Role of the thymus in streptococcal cell wall-induced arthritis and hepatic granuloma formation. Comparative studies of pathology and cell wall distribution in athymic and euthymic rats.
Systemic administration of an aqueous suspension of group A streptococcal cell wall fragments to susceptible rats induces acute and chronic polyarthritis, as well as noncaseating hepatic granulomas. To gain insight into the role of the thymus in the pathogenesis of this experimental model, pathologic responses and cell wall tissue distribution were compared in congenitally athymic rats (rnu/rnu) and their euthymic littermates (NIH/rnu). Within 24 h, both rat strains developed acute arthritis, characterized by polymorphonuclear leukocytic exudate in the synovium and joint spaces. This acute process was maximal at day 3 and gradually subsided. Beginning 2-3 wk after injection, the euthymic, but not the athymic, rats developed the typical exacerbation of arthritis, characterized by synovial cell hyperplasia with villus formation and T helper/inducer lymphocyte-rich mononuclear cell infiltration. This process eventually resulted in marginal erosions and destruction of periarticular bone and cartilage. Parallel development of acute and chronic hepatic lesions was observed. Bacterial cell wall antigen distribution and persistence were similar in the athymic and euthymic rats. Cell wall antigens were demonstrated in the cytoplasm of cells within subchondral bone marrow, synovium, liver, and spleen, coincident with the development of the acute lesions, and persisted in these sites, although in decreasing amounts, for the duration of the experiment. Our findings provide evidence that the acute and chronic phases of the experimental model are mechanistically distinct. The thymus and functional thymus derived-lymphocytes appear not to be required for the development of the acute exudative disease but are essential for the development of chronic proliferative and erosive disease. Induction of disease is dependent upon cell wall dissemination to and persistence in the affected tissues.


Disentangling the long-term effects of disturbance on soil biogeochemistry in a wet tropical forest ecosystem.
Climate change is increasing the intensity of severe tropical storms and cyclones (also referred to as hurricanes or typhoons), with major implications for tropical forest structure and function. These changes in disturbance regime are likely to play an important role in regulating ecosystem carbon (C) and nutrient dynamics in tropical and subtropical forests. Canopy opening and debris deposition resulting from severe storms have complex and interacting effects on ecosystem biogeochemistry. Disentangling these complex effects will be critical to better understand the long-term implications of climate change on ecosystem C and nutrient dynamics. In this study, we used a well-replicated, long-term (10 years) canopy and debris manipulation experiment in a wet tropical forest to determine the separate and combined effects of canopy opening and debris deposition on soil C and nutrients throughout the soil profile (1 m). Debris deposition alone resulted in higher soil C and N concentrations, both at the surface (0-10 cm) and at depth (50-80 cm). Concentrations of NaOH-organic P also increased significantly in the debris deposition only treatment (20-90 cm depth), as did NaOH-total P (20-50 cm depth). Canopy opening, both with and without debris deposition, significantly increased NaOH-inorganic P concentrations from 70 to 90 cm depth. Soil iron concentrations were a strong predictor of both C and P patterns throughout the soil profile. Our results demonstrate that both surface- and subsoils have the potential to significantly increase C and nutrient storage a decade after the sudden deposition of disturbance-related organic debris. Our results also show that these effects may be partially offset by rapid decomposition and decreases in litterfall associated with canopy opening. The significant effects of debris deposition on soil C and nutrient concentrations at depth (>50 cm), suggest that deep soils are more dynamic than previously believed, and can serve as sinks of C and nutrients derived from disturbance-induced pulses of organic matter inputs.


A mTurquoise-based cAMP sensor for both FLIM and ratiometric read-out has improved dynamic range.
FRET-based sensors for cyclic Adenosine Mono Phosphate (cAMP) have revolutionized the way in which this important intracellular messenger is studied. The currently prevailing sensors consist of the cAMP-binding protein Epac1, sandwiched between suitable donor- and acceptor fluorescent proteins (FPs). Through a conformational change in Epac1, alterations in cellular cAMP levels lead to a change in FRET that is most commonly detected by either Fluorescence Lifetime Imaging (FLIM) or by Sensitized Emission (SE), e.g., by simple ratio-imaging. We recently reported a range of different Epac-based cAMP sensors with high dynamic range and signal-to-noise ratio. We showed that constructs with cyan FP as donor are optimal for readout by SE, whereas other constructs with green FP donors appeared much more suited for FLIM detection. In this study, we present a new cAMP sensor, termed (T)Epac(VV), which employs mTurquoise as donor. Spectrally very similar to CFP, mTurquoise has about doubled quantum efficiency and unlike CFP, its fluorescence decay is strictly single-exponential. We show that (T)Epac(VV) appears optimal for detection both by FLIM and SE, that it has outstanding FRET span and signal-to-noise ratio, and improved photostability. Hence, (T)Epac(VV) should become the cAMP sensor of choice for new experiments, both for FLIM and ratiometric detection.


Management of Suspected Avian (H5N1) Influenza in a Non-pandemic Setting.
Avian (H5N1) influenza has been responsible for millions of wild bird and poultry deaths throughout the world. Sporadic human cases with a high mortality have occurred, almost exclusively in association with very close contact with sick, dying or dead birds. Appropriate management of suspected cases requires their prompt recognition via attention to travel and bird-exposure history. The early isolation, diagnosis and treatment of suspected cases as well as prompt involvement of the health protection unit should enable patients to be optimally managed with minimum risk to health care staff.


The small regulatory RNA molecule MicA is involved in Salmonella enterica serovar Typhimurium biofilm formation.
LuxS is the synthase enzyme of the quorum sensing signal AI-2. In Salmonella Typhimurium, it was previously shown that a luxS deletion mutant is impaired in biofilm formation. However, this phenotype could not be complemented by extracellular addition of quorum sensing signal molecules. Analysis of additional S. Typhimurium luxS mutants indicated that the LuxS enzyme itself is not a prerequisite for a wild type mature biofilm. However, in close proximity of the luxS coding sequence, a small RNA molecule, MicA, is encoded on the opposite DNA strand. Interference with the MicA expression level showed that a balanced MicA level is essential for mature Salmonella biofilm formation. Several MicA targets known to date have previously been reported to be implicated in biofilm formation in Salmonella or in other bacterial species. Additionally, we showed by RT-qPCR analysis that MicA levels are indeed altered in some luxS mutants, corresponding to their biofilm formation phenotype. We show that the S. Typhimurium biofilm formation phenotype of a luxS mutant in which the complete coding region is deleted, is dependent on the sRNA molecule MicA, encoded in the luxS adjacent genomic region, rather than on LuxS itself. Future studies are required to fully elucidate the role of MicA in Salmonella biofilm formation.


Automatic versus Choice-Dependent Value Representations in the Human Brain.
The subjective values of choice options can impact on behavior in two fundamentally different types of situations: first, when people explicitly base their actions on such values, and second, when values attract attention despite being irrelevant for current behavior. Here we show with functional magnetic resonance imaging (fMRI) that these two behavioral functions of values are encoded in distinct regions of the human brain. In the medial prefrontal cortex, value-related activity is enhanced when subjective value becomes choice-relevant, and the magnitude of this increase relates directly to the outcome and reliability of the value-based choice. In contrast, activity in the posterior cingulate cortex represents values similarly when they are relevant or irrelevant for the present choice, and the strength of this representation predicts attentional capture by choice-irrelevant values. Our results suggest that distinct components of the brain's valuation network encode value in context-dependent manners that serve fundamentally different behavioral aims.


Distribution and change of peroxynitrite in the guinea pig cochlea following noise exposure.
Nitric oxide (NO)-mediated pathology depends on the formation of reactive intermediates, such as the peroxynitrite (ONOO-). ONOO- can nitrate free tyrosine and tyrosine residues of proteins. Therefore, increases in tyrosine nitration reflect the amount of ONOO- produced by oxidative stress. The distribution of 3-nitrotyrosine (3-NT), an ONOO- marker, in the organ of corti and the cochlear lateral wall tissue from the guinea pig were examined using fluorescence immunohistochemistry. The immunoactivity of 3-NT in the normal guinea pig was compared with animals exposed to 122dBA broadband noise, 4 h/day, for 2 consecutive days. In the normal animals, 3-NT immunoreactivity was found in the outer hair cells (OHCs), inner hair cells (IHCs), pillar cells (PCs), spiral ganglion cells (SPCs) and the marginal cells of stria vascularis in the lateral wall. Sound exposure increased the 3-NT signal in all of the cells and resulted in extensive outer hair cell loss. A quantitative analysis of the 3-NT change in OHCs and marginal cells of lateral wall showed that immunolabeling was significant (P<0.01, n=10) in the noise exposure group compared with that of the control group. Anti-3-NT and propidium iodide double labeling showed that 3-NT was distributed mainly in the apical end of OHCs. In addition, 3-NT was distributed outside of the nucleus of the OHCs and marginal cells. In conclusion, the data indicate that noise exposure leads to a significant production of ONOO- in the cochlear lateral wall and organ of corti. This is consistent with the known increase of NO production by loud sound stress and suggests that NO-derived free radicals participate in the cochlear pathophysiology of noise-induced hearing loss.


Technical hazards of using nutritive mixtures in bags for cyclical intravenous nutrition: comparison with standard intravenous nutrition in 48 gastroenterological patients.
Three methods for dispensing nutritional solutions are compared in 48 patients with gastrointestinal diseases on intravenous nutrition during 3582 days. The protocol for intravenous nutrition applied by the nursing team and the solutions used were the same in the three groups. In group A standard bottles were used, while in group B, 31PVC-disposable bags were used--with fat emulsion included (group B1) or with fat excluded (group B2). When fat was excluded from the bags it was infused separately from a bottle. The mixtures were made under laminar flow by the nursing team who applied a strict protocol which included bacteriological testing. The infection rate observed in the bags was 0.046%. The rate of septic complications was not significantly reduced in group B2 or B1 compared with group A; the type of container used was therefore unimportant and the key was the aseptic handling of the intravenous solutions. The rate of mechanical complications, mainly due to catheter obstruction, was higher (p less than 0.001) when fat was included in the bags--that is, in group B1--than in groups B2 and A. For 26 patients a cyclical regime of intermittent feeding was easier to manage when bags were used. In group B, this system replaced the continuous method n 75% of all therapeutic days without adverse effect; it improved compliance and allowed ambulatory treatment. The use of cyclical feeding with separate fat infusions has further reduced the hazards of intravenous nutrition and allowed the development of a programme that can be implemented at home.


In vivo quantification and perturbation of Myc-Max interactions and the impact on oncogenic potential.
The oncogenic bHLH-LZ transcription factor Myc forms binary complexes with its binding partner Max. These and other bHLH-LZ-based protein-protein interactions (PPI) in the Myc-Max network are essential for the physiological and oncogenic activities of Myc. We have generated a genetically determined and highly specific protein-fragment complementation assay based on Renilla luciferase to analyze the dynamic interplay of bHLH-LZ transcription factors Myc, Max, and Mxd1 in vivo. We also applied this PPI reporter to quantify alterations of nuclear Myc-Max complexes in response to mutational events, competitive binding by the transcriptional repressor Mxd1, or perturbations by small-molecule Myc inhibitors, including recently identified potent PPI inhibitors from a Kröhnke pyridine library. We show that the specificity of Myc-Max PPI reduction by the pyridine inhibitors directly correlates with their efficient and highly specific potential to interfere with the proliferation of human and avian tumor cells displaying deregulated Myc expression. In a direct comparison with known Myc inhibitors using human and avian cell systems, the pyridine compounds reveal a unique inhibitory potential even at sub-micromolar concentrations combined with remarkable specificity for the inhibition of Myc-driven tumor cell proliferation. Furthermore, we show in direct comparisons using defined avian cell systems that different Max PPI profiles for the variant members of the Myc protein family (c-Myc, v-Myc, N-Myc, L-Myc) correlate with their diverse oncogenic potential and their variable sensitivity to the novel pyridine inhibitors.


Host nuclear genotype influences phenotype of a conditional mutualist symbiont.
Arthropods commonly carry maternally inherited intracellular bacterial symbionts that may profoundly influence host biology and evolution. The intracellular symbiont Rickettsia sp. nr. bellii swept rapidly into populations of the sweetpotato whitefly Bemisia tabaci in the south-western USA. Previous laboratory experiments showed female-bias and fitness benefits were associated with Rickettsia infection, potentially explaining the high frequencies of infection observed in field populations, but the effects varied with whitefly genetic line. Here, we explored whether host extranuclear or nuclear genes influenced the variation in the Rickettsia-host phenotype in two genetic lines of the whitefly host, each with Rickettsia-infected and uninfected sublines. Introgression between the Rickettsia-infected subline of one genetic line and the Rickettsia-uninfected subline of the other was used to create two new sublines, each with the maternally inherited extranuclear genetic lineages of one line (Rickettsia, two other symbionts and the mitochondria) and the nuclear genotype of the other. Performance assays comparing the original and new lines showed that in addition to Rickettsia, the interaction of Rickettsia infection with host nuclear genotype influenced development time and the sex ratio of the progeny, whereas the extranuclear genotype did not. Host nuclear genotype, but not extranuclear genotype, also influenced the titre of Rickettsia. Our results support the hypothesis that differences in host nuclear genotype alone may explain considerable within-population variation in host-symbiont phenotype and may contribute to the observed variation in Rickettsia-whitefly interactions worldwide.


A grounded theory approach to understand the cancer-coping process.
A focus on individualistic coping strategies such as positivity and fighting spirit has done little to inform the development of interventions for people affected by cancer. The aim of this study was to conduct an inquiry, which focused on the 'lived experience' and the social context within which communication about cancer occurs. A semi-structured interview schedule was used. These interviews were complemented by the Mini-International Neuropsychiatric Interview (M.I.N.I., Sheehan et al., 1998). Twenty participants with mixed diagnoses (13 female and seven male), ranging in age from 44 to 73 (Mean = 58.35, SD= 8.20) were interviewed. Consistent with Glaser's (1992) constant comparative method, research data were open coded, selectively coded, and then theoretically coded so that initial codes were collapsed into models and compared against existing literature for completeness. Fifty percent of participants met criteria for an Axis 1 diagnosis. Three models emerged from the research interview; the first provides a description of the way in which patients evaluate the quality of care they receive across the cancer pathway. Other models refer to the cancer-coping process in which participants describe successful and unsuccessful attempts at coping via communication and whereby coping with cancer is characterized by a series of losses accompanied by increased distress. A research focus on the social environment in which cancer is experienced provides considerable insight into the cancer-coping process. As such, interventions focused on understanding the social context in which patients experience cancer, and attempt to cope, may facilitate improved adjustment for those directly, and potentially indirectly, affected by cancer. Further research is required to develop interventions to address the unique needs of survivors.


Cryptic amyloidogenic regions in intrinsically disordered proteins: Function and disease association.
The amyloid conformation is considered a fundamental state of proteins and the propensity to populate it a generic property of polypeptides. Multiple proteome-wide analyses addressed the presence of amyloidogenic regions in proteins, nurturing our understanding of their nature and biological implications. However, these analyses focused on highly aggregation-prone and hydrophobic stretches that are only marginally found in intrinsically disordered regions (IDRs). Here, we explore the prevalence of cryptic amyloidogenic regions (CARs) of polar nature in IDRs. CARs are widespread in IDRs and associated with IDPs function, with particular involvement in protein-protein interactions, but their presence is also connected to a risk of malfunction. By exploring this function/malfunction dichotomy, we speculate that ancestral CARs might have evolved into functional interacting regions playing a significant role in protein evolution at the origins of life.


Factors associated with the acceptability of male circumcision among men in Jamaica.
To determine the prevalence of male circumcision (MC) among men in the western region of Jamaica, and to identify factors associated with acceptability of MC for self, infants (<1 year) and older sons (1-17 years). A cross-sectional, interviewer-administered questionnaire survey of 549 men aged 19-54 years was conducted in the western region of Jamaica. The survey included questions about the acceptance of MC for self, infants, and sons before and after an information session about the benefits of MC in preventing HIV/STI transmission. Logistic regression models were used to identify factors that were associated with acceptability of MC. Adjusted odds ratios (AOR) and 95% confidence intervals (CI) were calculated from the models. Fourteen percent of the men reported that they were circumcised. In the multivariable model, which adjusted for age, education, religion and income, there were increased odds of accepting MC for infants/sons among uncircumcised men who accepted MC for self (AOR=8.1; 95% CI = 4.1-15.9), believed they would experience more pleasure during sex if circumcised (AOR=4.0; 95% CI = 2.0-8.2), and reported having no concerns regarding MC (AOR=3.0; 95% CI = 1.8-4.8). Similarly, uncircumcised men who reported no concerns about MC or who believed that they would experience more pleasure during sex if circumcised were more likely to accept MC for self. Providing men with information about MC increased acceptance of MC for self, infants (<17 years) and sons (1-17 years). Since targeted education on the benefits of male circumcision for prevention of HIV/STI can be effective in increasing acceptability of MC, health professionals should be trained, and willing to discuss MC with men in healthcare facilities and in the community.


Impact of pupil transmission apodization on presbyopic through-focus visual performance with spherical aberration.
To investigate the impact on through-focus retinal image quality and visual performance of apodizing the pupil's transmission function in combination with extended depth of focus presbyopic corrections, such as spherical aberration (SA). Through-focus retinal image quality was determined theoretically for various magnitudes of pupil transmission apodization and Zernike primary SA (-0.5 to +0.5 μm) for a 4-mm pupil. The impact of pupil transmission apodization was also assessed psychophysically with a vision simulator equipped with a liquid crystal spatial light modulator for controlling pupil transmission. Through-focus visual acuity (VA) was measured with and without apodization in three cyclopleged subjects from distance to near with monochromatic light (550 nm) under two multifocal aberration conditions. Phase plates induced +0.2 and -0.2 μm of SA over a 4-mm artificial pupil. A baseline condition of zero SA was also included for comparison. The theoretical investigation showed that pupil transmission apodization significantly improved distance image quality in the presence of positive and negative SA. Retinal image quality at all target vergences for negative SA conditions was improved by apodization. Pupil transmission apodization improved through-focus VA by 0.1 to 0.2 logMAR at intermediate and near object distances for the zero and negative SA conditions. In the positive SA condition, apodization degraded VA by approximately 0.1 logMAR at intermediate object distances. Pupil transmission apodization had a significant impact on though-focus visual performance. Pupil transmission apodization affects through-focus retinal image quality by diminishing the relative contribution to the retinal image from the peripheral region of the wavefront aberration. Through-focus visual performance in presbyopic eyes with negative SA was improved due to pupil transmission apodization.


microRNA27a-3p mediates reduction of the Wnt antagonist sFRP-1 in systemic sclerosis.
Systemic Sclerosis (SSc) is an autoimmune connective tissue disease that leads to skin and lung fibrosis. The Wnt pathway is clearly elevated in SSc and is pro-fibrotic via activation of canonical Wnt signalling. sFRP-1 is a Wnt antagonist that acts as a negative regulator of Wnt signalling. We sought to measure the levels of serum sFRP-1 in early diffuse SSc patients compared to healthy controls and if this is regulated by microRNA27a-3p. Ten early diffuse SSc patients and healthy controls sera were taken and sFRP-1 quantified by ELISA. Skin biopsies were also taken in five SSc patients and controls. Fibroblasts were quantified for microRNA27-3p expression by Taqman qRT-PCR with an internal microRNA to normalize. 3'UTR luciferase assays were performed to confirm direct targets of microRNA27a-3p with microRNA overexpression. Fibroblasts were transfected with microRNA27a mimics or scramble controls and using ELISA sFRP-1 was quantified. Furthermore, Collagen, Axin-2, TIMP-1 and MMP-1 were measured. Serum sFRP-1 was significantly reduced in early diffuse SSc patients. We identified microRNA27a-3p-3p as regulating sFRP-1 in dermal fibroblasts. We found significantly elevated microRNA27a-3p in isolated dermal fibroblasts from SSc patients. We confirmed that sFRP-1 is a direct target of microRNA27a-3p through cloning of the 3'UTR into a luciferase vector. ECM genes were also upregulated by microRNA27a-3p-3p and the matrix-degrading enzyme MMP-1 was suppressed. Serum sFRP-1 is reduced in diffuse SSc patients and is regulated by microRNA27a-3p and this is a direct regulation. Modulation of microRNA27a-3p levels could mediate fibrosis regression.


Borealin: a novel chromosomal passenger required for stability of the bipolar mitotic spindle.
The chromosomal passenger complex of Aurora B kinase, INCENP, and Survivin has essential regulatory roles at centromeres and the central spindle in mitosis. Here, we describe Borealin, a novel member of the complex. Approximately half of Aurora B in mitotic cells is complexed with INCENP, Borealin, and Survivin; and Borealin binds Survivin and INCENP in vitro. A second complex contains Aurora B and INCENP, but no Borealin or Survivin. Depletion of Borealin by RNA interference delays mitotic progression and results in kinetochore-spindle misattachments and an increase in bipolar spindles associated with ectopic asters. The extra poles, which apparently form after chromosomes achieve a bipolar orientation, severely disrupt the partitioning of chromosomes in anaphase. Borealin depletion has little effect on histone H3 serine10 phosphorylation. These results implicate the chromosomal passenger holocomplex in the maintenance of spindle integrity and suggest that histone H3 serine10 phosphorylation is performed by an Aurora B-INCENP subcomplex.


Linkage-to-care Methods and Rates in U.S. Emergency Department-based HIV Testing Programs: A Systematic Literature Review Brief Report.
An increasing number of U.S. emergency departments (EDs) have implemented ED-based HIV testing programs since the Centers for Disease Control and Prevention issued revised HIV testing recommendations for clinical settings in 2006. In 2010, the National HIV/AIDS Strategy (NHAS) set an linkage-to-care (LTC) rate goal of 85% within 90 days of HIV diagnosis. LTC rates for newly diagnosed HIV-infected patients vary markedly by site, and many are suboptimal. The optimal approach for LTC in the ED setting remains unknown. The objective was to perform a brief descriptive analysis of the LTC methods practiced in EDs across the United States to determine the overall linkage rate of ED-based HIV testing programs. We conducted a systematic review of literature related to U.S. ED HIV testing in the adult population using PubMed, Embase, Web of Science, Scopus, and Cochrane. There were 333 articles were identified; 31 articles were selected after a multiphasic screening process. We analyzed data from the 31 articles to assess LTC methods and rates. LTC methods that involved physical escort of the newly diagnosed patient to an HIV/infectious disease (ID) clinic or interaction with a specialist health care provider at the ED were operationally defined as "intensive" LTC protocol. "Mixed" LTC protocol was defined as a program that employed intensive linkage only part of the coverage hours. All other forms of linkage was defined as "nonintensive" LTC protocol. An LTC rate of ≥85% was used to identify characteristics of ED-based HIV testing program associated with a higher LTC rate. There were 37 ED-based HIV testing programs in the 31 articles. The overall LTC rate was 74.4%. Regarding type of protocol, nine (24.3%) employed intensive LTC protocols, 25 (67.6%) nonintensive, two (5.4%) mixed, and one (2.7%) with unclear protocols. LTC rates for programs with intensive and nonintensive LTC protocols were 80.0 and 72.7%, respectively. Four (44.4%) with intensive protocols and nine (36.0%) with the nonintensive protocols had LTC rates > 85%. The linkage staff employed was different between ED programs. Among them, 25 (67.6%) programs used exogenous staff, 10 (27.0%) used the ED staff, and two had no information. All the programs in the nonintensive group utilized drop-in HIV/ID clinic or medical appointments while seven of nine of the programs in the intensive group physically escorted the patients to the initial medical intake appointment. There were no significant differences in characteristics of ED-based HIV testing programs between those with ≥85% LTC rate versus those with <85% within the intensive or nonintensive group. Intensive LTC protocols had a higher LTC rate and a higher proportion of programs that surpassed the >85% NHAS goal compared to nonintensive methods, suggesting that, when possible, ED-based HIV testing programs should adopt intensive LTC strategies to improve LTC outcomes. However, intensive LTC protocols most often required involvement of multidisciplinary non-ED professionals and external research funding. Our findings provide a foundation for developing best practices for ED-based HIV LTC programs.


Advances in the diagnosis, immunopathogenesis and therapies of IgM-anti-MAG antibody-mediated neuropathies.
Polyneuropathy with immunoglobulin M (IgM) monoclonal gammopathy is the most common paraproteinemic neuropathy, comprising a clinicopathologically and immunologically distinct entity. The clinical spectrum spans from distal paresthesias and mild gait imbalance to more severe sensory ataxia, with falls and a varying degree of distal sensorimotor deficits. In approximately 75% of patients, the monoclonal IgM immunoreacts with myelin-associated glycoprotein (MAG) and sulfoglucuronyl glycosphingolipid (SGPG), or other peripheral nerve glycolipids that serve as antigens. These antibodies are considered pathogenic because IgM and complement are deposited on the myelin sheath, splitting the myelin lamellae, while adoptive transfer of patients' IgM into susceptible host animals causes sensory ataxia and reproduces the human pathology. In spite of the apparently convincing pathogenicity of these antibodies, the response to immunotherapies remains suboptimal. Clorambuscil, cladibrine, cyclophospamide and intravenous immunoglobulin may help some patients but the benefits are minimal and transient. Open-label studies in >200 patients indicate that rituximab is helpful in 30-50% of these patients, even with long-term benefits, probably by suppressing IgM anti-MAG antibodies or inducing immunoregulatory T cells. Two controlled studies with rituximab did not however meet the primary endpoint, mostly because of the poor sensitivity of the scales used; they did however show statistical improvement in secondary endpoints and improved clinical functions in several patients. This review provides an overview of the clinical phenotypes and immunoreactivity of IgM to glycolipids or glycoproteins of peripheral nerve myelin, summarizes the progress on treatment with rituximab as a promising therapy, discusses the pitfalls of scales used, identifies possible biomarkers of response to therapy and highlights the promising new anti-B cell or target-specific immunotherapies.


Continuous passive motion and its effects on knee flexion after total knee arthroplasty in patients with knee osteoarthritis.
This study evaluated the effects of continuous passive motion (CPM) on accelerated flexion after total knee arthroplasty (TKA) and whether CPM application measures (i.e. initial angle and daily increment) are associated with functional outcomes. A retrospective investigation was conducted at the rehabilitation centre of a university-based teaching hospital. Patients who received CPM therapy immediately after TKA surgery were categorized into rapid-, normal-, and slow-progress groups according to their response to CPM during their acute inpatient stay. Knee pain, passive knee flexion, and knee function-measured using the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC)-were assessed preoperatively at discharge and at 3- and 6-month outpatient follow-up visits. A total of 354 patients were followed for 6 months after inpatient-stay discharge. The patients in the rapid-progress group (n = 119) exhibited significantly greater knee flexions than those in the slow-progress group did (n = 103) at the 3-month follow-up [mean difference (MD) = 10.3°, 95 % confidence interval (CI) 4.3°-16.3°, p < 0.001] and 6-month follow-up (MD = 10.9°, 95 % CI 6.3°-15.6°, p < 0.001). Significant WOMAC score differences between the rapid- and slow-progress groups were observed at the 3-month follow-up (MD = 7.2, 95 % CI 5.4-9.1, p < 0.001) and 6-month follow-up (MD = 16.1, 95 % CI 13.4-18.7, p < 0.001). CPM initial angles and rapid progress significantly predicted short- and long-term outcomes in knee flexion and WOMAC scores (p < 0.001). When CPM is used, early application with initial high flexion and rapid progress benefits knee function up to 6 months after TKA. II.


Evaluation of natural anthracene-derived compounds as antimitotic agents.
Plants that contain anthracene-derived compounds such as anthraquinones have been reported to act as anticancer besides their use for millennia to treat constipation, but the mechanism of action is still unfolding. Therefore we pursue this study to explore a new horizon in the anticancer property of these agents with relevance to mitotic arrest. To achieve this goal, the antimitotic activity of a series of naturally occurring anthracene-derived anthraquinones including anthrone, alizarin (1,2-dihydroxyanthraquinone), quinizarin (1,4-dihydroxyanthraquinone), rhein (4,5-dihydroxyanthraquinone-2-carboxylic acid), emodin (1,6,8-trihydroxy-3-methylanthraquinone), and aloe emodin (1,8-dihydroxy-3-hydroxymethylanthraquinone) were evaluated using Allium cepa root tips. Initial results revealed that the mitosis was inhibited after 3, 6, and 24 h, respectively, of incubation with 500, 250, and 125 ppm of each compound in a dose-dependent manner. Furthermore, alizarin at 500 ppm was proved to be the most active compound to arrest the mitosis after 24 h followed by emodin, aloe emodin, rhein, and finally quinizarin. Interestingly, this inhibition of mitosis was irreversible in root tips incubated with each compound at concentration of 500 ppm but not with 250 ppm or 125 ppm, where the roots regained their normal mitotic activity after 96 h post-incubation in water. This re-evaluation of an old remedy suggests that several bioactive anthraquinones possess promising anti-mitotic activity that may have the potential to be lead compounds for the development of a new class of multifaceted natural anticancer/antimitotic agents.


Kinetic and spectroscopic studies on the quercetin 2,3-dioxygenase from Bacillus subtilis.
Quercetin 2,3-dioxygenase from Bacillus subtilis (QueD) converts the flavonol quercetin and molecular oxygen to 2-protocatechuoylphloroglucinolcarboxylic acid and carbon monoxide. QueD, the only known quercetin 2,3-dioxygenase from a prokaryotic organism, has been described as an Fe2+-dependent bicupin dioxygenase. Metal-substituted QueDs were generated by expressing the enzyme in Escherichia coli grown on minimal media in the presence of a number of divalent metals. The addition of Mn2+, Co2+, and Cu2+ generated active enzymes, but the addition of Zn2+, Fe2+, and Cd2+ did not increase quercetinase activity to any significant level over a control in which no divalent ions were added to the media. The Mn2+- and Co2+-containing QueDs were purified, characterized by metal analysis and EPR spectroscopy, and studied by steady-state kinetics. Mn2+ was found to be incorporated nearly stoichiometrically to the two cupin motifs. The hyperfine coupling constant of the g = 2 signal in the EPR spectra of the Mn2+-containing enzyme showed that the two Mn2+ ions are ligated in an octahedral coordination. The turnover number of this enzyme was found to be in the order of 25 s(-1), nearly 40-fold higher than that of the Fe2+-containing enzyme and similar in magnitude to that of the Cu2+-containing quercertin 2,3-dioxygenase from Aspergillus japonicus. In addition, kinetic and spectroscopic data suggest that the catalytic mechanism of QueD is different from that of the Aspergillus quercetinases but similar to that proposed for the extradiol catechol dioxygenases. This study provides evidence that Mn2+ might be the preferred cofactor for this enzyme and identifies QueD as a new member of the manganese dioxygenase family.


Fibrillin immunoreactive fibers constitute a unique network in the human dermis: immunohistochemical comparison of the distributions of fibrillin, vitronectin, amyloid P component, and orcein stainable structures in normal skin and elastosis.
Fibrillin, a 350-kD glycoprotein, was recently localized to elastin-associated 10 nm microfibrils. Here, the distribution of fibrillin immunoreactivity was determined in normal skin in individuals of different ages and in lesions of solar elastosis or anetoderma. It was compared with the distribution of orcein-stainable fibers and with the immunoreactivities of vitronectin and amyloid P component. These glycoproteins are known to occur in conjunction with the orcein-stainable elastic fibers in adults, but not in the young. Fibrillin immunoreactivity was associated with orcein-stainable fibers in normal skin of both adults and the young. In addition, the fibrillin immunoreactive fiber network comprised fine fibers that were unstainable by orcein, anti-vitronectin, or anti-amyloid P component. Such fine fibers were especially abundant close to the dermal-epidermal junction zone. Immunoreactivities of anti-vitronectin and anti-amyloid P component were not always associated with fibrillin immunoreactivity but were consistently found to co-localize with orcein-stainable fibers in adults. This suggests vitronectin and amyloid P component to be associated with the amorphous elastin rather than with the microfibrils, although alternative interpretations are possible. In elastotic lesions, fibrillin immunoreactivity was generally fainter than that obtained using anti-vitronectin or anti-amyloid P component. In contrast, an extensive network of dermal fibers stained by anti-fibrillin, but not by anti-amyloid P component, anti-vitronectin, or orcein, was seen in an anetoderma lesion. In conclusion, fibrillin immunoreactivity is associated with a unique dermal network, which ultrastructurally is composed of microfibrils. These fibers are proposed to have an important structural and functional role in anchoring the dermal elastic fibers in the extracellular matrix and to the lamina densa.


Trends in incidence of oral and pharyngeal carcinoma in Florida: 1981-2008.
While the overall incidence rates of oral and pharyngeal squamous cell carcinoma (SCC) have decreased in the United States, there is evidence of increasing incidence at selected anatomic sites, particularly among younger adults. The objective of this study was to examine trends in incidence rates of oral and pharyngeal cancers in Florida. Using data from the Florida Cancer Data System, we examined the incidence of oral and pharyngeal carcinomas in Florida from 1981 through 2008. Factors of interest included sex, race, and trends over time. Percent change (PC) and annual percent change (APC) were computed to characterize trends over time. A total of 53,648 cases of oral or pharyngeal cancer were identified from 1981 through 2008. Significant increasing trends were observed only for pharyngeal cancers in males, with significant decreasing trends for pharyngeal cancer in females and oral cancer for both sexes. For tonsil and base of tongue cancers, increasing trends were detected for white males only. Further investigation among white males showed that, except for base of tongue cancer in the 20-44 age group, the incidence of both cancers increased across all age groups, with the largest increase for both sites found in the 45-64 age group. This study supports the finding of increasing incidence of SCC of the tonsil and base of tongue in males, in contrast to decreasing trends for most oral and pharyngeal carcinomas. However, we observed that this increase occurred in white males only and the most dramatic increase occurred in the 45-64 age group.


Inadvertent doping through supplement use by athletes: assessment and management of the risk in Australia.
Many athletes report using a wide range of special sports foods and supplements. In the present study of 77 elite Australian swimmers, 99% of those surveyed reported the use of these special preparations, with 94% of swimmers reporting the use of non-food supplements. The most popular dietary supplements were vitamin or mineral supplements (used by 94% of the group), herbal preparations (61%), and creatine (31%). Eighty-seven percent of swimmers reported using a sports drink or other energy-providing sports food. In total, 207 different products were reported in this survey. Sports supplements, particularly supplements presented as pills or other non-food form, are poorly regulated in most countries, with little assurance of quality control. The risk of an inadvertent "positive doping test" through the use of sports supplements or sports foods is a small but real problem facing athletes who compete in events governed by anti-doping rules. The elite swimmers in this survey reported that information about the "doping safety" of supplements was important and should be funded by supplement manufacturers. Although it is challenging to provide such information, we suggest a model to provide an accredited testing program suitable for the Australian situation, with targeted athlete education about the "sports safety" of sports supplements and foods.


Oesophageal transit of six commonly used tablets and capsules.
The oesophageal transit of six commonly used tablets and capsules containing barium sulphate was evaluated radiologically using fluoroscopy in 121 healthy volunteers. To determine the influence of the subject's position and the amount of water taken each subject swallowed three preparations while recumbent and standing and with 25 ml or 100 ml of water. Failure of swallowing (defined as oesophageal transit taking more than 90 seconds) occurred in 22% of 726 swallowings, but globus was complained of in only 33% of these. Sixty per cent of the volunteers had difficulty in taking one or more of the preparations. Many preparations adhered to the oesophageal membrane and started to disintegrate in the lower part of oesophagus. It is recommended that subjects should remain standing for at least 90 seconds after taking capsules or tablets and that all preparations should be taken with at least 100 ml of water. Small tablets are swallowed most easily. Liquid forms of medication (suspensions) should be considered for bedridden patients and those who have difficulty in swallowing.


The mitochondrial genome and psychiatric illness.
Psychiatric disorders are a leading cause of morbidity and mortality, yet their underlying pathophysiology remains unclear. Searches for a genetic cause of bipolar disorder, schizophrenia, and major depressive disorder have yielded inconclusive results. There is increasing interest in the possibility that defects in the mitochondrial genome may play an important role in psychiatric illness. We undertook a review of the literature investigating mitochondria and adult psychiatric disorders. MEDLINE, PsycINFO, and EMBASE were searched from their inception through September 2011, and the reference lists of identified articles were reviewed for additional studies. While multiple lines of evidence, including clinical, genetic, ultrastructural, and biochemical studies, support the involvement of mitochondria in the pathophysiology of psychiatric illness, many studies have methodological limitations and their findings have not been replicated. Clinical studies suggest that psychiatric features can be prominent, and the presenting features of mitochondrial disorders. There is limited but inconsistent evidence for the involvement of mitochondrial DNA haplogroups and mitochondria-related nuclear gene polymorphisms, and for mitochondrial ultrastructural and biochemical abnormalities in psychiatric illness. The current literature suggests that mitochondrial dysfunction and mitochondrial genetic variations may play an important role in psychiatric disorders, but additional methodologically rigorous and adequately powered studies are needed before definitive conclusions can be drawn.


Attitudes about complementary and alternative medicine did not predict outcome in a homeopathic proving trial.
A homeopathic proving states that when a homeopathic remedy is given to a healthy person, they will experience symptomatic effects specific to that remedy. The aim of this study was to assess if attitudes about complementary and alternative medicine (CAM) predicted proving outcome. An exploratory study nested in a double blinded randomized controlled proving trial of the remedy Belladonna C30. Two hundred and six (206) healthy subjects took part in the reproving trial of whom 29 (14%) proved. Subjects completed the Attitudes to Alternative Medicine Scale prior to participating in the proving trial (n = 206) and after the proving trial was completed (n = 77). The primary outcome was whether attitude to CAM predicted proving outcome. Attitudes about CAM at baseline did not predict proving outcome (chi2 = 35.34, df = 36, p = 0.50) and attitudes to CAM did not alter after taking part in the proving trial for either provers or non provers. Subjects classified as being provers in a proving trial did not have significantly different attitudes about CAM than nonprovers. The experience of proving the study remedy did not alter the prover's attitudes about CAM.


Expression of the Hsp23 chaperone during Drosophila embryogenesis: association to distinct neural and glial lineages.
In addition to their strong induction following stress, small heat shock proteins (Hsp) are also expressed during development in a wide variety of organisms. However, the precise identity of cell(s) expressing these proteins and the functional contribution of small heat shock proteins in such developmental context remain to be determined. The present study provides a detailed description of the Drosophila small heat shock protein Hsp23 expression pattern during embryogenesis and evaluates its functional contribution to central nervous system development. Throughout embryogenesis, Hsp23 is expressed in a stage-specific manner by a restricted number of neuronal and glial lineages of the central nervous system. Hsp23 is also detected in the amnioserosa and within a single lateral chordotonal organ. Its expression within the MP2 lineage does not require the presence of a functional midline nor the activity of the Notch signaling pathway. Transactivation assays demonstrate that transcription factors implicated in the differentiation of the midline also regulate hsp23 promoter activity. Phenotypic analysis of a transgenic line exhibiting loss of Hsp23 expression in the central nervous system suggests that Hsp23 is not required for development and function of this tissue. Likewise, its overexpression does not cause deleterious effects, as development remains unaffected. Based on the presented data, we suggest that the tightly regulated developmental expression of Hsp23 is not actively involved in cell differentiation and central nervous system development per se but rather reflects a putative role in preventive "pre-stress" neuroprotection or in non-vital process(es) common to the identified cell lineages.


High spectral efficient W-band optical/wireless system employing single-sideband single-carrier modulation.
With broader available bandwidth, W-band wireless transmission has attracted a lot of interests for future Giga-bit communication. In this article, we experimentally demonstrate W-band radio-over-fiber (RoF) system employing single-sideband single-carrier (SSB-SC) modulation with lower peak-to-average-power ratio (PAPR) than orthogonal frequency division multiplex (OFDM). To overcome the inter-symbol interference (ISI) of the penalty from uneven frequency response and SSB-SC modulation, frequency domain equalizer (FDE) and decision feedback equalizer (DFE) are implemented. We discuss the maximum available bandwidth of different modulation formats between SSB-SC and OFDM signals at the BER below forward error correction (FEC) threshold (3.8 × 10(-3)). Up to 50-Gbps 32-QAM SSB-SC signals with spectral efficiency of 5 bit/s/Hz can be achieved.


Unidirectional internuclear transfer of linked genes in heterokaryons of Candida albicans.
Nutritionally balanced heterokaryons of the naturally diploid, asexual yeast, Candida albicans are produced by fusing protoplasts of complementing auxotrophic strains. Spontaneous unidirectional internuclear transfers of an intact gene linkage group in established heterokaryons is demonstrated. Evidence is presented that a transfer event (i) typically involves a single chromosome which is added to the resident homologues of a recipient nucleus, (ii) can occur equally well in either direction between complementing nuclei and (iii) may encourage gene conversion at transferred loci in the recipient nucleus. The bearing of these findings on application of protoplast fusion to parasexual genetic analysis of C. albicans is discussed.


Inducible cell adhesion molecule 110 (INCAM-110) is an endothelial receptor for lymphocytes. A CD11/CD18-independent adhesion mechanism.
Inducible cell adhesion molecule 110 (INCAM-110) is a 110-kD glycoprotein expressed on cytokine-activated human vascular endothelial cells. mAb blocking studies indicate that INCAM-110 and intercellular adhesion molecule 1 (ICAM-1) independently support the adhesion of lymphocytes to activated human umbilical vein endothelial cell monolayers. Anti-CD11a/CD18 antibodies with anti-INCAM-110 mAb E1/6 produce greater inhibition of lymphocyte adhesion than either reagent alone, suggesting that INCAM-110 and LFA-1 are not an obligate receptor-ligand pair. Blood monocytes, but not polymorphonuclear leukocytes, also appear to bind endothelial INCAM-110. Endothelial expression of INCAM-110 is upregulated at sites of inflammation, suggesting a role in the recruitment of mononuclear leukocytes.


Aurones and derivatives as promising New Delhi metallo-β-lactamase (NDM-1) inhibitors.
Bacterial resistance is undoubtedly one of the main public health concerns especially with the emergence of metallo-β-lactamases (MBLs) able to hydrolytically inactivate β-lactam antibiotics. Currently, there are no inhibitors of MBLs in clinical use to rescue antibiotic action and the New Delhi metallo-β-lactamase-1 (NDM-1) is still considered as one of the most relevant targets for inhibitor development. Following a fragment-based strategy to find new NDM-1 inhibitors, we identified aurone as a promising scaffold. A series of 60 derivatives were then evaluated and two of them were identified as promising inhibitors with Ki values as low as 1.7 and 2.5 µM. Moreover, these two most active compounds were able to potentiate meropenem in in vitro antimicrobial susceptibility assays. The molecular modelling provided insights about their likely interactions with the active site of NDM-1, thus enabling further improvement in the structure of this new inhibitor family.


Milk sharing and formula feeding: Infant feeding risks in comparative perspective?
The advent of Internet forums that facilitate peer-to-peer human milk sharing has resulted in health authorities stating that sharing human milk is dangerous. There are risks associated with all forms of infant feeding, including breastfeeding and the use of manufactured infant formulas. However, health authorities do not warn against using formula or breastfeeding; they provide guidance on how to manage risk. Cultural distaste for sharing human milk, not evidenced-based research, supports these official warnings. Regulating bodies should conduct research and disseminate information about how to mitigate possible risks of sharing human milk, rather than proscribe the practice outright.


Young Drug Users: a Vulnerable Population and an Underutilized Resource in HIV/HCV Prevention.
The social networks of people who inject drugs (PWID) have long been studied to understand disease transmission dynamics and social influences on risky practices. We illustrate how PWID can be active agents promoting HIV, HCV, and overdose prevention. We assessed drug users' connections and interactions with others at risk for HIV/HCV in three cities: New York City (NYC), USA (n = 539); Pereira, Colombia (n = 50); and St. Petersburg, Russia (n = 49). In all three cities, the majority of participants' network members were of a similar age as themselves, yet connections across age groups were also present. In NYC, knowing any opioid user(s) older than 29 was associated with testing HCV-positive. In NYC and St. Petersburg, a large proportion of PWID engaged in intravention activities to support safer injection and overdose prevention; in Pereira, PWID injected, had sex, and interacted with other key groups at risk. People who use drugs can be active players in HIV/HCV and overdose risk- reduction; their networks provide them with ample opportunities to disseminate harm reduction knowledge, strategies, and norms to others at risk. Local communities could augment prevention programming by empowering drug users to be allies in the fight against HIV and facilitating their pre-existing health-protective actions.


First long-term behavioral records from Cuvier's beaked whales (Ziphius cavirostris) reveal record-breaking dives.
Cuvier's beaked whales (Ziphius cavirostris) are known as extreme divers, though behavioral data from this difficult-to-study species have been limited. They are also the species most often stranded in association with Mid-Frequency Active (MFA) sonar use, a relationship that remains poorly understood. We used satellite-linked tags to record the diving behavior and locations of eight Ziphius off the Southern California coast for periods up to three months. The effort resulted in 3732 hr of dive data with associated regional movements--the first dataset of its kind for any beaked whale--and included dives to 2992 m depth and lasting 137.5 min, both new mammalian dive records. Deep dives had a group mean depth of 1401 m (s.d. = 137.8, n = 1142) and duration of 67.4 min (s.d. = 6.9). The group mean time between deep dives was 102.3 min (s.d. = 30.8, n = 783). While the previously described stereotypic pattern of deep and shallow dives was apparent, there was considerable inter- and intra-individual variability in most parameters. There was significant diel behavioral variation, including increased time near the surface and decreased shallow diving at night. However, maximum depth and the proportion of time spent on deep dives (presumed foraging), varied little from day to night. Surprisingly, tagged whales were present within an MFA sonar training range for 38% of days locations were received, and though comprehensive records of sonar use during tag deployments were not available, we discuss the effects frequent acoustic disturbance may have had on the observed behaviors. These data better characterize the true behavioral range of this species, and suggest caution should be exercised when drawing conclusions about behavior using short-term datasets.


The cerebral cavernous malformation signaling pathway promotes vascular integrity via Rho GTPases.
Cerebral cavernous malformation (CCM) is a common vascular dysplasia that affects both systemic and central nervous system blood vessels. Loss of function mutations in the CCM2 gene cause CCM. Here we show that targeted disruption of Ccm2 in mice results in failed lumen formation and early embryonic death through an endothelial cell autonomous mechanism. We show that CCM2 regulates endothelial cytoskeletal architecture, cell-to-cell interactions and lumen formation. Heterozygosity at Ccm2, a genotype equivalent to that in human CCM, results in impaired endothelial barrier function. On the basis of our biochemical studies indicating that loss of CCM2 results in activation of RHOA GTPase, we rescued the cellular phenotype and barrier function in heterozygous mice with simvastatin, a drug known to inhibit Rho GTPases. These data offer the prospect for pharmacological treatment of a human vascular dysplasia with a widely available and safe drug.


Transient outward current in human ventricular myocytes of subepicardial and subendocardial origin.
In various mammalian species, shapes of action potentials vary within the cardiac wall because of differences in transient outward current (Ito). A prominent Ito exists in human ventricular myocytes, but cells have not been separated according to their original localization. Human ventricular myocytes were isolated from separated subepicardial and subendocardial tissue, and regional variations in Ito were studied. Ito was larger in subepicardial than subendocardial cells. Current density at +60 mV was 7.9 +/- 0.7 pA/pF (n = 28) in subepicardial cells and 2.3 +/- 0.3 pA/pF (n = 16) in subendocardial cells. When cells from explanted failing and nonfailing donor hearts were compared, Ito was not different in subepicardial cells; however, it was larger in subendocardial cells from nonfailing hearts. The potential of half-maximal activation (V0.5) was more positive in subendocardial cells (+25.6 +/- 3.5 mV, n = 15) than in subepicardial cells (+9.2 +/- 1.8 mV, n = 28). There was no difference in V0.5 between cells from failing and nonfailing hearts. Ito inactivation was similar in all cell types and independent of membrane depolarization (time constant [tau] = approximately 60 milliseconds at 22 degrees C). The potential of half-maximal steady-state inactivation was similar in all cell types. Recovery from inactivation of Ito was fast in subepicardial cells at -100 mV (tau = 24 +/- 4 milliseconds, n = 6), exceeding control values transiently (overshoot), and slow at -40 mV without overshoot (tau = 638 +/- 91 milliseconds, n = 6). In subendocardial cells, Ito recovered at -100 mV with a fast phase (tau = 25 milliseconds) and a slow phase (tau = 328 milliseconds), and recovery was not complete after 6 seconds at -100 mV. In conclusion, regional differences in Ito between subepicardial and subendocardial cells may have clinical implications with respect to rhythmic disturbance during heart failure.


Linezolid-resistant Staphylococcus aureus isolated from 2006 through 2008 at six hospitals in Japan.
Limited use of linezolid for treating methicillin-resistant Staphylococcus aureus (MRSA) infection was approved in Japan in 2006. We report here the status of linezolid-resistant MRSAs in Japan. Eleven linezolid-resistant clinical isolates from 11 patients at six hospitals were collected from 2006 through 2008. The minimal inhibitory concentration (MIC) of linezolid in these strains varied from 8 to 64 μg/ml. All strains had at least one G2576T mutation in the chromosomal gene(s) encoding domain V of the 23S ribosomal RNA (rRNA). Chromosomal DNA encoding five copies of the domain V region was analyzed by polymerase chain reaction (PCR). Strains with the linezolid MICs of 64, 32, 16, and 8 μg/ml had the G2576T mutation(s) in four, three (or four), two, and one copy of the 23S rRNA genes, respectively. These results suggest that the level of linezolid resistance seems to be roughly correlated with the number of mutations in the genes encoding 23S rRNA. DNA samples from all 11 strains were subjected to pulsed-field gel electrophoresis and were classified into seven independent clones having >92% identity. Among the 11 patients, five had been treated with linezolid and the remainder, in two hospitals, had no history of prior linezolid use. The results suggested possible nosocomial infections by linezolid-resistant MRSA.


Mutational characterization of the bile acid receptor TGR5 in primary sclerosing cholangitis.
TGR5, the G protein-coupled bile acid receptor 1 (GPBAR1), has been linked to inflammatory pathways as well as bile homeostasis, and could therefore be involved in primary sclerosing cholangitis (PSC) a chronic inflammatory bile duct disease. We aimed to extensively investigate TGR5 sequence variation in PSC, as well as functionally characterize detected variants. Complete resequencing of TGR5 was performed in 267 PSC patients and 274 healthy controls. Six nonsynonymous mutations were identified in addition to 16 other novel single-nucleotide polymorphisms. To investigate the impact from the nonsynonymous variants on TGR5, we created a receptor model, and introduced mutated TGR5 constructs into human epithelial cell lines. By using confocal microscopy, flow cytometry and a cAMP-sensitive luciferase assay, five of the nonsynonymous mutations (W83R, V178M, A217P, S272G and Q296X) were found to reduce or abolish TGR5 function. Fine-mapping of the previously reported PSC and UC associated locus at chromosome 2q35 in large patient panels revealed an overall association between the TGR5 single-nucleotide polymorphism rs11554825 and PSC (odds ratio = 1.14, 95% confidence interval: 1.03-1.26, p = 0.010) and UC (odds ratio = 1.19, 95% confidence interval 1.11-1.27, p = 8.5 x 10(-7)), but strong linkage disequilibrium precluded demarcation of TGR5 from neighboring genes. Resequencing of TGR5 along with functional investigations of novel variants provided unique insight into an important candidate gene for several inflammatory and metabolic conditions. While significant TGR5 associations were detected in both UC and PSC, further studies are needed to conclusively define the role of TGR5 variation in these diseases.


Expedient multi-step synthesis of organometallic complexes of Tc and Re in high effective specific activity. A new platform for the production of molecular imaging and therapy agents.
For over thirty years, instant labeling kits which involve no purification steps have been the only method used to prepare (99m)Tc radiopharmaceuticals for clinical studies. To address the limitations imposed by instant kits, which is hindering the development of molecularly targeted Tc- and Re-based imaging and therapy agents, a new strategy for the rapid multistep synthesis and purification of organometallic technetium-based molecular probes and corresponding rhenium-based therapeutic analogues was developed. Beginning with MO4(-) (M = (99m)Tc, (186/188)Re), the carbonyl precursor [M(CO)3(H2O)3](+) was synthesized in 3 min in quantitative yield in a microwave reactor. A dipicolyl ligand was added and the chelate complex was formed in high yield in 2 min using microwave heating at 150 degrees C. This was followed by a new purification strategy to remove unlabeled ligand which entailed using a copper resin/C18 solid phase extraction protocol giving the desired product in greater than 78% decay corrected yield (dcy). Conversion to the corresponding succinimidyl active ester was achieved following a 5 min microwave irradiation at 120 degrees C and C18 solid phase extraction purification in 60% dcy. A series of amides were prepared subsequently by microwave heating at 120 degrees C for 5 min producing the desired targets in greater than 86% dcy. The reported method represents a move away from traditional instant kits toward more versatile platform synthesis and purification technologies that are better suited for producing modern molecular imaging and therapy agents.


Increased portion sizes from energy-dense foods affect total energy intake at eating occasions in US children and adolescents: patterns and trends by age group and sociodemographic characteristics, 1977-2006.
Larger portion sizes of foods and beverages could affect overall energy intake at meals and promote overeating. We investigated trends in portion sizes of energy-dense foods and energy intakes at eating occasions in US children and adolescents. Four US nationally representative surveys from 1977 to 2006 were analyzed (n = 31,337). We measured trends in portion sizes (kcal, g, and mL) of selected foods [sugar-sweetened beverages (SSBs), salty snacks, desserts, French fries, burgers, pizzas, and Mexican fast foods] and energy intake (kcal) at eating occasions during which selected foods were consumed. Trends were reported by age group (2-6-, 7-12-, and 13-18-y-olds), sex, and socioeconomic status. In 2003-2006, the selected foods accounted for 38% of daily energy intake in 13-18-y-olds, 35% of the daily energy intake in 7-12-y-olds, and 28% of the daily energy intake in 2-6-y-olds. In all age groups, larger portion sizes of pizza coincided with higher energy intakes at eating occasions during which pizzas were consumed. In 7-12- and 13-18-y-olds, higher energy intakes at meals coincided with larger portion sizes of SSBs, French fries, or salty snacks. In all age groups, nonsignificant larger portions of Mexican fast foods were related to higher energy intakes at meals. Adolescent boys consumed larger portion sizes of the selected foods and had higher energy intakes at meals for all periods than did girls (P < 0.01). The percentage of kilocalories from pizza within a meal increased more sharply in non-Hispanic African Americans, in Hispanics, and in the group with a low household education than in the other groups. Adolescents are more susceptible to increased portion sizing than are younger children. The group of non-Hispanic African Americans and Hispanics and individuals with a lower education represents key concerns for public health policies.


Pulmonary Effects of Sustained Periods of High-G Acceleration Relevant to Suborbital Spaceflight.
AbstractBACKGROUND: Members of the public will soon be taking commercial suborbital spaceflights with significant Gx (chest-to-back) acceleration potentially reaching up to 6 Gx. Pulmonary physiology is gravity-dependent and is likely to be affected, which may have clinical implications for medically susceptible individuals.METHODS: During 2-min centrifuge exposures ranging up to 6 Gx, 11 healthy subjects were studied using advanced respiratory techniques. These sustained exposures were intended to allow characterization of the underlying pulmonary response and did not replicate actual suborbital G profiles. Regional distribution of ventilation in the lungs was determined using electrical impedance tomography. Neural respiratory drive (from diaphragm electromyography) and work of breathing (from transdiaphragmatic pressures) were obtained via nasoesophageal catheters. Arterial blood gases were measured in a subset of subjects. Measurements were conducted while breathing air and breathing 15 oxygen to simulate anticipated cabin pressurization conditions.RESULTS: Acceleration caused hypoxemia that worsened with increasing magnitude and duration of Gx. Minimum arterial oxygen saturation at 6 Gx was 86 1 breathing air and 79 1 breathing 15 oxygen. With increasing Gx the alveolar-arterial (A-a) oxygen gradient widened progressively and the relative distribution of ventilation reversed from posterior to anterior lung regions with substantial gas-trapping anteriorly. Severe breathlessness accompanied large progressive increases in work of breathing and neural respiratory drive.DISCUSSION: Sustained high-G acceleration at magnitudes relevant to suborbital flight profoundly affects respiratory physiology. These effects may become clinically important in the most medically susceptible passengers, in whom the potential role of centrifuge-based preflight evaluation requires further investigation.Pollock RD, Jolley CJ, Abid N, Couper JH, Estrada-Petrocelli L, Hodkinson PD, Leonhardt S, Mago-Elliott S, Menden T, Rafferty G, Richmond G, Robbins PA, Ritchie GAD, Segal MJ, Stevenson AT, Tank HD, Smith TG. Pulmonary effects of sustained periods of high-G acceleration relevant to suborbital spaceflight. Aerosp Med Hum Perform. 2021; 92(7):633641.


Relationships between serum free fatty acids and zinc, and attention deficit hyperactivity disorder: a research note.
The purpose of this study is to evaluate the relationships between serum free fatty acids (FFA) and zinc, and attention deficit hyperactivity disorder (ADHD). Forty eight children with ADHD (33 boys, 15 girls) were included in the patient group and 45 healthy volunteer children (30 boys, 15 girls) constituted the control group. The mean serum FFA level in the patient group was 0.176 +/- 0.102 mEq/L and in control group, 0.562 +/- 0.225 mEq/L (p < .001). The mean serum zinc level of patient group was 60.6 +/- 9.9 micrograms/dl and that of the control group, 105.8 +/- 13.2 micrograms/dl (p < .001). A statistically significant correlation was found between zinc and FFA levels in the ADHD group. These findings indicate that zinc deficiency may play a role in aetiopathogenesis of ADHD. Although we observed decreased FFA levels in ADHD cases, it is necessary to determine whether this condition is a principal cause of ADHD or is secondary to zinc deficiency.


Optical spatial intensity profiles for high order autocorrelation in fluorescence spectroscopy.
Interpretation of spatially resolved optical spectroscopies requires knowledge of the optical excitation and collection profiles of the experimental apparatus. This paper describes measurement of the relative norms of the spatial profile of a microscope- and laser-based optical system. The profile is given by the product of the spatial intensity of a focused laser beam and the point collection efficiency of the microscope. Experimental determination of the values of the norms is essential to the use of high order autocorrelation in fluorescence correlation spectroscopy to measure the concentrations and relative fluorescence yields of different fluorescent components (e.g., monomers and oligomers) in a multicomponent solution and also permits evaluation of theoretical models of the optical spatial intensity profile. In addition, the results may have applicability to high order autocorrelation in other optical spectroscopies, to confocal microscopy and to nonlinear optics in general.


Three-dimensional modeling of human placental terminal villi.
Placental transport is the main factor affecting the health and development of the fetus. Due to the placenta's geometrical and mathematical complexity, the structure-function relations of placental terminal villi have not been successfully modeled. Hence, a novel modeling approach is proposed. Computational models of four different specimens were generated from the three-dimensional reconstruction of confocal laser scanning microscopic image stacks. To evaluate the capabilities of the proposed methodology, stationary oxygen diffusion transport was calculated in the terminal villus volumes. The reconstructions automatically provided the spatial arrangement of the fetal capillaries inside the terminal villi. The surface and volume ratios between the fetal capillaries and the villus were also calculated, and the effects of model parameters on the placental diffusive capacity were assessed by parametric analysis. The potential of three-dimensional reconstructions combined with finite element analysis as a research tool for the human placenta was tested. The methodology herein could serve in the future as a simulation platform for complicated in vivo and in vitro scenarios.


Remotely-sensed active fire data for protected area management: eight-year patterns in the Manas National Park, India.
The Terai-Duar savanna and grasslands, which once extended along most of the Himalayan foothills, now only remain in a number of protected areas. Within these localities, grassland burning is a major issue, but data on frequency and distribution of fires are limited. Here, we analysed the incidence of active fires, which only occur during the dry season (Nov.-Mar.), within a significant area of Terai grasslands: the Manas National Park (MNP), India. We obtained locations of 781 fires during the 2000-2008 dry seasons, from the Fire Information for Resource Management System (FIRMS) that delivers global MODIS hotspot/fire locations using remote sensing and GIS technologies. Annual number of fires rose significantly from around 20 at the start of the study period to over 90 after 2002, with most (85%) detected between December and January. Over half of the fires occurred in tall grasslands, but fire density was highest in wetland and riverine vegetation, dry at the time. Most burning took place near rivers, roads and the park boundary, suggesting anthropogenic origins. A kernel density map of all recorded fires indicated three heavily burnt areas in the MNP, all within the tall grasslands. Our study demonstrates, despite some technical caveats linked to fire detection technology, which is improving, that remote fire data can be a practical tool in understanding fire concentration and burning temporal patterns in highly vulnerable habitats, useful in guiding management.


Perceiving temporal regularity in music: the role of auditory event-related potentials (ERPs) in probing beat perception.
The aim of this chapter is to give an overview of how the perception of a regular beat in music can be studied in humans adults, human newborns, and nonhuman primates using event-related brain potentials (ERPs). Next to a review of the recent literature on the perception of temporal regularity in music, we will discuss in how far ERPs, and especially the component called mismatch negativity (MMN), can be instrumental in probing beat perception. We conclude with a discussion on the pitfalls and prospects of using ERPs to probe the perception of a regular beat, in which we present possible constraints on stimulus design and discuss future perspectives.


Ethics in practice: the state of the debate on promoting the social value of global health research in resource poor settings particularly Africa.
Promoting the social value of global health research undertaken in resource poor settings has become a key concern in global research ethics. The consideration for benefit sharing, which concerns the elucidation of what if anything, is owed to participants, their communities and host nations that take part in such research, and the obligations of researchers involved, is one of the main strategies used for promoting social value of research. In the last decade however, there has been intense debate within academic bioethics literature seeking to define the benefits, the beneficiaries, and the scope of obligations for providing these benefits. Although this debate may be indicative of willingness at the international level to engage with the responsibilities of researchers involved in global health research, it remains unclear which forms of benefits or beneficiaries should be considered. International and local research ethics guidelines are reviewed here to delineate the guidance they provide. We reviewed documents selected from the international compilation of research ethics guidelines by the Office for Human Research Protections under the US Department of Health and Human Services. Access to interventions being researched, the provision of unavailable health care, capacity building for individuals and institutions, support to health care systems and access to medical and public health interventions proven effective, are the commonly recommended forms of benefits. The beneficiaries are volunteers, disease or illness affected communities and the population in general. Interestingly however, there is a divide between "global opinion" and the views of particular countries within resource poor settings as made explicit by differences in emphasis regarding the potential benefits and the beneficiaries. Although in theory benefit sharing is widely accepted as one of the means for promoting the social value of international collaborative health research, there is less agreement amongst major guidelines on the specific responsibilities of researchers over what is ethical in promoting the social value of research. Lack of consensus might have practical implications for efforts aimed at enhancing the social value of global health research undertaken in resource poor settings. Further developments in global research ethics require more reflection, paying attention to the practical realities of implementing the ethical principles in real world context.


Investigating the Spatiotemporal Variability and Driving Factors of Artificial Lighting in the Beijing-Tianjin-Hebei Region Using Remote Sensing Imagery and Socioeconomic Data.
With rapid urbanization and economic development, artificial lighting at night brings convenience to human life but also causes a considerable urban environmental pollution issue. This study employed the Mann-Kendall non-parametric test, nighttime light indices, and the standard deviation method to investigate the spatio-temporal characteristics of artificial lighting in the Beijing-Tianjin-Hebei region. Moreover, nighttime light imagery from the Defense Meteorological Satellite Program Operational Linescan System, socioeconomic data, and high-resolution satellite images were combined to comprehensively explore the driving factors of urban artificial lighting change. The results showed the following: (1) Overall, there was an increasing trend in artificial lighting in the Beijing-Tianjin-Hebei region, which accounted for approximately 56.87% of the total study area. (2) The change in artificial lighting in the entire area was relatively stable. The artificial lighting in the northwest area changed faster than that in the southeast area, and the areas where artificial lighting changed the most were Beijing, Tianjin and Tangshan. (3) The fastest growth of artificial lighting was in Chengde and Zhangjiakou, where the rates of increase were 334% and 251%, respectively. The spatial heterogeneity of artificial lighting in economically developed cities was higher than that in economically underdeveloped cities such as Chengde and Zhangjiakou. (4) Multi-source data were combined to analyse the driving factors of urban artificial lighting in the entire area. The Average Population of Districts under City (R2 = 0.77) had the strongest effect on artificial lighting. Total Passenger Traffic (R2 = 0.54) had the most non-obvious effect. At different city levels, driving factors varied with differences of economy, geographical location, and the industrial structures of cities. Urban expansion, transportation hubs, and industries were the major reasons for the significant change in nighttime light. Urban artificial lighting represents a trend of overuse closely related to nighttime light pollution. This study of artificial lighting contributes to the rational planning of urban lighting systems, the prevention and control of nighttime light pollution, and the creation of liveable and ecologically green cities.


Alzheimer's disease-like phosphorylation of the microtubule-associated protein tau by glycogen synthase kinase-3 in transfected mammalian cells.
Paired helical filaments (PHFs) are a characteristic pathological feature of Alzheimer's disease; their principal component is the microtubule-associated protein tau. The tau in PHFs (PHF-tau) is hyperphosphorylated, but the cellular mechanisms responsible for this hyperphosphorylation have yet to be elucidated. A number of kinases, including mitogen-activated protein (MAP) kinase, glycogen synthase kinase (GSK)-3 alpha, GSK-3 beta and cyclin-dependent kinase-5, phosphorylate recombinant tau in vitro so that it resembles PHF-tau as judged by its reactivity with a panel of antibodies capable of discriminating between normal tau and PHF-tau, and by a reduced electrophoretic mobility that is characteristic of PHF-tau. To determine whether MAP kinase, GSK-3 alpha and GSK-3 beta can also induce Alzheimer's disease-like phosphorylation of tau in mammalian cells, we studied the phosphorylation status of tau in primary neuronal cultures and transfected COS cells following changes in the activities of MAP kinase and GSK-3. Activating MAP kinase in cultures of primary neurons or transfected COS cells expressing tau isoforms did not increase the level of phosphorylation for any PHF-tau epitope investigated. But elevating GSK-3 activity in the COS cells by co-transfection with GSK-3 alpha or GSK-3 beta decreased the electrophoretic mobility of tau so that it resembled that of PHF-tau, and induced reactivity with eight PHF-tau-selective monoclonal antibodies. Our data indicate that GSK-3 alpha and/or GSK-3 beta, but not MAP kinase, are good candidates for generating PHF-type phosphorylation of tau in Alzheimer's disease. The involvement of other kinases in the generation of PHFs cannot, however, be eliminated. Our results suggest that aberrant regulation of GSK-3 may be a pathogenic mechanism in Alzheimer's disease.


A study to evaluate the acceptability, feasibility and impact of packaged interventions ("Diarrhea Pack") for prevention and treatment of childhood diarrhea in rural Pakistan.
Diarrhea remains one of the leading public health issues in developing countries and is a major contributor in morbidity and mortality in children under five years of age. Interventions such as ORS, Zinc, water purification and improved hygiene and sanitation can significantly reduce the diarrhea burden but their coverage remains low and has not been tested as packaged intervention before. This study attempts to evaluate the package of evidence based interventions in a "Diarrhea Pack" through first level health care providers at domiciliary level in community based settings. This study sought to evaluate the acceptability, feasibility and impact of diarrhea Pack on diarrhea burden. A cluster randomized design was used to evaluate the objectives of the project a union council was considered as a cluster for analysis, a total of eight clusters, four in intervention and four in control were included in the study. We conducted a baseline survey in all clusters followed by the delivery of diarrhea Pack in intervention clusters through community health workers at domiciliary level and through sales promoters to health care providers and pharmacies. Four quarterly surveillance rounds were conducted to evaluate the impact of diarrhea pack in all clusters by an independent team of Field workers. Both the intervention and control clusters were similar at the baseline but as the study progress we found a significant increase in uptake of ORS and Zinc along with the reduction in antibiotic use, diarrhea burden and hospitalization in intervention clusters when compared with the control clusters. We found that the Diarrhea Pack was well accepted with all of its components in the community. The intervention was well accepted and had a productive impact on the uptake of ORS and zinc and reduction in the use of antibiotics. It is feasible to deliver interventions such as diarrhea pack through community health workers in community settings. The intervention has the potential to be scaled up at national level.


P2Y receptors as regulators of lung endothelial barrier integrity.
Endothelial cells (ECs), forming a semi-permeable barrier between the interior space of blood vessels and underlying tissues, control such diverse processes as vascular tone, homeostasis, adhesion of platelets, and leukocytes to the vascular wall and permeability of vascular wall for cells and fluids. Mechanisms which govern the highly clinically relevant process of increased EC permeability are under intense investigation. It is well known that loss of this barrier (permeability increase) results in tissue inflammation, the hall mark of inflammatory diseases such as acute lung injury and its severe form, acute respiratory distress syndrome. Little is known about processes which determine the endothelial barrier enhancement or protection against permeability increase. It is now well accepted that extracellular purines and pyrimidines are promising and physiologically relevant barrier-protective agents and their effects are mediated by interaction with cell surface P2Y receptors which belong to the superfamily of G-protein-coupled receptors. The therapeutic potential of P2Y receptors is rapidly expanding field in pharmacology and some selective agonists became recently available. Here, we present an overview of recently identified P2Y receptor agonists that enhance the pulmonary endothelial barrier and inhibit and/or reverse endothelial barrier disruption.


Post-Synthetic Ligand Exchange in Zirconium-Based Metal-Organic Frameworks: Beware of The Defects!
Post-synthetic ligand exchange in the prototypical zirconium-based metal-organic framework (MOF) UiO-66 was investigated by in situ solution 1 H NMR spectroscopy. Samples of UiO-66 having different degrees of defectivity were exchanged using solutions of several terephthalic acid analogues in a range of conditions. Linker exchange only occurred in defect-free UiO-66, whereas monocarboxylates grafted at defect sites were found to be preferentially exchanged with respect to terephthalic acid over the whole range of conditions investigated. A 1:1 exchange ratio between the terephthalic acid analogue and modulator was observed, providing evidence that the defects had missing-cluster nature. Ex situ characterisation of the MOF powders after exchange corroborated these findings and showed that the physical-chemical properties of the MOF depend on whether the functionalisation occurs at defective sites or on the framework.


Bisphosphonates in the treatment of malignant bone disease.
Tumor-induced osteolysis or lytic bone disease is mediated by osteoclast activation. Osteoclasts can be activated directly by products produced by tumors or indirectly through other nonmalignant cells. By reducing osteoclastic activity, bisphosphonates inhibit bone resorption. Since these agents were shown effective in treating other diseases associated with increased bone resorption, including cancer-related hypercalcemia and Paget's disease of bone, studies have been initiated to explore the use of bisphosphonates in patients with osteolytic bone metastases. Recent large randomized double-blind studies show the efficacy of these agents in reducing skeletal complications in patients with bone metastases from both breast cancer and multiple myeloma.


Alterations in glucose metabolism in chick-embryo cells transformed by Rous sarcoma virus: intracellular levels of glycolytic intermediates.
Chick-embryo cells, transformed with Rous sarcoma virus, show enhanced rates of sugar transport and glycolysis. Determination of intracellular concentrations of glycolytic intermediates suggests that the enhanced glycolytic flux is due to increased activities of hexokinase (ATP:D-hexose 6-phosphotransferase, EC 18.104.22.168), phosphofructokinase, (ATP:D-fructose-1-phosphate 6-phosphotransferase, EC 18.104.22.168), and pyruvate kinase (ATP:pyruvate 2-O-phosphotransferase, EC 18.104.22.168), and not directly to the increased glucose transport. This conclusion is supported by the finding that the intracellular concentration of free glucose is decreased, rather than increased, in the transformed cells. The present observations suggest that the increased glycolytic flux is related to an increased rate of phosphorylation of glucose, and that hexokinase in the transformed cells is at least partly released from its normal control mechanism involving feedback inhibition by glucose-6-P.


The localized assembly of extracellular matrix integrin ligands requires cell-cell contact.
The assembly of an organism requires the interaction between different layers of cells, in many cases via an extracellular matrix. In the developing Drosophila larva, muscles attach in an integrin-dependent manner to the epidermis, via a specialized extracellular matrix called tendon matrix. Tiggrin, a tendon matrix integrin ligand, is primarily synthesized by cells distant to the muscle attachment sites, yet it accumulates specifically at these sites. Previous work has shown that the PS integrins are not required for tiggrin localization, suggesting that there is redundancy among tiggrin receptors. We have examined this by testing whether the PS2 integrin can recruit tiggrin to ectopic locations within the Drosophila embryo. We found that neither the wild type nor modified forms of the PS2 integrin, which have higher affinity for tiggrin, can recruit tiggrin to new cellular contexts. Next, we genetically manipulated the fate of the muscles and the epidermal muscle attachment cells, which demonstrated that muscles have the primary role in recruiting tiggrin to the tendon matrix and that cell-cell contact is necessary for this recruitment. Thus we propose that the inherent polarity of the muscle cells leads to a molecular specialization of their ends, and interactions between the ends produces an integrin-independent tiggrin receptor. Thus, interaction between cells generates an extracellular environment capable of nucleating extracellular matrix assembly.


Antimicrobial resistance in commensal opportunistic pathogens isolated from non-sterile sites can be an effective proxy for surveillance in bloodstream infections.
Antimicrobial resistance (AMR) surveillance in bloodstream infections (BSIs) is challenging in low/middle-income countries (LMICs) given limited laboratory capacity. Other specimens are easier to collect and process and are more likely to be culture-positive. In 8102 E. coli BSIs, 322,087 E. coli urinary tract infections, 6952 S. aureus BSIs and 112,074 S. aureus non-sterile site cultures from Oxfordshire (1998-2018), and other (55,296 isolates) rarer commensal opportunistic pathogens, antibiotic resistance trends over time in blood were strongly associated with those in other specimens (maximum cross-correlation per drug 0.51-0.99). Resistance prevalence was congruent across drug-years for each species (276/312 (88%) species-drug-years with prevalence within ± 10% between blood/other isolates). Results were similar across multiple countries in high/middle/low income-settings in the independent ATLAS dataset (103,559 isolates, 2004-2017) and three further LMIC hospitals/programmes (6154 isolates, 2008-2019). AMR in commensal opportunistic pathogens cultured from BSIs is strongly associated with AMR in commensal opportunistic pathogens cultured from non-sterile sites over calendar time, suggesting the latter could be used as an effective proxy for AMR surveillance in BSIs.


A case-control study of paternal occupational exposures and the risk of childhood sporadic bilateral retinoblastoma.
The risk factors for sporadic (ie, non-familial) retinoblastoma remain largely unknown. We examined the relationship between paternal occupational exposures from jobs held 10 years and 1 year prior to conception and the risk of sporadic bilateral retinoblastoma in children. Paternal occupational data were obtained for 198 incident cases diagnosed with sporadic bilateral retinoblastoma from January 1998 to May 2006 and 245 referral-based controls from the case child's relatives and friends who were matched to 135 of the cases on birth year. Industrial hygienists independently assigned exposure scores for nine agents. Adjusted ORs and 95% CIs were computed using logistic regression models, using the full sample of cases and controls as well as subset of cases with matched controls only. There was some indication of an elevated risk associated with paternal pesticide exposure in the 10 years prior to conception (OR=1.64; 95% CI 1.08 to 2.50) as well as in the year before conception (OR=2.12; 95% CI 1.25 to 3.61). However, results for pesticide exposure were inconsistent and varied by analysis approach. An increased risk was also observed for non-welding metal exposure during the 10 years prior to conception in the full (OR=1.35; 95% CI 0.86 to 2.12) and matched (OR=1.40; 95% CI 0.82 to 2.37) samples, but not in the year before conception. Exposure-response trends were observed for pesticides and non-welding metal exposures. Our findings suggest a potential role of paternal occupational exposures to non-welding metals and perhaps pesticides in the aetiology of childhood retinoblastoma.


Deeper penetration into tumor tissues and enhanced in vivo antitumor activity of liposomal paclitaxel by pretreatment with angiogenesis inhibitor SU5416.
The recently emerged concept of "vessel normalization" implies that judicious blockade of vascular endothelial growth factor (VEGF) signaling may transiently "normalize" the tumor vasculature, making it more suitable for tumor disposition of subsequently administered drugs. In this study, therefore, the effect of pretreatment with SU5416, a selective VEGF receptor-2 inhibitor, on tumor disposition and in vivo antitumor activity of polyethylene glycol (PEG)-modified liposomal paclitaxel (PL-PTX) was evaluated in Colon-26 solid tumor-bearing mice. To improve the solubility and in vivo disposition characteristics of SU5416, the inhibitor was formulated in PEGylated O/W emulsion (PE-SU5416). Pretreatment with PE-SU5416 significantly enhanced the in vivo antitumor effect of PL-PTX, although PE-SU5416 administration alone did not show any antitumor effect. Immunostaining for endothelial cells and pericytes demonstrated that the pretreatment with PE-SU5416 enhanced the pericyte coverage of the tumor vasculature. In addition, tumors treated with PE-SU5416 contained significantly smaller hypoxic regions compared with the nontreated control group, demonstrating that structural normalization of the tumor vasculature resulted in an improvement in tumor vessel functions, including oxygen supply. Furthermore, the pretreatment with PE-SU5416 increased the distribution of PEG liposomes and included PTX in the core region of the tumor, as well as conversely decreasing the ratio of their peripheral distribution. These results suggest that the structural and functional normalization of the tumor vasculature by the pretreatment with PE-SU5416 enabled liposomes to reach the deeper regions within tumor tissues, leading to more potent antitumor activity of PL-PTX.


Cardioprotective mechanism of omega-3 polyunsaturated fatty acids.
Omega-3 polyunsaturated fatty acids (PUFAs), such as eicosapentaenoic acid and docosahexaenoic acid, are widely regarded as cardioprotective. Several large-scale, randomized clinical trials have shown that dietary intake of omega-3 PUFAs improves the prognosis of patients with symptomatic heart failure or recent myocardial infarction. Therefore, dietary consumption of omega-3 PUFA is recommended in international guidelines for the general population to prevent the occurrence of cardiovascular diseases (CVDs). However, the precise mechanisms underlying the cardioprotective effects of omega-3 PUFAs are not fully understood. Omega-3 PUFAs can be incorporated into the phospholipid bilayer of cell membranes and can affect membrane fluidity, lipid microdomain formation, and signaling across membranes. Omega-3 PUFAs also modulate the function of membrane ion channels, such as Na and L-type Ca channels, to prevent lethal arrhythmias. Moreover, omega-3 PUFAs also prevent the conversion of arachidonic acid into pro-inflammatory eicosanoids by serving as an alternative substrate for cyclooxygenase or lipoxygenase, resulting in the production of less potent products. In addition, a number of enzymatically oxygenated metabolites derived from omega-3 PUFAs were recently identified as anti-inflammatory mediators. These omega-3 metabolites may contribute to the beneficial effects against CVDs that are attributed to omega-3 PUFAs.


Transulnar artery approach for percutaneous coronary intervention: an alternative route in a patient with challenging transfemoral access and hypoplastic radial artery.
Transradial coronary angiography and intervention has become a popular technique due to reduced local and bleeding complications, easier post-procedural care and patient preference. In certain patients, transradial access may not be possible due to various anatomical reasons. Usually, the transfemoral route is used for these patients. In a minority of patients, such as those with significant peripheral arterial disease or obesity, the transfemoral approach is challenging and is associated with an increased risk of local complications or bleeding. Transulnar arterial access, however, has recently been shown to be feasible and safe for both coronary angiography and intervention. The procedural success, advantages and complication rates for this procedure appear similar to those for the transradial approach. We describe a case of transulnar artery percutaneous coronary intervention in an obese patient with a hypoplastic radial artery.


Antigen-loaded pH-sensitive hydrogel microparticles are taken up by dendritic cells with no requirement for targeting antibodies.
Particle-based delivery of encapsulated antigens has great potential for improving vaccine constructs. In this study, we show that antigen-loaded, pH-sensitive hydrogel microparticles are taken up and presented by bone marrow-derived dendritic cells (BMDCs) in vitro and are taken up by dendritic cells (DCs) and monocytes in vivo. This uptake is irrespective of targeting antibodies. BMDCs in vitro and DCs in vivo also display upregulation of activation markers CD80 and CD86 when treated with microparticles, again with no difference in conjugated antibodies, even the agonistic CD40 antibody. We further show that these particles induce enhanced expansion of cytokine-producing CD8 T cells in response to challenge with ovalbumin-expressing vesicular stomatitis virus, in both an accelerated vaccination strategy using pre-loaded BMDCs and a traditional mouse immunization setting.