice.


Maternal factors regulating symmetry breaking and dorsal-ventral axis formation in the sea urchin embryo.
Specification of the main axes of polarity of the embryo is an essential process during embryonic development. In many species, this process is achieved by the localization of maternal factors into discrete regions of the egg. However, in other animals, like in amniotes and in echinoderms, the considerable plasticity of the early blastomeres seems to preclude the existence of maternal determinants and the mechanisms by which the radial symmetry of the egg is broken remain largely enigmatic. In this chapter, we review recent progress on the identification of maternal components involved in symmetry breaking and dorsal-ventral (D/V) axis formation of the sea urchin embryo. We will first review some key experiments on D/V axis formation from classical embryologists that provided evidence for a weak maternal D/V prepattern. We will then detail more recent molecular analyses that established the critical role played by Nodal signaling in allocating cell fates along the secondary axis and led to the discovery that maternal transcription factors such as the Sry-related HMG box B1 (SoxB1), the Octamer binding factor1/2 (Oct1/2), the T-cell factor/Lymphoid enhancer-binding factor (TCF/LEF) and the Erythroblastosis virus E26 Oncogene Homolog (ETS) domain transcriptional repressor Translocation-Ets-Leukemia virus protein (Yan/Tel) as well as maternal signaling molecules like Univin are essential for the initiation of nodal expression. Finally, we will describe recent advances that uncovered a role in symmetry breaking and dorsal-ventral axis orientation for the transforming growth factor beta (TGF-beta)-like factor Panda, which appears to be both necessary and sufficient for D/V axis orientation. Therefore, even in the highly regulative sea urchin embryo, the activity of localized maternal factors provides the embryo with a blueprint of the D/V axis.


Women in Free Clinics: An Assessment of Health-Related Quality of Life for Prevention and Health Education.
Understanding gender influences on health-related quality of life (HRQoL) is important to improve women's health when considering diseases that afflict women specifically. The target population of this study was uninsured female free clinic patients who are low socio-economic status and lack access to healthcare resources. Free clinics provide free or reduced fee healthcare to individuals who lack access to primary care and are socio-economically disadvantaged. While approximately half of free clinic patients are women, there is a paucity of comprehensive health-related data for female free clinic patients. US born English, non-US born English, and Spanish speaking female free clinic patients completed a self-administered survey using a standardized women's HRQoL measure in Fall 2014 (N = 389). Female free clinic patients reported lower HRQoL on all aspects of women's health compared to the US baseline scores, and were less likely to utilize preventive care including: mammograms, Pap smear, and HPV vaccination compared to the US general population. Spanish speakers reported a higher percentage of having had mammography and Pap smear, and heard about HPV compared to the other two groups. US born English speakers reported lower levels of HRQoL in vasomotor symptoms and sleep symptoms, and the lowest percentage of breast health and Pap smear screenings compared to non-US born English and Spanish speakers. Non-US born English speakers reported higher preference for female physician compared to US born English speakers and Spanish speakers. Free clinic female patients need preventative interventions and educational opportunities to improve their overall HRQoL.


Biomarkers in imaging: realizing radiology's future.
Modern pharmaceuticals and medical devices have provided substantial benefits to patients throughout the world. These benefits come at a high and increasing cost, with development of the typical pharmaceutical requiring 12 years and hundreds of millions of dollars before gaining U.S. Food and Drug Administration marketing approval. Appropriate use of imaging biomarkers--defined as anatomic, physiologic, biochemical, or molecular parameters detectable with imaging methods used to establish the presence or severity of disease--offer the prospect of smaller, less expensive, and more efficient preclinical studies and clinical trials. Scientists, government regulators, and industry have all recognized the potential of biomarkers in imaging. Although real, this promise can only be realized with the rigorous application of science to their use. Success is most likely when (a) the presence of an imaging marker is closely linked with the presence of a target disease; (b) detection and/or measurement of the biomarker is accurate, reproducible, and feasible over time; and (c) measured changes are closely linked to success or failure of the therapeutic effect of the product being evaluated. By applying this paradigm to the array of imaging modalities, the radiology community is poised to become a major force in preclinical and clinical evaluations of new medical treatments.


Evaluation of an anxiety-related phenotype in galanin overexpressing transgenic mice.
Understanding the role of neuropeptides in mediating emotional behaviors is an important avenue for discovering novel drug targets for anxiety disorders. A role for galanin in mediating anxiety-related behavior is suggested by the pattern of distribution in the CNS and the coexistence of galanin with norepinephrine in the locus coeruleus. Studies in rats have shown that central administration of galanin modulates anxiety-related behaviors, and galanin release blocks the proanxiety effects of noradrenergic activation in prestressed rats. To further investigate the role of galanin in anxiety behaviors, we conducted a comprehensive behavioral phenotyping of galanin overexpressing transgenic mice (GAL-tg). GAL-tg mice were normal on measures of general health, neurological reflexes, home cage social behaviors, sensory functions, motor coordination, and exploratory locomotor activity. In three separate tests for anxiety-related behaviors, the elevated plus-maze, light <--> dark exploration, and open field center time, GAL-tg mice showed no anxiety-like phenotype. GAL-tg mice and wild-type littermate controls were equally responsive to the anxiolytic effects of chlordiazepoxide (10 mg/kg) in the light <--> dark exploration test, indicating normal benzodiazepine receptor function in GAL-tg mice. Stimulation of noradrenergic cells via administration with an alpha2 adrenoreceptor antagonist, yohimbine (2.5 mg/kg), produced proanxiety effects in wild type mice in the light <--> dark exploration test, but not in the GAL-tg mice. These data suggest that galanin contributes to the modulation of anxiety states induced by high levels of noradrenergic activation, but is silent under less challenging situations. A specific role for galanin in extreme anxiety states represents an attractive target for the development of novel anxiolytic treatments.


Solution, solid state, and film properties of a structurally characterized highly luminescent molecular europium plastic material excitable with visible light.
The synthesis and X-ray crystal structure of the ligand L (4,7-dicarbazol-9-yl-[1,10]-phenanthroline) are reported, as well as those of the molecular complex, [Eu(tta)(3)(L)] (1), (tta = 2-thenoyl trifluoroacetylacetonate). Their photophysical properties have been investigated both in solution and in the solid state. It was shown that the ligands used for designing 1 are well-suited for sensitizing the Eu(III) ion emission, thanks to a favorable position of the triplet state as investigated in the Gd(III) complex [Gd(tta)(3)(L)], (2). The low local symmetry of the Eu(III) ion shown by the X-ray crystal structure of 1 is also revealed by luminescence spectroscopy. Because of interesting volatility and solubility properties, 1 is shown to behave as a real molecular material that can be processed both by thermal evaporation and from solution. When doped in poly(methylmethacrylate) (PMMA), 1 forms air-stable and highly red-emitting plastic materials that can be excited in a wide range of wavelengths from the UV to the visible part of the electromagnetic spectrum (250-560 nm). Absolute quantum yields of 80% have been obtained for films comprising 1-3% of 1. Ellipsometry measurements have been introduced to gain information on physical data of 1. They have been performed on thin films of 1 deposited by thermal evaporation and gave access to the refractive index, n, and the absorption coefficient, k, as a function of the wavelength. A value of 1.70 has been found for n at 633 nm. These thin films also show interesting air-stability.


A novel selective MAO-B inhibitor with neuroprotective and anti-Parkinsonian properties.
We previously reported that 1,3-bisbenzylimidazolium (DBZIM) bromide was neuroprotective for the dopaminergic system in Parkinson's disease (PD) models of rodent, however the underlying mechanism was unclear. We currently further confirmed that DBZIM ameliorated the Parkinsonian motor deficit and protected the nigrostriatal tract from the neurotoxicity of 1-methyl-4-(2'-methylphenyl)-1,2,3,6-tetrahydropyridine (2'-CH3-MPTP) in C57Bl/6 mice. The dopaminergic degeneration in the substantia nigra par compacta (SNc) and striatum was analyzed by immunohistochemistry while the monoamine oxidase B (MAO-B) inhibition effect of DBZIM was determined by enzyme kinetics. DBZIM was at least as effective as the clinically approved anti-PD drug, l-deprenyl (Selegiline), for both neuroprotection and correction of motor deficits. Mechanistically, DBZIM inhibited the specific activity of MAO-B in the striatum and C6 cells without affecting the protein expression. DBZIM directly inhibited the enzymatic activity of a purified MAO-B protein with an estimated Ki value from 780 to 940nM, in par with that of l-deprenyl (970nM). The physical interaction between DBZIM and MAO-B was proven by NMR analysis, with Kd around 21.5-46.8μM. Our binding and modelling data further illustrated that DBZIM is a mixed inhibitor with its binding to active site partially hindering the substrate binding. Therefore, inhibiting MAO-B is a major mechanism through which DBZIM confers neuroprotection for the dopaminergic neurons against 2'-CH3-MPTP toxicity. Remarkably, the post-lesion treatment with DBZIM provided greater anti-parkinsonian and neuroprotective effects than the l-deprenyl. The current study, together with our previous findings in a 6-OHDA PD model, demonstrated that DBZIM is a promising neuroprotectant for PD with anti-MAO-B property.


Minimum Inhibitory Concentrations of Vancomycin and Daptomycin Against Methicillin-resistant Staphylococcus Aureus Isolated from Various Clinical Specimens: A Study from South India.
Background Staphylococci are Gram-positive cocci arranged in clusters. They are colonized in humans and animals. Also, Staphylococcus aureus (S. aureus) is frequently associated with various superficial to deep-seated infections in humans. Due to the potential for easy transmission, Staphylococci are associated with both hospital-acquired and community-associated infections. Strains of S. aureus resistant to methicillin (MRSA) pose treatment challenges. In such cases, vancomycin is the treatment of choice. Due to the indiscriminate use of vancomycin, recently, we are seeing the emergence of vancomycin-intermediate sensitive S. aureus (VISA) and vancomycin-resistant S. aureus (VRSA). The present study aims to evaluate the minimum inhibitory concentrations (MICs) of vancomycin and daptomycin among MRSA strains isolated from human clinical specimens Methods The study included 115 MRSA isolates collected over 26 months from July 2010 to September 2012. The strains were isolated from pus, urine, wound swabs, catheters, blood, and sputum. The bacteria were acquired from different inpatient and outpatient departments of Prathima Institute of Medical Sciences, Karimnagar, Telangana, India. Kirby-Bauer disk diffusion method using cefoxitin was used to confirm the MRSA isolates. The agar dilution and the Epsilometer method (E-test) were used to test the MICs of MRSA isolates against vancomycin and daptomycin, respectively, by the standard procedures recommended by the clinical laboratory standards institute (CLSI). Results Of the 115 S. aureus isolates, seven (6.08%) strains were resistant to vancomycin (VRSA) and 53 (46.08%) were found to be VISA using the new CLSI breakpoints. The MIC of the daptomycin was found to be ≤1 µg/ml for all the MRSA isolates. Conclusion The study results depicted an increasing trend in the vancomycin MICs among the MRSA isolates. Several tested strains show MICs in the intermediate sensitive range (VISA). The daptomycin was effective against all the MRSA isolates.


Neurological outcome of newborns with neonatal seizures: a cohort study in a tertiary university hospital.
To describe the neurological outcome of newborns with seizures. Cohort study with newborns prospectively followed. Perinatal characteristics and etiological screening were related to outcome in a regression model. During the study 3,659 newborns were admitted and 2.7% were diagnosed as having seizures. Hypoxic ischemic encephalopathy (51%) was the etiology more frequently associated to seizures and also to postneonatal epilepsy (53%). In the follow up 25 died during the acute neonatal illness and 9 during the first years of life, 19 were diagnosed as having post neonatal epilepsy, 35 had developmental delay and 11 an association among this two comorbidities. A significant association between abnormal postnatal EEG and neuroimaging to developmental delay (p=0.014, p=0.026) was observed. The group of newborns that had seizures presented an increased risk of developing epilepsy compared to newborns from the same cohort without seizures (19.3/100 vs. 1.8/100, p<0.001). In this study neonatal seizures predominated in term newborns with perinatal asphyxia an elevated perinatal mortality and post neonatal morbidity was observed. The follow up showed an increased risk for developing postnatal epilepsy and developmental delay.


Stenting of the biliary tract in children.
We report on the technique and results of percutaneous transhepatic biliary drainage (PTBD) in children with obstructive jaundice. Three patients aged 8 - 15 years were treated, two of them for a benign and one for a malignant stricture. Endoscopic treatment was not possible and all the PTBD procedures were done under general anaesthesia. One of the children was treated with external-internal drainage, and the two others by insertion of a plastic endoprosthesis. There were no immediate complications. The PTBD had a good palliative effect in two cases, and in one case surgical treatment was necessary. We conclude that PTBD is a safe modality and that it can be used in children for the relief of obstructive jaundice.


Harmonic generation in a two-dimensional nonlinear quasi-crystal.
Second-harmonic generation in a two-dimensional nonlinear quasi-crystal is demonstrated for the first time to our knowledge. Temperature and wavelength tuning of the crystal reveal the uniformity of the pattern while angle tuning reveals the dense nature of the crystal's Fourier spectrum. These results compare well with theoretical predictions showing the excellent uniformity of the crystal and suggest that more-complicated nonlinear holograms should be possible.


Eddy covariance carbonyl sulphide flux measurements with a quantum cascade laser absorption spectrometer.
The trace gas carbonyl sulphide (COS) has lately received growing interest in the eddy covariance (EC) community due to its potential to serve as an independent approach for constraining gross primary production and canopy stomatal conductance. Thanks to recent developments of fast-response high-precision trace gas analysers (e.g. quantum cascade laser absorption spectrometers (QCLAS)), a handful of EC COS flux measurements have been published since 2013. To date, however, a thorough methodological characterisation of QCLAS with regard to the requirements of the EC technique and the necessary processing steps has not been conducted. The objective of this study is to present a detailed characterization of the COS measurement with the Aerodyne QCLAS in the context of the EC technique, and to recommend best EC processing practices for those measurements. Data were collected from May to October 2015 at a temperate mountain grassland in Tyrol, Austria. Analysis of the Allan variance of high-frequency concentration measurements revealed sensor drift to occur under field conditions after an averaging time of around 50 s. We thus explored the use of two high-pass filtering approaches (linear detrending and recursive filtering) as opposed to block averaging and linear interpolation of regular background measurements for covariance computation. Experimental low-pass filtering correction factors were derived from a detailed cospectral analysis. The CO2 and H2O flux measurements obtained with the QCLAS were compared against those obtained with a closed-path infrared gas analyser. Overall, our results suggest small, but systematic differences between the various high-pass filtering scenarios with regard to the fraction of data retained in the quality control and flux magnitudes. When COS and CO2 fluxes are combined in the so-called ecosystem relative uptake rate, systematic differences between the high-pass filtering scenarios largely cancel out, suggesting that this relative metric represents a robust key parameter comparable between studies relying on different post-processing schemes.


Occult choroidal neovascularization. Influence on visual outcome in patients with age-related macular degeneration. Macular Photocoagulation Study Group.
To determine whether the presence of occult choroidal neovascularization (CNV) influenced the anatomic and visual acuity outcomes in a randomized clinical trial of krypton red laser photocoagulation to treat juxtafoveal neovascular lesions in age-related macular degeneration. The fluorescein angiograms obtained at the baseline examination at tertiary retinal referral centers between April 1, 1981, and December 31, 1987, as part of the Macular Photocoagulation Study (MPS) Age-Related Macular Degeneration Study-Krypton Laser were evaluated retrospectively at the MPS Fundus Photograph Reading Center by two senior readers independently (with open adjudication of any differences) from 1992 to 1994. Criteria for classifying classic and occult CNV by the MPS Group were established in 1989, 2 years after the last patient had been assigned randomly to treatment or observation in the krypton laser study. Treatment coverage of classic and occult CNV, persistent CNV, recurrent CNV, and visual acuity from scheduled follow-up examinations for up to 5 years were analyzed for the absence or presence of occult CNV at baseline. The number of eyes with classic CNV but not occult CNV, classic and occult CNV, and occult CNV but no classic CNV were almost identical for the eyes assigned randomly to treatment or observation. Classic CNV almost always was covered completely with intense laser treatment; nevertheless, recurrent CNV developed in more than half of these eyes within 1 year after initial laser treatment. In contrast, in more than half of the eyes with occult CNV, more than 50% of the occult CNV was not covered with heavy laser treatment. Laser treatment was clearly beneficial for eyes with classic CNV but no occult CNV and almost equivalent to no treatment for eyes with classic and occult CNV. The few eyes with occult CNV but no classic CNV precluded conclusions about the value of treatment in this subgroup. These results strengthen previous reports that laser treatment is beneficial for eyes with juxtafoveal choroidal neovascular lesions when classic CNV is present, even though CNV often recurs. Treatment of classic CNV alone in eyes with classic and occult CNV was not beneficial in this study. Distinguishing classic CNV from occult CNV can aid in the selection of patients who will benefit most from laser treatment.


Enzyme-based resource allocated decomposition and landscape heterogeneity in the Florida Everglades.
Enzyme catalyzed reactions are generally considered the rate-limiting step in organic matter degradation and may be significantly influenced by the structure and composition of plant communities. Changes in these rates have the potential to effect long-term peat accumulation and influence the topography of a wetland ecosystem. To determine habitat influences on enzyme activities, we examined slough and sawgrass plots within enriched and reference phosphorus (P) sites in the Everglades. Assays were performed for the enzymes involved in carbon (C), nitrogen (N), and P cycling and lignin depolymerization. Enzyme activities were normalized and analyzed in terms of a resource allocation strategy. Plant composition was found to significantly alter the allocation of enzymatic resources due to varying substrate complexities. Potential decomposition in the slough was less influenced by lignin than in the sawgrass habitats. Additionally, an index relating hydrolytic and oxidative enzymes was significantly greater in the slough habitats, whereas C/N ratios were significantly lower. These indices suggest more favorable decomposition conditions and thus slower peat accretion within the slough communities, which may contribute to the development of elevation differences within the sawgrass ridge and slough topography of the Everglades.


Anakinra: a safe and effective first-line treatment in systemic onset juvenile idiopathic arthritis (SoJIA).
Systemic onset juvenile idiopathic arthritis (SoJIA) is a rare inflammatory disorder. It can result in disease and treatment-related disability. SoJIA is characterized by remitting fevers, evanescent rash, generalized lymphadenopathy, hepatomegaly/splenomegaly, and/or serositis. Non-responsiveness to standard therapy with corticosteroids and disease modifying antirheumatic drugs is not uncommon. IL-1β has been shown to be a main contributor to the pathogenesis of SoJIA. Anakinra, a recombinant IL-1β receptor antagonist, was shown to be effective in small cohorts of therapy-resistant adult and pediatric Still's patients. In order to assess the efficacy and safety of first-line anakinra treatment in SoJIA, we reviewed the charts of all SoJIA patients in our institution from 2005 to 2010, searching for first-line anakinra-treated patients. We report the clinical and laboratory course of four SoJIA patients. The mean follow-up was 13.5 (range: 2-50) months. Anakinra was started at doses from 1.5 to 4 mg/kg for a median duration of 3 (range: 3-18) months. Two patients responded to anakinra mono-therapy; two cases required corticosteroids. Normalized body temperatures and the absence of evanescent rashes were achieved after a median of 4 (range: 2-10) days. We did not see treatment-related adverse reactions other than local injection site inflammation. This is the first single-center series, reporting anakinra as first-line treatment in SoJIA. We show rapid efficacy of anakinra in early SoJIA with reduced treatment-related side effects. A subset of patients remains corticosteroid dependent. Further studies are warranted to follow larger cohorts and to assess long-term safety.


Low-depth genotyping-by-sequencing (GBS) in a bovine population: strategies to maximize the selection of high quality genotypes and the accuracy of imputation.
Genotyping-by-sequencing (GBS) has emerged as a powerful and cost-effective approach for discovering and genotyping single-nucleotide polymorphisms. The GBS technique was largely used in crop species where its low sequence coverage is not a drawback for calling genotypes because inbred lines are almost homozygous. In contrast, only a few studies used the GBS technique in animal populations (with sizeable heterozygosity rates) and many of those that have been published did not consider the quality of the genotypes produced by the bioinformatic pipelines. To improve the sequence coverage of the fragments, an alternative GBS preparation protocol that includes selective primers during the PCR amplification step has been recently proposed. In this study, we compared this modified protocol with the conventional two-enzyme GBS protocol. We also described various procedures to maximize the selection of high quality genotypes and to increase the accuracy of imputation. The in silico digestions of the bovine genome showed that the combination of PstI and MspI is more suitable for sequencing bovine GBS libraries than the use of single digestions with PstI or ApeKI. The sequencing output of the GBS libraries generated a total of 123,666 variants with the selective-primer approach and 272,103 variants with the conventional approach. Validating our data with genotypes obtained from mass spectrometry and Illumina's bovine SNP50 array, we found that the genotypes produced by the conventional GBS method were concordant with those produced by these alternative genotyping methods, whereas the selective-primer method failed to call heterozygotes with confidence. Our results indicate that high accuracy in genotype calling (>97%) can be obtained using low read-depth thresholds (3 to 5 reads) provided that markers are simultaneously filtered for genotype quality scores. We also show that factors such as the minimum call rate and the minor allele frequency positively influence the accuracy of imputation of missing GBS data. The highest accuracies (around 85%) of imputed GBS markers were obtained with the FIMPUTE program when GBS and SNP50 array genotypes were combined (80,190 to 100,297 markers) before imputation. We discovered that the conventional two-enzyme GBS protocol could produce a large number of high-quality genotypes provided that appropriate filtration criteria were used. In contrast, the selective-primer approach resulted in a substantial proportion of miscalled genotypes and should be avoided for livestock genotyping studies. Overall, our study demonstrates that carefully adjusting the different filtering parameters applied to the GBS data is critical to maximize the selection of high quality genotypes and to increase the accuracy of imputation of missing data. The strategies and results presented here provide a framework to maximize the output of the GBS technique in animal populations and qualified the PstI/MspI GBS assay as a low-cost high-density genotyping platform. The conclusions reported here regarding read-depth and genotype quality filtering could benefit many GBS applications, notably genome-wide association studies, where there is a need to increase the density of markers genotyped across the target population while preserving the quality of genotypes.


Local Anesthesia-Based Modified Cartilage "Inlay" Graft Myringoplasty for Repairing Large Perforation.
The objective of this study was to evaluate the graft success rate and hearing outcome of modified cartilage inlay graft technique for repairing large perforations under local anesthesia. Prospective case series. We performed a prospective study in 31 patients with large perforations who underwent endoscopic local anesthesia-based modified cartilage inlay graft technique with no raising of the squamous epithelium and no rimming of the perforation margins. The graft outcomes and complications were evaluated at 6 and 24 months postoperatively. Thirty-one patients (31 ears) were included in this study. The mean operation time was 42.3 ± 2.6 (ranged 36-51) minutes. The visual analog scale pain scores at intraoperative and 2 and 48 hours postoperatively were, respectively, 2.6 ± 0.6, 1.4 ± 0.2, and 0.6 ± 0.1. The graft success rate was 100.0% (31/31) at 6 months after surgery. However, only 25 (80.6%, 25/31) patients completed a 2-year follow-up and performed a temporal bone CT examination; re-perforation was seen in 2 patients, the graft success rate was 92.0% (23/25). CT revealed the well-developed pneumatization of the mastoids and middle ear in all the patients. Nevertheless, 1 (1/25, 8.0%) patient developed a graft epithelial inclusion cyst, which was removed by endoscope in the outpatient setting. The mean preoperative air bone gap (ABG) was 23.1 ± 4.3 dB, while the mean postoperative ABG after 6 months was 14.9 ± 2.6 dB; the difference between these values was significant (P < .05; Wilcoxon's signed-rank test), and the functional success rate was 93.5% (29/31). Endoscopic modified cartilage "inlay" graft technique with no raising the squamous epithelium and no rimming the perforation is a minimally invasive technique with a short operative time and satisfying success rate for closure of large perforations under local anesthesia.


[Intercentral relations of the electrical processes of the rabbit brain with a polarization dominant].
The dynamics of changes in intercentral relations of electrical activity of the sensorimotor and premotor zones of both hemispheres and the ventroposterolateral (VPL) nucleus of the left and right thalamus at formation of motor dominant under the action of the DC anode in the rabbit sensorimotor cortex was studied by the method of spectral-correlation analysis. It is shown that in the much less than dominant much greater than motor analyzer (the sensorimotor cortex and VPL) highly coherent connections of electrical processes are formed in the delta-range with conjugated lowering of biopotential connections between the structures of the motor analyzer of the much less than nondominant much greater than part of the brain. At the same time differently directed connections of electrical processes are formed between the structures of the motor analyzer, and between the premotor cortex and focus area. Thus, during formation of the much less than polarization much greater than dominant, a new structure of the intercentral relations of electrical processes is established not only in the much less than dominant much greater than but also in the other half of the brain.


Assessment of rheumatic mitral valve disease. Value of echocardiography in patients clinically suspected of predominant stenosis.
The value of echocardiography as compared with cardiac catheterisation was evaluated prospectively in 33 consecutive patients clinically suspected of predominant mitral stenosis. Patients with clinical signs of accompanying mitral regurgitation, no matter how severe, and patients with clinical findings indicating insignificant aortic valve disease were included. Critical mitral stenosis was defined by a valve area of less than or equal to 1 cm2. Severe mitral regurgitation was diagnosed by echocardiography on the basis of left ventricular dilatation (more than 3.2 cm/m2 at end-diastole) if not explained otherwise. Significant aortic valve disease was suspected in cases with aortic valve deformity and left ventricular dilatation or hypertrophy as defined by echocardiography. Mitral valve area by echocardiography correlated well with mitral valve area calculated from catheterisation data and a good interobserver correlation was found for echocardiographic measurement. Mitral stenosis, critical or non-critical, may mask significant coexistent valve lesions; echocardiography failed to discover severe mitral regurgitation requiring valve replacement in two patients with non-critical stenosis, and significant aortic regurgitation needing valve replacement was underestimated in one patient with critical mitral stenosis. A correct echocardiographic classification with respect to surgery, however, was obtained in: (1) all patients with clinically pure mitral stenosis (nine patients), and (2) all patients with combined mitral stenosis and regurgitation when either critical stenosis or severe regurgitation was found at echocardiography (12 patients). It thus appears that two out of three patients with mitral valve disease in whom the clinical findings indicate predominant stenosis can be correctly evaluated with the echocardiogram.


Progression of plasminogen activator inhibitor-1 and fibrinogen levels in relation to incident type 2 diabetes.
Several studies have shown that fibrinolytic and coagulation abnormalities as well as low-grade inflammation predict cardiovascular disease and type 2 diabetes. We studied in the Insulin Resistance Atherosclerosis Study the relation of incident diabetes to dynamic changes of plasminogen activator inhibitor-1 (PAI-1) and fibrinogen. After a follow-up of 5.2 years, diabetes developed in 140 (16.6%) of 843 individuals (57% women; mean age [range], 54.7 [40, 69] years) (converters versus nonconverters). Baseline and follow-up levels of PAI-1 and fibrinogen (demographically and smoking adjusted) were higher in converters versus nonconverters (mean [SE]): at baseline, 23.7 ng/mL (1.5) versus 14.5 (0.4) and 286.2 mg/dL (4.8) versus 273.6 (2.1); at follow-up, 45.3 ng/mL (3.2) versus 25.9 (0.8) and 292.0 mg/dL (5.6) versus 275.2 (2.5); all P<0.05. In a demographically and smoking-adjusted logistic regression model, the change in PAI-1 was related to incident diabetes (OR for a 1-SD change [CI], 1.75 [1.37, 2.22]; P<0.001) after adjusting for baseline PAI-1 levels. After further adjusting for insulin sensitivity (SI) or waist, change in PAI-1 remained significantly related to incident diabetes (OR, 1.66 [1.28, 2.15], and 1.64 [1.28, 2.10]; P<0.001). In contrast, change in fibrinogen was not significantly related to incident diabetes. Progression of PAI-1 levels over time, in addition to high baseline PAI-1 levels, is associated with incident diabetes. PAI-1 levels (but not fibrinogen) further increase with the rising glucose levels and the development of diabetes. These findings extend the current knowledge on the relation of fibrinolysis and coagulation abnormalities to the development of type 2 diabetes.


Effect on plasma insulin and plasma glucose of consuming white wine alone after a meal.
Rodent studies have highlighted the possibility that alcohol may promote a significant decrease in the level of glucose-stimulated plasma insulin concentration. The aim of this study was to investigate whether a similar alcohol-induced decrease in plasma insulin occurs in humans, by assessing the level of plasma insulin and plasma glucose when a moderate amount of commercially available bottled white wine is consumed alone after a meal. Eight nondiabetic men aged 19 to 22 years participated in this investigation. Participants were required to consume some food for 45 min before ingesting three standard units of white wine (30 g of alcohol) over 90 min. Plasma insulin and plasma glucose levels were assessed at regular 45-min intervals across the experimental period. The data showed a significant alcohol-induced decrease in the level of plasma insulin and a nonsignificant trend for a decrease in plasma glucose concentration in all participants after 15 g of alcohol had been consumed alone after a meal. These findings highlight the possibility that white wine, if consumed alone after a meal, may significantly alter energy utilization and possibly cause an alteration in glucose metabolism.


Development of microfluidic chip for entrapping tobacco BY-2 cells.
The tobacco BY-2 cell line has been used widely as a model system in plant cell biology. BY-2 cells are nearly transparent, which facilitates cell imaging using fluorescent markers. As cultured cells are drifted in the medium, therefore, it was difficult to observe them for a long period. Hence, we developed a microfluidic device that traps BY-2 cells and fixes their positions to allow monitoring the physiological activity of cells. The device contains 112 trap zones, with parallel slots connected in series at three levels in the flow channel. BY-2 cells were cultured for 7 days and filtered using a sieve and a cell strainer before use to isolate short cell filaments consisting of only a few cells. The isolated cells were introduced into the flow channel, resulting in entrapment of cell filaments at 25 out of 112 trap zones (22.3%). The cell numbers increased through cell division from 1 to 4 days after trapping with a peak of mitotic index on day 2. Recovery experiments of fluorescent proteins after photobleaching confirmed cell survival and permeability of plasmodesmata. Thus, this microfluidic device and one-dimensional plant cell samples allowed us to observe cell activity in real time under controllable conditions.


Effect of reduced graphene oxide on the solvent transport characteristics and sorption kinetics of fluoroelastomer nanocomposites.
The chemical resistivity of a fluoroelastomer forms the basis of its applicability in chemical industries as gloves and inner linings of storage tanks; in automotive and aircraft industries as linings of fuel hoses, etc. In this context, the study of the molecular transport of industrially important chemicals is of considerable interest. The present study depicts the solvent transport characteristics of fluoroelastomer/reduced graphene oxide nanocomposites (FKM/mRGO) in a homologous series of aromatic hydrocarbons. The solvent diffusion characteristics of the nanocomposites were analyzed in terms of mRGO loading, nature of solvents used, temperature etc. The parameters such as equilibrium solvent uptake, diffusivity, sorptivity, and permeability were analyzed and found to decrease with an increase in filler content. This is mainly attributed to the decrease in free volume with increasing concentration of mRGO due to the improved filler-elastomer interactions that hinder the solvent transport. Swelling parameters such as swelling coefficient and swelling index were analyzed to study the polymer-filler interactions. Diffusion studies with respect to temperatures were used to estimate the enthalpy, entropy and free energy change of the diffusion process. The molecular mass between crosslinks and hence the crosslink densities were calculated by using the experimental data and the reinforcement effect of mRGO was confirmed by the decrease in Mc values and increase in crosslink densities. Theoretical models, namely phanton and affine, were compared with the experimental values, which showed a moderate agreement with affine network models.


Role of NK cell subsets in organ-specific murine melanoma metastasis.
Tumor metastasis plays a major role in the morbidity and mortality of cancer patients. Among solid tumors that undergo metastasis, there is often a predilection to metastasize to a particular organ with, for example, prostate cancer preferentially metastasizing to bones and colon cancer preferentially metastasizing to the liver. Although many factors are thought to be important in establishing permissiveness for metastasis, the reasons for organ-specific predilection of each tumor are not understood. Using a B16 murine melanoma model, we tested the hypothesis that organ-specific NK cell subsets play a critical role in organ-specific metastasis of this tumor. Melanoma cells, given intravenously, readily colonized the lungs but not the liver. NK cell depletion (either iatrogenically or by using genetically targeted mice) resulted in substantial hepatic metastasis. Analysis of NK cell subsets, defined by the differential expression of a combination of CD27 and CD11b, indicated a significant difference in the distribution of NK cell subsets in the lung and liver with the mature subset being dominant in the lung and the immature subset being dominant in the liver. Several experimental approaches, including adoptive transfer, clearly indicated that the immature hepatic NK cell subset, CD27+ CD11b-, was protective against liver metastasis; this subset mediated its protection by a perforin-dependent cytotoxic mechanism. In contrast, the more mature NK cell subsets were more efficient at reducing pulmonary tumor load. These data indicate that organ-specific immune responses may play a pivotal role in determining the permissiveness of a given organ for the establishment of a metastatic niche.


Multiple Introductions and Predominance of Rotavirus Group A Genotype G3P[8] in Kilifi, Coastal Kenya, 4 Years after Nationwide Vaccine Introduction.
Globally, rotavirus group A (RVA) remains a major cause of severe childhood diarrhea, despite the use of vaccines in more than 100 countries. RVA sequencing for local outbreaks facilitates investigation into strain composition, origins, spread, and vaccine failure. In 2018, we collected 248 stool samples from children aged less than 13 years admitted with diarrheal illness to Kilifi County Hospital, coastal Kenya. Antigen screening detected RVA in 55 samples (22.2%). Of these, VP7 (G) and VP4 (P) segments were successfully sequenced in 48 (87.3%) and phylogenetic analysis based on the VP7 sequences identified seven genetic clusters with six different GP combinations: G3P[8], G1P[8], G2P[4], G2P[8], G9P[8] and G12P[8]. The G3P[8] strains predominated the season (n = 37, 67.2%) and comprised three distinct G3 genetic clusters that fell within Lineage I and IX (the latter also known as equine-like G3 Lineage). Both the two G3 lineages have been recently detected in several countries. Our study is the first to document African children infected with G3 Lineage IX. These data highlight the global nature of RVA transmission and the importance of increasing global rotavirus vaccine coverage.


Release of urinary extracellular vesicles in prostate cancer is associated with altered urinary N-glycosylation profile.
Nowadays, extracellular vesicles are of great interest in prostate cancer (PCa) research. Asparagine (N)-linked glycosylation could play a significant role in the pathological mechanism of these vesicles. We investigated if prostatic protein N-glycosylation profiles were related to urinary vesicle-associated prostate-specific antigen (PSA) extractability and if this parameter showed diagnostic potential for PCa. Urinary extracellular vesicles were visualised using transmission electron microscopy. Urinary extracellular vesicles extraction by means of n-butanol allowed determination of urinary vesicle-associated PSA extractability. Diagnostic value was assessed between benign prostate hyperplasia (BPH; n=122) and patients with PCa (n=85). Additionally, correlation with urine N-glycosylation was assessed. Urinary extracellular vesicles with a diameter of approximately 100 nm were more abundantly present in urine of patients with PCa versus patients with BPH resulting in a higher vesicle-associated PSA extraction ratio (p<0.001). Next, vesicle-associated PSA extraction ratio was correlated to biantennary core-fucosylation (p=0.003). Finally, vesicle-associated PSA extraction ratio proved beneficial in PCa diagnosis, next to serum PSA and the urinary glycosylation marker (p=0.021). The urinary vesicle-associated PSA extraction ratio is increased in PCa which is a direct result of the abundant presence of extracellular vesicles in urine of patients with PCa. The urinary vesicle-associated PSA extraction ratio was associated with changes in N-glycoforms and showed diagnostic potential. Further research is warranted to unravel the pathological link between N-glycosylation and extracellular vesicles in cancer, as well as to assess the prognostic value of this biomarker.


Modeling and Robust Control of a 5 DOF Model for Rowing Motion by Inverse Dynamics Method.
Competitive sailing requires efforts pertinent to physiological limitations and coordination between different parts of the body. Such coordination depends on the torques applied by muscles to the joints. This study aims to simulate the motion and provide a control law for the joint torques in order to track the desired motion paths. In this analytical study, an inverse dynamics based control is employed in order to simulate the motion by tracking the desired movement trajectories. First, the dynamics equations are obtained using Lagrange method for 5 degrees of freedom (5 DOF) model. In the following, a robust control scheme with inverse dynamics method based on the Proportional-Integral-Derivative (PID) approach is employed to track the desired joint angles obtained from the experiment. The simulation results demonstrate the performance of the proposed control method. Low settling times are achieved for the entire joint, which is appropriate in comparison with the time period of each cycle (3.75 s). Also, the maximum torques required to be applied to the joints are in physiological range. This study provided an appropriate model for the analysis of human movement in rowing sport. The model can also be cited in terms of basic biological theories in addition to practical computational uses in biomechanical engineering. Accordingly, the generated control signals can help to improve the interactive body movements during paddling and in designing robotic arms for automatic rowing.


Beta-glucuronidase and beta-glucosidase activity of Lactobacillus and Enterococcus isolated from human feces.
The domination of microorganisms characterized by excessive activity of the so-called fecal enzymes may be one of the reasons of the large intestine cancers. These enzymes are mainly those that belong to the hydrolase and reductase classes and their excessive activity may lead to disorders in the functioning of the digestive tract. The aim of tise research was to determine the activity of beta-glucuronidase and beta-glucosidase of Lactobacillus and Enterococcus strains isolated from the feces of healthy children, aged 1 and 8, and adults, aged 30 and 80. The analysis included 10 strains isolated from the feces of individuals in each of the age groups. beta-glucuronidase activity in the case of the isolates from children, depending on the strain, equaled from about 0.15 mM/h/mg of protein to 0.26 mM/h/mg of protein and was lower, respectively, by 52.35% and 57.81%, than the beta-glucosidase activity. Simultaneously, the activity of the Lactobacillus enzymes from children was 2.4 times higher, and in case of the isolates obtained from adults they were 4.6 and 2.7 times higher than the activity of the Entercoccus enzymes. The highest beta-glucuronidase activity was observed in Lactobacillus isolates coming from an 80-year-old subject. The differences between the activity of Enterococcus beta-glucuronidase isolated from the feces of 1 and 8 year old children were statistically insignificant. On the other hand, in the case of the subjects aged 30 and 8 the isolates were characterized by activity lower by, respectively. 48% and 37% than the isolates coming from children. The highest beta-glucosidase activity was discovered in the case of Lactobacillus and Enterococcus coming from children, which was higher by 32% than the activity of the isolates from adult persons. Therefore, it was determined that the activity of beta-glucuronidase of Lactobacillus strains isolated from feces from people aged 80 was the highest, and the isolates of the examined microorganisms coming from children were characterized by the highest beta-glucosidase activity.


Dropped-head syndrome resulting from injury to the central spinal cord at the upper cervical level.
There are many causes of paraspinal muscle weakness which give rise to the dropped-head syndrome. In the upper cervical spine the central portion of the spinal cord innervates the cervical paraspinal muscles. Dropped-head syndrome resulting from injury to the central spinal cord at this level has not previously been described. We report two patients who were treated acutely for this condition. Both presented with weakness in the upper limbs and paraspinal cervical musculature after a fracture of C2. Despite improvement in the strength of the upper limbs, the paraspinal muscle weakness persisted in both patients. One ultimately underwent cervicothoracic fusion to treat her dropped-head syndrome. While the cause of the dropped-head syndrome cannot be definitively ascribed to the injuries to the spinal cord, this pattern is consistent with the known patho-anatomical mechanisms of both injury to the central spinal cord and dropped-head syndrome.


Clinical manifestations of Lyme disease.
Lyme disease typically begins with a unique skin lesion, erythema chronicum migrans (ECM) (stage 1). Patients with this lesion may also have headache, meningeal irritation, mild encephalopathy, multiple annular secondary lesions, malar or urticarial rash, generalized lymphadenopathy and splenomegaly, migratory musculoskeletal pain, hepatitis, sore throat, non-productive cough, conjunctivitis, periorbital edema, or testicular swelling. After a few weeks to months (stage 2), about 15% of patients develop frank neurologic abnormalities, including meningitis, encephalitis, cranial neuritis (including bilateral facial palsy), motor or sensory radiculoneuritis, mononeuritis multiplex, or myelitis. At this time, about 8% of patients develop cardiac involvement--AV block, acute myopericarditis, cardiomegaly, or pancarditis. Throughout this stage, many patients continue to experience migratory musculoskeletal pain in joints, tendons, bursae, muscle, or bone. Months to years after disease onset (stage 3), about 60% of patients develop frank arthritis, which may be intermittent or chronic. Recently evidence suggests that Lyme disease may also be associated with chronic neurologic or skin involvement. Thus, Lyme disease occurs in stages with different clinical manifestations at each stage, but the course of the illness in each patient is highly variable.


Gelsolin sensitivity of microfilaments as a marker for muscle differentiation.
The ability of porcine smooth muscle gelsolin to sever actin filaments was used to study alterations in the organization of F-actin containing structures during skeletal myogenesis. In permeabilized fibroblasts and unfused myoblasts, gelsolin induced complete degradation of the actin cytoskeleton. After fusion of myoblasts to multinucleated myotubes, gelsolin removed a substantial amount of actin, revealing fibers with a sarcomere-like arrangement of gelsolin-insensitive actin. These fibrils were much thinner and had shorter sarcomeres than fully differentiated myofibrils. The proportion of gelsolin-resistant fibrils increased during differentiation, resulting in almost complete inertness of mature myofibrils. Fibrils isolated from adult muscle were also found nearly resistant to gelsolin. Extraction of tropomyosin and myosin in buffer of high ionic strength prior to gelsolin treatment reestablished the susceptibility to the severing protein, both in myotubes and isolated myofibrils. Only small remnants of phalloidin-stainable material were retained. We therefore conclude that during myotube differentiation either an increased interaction of actin with actin-binding proteins (e.g., myosin and tropomyosin), or the assembly of muscle-specific isoforms of these proteins protect the filaments against degradation by actin severing proteins.


Effects of synthetic and natural in vivo inhibitors of β-glucuronidase on azoxymethane-induced colon carcinogenesis in rats.
D-Glucaric acid is a non-toxic natural compound found in many fruits and vegetables. Our previous studies have shown that the β-glucuronidase inhibitor D-glucaro-1,4-lactone, an active metabolite of D-glucaric acid, inhibits chemically-induced tumorigenesis in rodents. D-Glucaro-1,4-lactone has a synthetic precursor, 2,5-di-O-acetyl-D-glucaro-1,4:6,3-dilactone or aceglatone (ACE), known as a postoperative prophylactic agent, and a natural precursor, D-glucurono-γ-lactone (GL). In the present study, we first examined the effect of ACE on the initiation phase of rat colon carcinogenesis induced by 15 mg/kg azoxymethane (AOM) administered 3 times by subcutaneous (s.c.) injection at weeks 1, 2, and 3 of a 5-week short-term experiment. ACE (0.5 or 2%) was administered as a dietary supplement for 5 weeks. At 5 weeks after the initiation of treatment, the formation of aberrant crypt foci (ACF) in the rat groups treated with AOM plus a 0.5 or 2% ACE diet was significantly reduced by 48.6 and 55.3%, respectively, compared to the group administered AOM alone. In a previous study, 0.5 and 2% ACE diets dispensed during AOM treatment had a tendency to decrease AOM-induced colonic tumor incidence. In the present long-term 36-week colon tumorigenesis experiment, GL (0.5 or 2%) administered via the diet during the initiation phase (starting 1 week before the first dose of AOM and ending 1 week after the 3rd dose) did not have any significant effects on tumor incidence. On the other hand, continued post-initiation treatment with ACE (0.5 and 2%) markedly reduced colonic tumor incidence by 70 and 80%, respectively. GL was effective to a similar extent (70% inhibition), but only at a concentration of 2%. We conclude that ACE inhibits the initiation and post-initiation stages of AOM-induced colon carcinogenesis, while GL affects only the post-initiation stages.


Orthographic and phonological preview benefits: parafoveal processing in skilled and less-skilled deaf readers.
Many deaf individuals do not develop the high-level reading skills that will allow them to fully take part into society. To attempt to explain this widespread difficulty in the deaf population, much research has honed in on the use of phonological codes during reading. The hypothesis that the use of phonological codes is associated with good reading skills in deaf readers, though not well supported, still lingers in the literature. We investigated skilled and less-skilled adult deaf readers' processing of orthographic and phonological codes in parafoveal vision during reading by monitoring their eye movements and using the boundary paradigm. Orthographic preview benefits were found in early measures of reading for skilled hearing, skilled deaf, and less-skilled deaf readers, but only skilled hearing readers processed phonological codes in parafoveal vision. Crucially, skilled and less-skilled deaf readers showed a very similar pattern of preview benefits during reading. These results support the notion that reading difficulties in deaf adults are not linked to their failure to activate phonological codes during reading.


Various strategies to improve efficacy of stem cell transplantation in multiple sclerosis: Focus on mesenchymal stem cells and neuroprotection.
Multiple sclerosis (MS) is an inflammatory demyelinating disease of the central nervous system (CNS) which predominantly affect young adults and undergo heavy socioeconomic burdens. Conventional therapeutic modalities for MS mostly downregulate aggressive immune responses and are almost insufficient for management of progressive course of the disease. Mesenchymal stem cells (MSCs), due to both immunomodulatory and neuroprotective properties have been known as practical cells for treatment of neurodegenerative diseases like MS. However, clinical translation of MSCs is associated with some limitations such as short-life engraftment duration, little in vivo trans-differentiation and restricted accessibility into damaged sites. Therefore, laboratory manipulation of MSCs can improve efficacy of MSCs transplantation in MS patients. In this review, we discuss several novel approaches, which can potentially enhance MSCs capabilities for treating MS.


Targeted therapies for renal cell carcinoma in Chinese patients: focus on everolimus.
Renal cell carcinoma (RCC) is the most common type of cancer arising from the kidney, with a male to female ratio of 2:1. The incidence of RCC is rising. In males, it was the seventh most common cancer in the People's Republic of China in 2012. RCC is resistant to radiotherapy and chemotherapy, but approximately 20% of patients with advanced RCC respond to immunotherapy. Novel therapies targeting angiogenesis and signaling pathways have been proven to be effective for advanced or metastatic RCC in Western countries. Due to the heterogeneity of RCC between races, it is necessary to have an overview of targeted therapies, especially everolimus, for patients with advanced RCC in the People's Republic of China. Three targeted therapeutic agents have been approved in Mainland China for the treatment of patients with advanced RCC, ie, two tyrosine kinase inhibitors (sorafenib and sunitinib) and one mammalian target of rapamycin (mTOR) inhibitor (everolimus). Compared with Western patients with advanced or metastatic RCC, Chinese patients with the same disease respond better to sorafenib and sunitinib as first-line targeted therapy, but sunitinib has a relatively higher risk of toxicity. Everolimus, an mTOR inhibitor that can be administered orally, is well tolerated and acceptable to Chinese patients. Everolimus has competitive advantages as second-line targeted treatment for Chinese patients with advanced RCC who are resistant to first-line tyrosine kinase inhibitors. Despite a lack of noninferiority when compared with sunitinib as first-line therapy, the sunitinib-everolimus paradigm is still recommended as standard therapy for patients with advanced RCC. Although most studies of targeted therapies for advanced RCC have obvious limitations, such as small sample size and retrospective design, up-to-date evidence indicates that everolimus would be an ideal agent as second-line targeted treatment for advanced or metastatic RCC in the People's Republic of China.


Multigenerational effects of a reduced balanced protein diet during the rearing and laying period of broiler breeders. 2. Zootechnical performance of the F1 broiler offspring.
Several studies in mammals focused on the maternal programming of the metabolism by epigenetic mechanisms, while currently, the consequences of a maternal dietary treatment on the offspring performance of farm animals are of particular interest for commercial purpose. In the present study, we investigated if the zootechnical performance of the progeny was altered by a maternal dietary treatment, being a lower dietary crude protein (CP) of the grandparent and/or parent generation. The multigenerational effects of a reduced maternal CP content were investigated by reducing the dietary CP level by 25% in rearing and laying diets of pure line A breeders. The F0 generation breeders were fed either control (C) or reduced balanced protein (RP) diets. The F1 breeder generation was constructed by dividing the F0 female progeny again over a C or RP diet, resulting in 4 dietary treatments in the F1 generation: C/C, C/RP, RP/C, and RP/RP (letters indicating the diets in, respectively, F0 and F1 generations). The offspring performance was evaluated by a zootechnical and nitrogen retention trial on C and low-protein (LP) broiler diets. For the C broiler diet, the C/RP and RP/RP offspring were characterized by a higher BW from d 35 until d 42 compared to the C/C progeny, whereas the RP/C offspring had an intermediate BW that did not differ from the other groups. A tendency (P = 0.067) towards a better nitrogen retention was observed for the offspring of breeders that received the RP diets in F0 and/or F1 generation compared to the C/C progeny. For the LP broiler diet, the C/RP (P = 0.021) and RP/C (P = 0.001) offspring had a higher BW compared to the C/C progeny during the entire grow-out period. In addition, the C/RP offspring were characterized by a lower FCR from d 28 onwards (P = 0.021). In conclusion, dietary treatments imposed on mother hens can have direct effects on the next generation, as well as indirect effects on multiple generations.


The roles of Sp1, Sp3, USF1/USF2 and NRF-1 in the regulation and three-dimensional structure of the Fragile X mental retardation gene promoter.
Expansion of a CGG.CCG-repeat tract in the 5'-untranslated region of the FMR1 (Fragile X mental retardation 1) gene causes its aberrant transcription. This produces symptoms ranging from premature ovarian failure and Fragile X associated tremor and ataxia syndrome to FMR syndrome, depending on the size of the expansion. The promoter from normal alleles shows four protein-binding regions in vivo. We had previously shown that in mouse brain extracts two of these sites are bound by USF1/USF2 (upstream stimulatory factors 1 and 2) heterodimers and NRF-1 (nuclear respiratory factor-1). We also showed that these sites are involved in the positive regulation of FMR1 transcription in neuronally derived cells. In the present study, we show that Sp1 (specificity protein 1) and Sp3 are also strong positive regulators of FMR1 promoter activity. We also show that, like Sp1 and E-box-binding proteins such as USF1 and USF2, NRF-1 causes DNA bending, in this case producing a bend of 57 degrees towards the major groove. The combined effect of the four protein-induced bends on promoter geometry is the formation of a highly compact arch-like structure in which the 5' end of the promoter is brought in close proximity to the 3' end. We had previously shown that while point mutations in the GC-boxes decrease promoter activity, deletion of either one of them leads to an increase in promoter activity. We can reconcile these observations with the positive effect of Sp1 and Sp3 if protein-induced bending acts, at least in part, to bring together distally spaced factors important for transcription initiation.


The Effectiveness of Technology-Supported Exercise Therapy for Low Back Pain: A Systematic Review.
Various technological systems have been developed to assist exercise therapy for low back pain. The aim of this systematic review was to provide an overview and to assess the effectiveness of the available technology-supported exercise therapy (TSET) programs for low back pain. The electronic databases Pubmed, Embase, Cochrane Central Register of Controlled Trials, PEDro, IEEE, and ACM were searched until January 2016. Randomized controlled trials (RCTs) using electronic technological systems simultaneously with exercise therapy for patients with low back pain were included. Twenty-five RCTs met the inclusion criteria. Seventeen studies involved patients with chronic low back pain, and electromyography biofeedback was the most prevalent type of technological support. This review shows that TSET seems to improve pain, disability, and quality of life for patients with low back pain, and that a standard treatment combined with an additional TSET program might be superior to a standard treatment alone. However, TSET seems not more effective compared to other interventions or a placebo intervention for improving these outcomes, which may partially be explained by the analytical approach of the current TSET-programs. For most technologies, only a limited number of RCTs are available, making it difficult to draw firm conclusions about the effectiveness of individual technological systems.


Intentional construction of high-performance SnO2 catalysts with a 3D porous structure for electrochemical reduction of CO2.
Herein, SnO2-NC (SnO2-nanocube) and SnO2-NF (SnO2-nanoflake) electro-catalysts featuring a large specific surface area and 3D porous structure were successfully constructed via acid etching and sulfurization-desulphurization methods, respectively. As catalysts for the electrochemical reduction of CO2, the faradaic efficiency (FHCOO-+CO = 82.4%, 91.5%, respectively) and partial current density (jHCOO-+CO = 10.7 and 11.5 mA cm-2, respectively) of SnO2-NCs and SnO2-NFs were enhanced in comparison with SnO2-NPs (SnO2-nanoparticles, FHCOO-+CO = 63.4%, jHCOO-+CO = 5.7 mA cm-2) at -1.0 V vs. RHE. The enhanced catalytic activity is attributed to their uniform 3D porous structure, high specific surface area and excellent wettability. Additionally, the morphology of SnO2-NCs and SnO2-NFs was largely preserved after electrolyzing for 12 h (after 12 h of electrolysis), indicating the effective buffering effect of the 3D structure in electrolysis. Naturally, the current density and faradaic efficiency of the SnO2-NC and SnO2-NF catalysts remained nearly unchanged after long-term stability measurements, revealing great stability.


Phase II trial of N,N-diethyl-2-[4-(phenylmethyl)phenoxy]ethanamine.HCl and doxorubicin chemotherapy in metastatic breast cancer: A National Cancer Institute of Canada clinical trials group study.
This multicenter phase II trial investigated the efficacy and toxicity of a combination of the novel intracellular histamine antagonist, N,N-diethyl-2-[4-(phenylmethyl)phenoxy]ethanamine.HCl (DPPE), and doxorubicin in patients with anthracycline-naïve metastatic breast cancer. Preclinical models and early single institutional studies suggested DPPE could potentiate the cytotoxicity of doxorubicin. Forty-two women, 32 to 77 years old (median, 59 years), with anthracycline-naïve metastatic breast cancer were treated. Patients may have had one previous regimen of nonanthracycline chemotherapy, either in the adjuvant or metastatic disease treatment setting. DPPE (6 mg/kg) was administered as an 80 minute intravenous infusion with doxorubicin (60 mg/m(2)) given intravenously over the last 20 minutes of the DPPE infusion. Patients were premedicated with an antiemetic and sedating regimen. The DPPE/doxorubicin treatment was given every 21 days for a maximum of seven cycles. All 42 patients were assessable. Overall, toxicity was comparable to that expected with doxorubicin alone, with the exception of DPPE-related motion sickness, mild hallucinations, and cerebellar signs at the time of the infusion. These CNS side effects were manageable in an ambulatory care setting, improved with subsequent cycles of treatment, and did not usually require hospitalization. Four patients developed febrile neutropenia. Thirty-five patients received four or more cycles of chemotherapy. The overall response rate was 52.5% (95% confidence interval, 36% to 68%), with 9.5% complete responses (n = 4), 43% partial responses (n = 18), and 38% of patients with stable disease (n = 16). The antitumour effects of DPPE/doxorubicin the 52.5% response rate seems encouraging, particularly in consideration of the fact that a recently reported randomized National Cancer Institute of Canada Clinical Trials Group trial using single-agent doxorubicin 60 mg/m(2) in one of the treatment arms achieved a 31% response rate. Thus, a randomized phase III trial of doxorubicin versus doxorubicin plus DPPE is being conducted in this clinical setting.


Systematic Evaluation of Key L-Carnitine Homeostasis Mechanisms during Postnatal Development in Rat.
The conditionally essential nutrient, L-carnitine, plays a critical role in a number of physiological processes vital to normal neonatal growth and development. We conducted a systematic evaluation of the developmental changes in key L-carnitine homeostasis mechanisms in the postnatal rat to better understand the interrelationship between these pathways and their correlation to ontogenic changes in L-carnitine levels during postnatal development. mRNA expression of heart, kidney and intestinal L-carnitine transporters, liver γ-butyrobetaine hydroxylase (Bbh) and trimethyllysine hydroxylase (Tmlh), and heart carnitine palmitoyltransferase (Cpt) were measured using quantitative RT-PCR. L-Carnitine levels were determined by HPLC-UV. Cpt and Bbh activity were measured by a spectrophotometric method and HPLC, respectively. Serum and heart L-carnitine levels increased with postnatal development. Increases in serum L-carnitine correlated significantly with postnatal increases in renal organic cation/carnitine transporter 2 (Octn2) expression, and was further matched by postnatal increases in intestinal Octn1 expression and hepatic γ-Bbh activity. Postnatal increases in heart L-carnitine levels were significantly correlated to postnatal increases in heart Octn2 expression. Although cardiac high energy phosphate substrate levels remained constant through postnatal development, creatine showed developmental increases with advancing neonatal age. mRNA levels of Cpt1b and Cpt2 significantly increased at postnatal day 20, which was not accompanied by a similar increase in activity. Several L-carnitine homeostasis pathways underwent significant ontogenesis during postnatal development in the rat. This information will facilitate future studies on factors affecting the developmental maturation of L-carnitine homeostasis mechanisms and how such factors might affect growth and development.


EUS-guided gastroenterostomy for gastric outlet obstruction: a comprehensive meta-analysis.
A comprehensive meta-analysis was performed to evaluate the efficacy and safety of endoscopic ultrasound-guided gastroenterostomy (EUS-GE) for benign and malignant gastric outlet obstruction (GOO). PubMed, Embase, Web of Science and Cochrane Library were searched to identify relevant studies. The primary outcomes evaluated technical success, clinical success, and adverse events (AEs). Twenty‑six studies with 1493 patients were included in this meta-analysis. The pooled rates of technical success, clinical success, and overall AEs of EUS-GE were 94.0%, 89.9%, and 13.1%, respectively. Eight studies were included in the subgroup meta-analysis for comparative evaluation of EUS-GE and surgical gastroenterostomy (SGE), while seven studies were for EUS-GE and enteral stenting (ES). Compared with SGE, the pooled odds ratios (ORs) of technical success, clinical success, and overall AEs of EUS-GE were 0.17 (p = .003), 1.42 (p = .40), and 0.15 (p < .00001), respectively. When compared with ES, the above corresponding pooled ORs were 0.55 (p = .11), 2.64 (p < .0001), and 0.41 (p = .01), respectively. Although it is technically challenging, this largest meta-analysis indicates that EUS‑GE has comparable and high technical and clinical success rates and hence a very effective minimally invasive procedure for GOO.


Effects of Chinese herbal mixture on performance, egg quality and blood biochemical parameters of laying hens.
The aim of the study was to evaluate the effect of Chinese herbal mixture (CHM) in laying hen diets on performance, egg quality and egg nutrition components, as well as selected blood biochemical parameters. A total of 270 Lohmann Brown laying hens, 24 week of age, were randomly distributed into three dietary treatments and fed for 6 week. The CHM was used at the levels of 0%, 0.5% and 1.0% in the diets of the control and two treatment groups respectively. Each dietary treatment was divided into five replicates, comprising of 18 hens each. Hens were housed in a 3-tier battery system. Feed and water were provided ad libitum. Blood samples and eggs were collected at the end of the experiment. The results showed that dietary treatments did not affect egg weight, mortality, eggshell thickness, albumen height, Haugh units, eggshell-breaking strength and egg-shape index. Dietary supplementation of CHM significantly improved (p < 0.05) laying rate and the lowest feed conversion ratio were observed with 1.0% CHM supplementation diets. CHM supplementation decreased (p < 0.05) cracked-egg rate, egg yolk cholesterol, blood serum levels of cholesterol, triglyceride, low-density lipoprotein cholesterol (LDL-C) and alanine aminotransferase (ALT) and increased blood serum levels of high-density lipoprotein cholesterol (HDL-C). No differences in serum total protein, serum albumen, serum glucose and aspartate amino transferase (AST) were observed in hens fed CHM. In conclusion, dietary supplementation with CHM improved egg production and egg quality and reduced serum cholesterol concentrations in laying hens. Dietary supplementation of 1.0% CHM in layer diets may be a feasible means of producing eggs with lower cholesterol and higher egg yolk phospholipid contents for health conscious consumers.


Neurologic complications following bone marrow transplantation for sickle cell disease.
A boy with sickle cell anemia underwent bone marrow transplantation (BMT). He was normal on neurological examination, but had radiologic evidence of an old left frontal lobe infarct, multiple cerebral vascular stenoses and moyamoya collaterals. After BMT he developed seizures with extension of the infarct and subarachnoid hemorrhage. One year later angiography revealed worsening stenosis of the M1 segments of both middle cerebral arteries. At that time an increase in von Willebrand's factor with decreased large molecular weight multimers (LvWF) was observed. We speculate that LvWF dependent, shear-induced platelet aggregation, together with endothelial damage may have contributed to the development of neurologic complications in this patient.


Changes in functioning of mesolimbic incentive processing circuits during the premenstrual phase.
The premenstrual phase of the menstrual cycle is associated with marked changes in normal and abnormal motivated behaviors. Animal studies suggest that such effects may result from actions of gonadal hormones on the mesolimbic dopamine (DA) system. We therefore investigated premenstrual changes in reward-related neural activity in terminal regions of the DA system in humans. Twenty-eight healthy young women underwent functional magnetic resonance imaging on 2 days during the menstrual cycle, once during the late follicular phase and once during the premenstrual phase, in counterbalanced order. Using a modified version of the monetary incentive delay task, we assessed responsiveness of the ventral striatum to reward anticipation. Our results show enhanced ventral striatal responses during the premenstrual as compared to the follicular phase. Moreover, this effect was most pronounced in women reporting more premenstrual symptoms. These findings provide support for the notion that changes in functioning of mesolimbic incentive processing circuits may underlie premenstrual changes in motivated behaviors. Notably, increases in reward-cue responsiveness have previously been associated with DA withdrawal states. Our findings therefore suggest that the sharp decline of gonadal hormone levels in the premenstrual phase may trigger a similar withdrawal-like state.


Oligodendroglial cell behaviour in traumatic oedematous human cerebral cortex: a light and electron microscopic study.
Cortical biopsies of 12 patients with traumatic brain injuries have been used in the present study to examine oligodendroglial cell changes and reactivity. The samples were processed for light and transmission electron microscopy. Four main types of oligodendrocyte populations have been found: resting or unreactive oligodendrocytes, reactive oligodendrocytes, anoxic-ischaemic oligodendroglial cells and hyperthrophic phagocytic oligodendrocytes. The unreactive or resting oligodendrocyte type exhibited a fusiform or elongated shape, a clear or dense band of scarce perikaryal cytoplasm and a nucleus with peripheral heterororomatin masses. Clear or dense reactive oligodendrocytes showed increased amount of perikaryal citoplasm, dilated endoplasmic reticulum and nuclear envelope, numerous clear, oedematous mitochondria and dense bodies. These oligodendrocytes appeared associated with degenerated myelinated axons. Anoxic-ischaenmic oligodendrocytes showed lacunar enlargement of endoplasmic reticulum, dilated Golgi complex and enlargement and disassembly of nuclear envelope. They appeared also in contact with degenerated myelinated axons. Hypertrophic phagocytic oligodendrocytes were observed engulfing the associated degenerated myelinated axons, invading the myelin sheath, separating the myelin lamellae and exerting myelinolitic effects. Oligodendroglialpseudopodic expansions were observed phagocyting the axoplasmic matrix and leaving a huge vacuolar axoplasmic space. The vasogenic and cytotoxic components of traumatic brain oedema are discussed in relation with the oligodendroglial cell changes and reactivity.


Topogenic analysis of the human immunodeficiency virus type 1 envelope glycoprotein, gp160, in microsomal membranes.
The orientation in cellular membranes of the 856 amino acid envelope glycoprotein precursor, gp160, of human immunodeficiency virus type 1 was investigated in vitro. Variants of the env gene were transcribed using the bacteriophage SP6 promoter, translated using a rabbit reticulocyte lysate, and translocated into canine pancreatic microsomal membranes. Immunoprecipitation studies of gp160 variants using antibodies specific for various gp160-derived polypeptides provided evidence that the external (cell surface) domain of gp160 begins at the mature amino terminus of the protein and continues through amino acid 665. A stop-transfer sequence (transmembrane domain) was identified in a hydrophobic region COOH-terminal to amino acid 665 and NH2-terminal to amino acid 732. Protease protection experiments demonstrated that gp160 possesses a single cytoplasmic domain COOH-terminal to residue 707. Membrane extraction studies using carbonate buffer provided evidence that the 29 amino acid hydrophobic domain (residues 512-541) of gp160 was unable to serve as a stop-transfer sequence. Finally, we propose that the cytoplasmic tail of gp160 forms a secondary association with the microsomal membranes.


Oncogenesis by interspecific interaction of malignant murine and non-malignant hamster cells in vitro.
A clone of Cloudman S91 murine melanoma was fused in vitro with non-malignant hamster cheek pouch cells by means of lysolecithin, and the putative hybrid progeny cells, HCP-MM, were found to be highly malignant in hamster, but not in appropriate mice. A malignant clone of HCP-MM cells was shown to have hamster species-specific surface antigens (as demonstrated by immunofluorescence and the cytotoxic antibody) and hamster-like lactate dehydrogenase and NAD-dependent malate dehydrogenase isoenzyme profiles. Nevertheless, chromosomes similar to those of both murine and hamster parental cells could be distinguished in cells of this malignant clone and in hamster tumor grafts by the method of trypsin-Giemsa banding. A majority of the murine chromosomes, however, appeared to be lost. This study indicates that a murine melanoma previously found untransplantable in hamsters could produce a highly malignant and lethal tumor for hamsters after being mixed in vitro with non-malignant hamster cells, in the presence of a fusing chemical. It is not as yet certain whether the production of transformed cells in vitro and of highly malignant tumors in the hamster (both with predominantly hamster properties) required heterosynkarion formation between the murine melanoma and hamster cheek pouch cells. Nevertheless, our results suggest that the presence of the murine melanoma, and possibly the interaction of its genome with non-malignant hamster cells, was implicated in this process.


Constant activation of the RAF-MEK-ERK pathway as a diagnostic and therapeutic target in hairy cell leukemia.
The BRAF-V600E mutation defines genetically hairy cell leukemia among B-cell leukemias and lymphomas. In solid tumors, BRAF-V600E is known to aberrantly activate the oncogenic MEK-ERK pathway, and targeted BRAF and/or MEK inhibitors have shown remarkable efficacy in clinical trials in melanoma patients. However, the MEK-ERK pathway status in hairy cell leukemia has not been thoroughly investigated. We assessed phospho-ERK expression in 37 patients with hairy cell leukemia and 44 patients with neoplasms mimicking hairy cell leukemia (40 splenic marginal zone lymphoma, 2 hairy cell leukemia-variant and 2 splenic lymphoma/leukemia unclassifiable) using immunohistochemistry on routine biopsies and/or Western blotting on purified leukemic cells, and correlated the phospho-ERK status with the BRAF-V600E mutation status. Besides confirming the constant presence of BRAF-V600E in all patients with hairy cell leukemia, we observed ubiquitous phospho-ERK expression in this malignancy. Conversely, all 44 cases with neoplasms mimicking hairy cell leukemia were devoid of BRAF-V600E and none expressed phospho-ERK. Furthermore, the two exceptionally rare cases of non-hairy cell leukemia unclassifiable chronic B-cell neoplasms previously reported to be BRAF-V600E(+) on allele-specific polymerase chain reaction lacked phospho-ERK expression as well, suggesting the presence of the mutation in only a small part of the leukemic clone in these cases. In conclusion, our findings support the use of phospho-ERK immunohistochemistry in the differential diagnosis between hairy cell leukemia and its mimics, and establish the MEK-ERK pathway as a rational therapeutic target in this malignancy.


Sequential Chorioretinal Changes in Presumed Ocular Histoplasmosis Syndrome Analyzed Using Spectral Domain Optical Coherence Tomography.
To analyze sequential chorioretinal changes in presumed ocular histoplasmosis syndrome (POHS) using volumetric spectral-domain optical coherence tomography (SD-OCT). A total of 28 treatment-naive eyes of 20 POHS patients were characterized as: no choroidal neovascular membrane (CNVM) at initial SD-OCT but CNVM development during follow-up (primary group); CNVM at initial SD-OCT (progressive group); and contralateral asymptomatic eyes (control group). The primary group (4/28 eyes) initially demonstrated external limiting membrane (ELM), ellipsoid zone and retinal pigment epithelium (RPE)/Bruch's disruption in 100%, and pigment epithelial detachment (PED) in 75% of eyes. CNVM developed in 100% of eyes during follow-up. Progressive group (8/28 eyes) showed CNVM in 100% of eyes. Control group (16/28 eyes) showed ELM disruption in two and PED in three additional eyes on follow-up from initial analysis. SD-OCT is a useful tool to sequentially follow eyes with POHS for subtle chorioretinal changes that may suggest impending CNVM development, and thus benefit from closer clinical monitoring.


The impact of the menopause transition on the health and wellbeing of women living with HIV: A narrative review.
Improvements in survival due to advances in antiretroviral therapy (ART) have led to a shift in the age distribution of those receiving HIV care, with increasing numbers of women living with HIV (WLHIV) reaching menopausal age. We present a narrative literature review of 26 studies exploring the menopause transition in WLHIV, focusing on: (1) natural history (2) symptomatology and management, and (3) immunologic and virologic effects. Data are conflicting on the association between HIV and earlier age at menopause, and the role of HIV-specific factors such as HIV viral load and CD4 count. There are some data to suggest that WLHIV experience more vasomotor and psychological symptoms during the menopause than HIV-negative women, and that uptake of hormone replacement therapy by WLHIV is comparatively low. There is no evidence that menopause affects either CD4 count or response to ART, although there may be increased immune activation in older WLHIV. We conclude that menopause in WLHIV is a neglected area of study. Specific information gaps include qualitative studies on experiences of reproductive ageing; data on the impact of the menopause on women's quality of life and ability to adhere to health-sustaining behaviors; as well as studies investigating the safety and efficacy of pharmacological and psychosocial interventions. There is likely to be a burden of unmet health need among this growing population, and better data are required to inform optimal provision of care, supporting WLHIV to maintain their health and wellbeing into their post-reproductive years.


Assessment and significance of mediastinal bulk in Hodgkin's disease: comparison between computed tomography and chest radiography.
In Hodgkin's disease (HD), mediastinal bulk is currently defined from chest radiograph (CXR) measurements as a ratio of the maximum transverse mass diameter to the internal thoracic diameter at T5/6 level > or = 0.33. We evaluated how computed tomographic (CT) measurements of bulk correspond to those obtained from the CXR and correlated nodal mass long axis diameter with freedom from progression. Ninety-five adult patients who had a CXR thoracic ratio of greater than 0.3 and a CT scan within 28 days of the CXR were included in the study, provided that both investigations were performed before the start of treatment. Measurements of the widest mediastinal diameter and internal thoracic diameter were made on both CXR and CT scan. The thoracic ratio (TR) was calculated for each modality and compared using paired t tests. The longest diameter of the largest individual nodal mass (LIM(CT)) was also measured from the CT and correlated with freedom from progression using Cox regression. There was excellent correlation between CT and CXR for measurement of TR, with TR(CT) greater than TR(CXR) (mean difference of 2%). A TR(CT) of 0. 35 was found to be equivalent to a TR(CXR) of 0.33. No single measurement of nodal size correlated with the current definition of bulk. However LIM(CT) greater than 10 cm did correlate with increased risk of progressive HD (P =.03), even after adjustment for other prognostic variables (chemotherapy regimen and Hasenclever Prognostic Index). Excellent correlation was observed between assessment of TR by CXR and CT scan. The longest diameter of the LIM(CT) greater than 10 cm was found to be associated with an increased risk of disease progression.