Serum galectin-1 in patients with multiple myeloma: associations with survival, angiogenesis, and biomarkers of macrophage activation.
Galectin-1 (Gal-1) is known to regulate cell signaling within the immune system and may be a target for new anticancer immune therapy. In patients with chronic lymphocytic leukemia (CLL) and classical Hodgkin lymphoma (cHL), high levels of Gal-1 within the tumor microenvironment were associated with worse disease state or poor outcome. Gal-1 can be secreted from cells by an unknown mechanism, and levels in blood samples were associated with high tumor burden and worse disease state in cHL and CLL patients. However, serum levels of Gal-1 have never been investigated in patients with multiple myeloma (MM). We measured serum Gal-1 levels in samples from patients with treatment demanding MM at the time of diagnosis (n=102) and after treatment (n=24) and examined associations of serum Gal-1 with clinicopathological information obtained from patient medical records, as well as data on bone marrow angiogenesis and the macrophage activation biomarkers soluble CD163 (sCD163) and soluble mannose receptor. Serum Gal-1 levels were not elevated in patients with MM at diagnosis compared with healthy donors (median values 8.48 vs 11.93 ng/mL, P=0.05), which is in contrast to results in cHL and CLL. Furthermore, Gal-1 levels did not show association with bone marrow angiogenesis, clinicopathological parameters, overall survival, or response to treatment. There was a statically significant association between Gal-1 and sCD163 levels (R=0.24, P=0.02), but not with soluble mannose receptor (P=0.92). In conclusion, our results indicate that Gal-1 is not an important serum biomarker in MM, which is in contrast to data from patients with cHL and CLL. However, the association with sCD163 is in line with previous data showing that Gal-1 may be involved in alternative (M2-like) activation of macrophages.


Cytological and deoxyribonucleic acid-deoxyribonucleic acid hybridization studies on lactobacillus isolates from San Francisco sourdough.
Molecular taxonomic and electron microscopy studies were performed on four bacterial isolates obtained from different sources of San Francisco sourdough (SD). These bacteria were first isolated by Kline and Sugihara who tentatively described them as a previously unreported species of heterofermentative Lactobacillus; they suggested the name Lactobacillus sanfrancisco. The guanine plus cytosine base composition (%GC) of the deoxyribonucleic acid (DNA) ranged from 38 to 40%. The possible genetic relatedness of these SD isolates to five known species of Lactobacillus with comparable GC contents was assessed in the present work by means of DNA-DNA hybridization competition experiments. Little or no DNA homology was observed between the SD bacteria and the known species. The SD bacteria exhibited a high degree of homology (>88%) among themselves, suggesting that the four isolates were identical taxonomically. Also, the electron photomicrographs revealed structures similar to those of gram-positive bacilli. Accordingly, since these SD isolates have the characteristic phenotypic and morphological properties of the genus Lactobacillus and are not related genetically to any known species, the tentative characterization by the above workers of these isolates as a new species is substantiated.


Manufacturing feature recognition toward integration with process planning.
Process planning plays a key role by linking CAD and CAM. Its front-end is feature recognition, but feature recognition research has not been in accord with the requirements of process planning. This paper presents an effort for integrating the two activities: feature-based machining sequence generation primarily based on tool capabilities. The system recognizes only manufacturable features by consulting the tool database, and simultaneously constructs dependencies among the features. Then, the A* algorithm is used to search for an optimal machining sequence by the aid of the feature dependencies and a manufacturing cost function.


Heterogeneous nuclear ribonucleoprotein A2 participates in the replication of Japanese encephalitis virus through an interaction with viral proteins and RNA.
Japanese encephalitis virus (JEV) is a mosquito-borne flavivirus that is kept in a zoonotic transmission cycle between pigs and mosquitoes. JEV causes infection of the central nervous system with a high mortality rate in dead-end hosts, including humans. Many studies have suggested that the flavivirus core protein is not only a component of nucleocapsids but also an important pathogenic determinant. In this study, we identified heterogeneous nuclear ribonucleoprotein A2 (hnRNP A2) as a binding partner of the JEV core protein by pulldown purification and mass spectrometry. Reciprocal coimmunoprecipitation analyses in transfected and infected cells confirmed a specific interaction between the JEV core protein and hnRNP A2. Expression of the JEV core protein induced cytoplasmic retention of hnRNP A2 in JEV subgenomic replicon cells. Small interfering RNA (siRNA)-mediated knockdown of hnRNP A2 resulted in a 90% reduction of viral RNA replication in cells infected with JEV, and the reduction was cancelled by the expression of an siRNA-resistant hnRNP A2 mutant. In addition to the core protein, hnRNP A2 also associated with JEV nonstructural protein 5, which has both methyltransferase and RNA-dependent RNA polymerase activities, and with the 5'-untranslated region of the negative-sense JEV RNA. During one-step growth, synthesis of both positive- and negative-strand JEV RNAs was delayed by the knockdown of hnRNP A2. These results suggest that hnRNP A2 plays an important role in the replication of JEV RNA through the interaction with viral proteins and RNA.


Oral region homologies in paleozoic crinoids and other plesiomorphic pentaradial echinoderms.
The phylogenetic relationships between major groups of plesiomorphic pentaradial echinoderms, the Paleozoic crinoids, blastozoans, and edrioasteroids, are poorly understood because of a lack of widely recognized homologies. Here, we present newly recognized oral region homologies, based on the Universal Elemental Homology model for skeletal plates, in a wide range of fossil taxa. The oral region of echinoderms is mainly composed of the axial, or ambulacral, skeleton, which apparently evolved more slowly than the extraxial skeleton that forms the majority of the body. Recent phylogenetic hypotheses have focused on characters of the extraxial skeleton, which may have evolved too rapidly to preserve obvious homologies across all these groups. The axial skeleton conserved homologous suites of characters shared between various edrioasteroids and specific blastozoans, and between other blastozoans and crinoids. Although individual plates can be inferred as homologous, no directly overlapping suites of characters are shared between edrioasteroids and crinoids. Six different systems of mouth (peristome) plate organization (Peristomial Border Systems) are defined. These include four different systems based on the arrangement of the interradially-positioned oral plates and their peristomial cover plates, where PBS A1 occurs only in plesiomorphic edrioasteroids, PBS A2 occurs in plesiomorphic edrioasteroids and blastozoans, and PBS A3 and PBS A4 occur in blastozoans and crinoids. The other two systems have radially-positioned uniserial oral frame plates in construction of the mouth frame. PBS B1 has both orals and uniserial oral frame plates and occurs in edrioasterid and possibly edrioblastoid edrioasteroids, whereas PBS B2 has exclusively uniserial oral frame plates and is found in isorophid edrioasteroids and imbricate and gogiid blastozoans. These different types of mouth frame construction offer potential synapomorphies to aid in parsimony-based phylogenetics for exploring branching order among stem groups on the echinoderm tree of life.


Movement amplitude on the Functional Re-adaptive Exercise Device: deep spinal muscle activity and movement control.
Lumbar multifidus (LM) and transversus abdominis (TrA) show altered motor control, and LM is atrophied, in people with low-back pain (LBP). The Functional Re-adaptive Exercise Device (FRED) involves cyclical lower-limb movement against minimal resistance in an upright posture. It has been shown to recruit LM and TrA automatically, and may have potential as an intervention for non-specific LBP. However, no studies have yet investigated the effects of changes in FRED movement amplitude on the activity of these muscles. This study aimed to assess the effects of different FRED movement amplitudes on LM and TrA muscle thickness and movement variability, to inform an evidence-based exercise prescription. Lumbar multifidus and TrA thickness of eight healthy male volunteers were examined using ultrasound imaging during FRED exercise, normalised to rest at four different movement amplitudes. Movement variability was also measured. Magnitude-based inferences were used to compare each amplitude. Exercise at all amplitudes recruited LM and TrA more than rest, with thickness increases of approximately 5 and 1 mm, respectively. Larger amplitudes also caused increased TrA thickness, LM and TrA muscle thickness variability and movement variability. The data suggests that all amplitudes are useful for recruiting LM and TrA. A progressive training protocol should start in the smallest amplitude, increasing the setting once participants can maintain a consistent movement speed, to continue to challenge the motor control system.


Evaluation of the effects of and earliest response rate to anti-D treatment in children with chronic idiopathic thrombocytopenic purpura: a pilot study.
In this pilot study, 30 (14 male, 16 female; median age: 8 years, range: 2-18) chronic non-splenectomized idiopathic thrombocytopenic purpura (ITP) patients with Rh+ blood group and their 49 attacks were evaluated after intravenous (i.v.) anti-D (WinRho SDF, Cangene Corporation, Winnipeg, MB, Canada) treatment at a dose of 50 microg/kg x 3 days (n = 21 cases; 35 attacks) or a single dose of 75 microg/kg (n = 9 cases; 14 attacks) to define the hemostatic dose of anti-D. Five of 30 patients (22/49 attacks) were resistant to steroid, intravenous immunoglobulin (IVIG) and vincristine treatment. Hemoglobin (Hb), white blood cells (WBC), platelets (plt) and reticulocytes (ret) were evaluated before and after treatment during the follow-up in sequences on the 1st, 7th, 14th and 21st days after anti-D treatment if the patients had no symptom. All patients, even the resistant ones, experienced an increase in plt count to provide protection from bleeding (> or = 20 x 10(9)/L in patients with symptoms, > or = 10 x 10(9)/L in patients without symptoms). The plt responses of one resistant and five non-resistant patients treated with a single 75 microg/kg dose of i.v. anti-D in 8 attacks were monitored at the 2nd, 4th, 8th, 24th and 48th hours of the treatment. A protective plt level was attained within 2 hours in 6 attacks of five non-resistant cases and in 24 hours in the remaining 2 attacks of one resistant case. This pilot study suggests that anti-D treatment in ITP patients is effective and can increase plt to a level adequate enough to protect from hemorrhage within 2 hours, when given in a 75 microg/kg dose. A few adverse events (i.e. chills, hemolysis and hemoglobinuria) resolved without intervention.


Variability in small bowel histopathology reporting between different pathology practice settings: impact on the diagnosis of coeliac disease.
Coeliac disease (CD) diagnosis requires the detection of characteristic histological alterations of small bowel mucosa, which are prone to interobserver variability. This study evaluated the agreement in biopsy interpretation between different pathology practice types. Biopsies from community hospitals (n=46), university hospitals (n=18) and commercial laboratories (n=38) were blindly assessed by a pathologist at our institution for differences in histopathology reporting and agreement in diagnosis of CD and degree of villous atrophy (VA) by κ analysis. Agreement for primary diagnosis was very good between this institution and university hospitals (κ=0.888), but moderate compared with community hospitals (κ=0.465) or commercial laboratories (κ=0.419). Diagnosis differed in 26 (25%) cases, leading to a 20% increase in CD diagnosis after review. Among those diagnosed with CD by both institutions (n=49), agreement in degree of VA was fair (κ=0.292), with moderate agreement between the authors and commercial laboratories (κ=0.500) and fair with university hospitals (κ=0.290) or community hospitals (κ=0.211). The degree of VA was upgraded in 27% and downgraded in 2%. Within different Marsh score categories, agreement was poor (κ<0.0316) for scores 1 and 2, both missed at other centres, and fair or moderate for scores 3a and 3b. Information regarding degree of VA and intraepithelial lymphocytosis was lacking in 26% and 86% of reports and non-quantifiable descriptors, eg, 'blunting' or 'marked atrophy' were prevalent. CD-related histological changes are underdiagnosed in community-based hospitals and commercial pathology laboratories. Because incorrect biopsy interpretation can cause underdiagnosis of CD, greater CD awareness and uniformity in small bowel biopsy reporting is required among pathologists.


prewas: data pre-processing for more informative bacterial GWAS.
While variant identification pipelines are becoming increasingly standardized, less attention has been paid to the pre-processing of variants prior to their use in bacterial genome-wide association studies (bGWAS). Three nuances of variant pre-processing that impact downstream identification of genetic associations include the separation of variants at multiallelic sites, separation of variants in overlapping genes, and referencing of variants relative to ancestral alleles. Here we demonstrate the importance of these variant pre-processing steps on diverse bacterial genomic datasets and present prewas, an R package, that standardizes the pre-processing of multiallelic sites, overlapping genes, and reference alleles before bGWAS. This package facilitates improved reproducibility and interpretability of bGWAS results. prewas enables users to extract maximal information from bGWAS by implementing multi-line representation for multiallelic sites and variants in overlapping genes. prewas outputs a binary SNP matrix that can be used for SNP-based bGWAS and will prevent the masking of minor alleles during bGWAS analysis. The optional binary gene matrix output can be used for gene-based bGWAS, which will enable users to maximize the power and evolutionary interpretability of their bGWAS studies. prewas is available for download from GitHub.


Plasma membrane aquaporin activity can affect the rate of apoptosis but is inhibited after apoptotic volume decrease.
Apoptosis is characterized by a conserved series of morphological events beginning with the apoptotic volume decrease (AVD). This study investigated a role for aquaporins (AQPs) during the AVD. Inhibition of AQPs blocked the AVD in ovarian granulosa cells undergoing growth factor withdrawal and blocked downstream apoptotic events such as cell shrinkage, changes in the mitochondrial membrane potential, DNA degradation, and caspase-3 activation. The effects of AQP inhibition on the AVD and DNA degradation were consistent in thymocytes and with two additional apoptotic signals, thapsigargin and C(6)-ceramide. Overexpression of AQP-1 in Chinese hamster ovary (CHO-AQP-1) cells enhanced their rate of apoptosis. The AVD is driven by loss of K(+) from the cell, and we hypothesize that after the AVD, AQPs become inactive, which halts further water loss and allows K(+) concentrations to decrease to levels necessary for apoptotic enzyme activation. Swelling assays on granulosa cells, thymocytes, and CHO-AQP-1 cells revealed that indeed, the shrunken (apoptotic) subpopulation has very low water permeability compared with the normal-sized (nonapoptotic) subpopulation. In thymocytes, AQP-1 is present and was shown to colocalize with the plasma membrane receptor tumor necrosis factor receptor-1 (TNF-R1) both before and after the AVD, which suggests that this protein is not proteolytically cleaved and remains on the cell membrane. Overall, these data indicate that AQP-mediated water loss is important for the AVD and downstream apoptotic events, that the water permeability of the plasma membrane can control the rate of apoptosis, and that inactivation after the AVD may help create the low K(+) concentration that is essential in apoptotic cells. Furthermore, inactivation of AQPs after the AVD does not appear to be through degradation or removal from the cell membrane.


[Inherent efficiency of stress-limiting systems as a factor of the resistance to stress-induced disorders].
A hypothesis is substantiated in accordance to which a resistance of an organism to stress damages depends upon genetically determined peculiarities of its regulatory stress-limiting systems that restrict stress reaction and its detrimental effects. A comparison of differences between the stress resistance and the activity of the stress-limiting systems (dopaminergic, serotoninergic, nitric oxide and heat shock proteins systems) in rats of two strains August and Wistar indicates that the higher hereditary activity of mentioned systems is associated with the higher resistance to acute emotional stress; and the lower hereditary activity of these systems associated with the lower resistance to this stress. At the same time the adaptation to repeated non-damaging exposures to stressor aimed to rise the stress resistance leads to opposite results in rats of the mentioned strains. In the animals with the higher hereditary resistance to acute stress (August rats) the adaptation reduces this resistance. In the animals with the lower hereditary stress resistance (Wistar rats) that sort of adaptation really rises this resistance. That is determined by changes in activity of the stress-limiting systems during repeated stress exposures. In the animals with the higher hereditary activity of the stress-limiting systems the adaptation reduces the activity of these systems. In the animals with the lower hereditary activity of the stress-limiting systems the adaptation rises this activity and the resistance of these animals to stress damages.


Food protein-induced enterocolitis syndrome caused by solid food proteins.
Infantile food protein-induced enterocolitis syndrome (FPIES) is a severe, cell-mediated gastrointestinal food hypersensitivity typically provoked by cow's milk or soy. Solid foods are rarely considered a cause. To describe the clinical characteristics and natural history of FPIES provoked by solid foods. Patients with FPIES induced by solid foods were identified and their clinical course compared with a control group with FPIES caused by cow's milk and/or soy evaluated over the same time period. Fourteen infants with FPIES caused by grains (rice, oat, and barley), vegetables (sweet potato, squash, string beans, peas), or poultry (chicken and turkey) were identified. Symptoms were typical of classical FPIES with delayed (median: 2 hours) onset of vomiting, diarrhea, and lethargy/dehydration. Eleven infants (78%) reacted to >1 food protein, including 7 (50%) that reacted to >1 grain. Nine (64%) of all patients with solid food-FPIES also had cow's milk and/or soy-FPIES. Initial presentation was severe in 79% of the patients, prompting sepsis evaluations (57%) and hospitalization (64%) for dehydration or shock. The diagnosis of FPIES was delayed, after a median of 2 reactions (range: 2-5). Thirty patients with typical cow's milk- and/or soy-FPIES were identified for comparison. Overall, 48% of the 44 infants with FPIES were reactive to >1 food protein, and the risk for multiple food hypersensitivity approached 80% in the infants with solid food or soy-induced FPIES. None of the patients developed FPIES to maternally ingested foods while breastfeeding unless the causal food was fed directly to the infant. Cereals, vegetables, and poultry meats, typically regarded as of low allergenic potential, must be considered in the evaluation of FPIES, particularly in infants previously diagnosed with FPIES to cow's milk or soy, and as an initial cause in patients who have been exclusively breastfed. Infants with FPIES are at risk for multiple dietary protein hypersensitivities during an apparent period of immunologic susceptibility. Pediatricians should consider FPIES in the differential diagnosis of shock and sepsis.


Quercetin and kaempferol suppress immunoglobulin E-mediated allergic inflammation in RBL-2H3 and Caco-2 cells.
We investigated the inhibitory effects of quercetin and kaempferol treatment on the suppression of immunoglobulin E (IgE)-mediated allergic responses in relation to intestinal epithelium barrier function in RBL-2H3 and Caco-2 cells. RBL-2H3 cells as a model of intestinal mucosa mast cells were treated with flavonols followed by IgE-anti-dinitrophenyl sensitization. The extent of degranulation and the release of pro-inflammatory cytokines were measured. Caco-2 cells were stimulated with interleukin (IL)-4 or IgE-allergen with or without flavonol pretreatment and changes in the expression of CD23 mRNA and mitogen-activated protein kinase (MAPK), and chemokine release were determined. Flavonols inhibited the secretion of allergic mediators in RBL-2H3 cells and suppressed the CD23 mRNA expression and p38 MAPK activation in IL-4 stimulated Caco-2 cells. Flavonols also suppressed IgE-OVA induced extra signal-regulated protein kinase (ERK) activation and chemokine release. Quercetin and kaempferol effectively suppressed the development of IgE-mediated allergic inflammation of intestinal cell models.


Fifty Men, 3510 Marathons, Cardiac Risk Factors, and Coronary Artery Calcium Scores.
Many male marathon runners have elevated coronary artery calcium (CAC) scores despite high physical activity. We examined the association between CAC scores, cardiovascular risk factors, and lifestyle habits in long-term marathoners. We recruited men who had run one or more marathons annually for 25 consecutive years. CAC was assessed using coronary computed tomography angiography. Atherosclerotic cardiovascular disease risk factors were measured with a 12-lead ECG, serum lipid panel, height, weight, resting blood pressure and heart rate, and a risk factor questionnaire. Fifty males, mean age 59 ± 0.9 yr with a combined total of 3510 marathons (median = 58.5, range = 27-171), had a mean BMI of 22.44 ± 0.4 kg·m, HDL and LDL cholesterols of 58 ± 1.6 and 112 ± 3.7 mg·dL, and CAC scores from 0 to 3153. CAC scores varied from 0 in 16 runners to 1-100 in 12, 101-400 in 12, and >400 in 10. There was no statistical difference in the number of marathons run between the four groups. Compared with marathoners with no CAC, marathoners with moderate and extensive CAC were older (P = 0.002), started running at an older age (P = 0.003), were older when they ran their first marathon (P = 0.006), and had more CAD risk factors (P = 0.005), and marathoners with more CAC had higher rates of previous tobacco use (P = 0.002) and prevalence of hyperlipidemia (P = 0.01). Among experienced males who have run marathons for 26-34 yr and completed between 27 and 171 marathons, CAC score is related to CAD risk factors and not the number of marathons run or years of running. This suggests that among long-term marathoners, more endurance exercise is not associated with an increased risk of CAC.


A nociceptive-intensity-dependent role for hydrogen sulphide in the formalin model of persistent inflammatory pain.
The present study investigated the hypothesis that hydrogen sulfide (H2S) is pro-nociceptive in the formalin model of persistent inflammatory pain in the adult rat. Hind paw injection of formalin evoked a concentration-dependent increase in the hind paw concentration of H2S. Increased concentration of H2S was found in homogenates prepared from hind paws injected with 5% (but not 1.25%) formalin. Correspondingly, animal nociceptive flinching and hind paw edema were maximal with 5% formalin. Both nociceptive flinching and hind paw edema induced by injection of 5% formalin were attenuated by pretreatment with DL-propargylglycine (PPG; 50 mg/kg, i.p.) which is an inhibitor of the H2S synthesizing enzyme cystathionine-gamma-lyase (CSE). The effect of pretreatment with PPG was selective and the drug did not influence animal behavior or hind-paw edema with injection of 1.25% formalin. Furthermore, PPG pretreatment attenuated the induction of c-Fos in spinal laminae I-II following injection of 5% formalin. In contrast, co-injection of 1.25% formalin with sodium hydrogen sulfide (NaHS; 1 nmol/0.1 ml), a H2S donor, into the hind paw increased animal nociceptive behavior. Collectively, these findings show that the effect of peripheral H2S in the pathogenesis of inflammatory pain depends, at least in part, on the nociceptive intensity level.


Reversion of PB2-627E to -627K during replication of an H5N1 Clade 2.2 virus in mammalian hosts depends on the origin of the nucleoprotein.
H5N1 highly pathogenic avian influenza viruses (HPAIV) of clade 2.2 spread from Southeast Asia to Europe. Intriguingly, in contrast to all common avian strains specifying glutamic acid at position 627 of the PB2 protein (PB2-627E), they carry a lysine at this position (PB2-627K), which is normally found only in human strains. To analyze the impact of this mutation on the host range of HPAIV H5N1, we altered PB2-627K to PB2-627E in the European isolate A/Swan/Germany/R65/2006 (R65). In contrast to the parental R65, multicycle growth and polymerase activity of the resulting mutant R65-PB2(K627E) were considerably impaired in mammalian but not in avian cells. Correspondingly, the 50% lethal dose (LD₅₀) in mice was increased by three orders of magnitude, whereas virulence in chicken remained unchanged, resulting in 100% lethality, as was found for the parental R65. Strikingly, R65-PB2(K627E) reverted to PB2-627K after only one passage in mice but did not revert in chickens. To investigate whether additional R65 genes influence reversion, we passaged R65-PB2(K627E) reassortants containing genes from A/Hong Kong/156/97 (H5N1) (carrying PB2-627E), in avian and mammalian cells. Reversion to PB2-627K in mammalian cells required the presence of the R65 nucleoprotein (NP). This finding corresponds to results of others that during replication of avian strains in mammalian cells, PB2-627K restores an impaired PB2-NP association. Since this mutation is apparently not detrimental for virus prevalence in birds, it has not been eliminated. However, the prompt reversion to PB2-627K in MDCK cells and mice suggests that the clade 2.2 H5N1 HPAIV may have had a history of intermediate mammalian hosts.


The role of epidemiology in disaster response policy development.
Disasters expose the general population and responders to a range of potential contaminants and stressors which may harm physical and mental health. This article addresses the role of epidemiology in informing policies after a disaster to mitigate ongoing exposures, provide care and compensation, and improve preparedness for future disasters. The World Trade Center disaster response is used as a case study. We examine how epidemiologic evidence was used to shape postdisaster policy and identify important gaps in early research. In the wake of World Trade Center attacks, epidemiologic research played a key role in identifying and characterizing affected populations, assessing environmental exposures, quantifying physical and mental health impacts, and producing evidence to ascribe causation. However, most studies suffered from methodological challenges, including delays, selection biases, poor exposure measurement, and nonstandardized outcomes. Gaps included measuring unmet health needs and financing coverage, as well as coordination across longitudinal cohorts of studies for rare conditions with long latency, such as cancer. Epidemiologists can increase their impact on evidence-based policymaking by ensuring core mechanisms are in place before a disaster to mount monitoring of responders and other affected populations, improve early exposure assessment efforts, identify critical gaps in scientific knowledge, and coordinate communication of scientific findings to policymakers and the public.


The murine Sim-2 gene product inhibits transcription by active repression and functional interference.
The Drosophila single-minded (Dsim) gene encodes a master regulatory protein involved in cell fate determination during midline development. This protein is a member of a rapidly expanding family of gene products possessing basic helix-loop-helix (bHLH) and hydrophobic PAS (designated a conserved region among PER, ARNT [aryl hydrocarbon receptor nuclear translocator] and SIM) protein association domains. Members of this family function as central transcriptional regulators in cellular differentiation and in the response to environmental stimuli such as xenobiotics and hypoxia. We have previously identified a murine member of this family, called mSim-2, showing sequence homology to the bHLH and PAS domains of Dsim. Immunoprecipitation experiments with recombinant proteins indicate that mSIM-2 associates with the arnt gene product. In the present work, by using fine-structure mapping we found that the HLH and PAS motifs of both proteins are required for optimal association. Forced expression of GAL4/mSIM-2 fusion constructs in mammalian cells demonstrated the presence of two separable repression domains within the carboxy terminus of mSIM-2. We found that mSIM-2 is capable of repressing ARNT-mediated transcriptional activation in a mammalian two-hybrid system. This effect (i) is dependent on the ability of mSIM-2 and ARNT to heterodimerize, (ii) is dependent on the presence of the mSIM-2 carboxy-terminal repression domain, and (iii) is not specific to the ARNT activation domain. These results suggest that mSIM-2 repression activity can dominantly override the activation potential of adjacent transcription factors. We also demonstrated that mSIM-2 can functionally interfere with hypoxia-inducible factor 1alpha (HIF-1alpha)/ARNT transcription complexes, providing a second mechanism by which mSIM-2 may inhibit transcription.


Infectious disease in the Pleistocene: Old friends or old foes?
The impact of endemic and epidemic disease on humans has traditionally been seen as a comparatively recent historical phenomenon associated with the Neolithisation of human groups, an increase in population size led by sedentarism, and increasing contact with domesticated animals as well as species occupying opportunistic symbiotic and ectosymbiotic relationships with humans. The orthodox approach is that Neolithisation created the conditions for increasing population size able to support a reservoir of infectious disease sufficient to act as selective pressure. This orthodoxy is the result of an overly simplistic reliance on skeletal data assuming that no skeletal lesions equated to a healthy individual, underpinned by the assumption that hunter-gatherer groups were inherently healthy while agricultural groups acted as infectious disease reservoirs. The work of van Blerkom, Am. J. Phys. Anthropol., vol. suppl 37 (2003), Wolfe et al., Nature, vol. 447 (2007) and Houldcroft and Underdown, Am. J. Phys. Anthropol., vol. 160, (2016) has changed this landscape by arguing that humans and pathogens have long been fellow travelers. The package of infectious diseases experienced by our ancient ancestors may not be as dissimilar to modern infectious diseases as was once believed. The importance of DNA, from ancient and modern sources, to the study of the antiquity of infectious disease, and its role as a selective pressure cannot be overstated. Here we consider evidence of ancient epidemic and endemic infectious diseases with inferences from modern and ancient human and hominin DNA, and from circulating and extinct pathogen genomes. We argue that the pandemics of the past are a vital tool to unlock the weapons needed to fight pandemics of the future.


Comparing the microbial risks associated with household drinking water supplies used in peri-urban communities of Phnom Penh, Cambodia.
Most Cambodians lack access to a safe source of drinking water. Piped distribution systems are typically limited to major urban centers in Cambodia, and the remaining population relies on a variety of surface, rain, and groundwater sources. This study examines the household water supplies available to Phnom Penh's resettled peri-urban residents through a case-study approach of two communities. A quantitative microbial risk assessment is performed to assess the level of diarrheal disease risk faced by community members due to microbial contamination of drinking water. Risk levels found in this study exceed those associated with households consuming piped water. Filtered and boiled rain and tank water stored in a kettle, bucket/cooler, bucket with spigot or a 500 mL bottle were found to provide risk levels within one order-of-magnitude to the piped water available in Phnom Penh. Two primary concerns identified are the negation of the risk reductions gained by boiling due to prevailing poor storage practices and the use of highly contaminated source water.


Community-acquired staphylococcal musculoskeletal infection in infants and young children: necessity of contrast-enhanced MRI for the diagnosis of growth cartilage involvement.
Previous studies have reported that contrast-enhanced sequences do not increase the sensitivity of MRI for the diagnosis of pediatric osteomyelitis and are not needed in the absence of edema on unenhanced MRI sequences. Invasive skeletal infections due to community-acquired Staphylococcus aureus are increasingly encountered in infants and young children and have a proclivity for involvement of both the unossified growth cartilage and the metadiaphyseal bone marrow of the extremities. The study objective is to assess the diagnostic efficacy of contrast-enhanced and unenhanced MRI sequences for the diagnosis of community-acquired S. aureus extremity skeletal infection in infants and young children. A retrospective review was conducted of the clinical charts and imaging studies of patients younger than 18 months diagnosed with invasive community-acquired S. aureus skeletal infections from 2001 to 2009 at a large children's hospital. Sensitivity was calculated for the detection of skeletal infection on contrast-enhanced and unenhanced MRI sequences. The p values were calculated using the Fisher exact score method. The kappa value for interobserver reliability was determined. Community-acquired S. aureus skeletal infections were noted in 34 extremity sites in 25 patients, five of whom had more than one site of disease. The affected skeletal sites were metaphyseal or metadiaphyseal bone marrow only in 16 cases (47%), unossified growth cartilage only in nine cases (26%), and both the unossified growth cartilage and metaphyseal or metadiaphyseal bone marrow in nine cases (26%). In seven of the nine cases of isolated involvement of the unossified growth cartilage, the cartilage appeared normal on unenhanced sequences and the diagnosis was made only by the demonstration of hypoenhancing or nonenhancing foci in the cartilage after gadolinium-based contrast agent administration. In five of the nine cases of infection of both the unossified growth cartilage and metaphyseal or metadiaphyseal bone marrow, neither the cartilage nor bone marrow appeared abnormal on unenhanced sequences. Therefore, 12 cases of skeletal infection would have been missed without the inclusion of contrast-enhanced sequences. Follow-up extremity radiographs were available for 10 patients, eight (80%) of whom exhibited growth disturbances. Skeletal infection caused by community-acquired S. aureus in infants and young children manifests differently than in older children, including a propensity for involvement of the unossified growth cartilage and potentially occult nature of both cartilage and bone marrow involvement on unenhanced MRI sequences.


"Sicca complex" in liver disease.
Sixty-three patients with liver disease were studied for the presence of the components of Sjögren's syndrome. The "sicca complex" (that is, patients without arthritis) was detected in 42% of patients with active chronic hepatitis, 72% with primary biliary cirrhosis, and 38% with cryptogenic cirrhosis. One patient with active chronic hepatitis and one with primary biliary cirrhosis had rheumatoid arthritis. No evidence of Sjögren's syndrome was detected in seven patients with alcoholic cirrhosis. It is suggested that the sicca complex and autoimmune liver disease may be part of a systemic disorder in which immunological mechanisms are concerned in the pathogenesis.


Point-of-Care Ocular Ultrasound for the Diagnosis of Retinal Detachment: A Systematic Review and Meta-Analysis.
Ocular complaints are common presentations to the emergency department (ED). Among these, retinal detachment can cause significant vision loss if not rapidly diagnosed and referred for appropriate treatment. Point-of-care ultrasound has been suggested to identify the diagnosis rapidly when the ocular examination is limited or the ophthalmology service is not readily available. However, prior studies were limited by small sample sizes, resulting in wide ranges of potential accuracy. The primary outcome for this review was to determine the test characteristics of point-of-care ocular ultrasound for the diagnosis of retinal detachment. PubMed, CINAHL, Scopus, LILACS, the Cochrane Database of Systematic Reviews, the Cochrane Central Register of Controlled Trials, and bibliographies of selected articles were assessed for all prospective and randomized controlled trials assessing the accuracy of point-of-care ultrasound for identifying retinal detachment. Data were dual extracted into a predefined worksheet and quality analysis was performed using the QUADAS-2 tool. Data were summarized and a meta-analysis was performed with planned subgroup analyses by location and provider specialty. This review was registered with PROSPERO CRD42018097288. There was no funding for this review. Eleven studies (n = 844 patients) were identified. Overall, ultrasound was 94.2% (95% confidence interval [CI] = 78.4% to 98.6%) sensitive and 96.3% (95% CI = 89.2% to 98.8%) specific for the diagnosis of retinal detachment with a positive likelihood ratio of 25.2 (95% CI = 8.1 to 78.0) and a negative likelihood ratio of 0.06 (95% CI = 0.01 to 0.25). Subgroup analysis found that ultrasound was more accurate among ED patients, but was not significantly different when performed by ED or non-ED providers. Point-of-care ocular ultrasound is sensitive and specific for the diagnosis of retinal detachment. Future studies should determine the ideal training protocol and the influence of color Doppler and contrast-enhanced ultrasound on diagnostic accuracy.


Palm tocotrienols protect ApoE +/- mice from diet-induced atheroma formation.
We evaluated the effects of vitamin E and beta-carotene on apolipoprotein (apo)E +/- female mice, which develop atherosclerosis only when fed diets high in triglyceride and cholesterol. Mice were fed a nonpurified control diet (5.3 g/100 g triglyceride, 0.2 g/100 g cholesterol), an atherogenic diet alone (15.8 g/100 g triglyceride, 1.25 g/100 g cholesterol, 0.5 g/100 g Na cholate) or the atherogenic diet supplemented with either 0.5 g/100 g (+)-alpha-tocopherol (mixed isomers); 0.5 g/100 g palm tocopherols (palm-E; 33% alpha-tocopherol, 16.1% alpha-tocotrienol, 2.3% beta-tocotrienol, 32.2% gamma-tocotrienol, 16.1% delta-tocotrienol); 1.5 g/100 g palm-E; or 0.01 g/100 g palm-carotenoids (58% beta-carotene, 33% alpha-carotene, 9% other carotenoids). Compared with mice fed the control diet, plasma cholesterol was fourfold greater in mice fed the atherogenic diet. Mice fed the 1.5 g/100 g palm-E supplement had 60% lower plasma cholesterol than groups fed the other atherogenic diets. Mice fed the atherogenic diet had markedly higher VLDL, intermediate density lipoprotein (IDL) and LDL cholesterol and markedly lower HDL cholesterol than the controls. Lipoprotein patterns in mice supplemented with alpha-tocopherol or palm carotenoids were similar to those of the mice fed the atherogenic diet alone, but the pattern in mice supplemented with 1. 5 g/100 g palm-E was similar to that of mice fed the control diet. In mice fed the atherogenic diet, the hepatic cholesterol plus cholesterol ester concentration was 4.4-fold greater than in mice fed the control diet. Supplementing with 1.5 g/100 g palm-E lowered hepatic cholesterol plus cholesterol ester concentration 66% compared with the atherogenic diet alone. Mice fed the atherogenic diet had large atherosclerotic lesions at the level of the aortic valve. With supplements of 0.5 g/100 g palm-E or 1.5 g/100 g palm-E, the size of the lesions was 92 or 98% smaller, respectively. The 0.5 g/100 g alpha-tocopherol and palm carotenoid supplements had no effect. Supplements did not alter mRNA abundance for apolipoproteins A1, E, and C3. The beneficial effect of tocotrienols on atherogenesis, the plasma lipoprotein profile and accumulation of hepatic cholesterol esters cannot be attributed to their antioxidant properties.


Familiarity Breeds Strategy: In Silico Untangling of the Molecular Complexity on Course of Autoimmune Liver Disease-to-Hepatocellular Carcinoma Transition Predicts Novel Transcriptional Signatures.
Autoimmune liver diseases (AILD) often lead to transformation of the liver tissues into hepatocellular carcinoma (HCC). Considering the drawbacks of surgical procedures in such cases, need of successful non-invasive therapeutic strategies and treatment modalities for AILD-associated-HCC still exists. Due to the lack of clear, sufficient knowledge about factors mediating AILD-to-HCC transition, an in silico approach was adopted to delineate the underlying molecular deterministic factors. Parallel enrichment analyses on two different public microarray datasets (GSE159676 and GSE62232) pinpointed the core transcriptional regulators as key players. Correlation between the expression kinetics of these transcriptional modules in AILD and HCC was found to be positive primarily with the advancement of hepatic fibrosis. Most of the regulatory interactions were operative during early (F0-F1) and intermediate fibrotic stages (F2-F3), while the extent of activity in the regulatory network considerably diminished at late stage of fibrosis/cirrhosis (F4). Additionally, most of the transcriptional targets with higher degrees of connectivity in the regulatory network (namely DCAF11, PKM2, DGAT2 and BCAT1) may be considered as potential candidates for biomarkers or clinical targets compared to their low-connectivity counterparts. In summary, this study uncovers new possibilities in the designing of novel prognostic and therapeutic regimen for autoimmunity-associated malignancy of liver in a disease progression-dependent fashion.


Xylobiose treatment triggers a defense-related response and alters cell wall composition.
Plant cell wall-derived oligosaccharides, i.e., damage-associated molecular patterns (DAMPs), could be generated after pathogen attack or during normal plant development, perceived by cell wall receptors, and can alter immunity and cell wall composition. Therefore, we hypothesised that xylo-oligosaccharides (XOS) could act as an elicitor and trigger immune responses. To test this, we treated Arabidopsis with xylobiose (XB) and investigated different parameters. XB-treatment significantly triggered the generation of reactive oxygen species (ROS), activated MAPK protein phosphorylation, and induced callose deposition. The combination of XB (DAMP) and flg22 a microbe-associated molecular pattern (MAMP) further enhanced ROS response and gene expression of PTI marker genes. RNA sequencing analysis revealed that more genes were differentially regulated after 30 min compared to 24 h XB-treated leaves, which correlated with ROS response. Increased xylosidase activity and soluble xylose level after 30 min and 3 h of XB-treatment were observed which might have weakened the DAMP response. However, an increase in total cell wall sugar and a decrease in uronic acid level was observed at both 30 min and 24 h. Additionally, arabinose, rhamnose, and xylose levels were increased in 30 min, and glucose was increased in 24 h compared to mock-treated leaves. The level of jasmonic acid, abscisic acid, auxin, and cytokinin were also affected after XB treatment. Overall, our data revealed that the shortest XOS can act as a DAMP, which triggers the PTI response and alters cell wall composition and hormone level.


Total Knee Arthroplasty Using Computer-Assisted Navigation in a Patient with Nail-Patella Syndrome: A Case Report.
Nail-patella syndrome (NPS) is a genetic disorder causing anatomical abnormalities about the knee, including significant patellar hypoplasia. We present a case of a patient with NPS and severe knee osteoarthritis undergoing computer-assisted total knee arthroplasty (TKA). Several intraoperative anatomical challenges were appreciated. Postoperatively, the patient developed arthrofibrosis requiring manipulation; however, his final outcome was favorable. Computer assistance may improve precision in patients with NPS undergoing TKA, but surgeons must be aware of the associated anatomic abnormalities and potentially increased risk of arthrofibrosis. Patellar resurfacing is often not feasible because of lack of bone stock.


Analysis of figure recognition on the modified MT Beta-86 test for adaptation: description and discussion of methodological aspects.
The purpose of this study was to analyze the recognition of figures from the modified MT Beta-86 test for subsequent refinement and adaptation of this language assessment instrument. This study was organized into two parts. The first part of the study involved 61 subjects with educational level higher than nine years. Naming of 266 visual stimuli was elicited and concordance among participants on figure naming was also determined. Concordance of responses was assessed using Cochrane's test at a 5% level of significance. Some visual stimuli were subsequently re-designed and images were replaced. The second part of this study involved the re-testing of the new images among 110 individuals ranging in age from 18 to 75 years of both genders, stratified into four educational bands (A: 1-4 years; B: 5-8 years; C: 9-11 years; and D: more than 12 years). Stimuli with concordance level of 70% or higher were considered adequate. In part A of the study, from the index of agreement of naming, the figures considered suitable for inclusion in the adapted version were selected. In part B, all the figures had a response concordance level over 70%. Figures in the Modified MT Beta-86 test suitable for retention in the new version of the instrument were identified. Analysis of low sample recognition was used to identify boards for replacement, and those for retention, in the new version of the test.


Diabetic ketoacidosis promotes a prothrombotic state.
Cerebrovascular accidents are one of the life-threatening complications of diabetic ketoacidosis (DKA) in children and adolescents. Our objective was to evaluate the effect of DKA and its treatment on factors known to affect thrombotic activity (protein C; protein S; von Willebrand factor, fibrinogen; homocysteine; and folate) by comparing seven adolescents with DKA prior to treatment and at 6, 24, and 120 hours after initiation of treatment. We found that protein C activity was significantly decreased by DKA, but normalized slowly following treatment. Free protein S was low throughout the study. Protein C antigen and protein S antigen showed varying degrees ofchange within the first 24 hours, but remained in the normal range, with the exception of the initial value of protein C antigen, which was elevated. von Willebrand factor (vWF) antigen and vWF activity were both significantly increased prior to treatment, but decreased with treatment. However, vWF activity remained elevated at 120 hours. Fibrinogen concentrations showed no significant changes throughout the study. Homocysteine was significantly decreased prior to treatment and increased with the initiation of treatment Folate was significantly increased prior to treatment, and decreased to high normal levels. The increased vWF and the decreased levels of protein C activity and of free protein S support the hypothesis that DKA and its treatment results in a prothrombotic state and activation of the vascular endothelium, which, in turn, predispose to cerebrovascular accidents.


Bridging to cardiac transplantation with mechanical circulatory support.
During the last decade, it has been clearly demonstrated that mechanical assist devices are capable of supporting patients in the interim before cardiac transplantation. The use of these devices prior to transplantation has provided an important window of opportunity to evaluate critically the devices and their potential for adaptation to permanent systems. The overall survival rates in the bridge-to-transplant patient population have improved. This is in part due to lessons learned in the past several years with regard to better patient selection and applicability of the devices. This review focuses on selected papers published in the past year that have made significant contributions to the evolution of mechanical support.


Efficacy of Task Oriented Exercise Program Based on Ergonomics on Cobb's Angle and Pulmonary Function Improvement in Adolescent Idiopathic Scoliosis- A Randomized Control Trial.
Scoliosis is lateral curvature of the spine greater than 100 accompanied by vertebral rotation. The prime risk factors for curve progression are a large curve magnitude, skeletal immaturity and female gender. The curve progression can be recorded by measuring the curve magnitude using the Cobb's method on radiographs. To assess the effect of task oriented exercises based on ergonomics on Cobb's angle and pulmonary functions on one year outcome of Adolescent Idiopathic Scoliosis(AIS) (defined as curves < 150 for thoracolumbar region and < 200 for thoracic region). A total of 36 patients were enrolled in the study and 18 patients in each group (experimental and control group) were allocated randomly. The subjects in control group underwent spinal strengthening exercises, active self-correction and breathing exercises, whereas subjects in the experimental group followed task oriented exercises based on ergonomics in addition to exercises for conventional group for one year. Chi square test was used to compare the categorical/ dichotomous variables between the groups. Unpaired t-test was used to compare continuous variables between the groups at pre and post intervention. Paired t-test was used to compare the changes in continuous variables from pre to post intervention within the group. Forced Vital Capacity (FVC) was found to be significantly (p=0.001) higher in experimental group (2.68±0.37) than control group (2.20±0.41) at post-intervention. Force Expritatory Volume 1 (FEV1) (p=0.01) and Vital Capacity (VC) (p=0.002) were also found to be significantly higher in experimental group compared to control group at postintervention. Also, there was significant (p=0.001) mean reduction in Cobb's angle from pre to post intervention in both the groups being higher in Experimental group than control group. The task oriented exercise protocol benefited patients with AIS which had a significant improvement of their pulmonary functions and Cobb's angle.


Liability insurance impact uncertain.
Practice parameters are here to stay and will have, in fact already have had, a dramatic effect upon the practice of medicine. The effect of practice parameters upon professional liability and professional liability carriers is uncertain.


[Assimilation of rhythm by the isolated dog heart during gradual raising of stimulation frequency].
An ability for a forestalling regulation of contractility of the heart with calculation of the tendency of rhythm increasing was revealed under a gradual increasing of heart rhythm. A forestalling regulation of heart contractility occurs with rhythm assimilation at the cell level of the heart and irrespective of the influence of Frank-Starling law and neurohumoral factors on the work of the heart. A 5-10% increasing of heart rhythm is characterized by optimal rhythm assimilation. A 15-40% increasing of heart rhythm is not optimal and results in transformation of the rhythm. The following sequence of events take place in the process of transition from rhythm assimilation to rhythm transformation under a gradual increasing of heart rhythm: rhythm assimilation--rhythm by mechanical function--incomplete rhythm assimilation by electrical function-transformation of rhythm by electrical function.


Equine coronavirus: An emerging enteric virus of adult horses.
Equine coronavirus (ECoV) is an emerging virus associated clinically and epidemiologically with fever, depression, anorexia and less frequently colic and diarrhoea in adult horses. Sporadic cases and outbreaks have been reported with increased frequency since 2010 from Japan, the USA and more recently from Europe. A faeco-oral transmission route is suspected and clinical or asymptomatic infected horses appear to be responsible for direct and indirect transmission of ECoV. A presumptive clinical diagnosis of ECoV infection may be suggested by clinical presentation, haematological abnormalities such as leucopenia due to lymphopenia and/or neutropenia. Confirmation of ECoV infection is provided by specific ECoV nucleic acid detection in faeces by quantitative PCR (qPCR) or demonstration of coronavirus antigen by immunohistochemistry or electron microscopy in intestinal biopsy material obtained ante or post mortem. The disease is generally self-limiting and horses typically recover with symptomatic supportive care. Complications associated with disruption of the gastrointestinal barrier have been reported in some infected horses and include endotoxaemia, septicaemia and hyperammonaemia-associated encephalopathy. Although specific immunoprophylactic measures have been shown to be effective in disease prevention for closely-related coronaviruses such as bovine coronavirus (BCoV), such strategies have yet not been investigated for horses and disease prevention is limited to basic biosecurity protocols. This article reviews current knowledge concerning the aetiology, epidemiology, clinical signs, diagnosis, pathology, treatment and prevention of ECoV infection in adult horses.


Impaired Relaxation and Reduced Lusitropic Reserve in Atrial Myocardium in the Obese Patients.
Background: Obesity can influence the structure and function of the atrium, but most studies focused on the relationship of body mass index (BMI) and overt left atrium (LA) dysfunction as assessed by clinical imaging. We combined the assessment of right atrium (RA) function in vivo and in vitro in obese and non-obese patients scheduled for elective cardiac surgery. Methods: Atrial structure and function were quantified pre-operatively by echocardiography. RA tissue removed for the establishment of extracorporeal support was collected and RA trabeculae function was quantified in vitro at baseline and with adrenergic stimulation (isoproterenol). Fatty acid-binding protein 3 (FABP3) was quantified in RA tissue. Results were stratified according to the BMI of the patients. Results: About 76 patients were included pre-operatively for the echocardiographic analysis. RA trabeculae function at baseline was finally quantified from 46 patients and RA function in 28 patients was also assessed with isoproterenol. There was no significant correlation between BMI and the parameters of atrial function measured by the clinical echocardiography. However, in vitro measurements revealed a significant correlation between BMI and a prolonged relaxation of the atrial myocardium at baseline, which persisted after controlling for the atrial fibrillation and diabetes by the partial correlation analysis. Acceleration of relaxation with isoproterenol was significantly lower in the obese group (BMI ≥ 30 kg/m2). As a result, relaxation with adrenergic stimulation in the obese group remained significantly higher compared to the overweight group (25 kg/m2 ≤ BMI < 30 kg/m2, p = 0.027) and normal group (18.5 kg/m2 ≤ BMI < 25 kg/m2, p = 0.036). There were no differences on impacts of the isoproterenol on (systolic) developed force between groups. The expression of FABP3 in the obese group was significantly higher compared to the normal group (p = 0.049) and the correlation analysis showed the significant correlations between the level of FABP3 in the RA trabeculae function. Conclusion: A higher BMI is associated with the early subclinical changes of RA myocardial function with the slowed relaxation and reduced adrenergic lusitropy.


[Shoulder cartilage regeneration : Indications, possibilities, surgical implementation].
Asymptomatic cartilage lesions of the shoulder are frequent. Symptomatic lesions are treated analogously to other hyaline cartilage bearing joints and can be treated arthroscopically in the vast majority of cases. The therapeutic options can be subdivided into resection, stimulation and transplantation of the defect cartilage. There are only a few reports about outcome after cartilage restoration surgery in the literature, and microfracturing is the surgical technique that has been most investigated.


Microglia-derived interleukin-6 and leukaemia inhibitory factor promote astrocytic differentiation of neural stem/progenitor cells.
Neural stem/progenitor cells (NSPCs) proliferate and differentiate depending on their intrinsic properties and local environment. It has been recognized that astrocytes promote neurogenic differentiation of NSPCs, suggesting the importance of cell-cell interactions between glial cells and NSPCs. Recent studies have demonstrated that microglia, one type of glial cells, play an important role in neurogenesis. However, little is known about how activated microglia control the proliferation and differentiation of NSPCs. In this study, we investigated the possibility that microglia-derived soluble factors regulate the behaviour of NSPCs. To this end, NSPCs and microglial cultures were obtained from rat embryonic day 16 subventricular zone (SVZ) and rat postnatal 1 day cortex, respectively, and the conditioned medium from microglia was prepared. Microglial-conditioned medium had no significant effect on the proliferation of NSPCs. In contrast, it increased the percentage of cells positive for a marker of astrocytes, glial fibrillary acidic protein (GFAP) during differentiation. The induction of astrocytic differentiation by microglial-conditioned medium was reduced by the inhibition of the Janus kinase/signal transducer and activation of transcription (JAK/STAT) and mitogen-activated protein kinase (MAPK) pathways. Furthermore, microglia-derived interleukin (IL)-6 and leukaemia inhibitory factor (LIF) were identified as essential molecules for this astrocytic differentiation using neutralizing antibodies and recombinant cytokines. Our results suggest that microglia as well as astrocytes contribute to the integrity of the local environment of NSPCs, and at least IL-6 and LIF released by activated microglia promote astrocytic differentiation of NSPCs via the activation of the JAK/STAT and MAPK pathways.


Vitreous mediators in retinal hypoxic diseases.
The causes of retinal hypoxia are many and varied. Under hypoxic conditions, a variety of soluble factors are secreted into the vitreous cavity including growth factors, cytokines, and chemokines. Cytokines, which usually serve as signals between neighboring cells, are involved in essentially every important biological process, including cell proliferation, inflammation, immunity, migration, fibrosis, tissue repair, and angiogenesis. Cytokines and chemokines are multifunctional mediators that can direct the recruitment of leukocytes to sites of inflammation, promote the process, enhance immune responses, and promote stem cell survival, development, and homeostasis. The modern particle-based flow cytometric analysis is more direct, stable and sensitive than the colorimetric readout of the conventional ELISA but, similar to ELISA, is influenced by vitreous hemorrhage, disruption of the blood-retina barrier, and high serum levels of a specific protein. Finding patterns in the expression of inflammatory cytokines specific to a particular disease can substantially contribute to the understanding of its basic mechanism and to the development of a targeted therapy.


Experimental and computational investigation on the gas phase reaction of ethyl formate with Cl atoms.
The rate coefficient for the gas-phase reaction of Cl atoms with ethyl formate was measured over the temperature range of 268-343 K using relative rate methods, with ethyl chloride as a reference compound. The temperature dependent relative rate coefficients for the ethyl formate + Cl reaction were measured, and the modified Arrhenius expression kethyl formate(268-343) = (2.54 ± 0.57) × 10(-23) T(4.1) exp {-(981 ± 102)/T} cm(3) molecule(-1) s(-1) was obtained with 2σ error limits. The room temperature rate coefficient for the title reaction is (9.84 ± 0.79) × 10(-12) cm(3) molecule(-1) s(-1), which is in good agreement with reported values. To complement the experimental measurement, computational methods were used to calculate the rate coefficient for the ethyl formate + Cl reaction atoms using canonical variational transition state theory (CVT) with small curvature tunneling (SCT) and the CCSD (T)/cc-pVDZ//M062X/6-31+g(d,p) level of theory. The temperature dependent Arrhenius expression was obtained to be 2.97 × 10(-18) T(2.4) exp[-(390/T)] cm(3) molecule(-1) s(-1) over the temperature range of 200-400 K. The thermodynamic parameters and branching ratio were calculated. Also, the atmospheric lifetime and global warming potentials (GWPs) were calculated for ethyl formate.


Anticoagulant treatment in COVID-19: a narrative review.
The actual Coronavirus Disease (COVID 19) pandemic is due to Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a member of the coronavirus family. Besides the respiratory involvement, COVID 19 patients frequently develop a pro-coagulative state caused by virus-induced endothelial dysfunction, cytokine storm and complement cascade hyperactivation. It is common to observe diffuse microvascular thrombi in multiple organs, mostly in pulmonary microvessels. Thrombotic risk seems to be directly related to disease severity and worsens patients' prognosis. Therefore, the correct understanding of the mechanisms underlying COVID-19 induced prothrombotic state can lead to a thorough assessment of the possible management strategies. Hence, we review the pathogenesis and therapy of COVID 19-related thrombosis disease, focusing on the available evidence on the possible treatment strategies and proposing an algorithm for the anticoagulation strategy based on disease severity.


A quality assessment tool for tuberculosis control activities in resource limited settings.
Tuberculosis (TB) is a significant problem, infecting nearly 9 million new patients per year and killing about 2 million a year. The primary means with which to affect TB globally are to decrease transmission locally, mainly by effective identification, diagnosis, and treatment of infectious TB patients. Therefore, quality assurance of TB control efforts at the local level is essential. This study describes the creation of a data extraction tool for retrospective chart review based on the International Standards for TB Care, 2009 for the assessment of TB control programs located in resource limited settings. The tool was field tested at a rural mission hospital in central Kenya. Results were used by host site staff to develop a quality improvement plan. The process prompted revision of the tool to clarify questions and answers. This is a tool that can be used in resource limited settings for data collection to assess the quality of TB care and to inform the design, implementation, and further assessment of future quality improvement initiatives.


Controlled precipitation for enhanced dissolution rate of flurbiprofen: development of rapidly disintegrating tablets.
The aim of this work was to investigate the potential of controlled precipitation of flurbiprofen on solid surface, in the presence or absence of hydrophilic polymers, as a tool for enhanced dissolution rate of the drug. The work was extended to develop rapidly disintegrated tablets. This strategy provides simple technique for dissolution enhancement of slowly dissolving drugs with high scaling up potential. Aerosil was dispersed in ethanolic solution of flurbiprofen in the presence and absence of hydrophilic polymers. Acidified water was added as antisolvent to produce controlled precipitation. The resultant particles were centrifuged and dried at ambient temperature before monitoring the dissolution pattern. The particles were also subjected to FTIR spectroscopic, X-ray diffraction and thermal analyses. The FTIR spectroscopy excluded any interaction between flurbiprofen and excipients. The thermal analysis reflected possible change in the crystalline structure and or crystal size of the drug after controlled precipitation in the presence of hydrophilic polymers. This was further confirmed by X-ray diffraction. The modulation in the crystalline structure and size was associated with a significant enhancement in the dissolution rate of flurbiprofen. Optimum formulations were successfully formulated as rapidly disintegrating tablet with subsequent fast dissolution. Precipitation on a large solid surface area is a promising strategy for enhanced dissolution rate with the presence of hydrophilic polymers during precipitation process improving the efficiency.


Ligand targeting and peptide functionalized polymers as non-viral carriers for gene therapy.
Polymeric gene carriers have been developed to deliver therapeutic genes for treating various genetic diseases. They may diminish the problems related with viral vectors in terms of safety, immunogenicity and mutagenesis. But inefficient endosomal release, cytoplasmic transport and nuclear entry are the main limiting issues in the usage of polymeric carriers. Different strategies have been proposed to functionalize gene carriers for individually overcoming these barriers. Towards this aim, various polymeric carriers have been developed and further modified with certain ligands and peptides to achieve properties like good cargo DNA protection, excellent colloidal stability, high cellular uptake efficiency, efficient endo/lysosome escape, efficient import into the nucleus and specificity to target cells. These multifunctional polymeric carriers are beneficial for efficient gene delivery. In this review, we provide a comprehensive summary of the interaction between gene and polymeric carriers and the mechanism and challenges of gene carriers. We also highlight and discuss the recent developments and advancements in the synthesis of ligand targeting and peptide functionalized gene carriers.


Spectroscopic and binding studies on the stereoselective interaction of tyrosine with horseradish peroxidase and lactoperoxidase.
The interaction of a series of derivatives of tyrosine with horseradish peroxidase (HRP) and lactoperoxidase (LPO) was studied by using optical difference spectroscopy, c.d. and proton n.m.r. spectroscopy in order to reveal differences in the mode of binding of L-tyrosine and D-tyrosine, which are substrates of but react at different rates with the two peroxidases, to HRP and LPO. All the donor molecules form 1:1 complexes with HRP and LPO, but they display a range of affinities for the enzymes. Whereas D-tyrosine binds to HRP more strongly than does L-tyrosine, the opposite holds for the binding to LPO. The distances of the protons of bound tyrosine molecules from the haem iron atoms of HRP and LPO indicate that the site of binding of these substrates is the same as that of simple phenols. This involves the interaction of the phenol nucleus with a protein tyrosine residue [Sakurada, Takahashi & Hosoya (1986) J. Biol. Chem. 261, 9657-9662; Modi, Behere & Mitra (1989) Biochim. Biophys. Acta 996, 214-225]. However, for the present substrates the additional interaction of the carboxylate group with a protein residue (probably an arginine residue) provides further stabilization for the adducts HRP-D-tyrosine and LPO-L-tyrosine with respect to the corresponding complexes with the opposite enantiomers. The differences in the mode of binding of L-tyrosine and D-tyrosine to HRP and LPO is thus determined by the fact that the spatial arrangement of the interacting protein residues can recognize the chirality of the C(alpha)-CO2- and C(beta)-C6H4OH attachment bonds of the substrates.


The fatal cafe coronary. Foreign-body airway obstruction.
Demographic features and predisposing factors to fatal food asphyxiation (cafe coronary) were determined by analyzing 141 such fatalities over a 20-year period. Old age, poor dentition, and alcohol consumption were expected findings. Other predisposing factors included institutionalization in long-term medical care facilities, sedative drugs, and natural diseases, most notably parkinsonism. Since institutionalized patients tended to asphyxiate on soft, friable, or snack-like foods, future improvements in rescue techniques should take this into consideration. The incidence of 0.66 per 100,000 population has remained unchanged over the two decades studied. Observers were present at the time of the fatal incident in 85% of the cases. In only a third, the food was lodged ion a supraglottic location. These data indicate that continued efforts toward greater awareness of the cafe coronary syndrome still are needed. Special attention should be directed to the predisposing role of institutionalization, sedative drugs, and natural diseases.


NKT cell development in the absence of the autoimmune regulator gene (Aire).
Autoimmune regulator gene (Aire)-deficient mice develop an array of autoimmune lesions that reflect failures of immune tolerance. Negative selection is clearly compromised in these mice, but there is evidence to suggest that other mechanisms of tolerance might also be affected, including a possible impairment of regulatory T cell (Treg) development. Studies to date have failed to demonstrate any significant impact on the development or function of the FOXP3+ Treg compartment, but NKT cells represent a distinct regulatory cell lineage that also develop in the thymus and which are known to influence self-tolerance. Aire-related defects coincide with NKT cell deficiencies in a number of animal models, but the direct consequence of Aire-deficiency on NKT cell development has not been established. In this study, we demonstrate that the frequency, distribution and cytokine production of NKT cells and their subsets is principally normal in Aire-deficient mice. We conclude that Aire has little or no effect on regulatory T cell development in general and NKT cells in particular.


Postcritical behavior of a gelling system.
The formation of a gel in a disperse coagulating system, wherein binary coagulation governs the temporal changes of the particle mass spectrum is studied for the coagulation kernel proportional to K(g,l)=gl with g,l being the numbers of monomers in the coalescing pair of particles. This model is known to reveal the sol-gel transition, i.e., the formation of one giant cluster with the mass comparable to the total mass of the whole system. This paper reports on some exact results that are valid for arbitrary initial particle mass spectra. The particle spectrum is shown to be expressed through a contour integral of the exponentiated initial generating function of the particle mass spectrum. The exact formula for the gel mass is derived for the scenario of passive gelation. Two scenarios of gelation, where the gel is either active or passive, are considered.


[Disseminated mycobacteriosis associated with Mycobacterium avium-intracellular (MAI) complex in HIV infected patients].
Different types of samples from a 27-year-old female patient who was diagnosed AIDS after presenting an acute respiratory picture caused by Pneumocistis carinii, in 1997, were studied. From that moment on, the patient continued having respiratory pictures repeatedly, until she was admitted at "Pedro Kouri" Institute of Tropical Medicine with a chronic diarrheical picture, significant loss of weight and extreme anorexia, together with a constant wet cough, fever, etc. The necessary complementary tests were made and a hyperergic Mantoux and very low levels of CD4 cells (under 200 cells/m3) outstood. Microbiological studies were also conducted with samples from sputum, hemocultures and cerebrospinal fluid to discard the presence of acid-alcohol resistant bacilli (AARB). As a result of these studies, the repeated isolation was obtained in all types of worked samples from a slow-growth, creamy and non pigmented strain corresponding to Runyon's group III, which was classified as from the Mycobacterium avium-intracellulare complex. The patient was treated with antibacillary drugs without results, since she dies 4 weeks later due to the total dissemination of this infection. It should be stressed that it is the first case reported and microbiologically diagnosed of a generalized mycobacteriosis in a HIV positive patient due to Mycobacterium avium-intracellulare.


Chebyshev series for designing RF pulses employing an optimal control approach.
Magnetic resonance imaging (MRI) provides bidimensional images with high definition and selectivity. Selective excitations are achieved applying a gradient and a radio frequency (RF) pulse simultaneously. They are modeled by the Bloch differential equation, which has no closed-form solution. Most methods for designing RF pulses are derived from approximation of this equation or are based on iterative optimization methods. The approximation methods are only valid for small tip angles and the optimization-based algorithms yield better results, but they are computationally intensive. To improve the solutions and to reduce processing time, a method for designing RF pulses using a pseudospectral approach is presented. The Bloch equation is expanded in Chebyshev series, which can be solved using a sparse linear algebraic system. The method permits three different formulations derived from the optimal control theory, minimum distance, minimum energy, or minimum time, which are solved as algebraic constrained minimization problems. The results were validated through simulated and real experiments of 90 degrees and 180 degrees RF pulses. They show improvements compared to the corresponding solutions obtained using the Shinnar-Le Roux method. The minimum time formulation produces the best performance for 180 degrees pulses, reducing the excitation length in 4% and the RF pulse energy in 3%.


Repair of medial meniscus posterior root tear is effective for root healing and cartilage regeneration in opening wedge high tibial osteotomy.
This study aimed to determine whether the repair of a medial meniscus posterior root tear (MMPRT) is effective for MMPRT healing, cartilage regeneration, and clinical outcomes in opening wedge high tibial osteotomy (OWHTO). This retrospective study included 80 patients who underwent OWHTO and subsequent second-look arthroscopy. The patients were divided into OWHTO-with-MMPRT-repair (n = 40) and OWHTO alone (n = 40) groups, and the healing rates (complete/partial/failure) were compared. Each group was further divided into over- and under-corrected subgroups to compare healing rates. The International Cartilage Repair Society (ICRS) grade, cartilage defect size, Koshino stage, ICRS cartilage repair assessment score of the medial femoral condyle (MFC), and International Knee Documentation Committee (IKDC) scores between the OWHTO-with-MMPRT-repair and OWHTO alone groups were compared according to whether microfracture was performed on the MFC. The overall healing rate of the MMPRT was higher in the OWHTO-with-MMPRT-repair group than that in the OWHTO alone group (P < 0.001). In addition, in the subgroup analysis, no difference in the MMPRT healing rate between the over-correction and under-correction groups when MMPRT repair was performed (n.s). In contrast, without MMPRT repair, the healing rate was lower in the under-correction group than that in the over-correction group (P = 0.03). Cartilage regeneration of the OWHTO-with-MMPRT-repair group was superior to that of the OWHTO alone group (P < 0.05). The IKDC subjective scores of the OWHTO-with-MMPRT-repair and OWHTO alone groups were 34.5 and 33.1 before surgery (n.s) and 50 and 47.2 at one year after surgery, respectively (n.s). These differences between the two groups for cartilage regeneration and IKDC subjective scores showed the same pattern regardless of microfractures. MMPRT repair during OWHTO might improve MMPRT healing, even with under-correction, and cartilage regeneration of MFC, regardless of microfracture. However, OWHTO with MMPRT repair might not improve short-term clinical outcomes compared to OWHTO alone. Further randomized clinical trials are necessary. III, Retrospective cohort study.


p53 is essential for chemotherapy-induced hair loss.
Anticancer drugs stimulate apoptosis in the hair follicles (HF) and cause hair loss, the most common side effect of chemotherapy. In a mouse model for chemotherapy-induced hair loss, we demonstrate that p53 is essential for this process: in contrast to wild-type mice, p53-deficient mice show neither hair loss nor apoptosis in the HF keratinocytes that maintained active proliferation after cyclophosphamide treatment. HF in p53 mutants are characterized by down-regulation of Fas and insulin-like growth factor-binding protein 3 and by increased expression of Bcl-2. These observations indicate that local pharmacological inhibition of p53 may be useful to prevent chemotherapy-associated hair loss.


The Sleeping Beauty transposon system for clinical applications.
Extensive efforts have been made to establish efficient and safe gene delivery protocols that could meet demanding expectations of a successful gene therapy. The Sleeping Beauty (SB) transposon system combines simplicity and inexpensive manufacture offered by plasmid-based vector formulation with integrative features exhibited by some viral vectors. Activated after over ten million years of silent genomic existence, the SB transposable element entered the 21st century as a potent technology for a broad range of applications in genome engineering, including gene therapy. Beneficially for gene therapy purposes, the SB system has been demonstrated to enable persistent expression of therapeutic genes followed by restoration of homeostasis in a variety of disease models. Importantly, this non-viral gene delivery vehicle is postulated to constitute a relatively safe vector system, because it lacks a preference for inserting into transcription units and their upstream regulatory regions, thereby minimizing genotoxic risks that might be associated with vector integration. Further evolution and wide, comprehensive preclinical testing of the SB transposon system in the context of several disease models is expected to further refine this valuable technology matched by enhanced biosafety towards disease treatment.


Phylogenetic Analysis and Molecular Typing of Trichothecene-Producing Fusarium Fungi from Russian Collections.
We performed a three-locus phylogenetic analysis of Fusarium strains presumably capable of trichothecene production, which were deposited in the Russian national collections. The intra- and interspecific polymorphism of partial sequences of the translation elongation factor 1 alpha (TEF1α) gene and two genes from the trichothecene cluster TRI5 and TRI14 was studied. A study of 60 strains of different origins using DNA markers confirmed, and in the case for several strains, clarified their taxonomic characteristics. As a result, a strain of F. commune (F-900) was identified in Russia for the first time. Furthermore, the strain F-846 proved to be phylogenetically distinct from any of the known Fusarium species. F. equiseti strains from Northwest Russia were found to belong to the North European group (I), whereas a strain from the North Caucasus - to the South European one (II). Partial TRI14 sequences from 9 out of 12 species were determined for the first time. Their comparative analysis demonstrated a relatively high level of intraspecific variability in F. graminearum and F. sporotrichioides, but no correlation between the sequence polymorphism and the geographic origin of the strains or their chemotype was found. Specific chemotypes of trichothecene B producers were characterized using two primer sets. The chemotyping results were verified by HPLC.


Whole body CT versus selective radiological imaging strategy in trauma: an evidence-based clinical review.
Trauma patients often present with injuries requiring resuscitation and further evaluation. Many providers advocate for whole body computed tomography (WBCT) for rapid and comprehensive diagnosis of life-threatening injuries. Evaluate the literature concerning mortality effect, emergency department (ED) length of stay, radiation, and incidental findings associated with WBCT. Physicians have historically relied upon history and physical examination to diagnose life-threatening injuries in trauma. Diagnostic imaging modalities including radiographs, ultrasound, and computed tomography have demonstrated utility in injury detection. Many centers routinely utilize WBCT based on the premise this test will improve mortality. However, WBCT may increase radiation and incidental findings when used without considering pre-test probability of actionable traumatic injuries. Studies supporting WBCT are predominantly retrospective and incorporate trauma scoring systems, which have significant design weaknesses. The recent REACT-2 trial randomized trauma patients with high index of suspicion for actionable injuries to WBCT versus selective imaging and found no mortality difference. Additional prospective trials evaluating WBCT in specific trauma subgroups (e.g. polytrauma) are needed to evaluate benefit. In the interim, the available data suggests clinicians should adopt a selective imaging strategy driven by history and physical examination. While observational data suggests an association between WBCT and a benefit in mortality and ED length of stay, randomized controlled data suggests no mortality benefit to this diagnostic tool. The literature would benefit from confirmatory studies of the use of WBCT in trauma sub-groups to clarify its impact on mortality for patients with specific injury patterns.


Elevated NT-proBNP and coronary calcium score in relation to coronary artery disease in asymptomatic type 2 diabetic patients with elevated urinary albumin excretion rate.
Elevated plasma N-terminal (NT)-proBNP levels and coronary calcium score (CCS) not only predicts myocardial ischaemia and coronary artery stenosis but also adverse cardiovascular events and mortality in type 2 diabetic patients with an increased urinary albumin excretion rate (UAER), whereas low levels are associated with low frequency of coronary artery disease (CAD) and good prognosis. The underlying causes of poor prognosis in patients with elevated NT-proBNP are not known; thus, we investigated the role of putative asymptomatic CAD in type 2 diabetic patients with UAER >30 mg/24 h and elevated P-NT-proBNP and/or CCS. We identified 200 type 2 diabetic patients without known CAD and with normal creatinine levels. Patients with P-NT-proBNP >45.2 ng/L (the median P-NT-proBNP value in this cohort and in accordance with our previous findings) and/or CCS ≥ 400 were stratified as high-risk patients for CAD (n = 133) and all other patients as low-risk patients (n = 67). High-risk patients were examined by myocardial perfusion imaging (MPI; n = 109) and/or computer tomography angiography (n = 20) and/or coronary angiography (CAG; n = 86). All patients received intensive mulitifactorial intervention. In 70 of 133 (53%) high-risk patients, significant CAD was demonstrated by MPI and/or CAG, corresponding to 35% (70/200) of the total cohort. Among high-risk patients, CCS but not P-NT-proBNP was paralleled by increased prevalence of significant CAD and in the 86 patients where CAG was performed, a CCS <100 had a negative predictive value for coronary artery stenosis of 94% (P = 0.04). Our study revealed that >50% of asymptomatic type 2 diabetic patients with UAER >30 mg/24 h had significant CAD based on risk stratification with P-NT-proBNP and CCS. This provides some explanation to the previously reported poor prognosis in these asymptomatic patients. Optimized cardio protective treatment in these patients is warranted.


Intestinal myofibroblasts produce nitric oxide in response to combinatorial cytokine stimulation.
Inflammatory bowel disease (IBD) patients display elevated levels of intraluminal nitric oxide (NO). NO can react with other molecules to form toxic compounds, which has led to the idea that NO may be an important mediator of IBD. However, the cellular source of NO and how its production is regulated in the intestine are unclear. In this study we aimed to determine if intestinal myofibroblasts produce NO in response to the IBD-associated cytokines IL-1β, TNFα, and IFNγ. Intestinal myofibroblasts were isolated from mice and found to express inducible nitric oxide synthase (iNOS) mRNA, but not endothelial NOS or neuronal NOS. Individual treatment of myofibroblasts with IL-1β, TNFα, or IFNγ had no effect on NO production, but stimulation with combinations of these cytokines synergistically increased iNOS mRNA and protein expression. Treatment with TNFα or IFNγ increased cell surface expression of IFNγRI or TNFRII, respectively, suggesting that these cytokines act in concert to prime NO production by myofibroblasts. Impairment of NF-κB activity with a small molecule inhibitor was sufficient to prevent increased expression of IFNγRI or TNFRII, and inhibition of Akt, JAK/STAT, or NF-κB blocked nearly all NO production induced by combinatorial cytokine treatment. These data indicate that intestinal myofibroblasts require stimulation by multiple cytokines to produce NO and that these cytokines act through a novel pathway involving reciprocal cytokine receptor regulation and signaling by Akt, JAK/STAT, and NF-κB.


Development and evaluation of microdevices for studying anisotropic biaxial cyclic stretch on cells.
Mechanical effects on cells have received more and more attention in the studies of tissue engineering, cellular pathogenesis, and biomedical device design. Anisotropic biaxial cyclic stress, reminiscent of the in vivo cellular mechanical environment, may promise significant implications for biotechnology and human health. We have designed, fabricated and characterized a microdevice that imparts a variety of anisotropic biaxial cyclic strain gradients upon cells. The device is composed of an elastic membrane with microgroove patterns designed to associate cell orientation axes with biaxial strain vectors on the membrane and a Flexcell stretcher with timely controlled vacuum pressure. The stretcher generates strain profile of anisotropic biaxial microgradients on the membrane. Cell axes determined by the microgrooves are associated with the membrane strain profile to impose proper biaxial strains on cells. Using vascular smooth muscle cells as a cell model, we demonstrated that the strain anisotropy index of a cell was likely one of the determinant mechanical factors in cell structural and functional adaptations. The nuclear shape and cytoskeleton structure of smooth muscle cells were influenced by mechanical loading, but were not significantly affected by the strain anisotropy. However, cell proliferation has profound responses to strain anisotropy.


Biocompatibility of submicron Bioglass® powders obtained by a top-down approach.
In this study in vitro bioactivity and biocompatibility of two submicron 45S5 Bioglass® powders obtained by topdown processing have been evaluated and are compared to the as-received powder. Both submicron powders exhibited flake-like morphologies with lateral extensions of only a few microns; the flake thickness accounted for a few tens of nanometers. Enhanced in vitro bioactivity was found for the comminuted powders upon immersion in simulated body fluid. In vitro biocompatibility was evaluated by incubation of MG-63 osteoblast-like cells with various amounts (0–200 μg/mL) of the glass powders. Neither LDH-activity nor mitochondrial activity (WST-8) tests indicated cell toxicity. Increased mitochondrial activity was found for the submicron powders: incubation with high amounts revealed up to a threefold increase of osteoblast activity (ALP-activity). An overgrowth of the formed mineralized phase with phenotypical MG-63 cells was found by staining only for the submicron glasses. A distance ring is formed for the as-received powder. Superior bioactivity markers are found for shorter process times, that is, lower mass specific surface areas. This is attributed to the formation of carbonates during the comminution process.


Evaluating and Strengthening the Evidence for Nutritional Bone Research: Ready to Break New Ground?
A healthy diet is essential to attain genetically determined peak bone mass and maintain optimal skeletal health across the adult lifespan. Despite the importance of nutrition for bone health, many of the nutritional requirements of the skeleton across the lifespan remain underexplored, poorly understood, or controversial. With increasingly aging populations, combined with rapidly changing diets and lifestyles globally, one anticipates large increases in the prevalence of osteoporosis and incidence of osteoporotic fractures. Robust, transparent, and reproducible nutrition research is a cornerstone for developing reliable public health recommendations to prevent osteoporosis and osteoporotic fractures. However, nutrition research is often criticized or ignored by healthcare professionals due to the overemphasis of weak science, conflicting, confusing or implausible findings, industry interests, common misconceptions, and strong opinions. Conversely, spurious research findings are often overemphasized or misconstrued by the media or prominent figures especially via social media, potentially leading to confusion and a lack of trust by the general public. Recently, reforms of the broader discipline of nutrition science have been suggested and promoted, leading to new tools and recommendations to attempt to address these issues. In this perspective, we provide a brief overview of what has been achieved in the field on nutrition and bone health, focusing on osteoporosis and osteoporotic fractures. We discuss what we view as some of the challenges, including inherent difficulties in assessing diet and its change, disentangling complex interactions between dietary components and between diet and other factors, selection of bone-related outcomes for nutrition studies, obtaining evidence with more unbiased designs, and perhaps most importantly, ensuring the trust of the public and healthcare professionals. This perspective also provides specific recommendations and highlights new developments and future opportunities for scientists studying nutrition and bone health. © 2021 American Society for Bone and Mineral Research (ASBMR).


Determination of piceid and resveratrol in Spanish wines deriving from Monastrell (Vitis vinifera L.) grape variety.
The presence of stilbenes in wine is becoming an important issue due to their claimed relation to a low incidence in coronary diseases and their increasing implication as cancer chemopreventive and neuroprotective agents. Total resveratrol content, quantified as glucoside and aglycone forms of resveratrol, has been determined in a survey of 45 Monastrell monovarietal Spanish red wine types (around 135 wine samples), belonging to Alicante and Bullas appellations. The average between ratio glucoside/aglycone forms of resveratrol in these wines was considerably high, ranging from 82 to 91% of resveratrol in its glycosidic form. This characteristic was observed in a high percentage of the studied wines, which were made under different winemaking procedures, and from different vintages (1995-2002). In addition, wines made using macerative fermentations with double amount of solid parts ("doble pasta") reached the highest levels of total stilbene content expressed as resveratrol equivalent, i.e., 30 mg/L (average of 18.8 mg/L). It can be concluded that high resveratrol glucoside concentration and low free isomer content can be considered characteristics of the Monastrell variety, as it happens to red wines deriving from other varieties grown at warm climates. This fact, also observed for other French and Portuguese red varieties, might play an important role in food habits involving these types of wines.


Influenza vaccines for preventing acute otitis media in infants and children.
Acute otitis media (AOM) is one of the most common infectious diseases in children. It has been reported that 64% of infants have an episode of AOM by the age of six months and 86% by one year. Although most cases of AOM are due to bacterial infection, it is commonly triggered by a viral infection. In most children AOM is self limiting, but it does carry a risk of complications. Since antibiotic treatment increases the risk of antibiotic resistance, influenza vaccines might be an effective way of reducing this risk by preventing the development of AOM. To assess the effectiveness of influenza vaccine in reducing the occurrence of acute otitis media in infants and children. We searched the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, Embase, CINAHL, LILACS, Web of Science, the WHO International Clinical Trials Registry Platform, and ClinicalTrials.gov (15 February 2017). We also searched the reference lists of included studies to identify any additional trials. Randomised controlled trials comparing influenza vaccine with placebo or no treatment in infants and children aged younger than six years. We included children of either sex and of any ethnicity, with or without a history of recurrent AOM. Two review authors independently screened studies, assessed trial quality, and extracted data. We performed statistical analyses using the random-effects and fixed-effect models and expressed the results as risk ratio (RR), risk difference (RD), and number needed to treat for an additional beneficial outcome (NNTB) for dichotomous outcomes, with 95% confidence intervals (CI). We included 11 trials (6 trials in high-income countries and 5 multicentre trials in high-, middle-, and low-income countries) involving 17,123 children aged 6 months to 6 years. Eight trials recruited participants from a healthcare setting. Ten trials (and all four trials that contributed to the primary outcome) declared funding from vaccine manufacturers. Four trials reported adequate allocation concealment, and 10 trials reported adequate blinding of participants and personnel. Attrition was low for eight trials included in the analysis.The primary outcome showed a small reduction in at least one episode of AOM over at least six months of follow-up (4 trials, 3134 children; RR 0.84, 95% CI 0.69 to 1.02; RD -0.04, 95% CI -0.08 to -0.00; NNTB 25, 95% CI 12.5 to 100; low-quality evidence).The subgroup analyses (i.e. number of courses and types of vaccine administered) showed no differences.There was a reduction in the use of antibiotics in vaccinated children (2 trials, 1223 children; RR 0.70, 95% CI 0.59 to 0.83; RD -0.11, 95% CI -0.16 to -0.06; moderate-quality evidence).We were unable to demonstrate whether there was any difference in the utilisation of health care. The use of influenza vaccine resulted in a significant increase in fever (7 trials, 10,615 children; RR 1.15, 95% CI 1.06 to 1.24; RD 0.02, 95% CI 0.00 to 0.04; low-quality evidence), rhinorrhoea (6 trials, 10,563 children; RR 1.17, 95% CI 1.07 to 1.29; RD 0.09, 95% CI 0.01 to 0.16; low-quality evidence), but no difference in pharyngitis. No major adverse events were reported.Differing from the protocol, the original publication of the review included a subgroup analysis of AOM episodes by season, and the secondary outcome 'types of influenza vaccine' was changed to a subgroup analysis. For this update, we removed the subgroup analyses for trial setting, season, and utilisation of health care due to the small number of trials involved. We removed Belshe 2000 from primary and secondary outcomes (courses of vaccine and types of vaccine) because it reported episodes of AOM per person. We did not perform a subgroup analysis by type of adverse event. We have reported each type of adverse event as a separate analysis. Influenza vaccine results in a small reduction in AOM. The observed reduction in the use of antibiotics needs to be considered in light of current recommended practices aimed at avoiding antibiotic overuse. Safety data from these trials were limited. The benefits may not justify the use of influenza vaccine without taking into account the vaccine efficacy in reducing influenza and safety data. We judged the quality of the evidence to be low to moderate. Additional research is needed.


Handling baselines in repeated measures analyses with missing data at random.
In longitudinal clinical studies, after randomization at baseline, subjects are followed for a period of time for development of symptoms. A mixed model for repeated measures (MMRM) can be used to analyze data from such studies. Fitzmaurice et al. (2004) outlined five approaches for handling baseline responses in an MMRM analysis. They are: (1) Retain the baselines as part of the outcome vector and make no assumptions about group differences in the mean response at baseline. (2) Retain the baselines as part of the outcome vector and assume the group means are equal at baseline. (3) Subtract the baselines from all of the remaining post-baseline responses, and analyze the differences from baseline. (4) Use the baselines as a covariate in the analysis of the post-baseline responses, assuming homogeneous regression slopes. (5) Use the baselines as a covariate in the analysis of the post-baseline responses, allowing different regression slopes. In this paper, we evaluate these five approaches in the presence of data missing at random. We evaluate the approaches based on the bias of the estimate and the coverage accuracy of the confidence interval. The results suggest that strategies 2 and 5 are recommended.