Structural and functional adaptations to extreme temperatures in psychrophilic, mesophilic, and thermophilic DNA ligases.
Psychrophiles, host of permanently cold habitats, display metabolic fluxes comparable to those exhibited by mesophilic organisms at moderate temperatures. These organisms have evolved by producing, among other peculiarities, cold-active enzymes that have the properties to cope with the reduction of chemical reaction rates induced by low temperatures. The emerging picture suggests that these enzymes display a high catalytic efficiency at low temperatures through an improved flexibility of the structural components involved in the catalytic cycle, whereas other protein regions, if not implicated in catalysis, may be even more rigid than their mesophilic counterparts. In return, the increased flexibility leads to a decreased stability of psychrophilic enzymes. In order to gain further advances in the analysis of the activity/flexibility/stability concept, psychrophilic, mesophilic, and thermophilic DNA ligases have been compared by three-dimensional-modeling studies, as well as regards their activity, surface hydrophobicity, structural permeability, conformational stabilities, and irreversible thermal unfolding. These data show that the cold-adapted DNA ligase is characterized by an increased activity at low and moderate temperatures, an overall destabilization of the molecular edifice, especially at the active site, and a high conformational flexibility. The opposite trend is observed in the mesophilic and thermophilic counterparts, the latter being characterized by a reduced low temperature activity, high stability and reduced flexibility. These results strongly suggest a complex relationship between activity, flexibility and stability. In addition, they also indicate that in cold-adapted enzymes, the driving force for denaturation is a large entropy change.


Sequence requirements for synthetic peptide-mediated translocation to the nucleus.
The abilities of 18 synthetic peptides to target a carrier protein to the nucleus following microinjection into the cytoplasm of HeLa cells were determined. Eight of the sequences chosen for synthesis were based on published nuclear targeting regions as determined by gene fusion and deletion experiments. Six of these sequences were found to be effective when mimicked by a synthetic peptide and conjugated to a carrier protein. One additional peptide was based on a region of lamin L1, a nuclear protein from Xenopus laevis, in which the nuclear targeting region had not been previously investigated. This peptide was also able to target a carrier protein to the nucleus. Eight other peptides which resemble the known targeting signals had little or no nuclear targeting ability. Peptides which were able to target a carrier protein to the nucleus did so within 45 min of injection into the cytoplasm. Two peptides with little or no apparent nuclear targeting ability after 45 min were examined for longer times as well. No increase in nuclear accumulation was observed between 45 min and 4 h after cytoplasmic injection. Comparison of the sequences which were effective at nuclear targeting with those that were not revealed a possible consensus sequence for peptide-mediated nuclear transport.


An expectation-maximization algorithm based Kalman smoother approach for single-trial estimation of event-related potentials.
This paper applies an expectation-maximization (EM) based Kalman smoother (KS) approach for single-trial event-related potential (ERP) estimation. Existing studies assume a Markov diffusion process for the dynamics of ERP parameters which is recursively estimated by optimal filtering approaches such as Kalman filter (KF). However, these studies only consider estimation of ERP state parameters while the model parameters are pre-specified using manual tuning, which is time-consuming for practical usage besides giving suboptimal estimates. We extend the KF approach by adding EM based maximum likelihood estimation of the model parameters to obtain more accurate ERP estimates automatically. We also introduce different model variants by allowing flexibility in the covariance structure of model noises. Optimal model selection is performed based on Akaike Information Criterion (AIC). The method is applied to estimation of chirp-evoked auditory brainstem responses (ABRs) for detection of wave V critical for assessment of hearing loss. Results shows that use of more complex covariances are better estimating inter-trial variability.


Adipose-derived stem cell therapy for intervertebral disc regeneration: an in vitro reconstructed tissue in alginate capsules.
The degenerative pathologies of the intervertebral disc have a remarkable social impact in the industrialized countries and can provide serious disabilities in the population. The current treatment consists of conservative treatments (such as symptomatic pharmacological therapies and physiokinetic therapy) and surgical treatments (intervertebral fusion, total disc replacement, nucleus pulposus (NP) replacement, or surgical exeresis). Recent advances in cell therapy foresee the possibility of regenerating the damaged disc; the autologous disc tissue can be withdrawn, in vitro regenerated, and re-implanted. The aim of this work was to verify whether autologous adipose-derived adult stem cells can improve the quality of an in vitro reconstructed nucleus pulposus tissue. A three-dimensional (3D) co-culture of NP cells and adipose tissue non-adipocyte fraction cells (nAFs) was assessed in a previously developed alginate 3D culture system following the good manufacturing practice guidelines to ensure patient safety for clinical studies. Morphological investigation of cultured and co-cultured cells was performed using transmission electron microscopy and immunofluorescence for collagen type I, aggrecan, CD90, CD34, and vimentin. Results indicate that co-culture of NP and nAFs improves the quality of the in vitro reconstructed tissue in term of extracellular matrix production and 3D cell organization. Technological resources are available for NP cell encapsulation intended for regenerating the intervertebral disc.


Nonassociative learning as gated neural integrator and differentiator in stimulus-response pathways.
Nonassociative learning is a basic neuroadaptive behavior exhibited across animal phyla and sensory modalities but its role in brain intelligence is unclear. Current literature on habituation and sensitization, the classic "dual process" of nonassociative learning, gives highly incongruous accounts between varying experimental paradigms. Here we propose a general theory of nonassociative learning featuring four base modes: habituation/primary sensitization in primary stimulus-response pathways, and desensitization/secondary sensitization in secondary stimulus-response pathways. Primary and secondary modes of nonassociative learning are distinguished by corresponding activity-dependent recall, or nonassociative gating, of neurotransmission memory. From the perspective of brain computation, nonassociative learning is a form of integral-differential calculus whereas nonassociative gating is a form of Boolean logic operator--both dynamically transforming the stimulus-response relationship. From the perspective of sensory integration, nonassociative gating provides temporal filtering whereas nonassociative learning affords low-pass, high-pass or band-pass/band-stop frequency filtering--effectively creating an intelligent sensory firewall that screens all stimuli for attention and resultant internal model adaptation and reaction. This unified framework ties together many salient characteristics of nonassociative learning and nonassociative gating and suggests a common kernel that correlates with a wide variety of sensorimotor integration behaviors such as central resetting and self-organization of sensory inputs, fail-safe sensorimotor compensation, integral-differential and gated modulation of sensorimotor feedbacks, alarm reaction, novelty detection and selective attention, as well as a variety of mental and neurological disorders such as sensorimotor instability, attention deficit hyperactivity, sensory defensiveness, autism, nonassociative fear and anxiety, schizophrenia, addiction and craving, pain sensitization and phantom sensations, etc.


Immune Responses Induced at One Hour Post Cataract Surgery Wounding of the Chick Lens.
While the lens is an avascular tissue with an immune-privileged status, studies have now revealed that there are immune responses specifically linked to the lens. The response to lens injury, such as following cataract surgery, has been shown to involve the activation of the resident immune cell population of the lens and the induction of immunomodulatory factors by the wounded epithelium. However, there has been limited investigation into the immediate response of the lens to wounding, particularly those induced factors that are intrinsic to the lens and its associated resident immune cells. Using an established chick embryo ex vivo cataract surgery model has made it possible to determine the early immune responses of this tissue to injury, including its resident immune cells, through a transcriptome analysis. RNA-seq studies were performed to determine the gene expression profile at 1 h post wounding compared to time 0. The results provided evidence that, as occurs in other tissues, the resident immune cells of the lens rapidly acquired a molecular signature consistent with their activation. These studies also identified the expression of many inflammatory factors by the injured lens that are associated with both the induction and regulation of the immune response.


Analysis of family history data for evidence of non-Mendelian inheritance resulting from vertical transmission.
A number of infections of man, as well as of other animal and plant species, are heavily dependent upon 'vertical transmission'--that is direct parent-to-progeny transfer--for their maintenance in host populations. Such vertical transmission may be considered as a form of inheritance. It is usually non-Mendelian. This paper discusses the implications of such inheritance for the distribution of disease in families. A method is described for making quantitative predictions of prevalence rates of infection and of disease within different classes of relatives of either infected or uninfected probands. It is pointed out that, whereas a maternal line excess is to be expected among relatives of positive probands, the opposite should be found in families of negative probands. Expected differences between maternal and paternal line prevalence rates of observable disease decline rapidly with distance of relationship from the proband, and are greatly reduced by diagnostic insensitivity (analogous to penetrance). The implications of this analytic method for the design of family history studies are discussed. Published data on familial breast cancer are reviewed, and found to show no evidence that this condition is associated with a non-integrated vertically transmitted agent.


Dendritic-cell-based therapeutic cancer vaccines.
The past decade has seen tremendous developments in novel cancer therapies through the targeting of tumor-cell-intrinsic pathways whose activity is linked to genetic alterations and the targeting of tumor-cell-extrinsic factors, such as growth factors. Furthermore, immunotherapies are entering the clinic at an unprecedented speed after the demonstration that T cells can efficiently reject tumors and that their antitumor activity can be enhanced with antibodies against immune-regulatory molecules (checkpoint blockade). Current immunotherapy strategies include monoclonal antibodies against tumor cells or immune-regulatory molecules, cell-based therapies such as adoptive transfer of ex-vivo-activated T cells and natural killer cells, and cancer vaccines. Herein, we discuss the immunological basis for therapeutic cancer vaccines and how the current understanding of dendritic cell and T cell biology might enable the development of next-generation curative therapies for individuals with cancer.


Altered monoamine system activities after prenatal and adult stress: A role for stress resilience?
Prenatal stress (PNS) and stress in adulthood are risk factors for development of major depressive disorder. The present study aimed to 1) confirm previous neuroendocrine and behavioral changes induced by PNS, and 2) to characterize the effect of early- and late life stress on the in vivo activity of monoamine systems. Gestational dams were restrained thrice daily under bright illumination from gestational day (GD)11-20. Behavior and neuroendocrine responses to the forced swim test (FST) were determined in adult (50-80 days) offspring, and electrophysiological single unit recordings of dorsal raphe nucleus serotonin (5-HT), ventral tegmental area dopamine (DA) and locus coeruleus norepinephrine (NE) neurons were obtained at baseline and 24h after the FST. Gestational dams did not habituate to chronic restraint stress, and PNS reduced the birth weight of offspring. In adulthood, swim stress elevated CORT levels longer in PNS animals, while it had no effect on swim behaviors. Baseline firing activity of 5-HT neurons was decreased in PNS animals, while the firing activity of NE and DA neurons was increased. Swim stress had no effect on the firing on 5-HT neurons, but normalized the firing activity of catecholamine neurons in PNS animals. The present data confirm previously established effects on neuroendocrine and physiological measures, and demonstrate an altering effect of PNS and stress on monoamine system activities in adulthood. Since PNS did not result in a depressive-like phenotype, these central changes following PNS might play reflect adaptive changes contributing to stress resilience in adulthood.


The structure of signal peptides from bacterial lipoproteins.
Statistical analysis of lipoprotein and non-lipoprotein signal peptides reveals that the two classes differ significantly only in the region close to the signal peptidase cleavage site. This region is apolar and has the consensus sequence LA(G,A) decreases C in the lipoproteins, but is polar and has small, uncharged residues in positions -3 and -1 in the non-lipoproteins. A simple search for matches to the lipoprotein consensus cleavage site suffices to discriminate between the two groups with close to 100% reliability.


Male involvement and maternal health outcomes: systematic review and meta-analysis.
The developing world accounts for 99% of global maternal deaths. Men in developing countries are the chief decision-makers, determining women's access to maternal health services and influencing their health outcomes. At present, it is unclear whether involving men in maternal health can improve maternal outcomes. This systematic review and meta-analysis aimed to investigate the impact of male involvement on maternal health outcomes of women in developing countries. Four electronic databases and grey literature sources were searched (up to May 2013), together with reference lists of included studies. Two reviewers independently screened and assessed the quality of studies based on prespecified criteria. Measures of effects were pooled and random effect meta-analysis was conducted, where possible. Fourteen studies met the inclusion criteria. Male involvement was significantly associated with reduced odds of postpartum depression (OR=0.36, 95% CI 0.19 to 0.68 for male involvement during pregnancy; OR=0.34, 95% CI 0.19 to 0.62 for male involvement post partum), and also with improved utilisation of maternal health services (skilled birth attendance and postnatal care). Male involvement during pregnancy and at post partum appeared to have greater benefits than male involvement during delivery. Male involvement is associated with improved maternal health outcomes in developing countries. Contrary to reports from developed countries, there was little evidence of positive impacts of husbands' presence in delivery rooms. However, more rigorous studies are needed to improve this area's evidence base.


Discovering key regulatory mechanisms from single-factor and multi-factor regulations in glioblastoma utilizing multi-dimensional data.
Glioblastoma (GBM) is the most common malignant brain cancer in adults. Investigating the regulatory mechanisms underlying GBM is effective for the in-depth study of GBM. The Cancer Genome Atlas (TCGA) project is producing large-scale data and makes the comprehensive study of the diverse regulatory mechanisms underlying GBM possible. Although there have been research studies on GBM with large-scale data, distinguishing different regulatory mechanisms and identifying the key regulation types remain challenging. In this study, we integrated multi-dimensional data of differentially expressed genes in GBM: copy number variation (CNV), gene expression, miRNA expression and methylation, by performing partial correlation analysis with the Lasso technique. Our results showed that there were single-factor and multi-factor regulatory mechanisms in GBM. In further analysis of the regulation subtypes, we discovered that single-factor and multi-factor regulations are potentially distinct in functionality. Moreover, multi-factor regulations especially the key regulation subtypes may be more relevant to GBM and affect many GBM-related genes such as ERBB2 and MAPK1. This study not only verifies the utility of multi-dimensional data integration into GBM research but also distinguishes the key multi-factor regulatory subtypes that may drive pathogenesis of GBM from various regulatory mechanisms.


CD4 T cell control of acute and latent murine gammaherpesvirus infection requires IFNgamma.
Murine gammaherpesvirus 68 (gammaHV68, MHV-68)-specific CD4 T cells control gammaHV68 infection by reducing the frequency of latently infected cells and by inhibiting viral replication. We have previously demonstrated that CD4 T cells do not require CD8 T or B cells to control gammaHV68 replication, demonstrating a helper-independent activity of CD4 T cells during gammaHV68 infection. The effector mechanism(s) required for this helper-independent function of CD4 T cells and for the inhibition of the establishment of latency by CD4 T cells are not known. Since IFNgamma has been previously shown to be important for control of acute, latent, and persistent gammaHV68 infection, we tested the hypothesis that CD4 T cells require IFNgamma to limit gammaHV68 latency and replication. We utilized a previously described system in which T cell receptor (TCR) transgenic T cells (DO.11.10) and a recombinant virus (gammaHV68.OVA) allow for evaluation of high numbers of virus-specific CD4 T cells during both acute and latent infection. We show here that virus-specific CD4 T cells require IFNgamma for their anti-viral function in both acute and latent gammaHV68 infection. We additionally show that an in vitro derived T helper type 1 (TH1) CD4 T cell clone, which produces IFNgamma, inhibits gammaHV68 replication after adoptive transfer into RAG mice. Together, data presented here demonstrate that both CD4 T cell-mediated helper-independent control of gammaHV68 replication and inhibition of the establishment of gammaHV68 latency require IFNgamma.


Ginkgo biloba extract EGb 761 in dementia: intent-to-treat analyses of a 24-week, multi-center, double-blind, placebo-controlled, randomized trial.
In 1996, Kanowski et al. reported about the beneficial effects of ginkgo biloba special extract EGb 761 (240 mg/day) in outpatients with pre-senile and senile primary degenerative dementia of the Alzheimer type (DAT) and multi-infarct dementia (MID) of mild to moderate severity. The comparison of the results of this double-blind, placebo-controlled, randomized, multi-center study with other dementia studies is hampered by the fact that only the responder analysis of the per-protocol (PP) population, which was pre-specified in the protocol as confirmatory analysis, has been published in detail so far. Moreover, cognitive functioning was measured using the Syndrom-Kurztest (SKT), whereas results of other studies are based on the Alzheimer's Disease Assessment Scale-Cognitive Subscale (ADAS-cog). Therefore, the conventional intention-to-treat (ITT) analysis of this study is provided with an estimation of ADAS-cog scores based on measured SKT scores. After 24 weeks of treatment, the ITT analysis of the SKT and estimated ADAS-cog scores revealed a mean decrease in the total score by -2.1 (95 % CI: -2.7; -1.5) points and -2.7 (95 % CI: -3.5; -1.9) points, respectively, for the EGb 761 group, which indicates an improvement in cognitive function. On the contrary, the placebo group exhibited only a minimal change of -1.0 (95 % CI: -1.6; -0.3) and -1.3 (95 % CI: -2.0; -0.4) points, respectively. The changes from baseline differed significantly between treatment groups by 1.1 (SKT) and 1.4 (estimated ADAS-cog) points, respectively (P = 0.01). The Clinical Global Impression of Change (CGI, Item 2) favored the EGb 761 group with a mean difference of 0.4 points (P = 0.007). Changes in the rating related to activities of daily living (Nürnberger-Alters-Beobachtungs-Skala, NAB) showed a favorable trend for EGb 761R. A subgroup analysis regarding patients with DAT yielded comparable results. Using a decrease of at least 4 points on the estimated ADAS-cog scores as cutoff criterion for treatment response, 35 % of EGb 761-treated patients were considered responders versus only 19 % for the placebo group (P = 0.01). The results of this ITT analysis substantiate the outcomes previously obtained with a responder analysis of the per-protocol population and confirm that EGb 761 improves cognitive function in a clinically relevant manner in patients suffering from dementia. The therapeutic effect is in line with the outcome of another EGb 761 study conducted in the U.S.


Pseudomonas cytochrome C-551 peroxidase. A purification procedure and study of CO-binding kinetics.
A procedure is described for the purification of cytochrome c peroxidase from Pseudomonas aeruginosa involving extraction by sonication, followed by acid precipitation and chromatography on only two types of gel. The final preparation had a purity ratio A407/A280 of 4.2, and was found to be essentially pure by isoelectric focusing. The enzyme was shown to be unstable during degassing under vacuum except in the presence of detergent. The kinetics of CO binding to dithionite-reduced peroxidase were studied with stopped-flow and flash-photolysis techniques, and the results obtained between pH 5 and 7 suggest the existence of two forms of dithionite-reduced enzyme in slow equilibrium.


Extracellular potassium modifies the structure of kidney epithelial cells in culture.
In animals fed a K-deficient diet, alterations in kidney cell structure and function occur in association with changes in the ionic composition of the extracellular fluid. The hypothesis that the extracellular K concentration mediates these changes in renal tissue was tested in cultures of monkey kidney epithelial cells (BSC-1 line) by reducing the K concentration of the culture medium from the control value of 5.4 to 3.2 mM. Exposure of BSC-1 cells to low-K medium raised the maximal rate of uptake for L-glutamic acid by 39% without a change in apparent Km. To determine whether this alteration in plasma membrane function had a structural correlate, studies of the cell surface were performed using scanning and transmission electron microscopy. Morphometric analysis of scanning electron micrographs revealed that the number of microvilli per cell per unit surface area was 45% greater in cells exposed to low-K medium for 3 min than those exposed to control medium. This observation was confirmed by transmission electron microscopy. The results indicate that an alteration in the extracellular K concentration per se can modify specific structural and functional characteristics of kidney epithelial cells.


Insulin-induced oedema in a patient with diabetes mellitus complicated by ketoacidosis.
In this article we reported a recent case of a 15-year-old grossly underweight (29 kg) Nigerian girl diagnosed of type 1 diabetes mellitus four years ago and who defaulted from follow up but presented with diabetic ketoacidosis. Glycaemic control was poor because of poor compliance. On the 5th day on admission, a non tender pitting oedema without skin discolouration developed over the ankles. The natural history of insulin-induced oedema was observed in this patient since the oedema resolved seven days later without any specific therapy, such as administration of diuretics. The major causes of generalized oedema in childhood and adolescence, such as kwashiorkor, nephrotic syndrome, liver cirrhosis, congestive heart failure and acute glumerulonephritis were excluded by findings from the history, physical examination and relevant laboratory investigations. Having excluded these major causes of oedema, the obvious conclusion was that the insulin therapy was the cause of the oedema observed in our patient. The aim of this article is to review existing medical literature on the subject of insulin-induced oedema and raise the awareness of clinicians on the subject. In conclusion, insulin-induced oedema should be considered in the differential diagnosis of oedema in children and adolescents with type 1 diabetes mellitus complicated by ketoacidosis, particularly if they are underweight.


Interdental oral hygiene interventions elicit varying compositional microbiome changes in naturally occurring gingivitis: Secondary data analysis from a clinical trial.
To evaluate the effect of different oral irrigators on the sub-gingival microbiome composition in patients with naturally occurring plaque-induced gingivitis. Sub-gingival plaque was collected from adults participating in a clinical trial assessing the efficacy of oral hygiene with two different oral irrigators (Waterpik Water Flosser [Group 1] and Oral-B Water Flosser [Group 2]) versus dental flossing (Group 3) for microbiome analysis. Plaque samples were reflective of naturally occurring plaque-induced gingivitis at baseline and of gingival health at the endpoint (4 weeks). Clinical measures of gingival inflammation were collected, and the sub-gingival microbiome was analysed by 16S rRNA sequencing to identify amplicon sequence variants. Oral hygiene instruction with self-performed manual toothbrushing and water-jet irrigation led to significant reductions in inflammation for all groups; both oral irrigators outperformed flossing in bleeding-on-probing reduction (p < .001). Microbiome diversity of sub-gingival plaque remained relatively stable over time, but significant changes were noted in certain taxa, consistent with increases in the relative abundance of commensals and reductions in late colonizers and periodontal pathogens in the water-jet groups. Reduction in gingival inflammation at 4 weeks within the water-jet groups is accompanied by slight but critical changes in microbiome composition. Although biodiversity does not substantially change within 4 weeks during the resolution of naturally induced gingivitis, significant relative increases in commensal early colonizers such as Streptococcus, Veillonella and Fusobacterium were accompanied by a shift towards a less anaerobic microbiota associated with return to health. These changes were contingent upon the type of interdental hygiene, with Group 1 exhibiting more significant alterations in microbiome composition towards a periodontal-health-compatible community.


Rare case of sclerosing mesenteritis and low grade follicular lymphoma.
An unusual case of long standing sclerosing mesenteritis; initially presented with recurrent abdominal pain and a mesenteric mass with surrounding fat oedema and stranding with a pseudocapsule and fat ring sign were clearly visualised on the initial computed tomography scan. Laparotomy showed diffuse thickening at the root of the mesentery and histology from this specimen revealed fat necrosis and reactive lymphoid tissue consistent with sclerosing mesenteritis. Initial treatment with steroids and tamoxifen relieved the symptoms and the mass. He was maintained on tamoxifen. Three years later he developed a recurrence of his symptoms and abdominal mass that responded to a course of steroids. Two years following this, he developed a follicular Hodgkin's lymphoma.


[Deliberate heart arrest in man and its possible mechanism].
A case of deliberate 7-second heart arrest is described. It is assumed that the effect is the result of voluntary self-irritation of peritoneal receptors by means of rhythmical diaphragm muscle contractions realized in conditions of preliminary breathing arrest.


Advancing Antiamyloidogenic Activity by Fine-Tuning Macromolecular Topology.
Amyloid β peptide can aggregate into thin β-sheet fibrils or plaques deposited on the extracellular matrix, which is the hallmark of Alzheimer's disease. Multifunctional macromolecular structures play an important role in inhibiting the aggregate formation of amyloidogenic materials and thus are promising candidates with antiamyloidogenic characteristics for the development of next-generation therapeutics. In this study, we evaluate how small differences in the dendritic topology of these structures influence their antiamyloidogenic activity by the comparison of "perfectly dendritic" and "pseudodendritic" macromolecules, both decorated with mannose units. Their compactness, the position of surface units, and the size of glyco-architectures influence their antiamyloidogenic activity against Aβ 40, a major component of amyloid plaques. For the advanced analysis of the aggregation of the Aβ peptide, we introduce asymmetric flow field flow fractionation as a suitable method for the quantification of large and delicate structures. This alternative method focuses on the quantification of complex aggregates of Aβ 40 and glycodendrimer/glyco-pseudodendrimer over different time intervals of incubation, showing a good correlation to ThT assay and CD spectroscopy results. Kinetic studies of the second-generation glyco-pseudodendrimer revealed maximum inhibition of Aβ 40 aggregates, verified with atomic force microscopy. The second-generation glyco-pseudodendrimer shows the best antiamyloidogenic properties confirming that macromolecular conformation in combination with optimal functional group distribution is the key to its performance. These molecular properties were validated and confirmed by molecular dynamics simulation.


Decellularized human cornea for reconstructing the corneal epithelium and anterior stroma.
In this project, we strived to develop a decellularized human cornea to use as a scaffold for reconstructing the corneal epithelium and anterior stroma. Human cadaver corneas were decellularized by five different methods, including detergent- and nondetergent-based approaches. The success of each method on the removal of cells from the cornea was investigated. The structural integrity of decellularized corneas was compared with the native cornea by electron microscopy. The integrity of the basement membrane of the epithelium was analyzed by histology and by the expression of collagen type IV, laminin, and fibronectin. Finally, the ability of the decellularized corneas to support the growth of human corneal epithelial cells and fibroblasts was assessed in vitro. Corneas processed using Triton X-100, liquid nitrogen, and poly(ethylene glycol) resulted in incomplete removal of cellular material. Corneas processed with the use of sodium dodecyl sulfate (SDS) or with sodium chloride (NaCl) plus nucleases successfully removed all cellular material; however, only the NaCl plus nuclease treatment kept the epithelial basement membrane completely intact. Corneas processed with NaCl plus nuclease supported both fibroblast and epithelial cell growth in vitro, while corneas treated with SDS supported the growth of only fibroblasts and not epithelial cells. Decellularized human corneas provide a scaffold that can support the growth of corneal epithelial cells and stromal fibroblasts. This approach may be useful for reconstructing the anterior cornea and limbus using autologous cells.


A least squares quantization table method for direct reconstruction of MR images with non-Cartesian trajectory.
The direct Fourier transform method is a straightforward solution with high accuracy for reconstructing magnetic resonance (MR) images from nonuniformly sampled k-space data, given that the optimal density compensation function is selected and the underlying magnetic field is sufficiently uniform. The computation however is very time-consuming, making it impractical especially for large-size images. In this paper, the least squares quantization table (LSQT) method is proposed to accelerate the direct Fourier transform computation, similar to the recently proposed methods such as using look-up table (LUT) or equal-phase-line (EPL). With LSQT, all the image pixels are first classified into several groups where the Lloyd-Max quantization scheme is used to ensure the minimal classification error. The representative value of each group is stored in a small-size LSQT in advance to reduce the computational load. The pixels in the same group receive the same contribution, which is calculated only once for each group instead of for each pixel, resulting in the reduction of computation because the number of groups is far smaller than the number of pixels. Finally, each image pixel is mapped into the nearest group and its representative value is used to reconstruct the image. The experimental results show that the LSQT method requires far smaller memory size than the LUT method and fewer multiplication operations than the LUT and EPL methods. Moreover, the LSQT method can perform large-size reconstructions that achieve comparable or higher accuracy as compared to the EPL and gridding methods when the appropriate parameters are given. The inherent parallel structure also makes the LSQT method easily adaptable to a multiprocessor system.


Lipid bilayer regulation of membrane protein function: gramicidin channels as molecular force probes.
Membrane protein function is regulated by the host lipid bilayer composition. This regulation may depend on specific chemical interactions between proteins and individual molecules in the bilayer, as well as on non-specific interactions between proteins and the bilayer behaving as a physical entity with collective physical properties (e.g. thickness, intrinsic monolayer curvature or elastic moduli). Studies in physico-chemical model systems have demonstrated that changes in bilayer physical properties can regulate membrane protein function by altering the energetic cost of the bilayer deformation associated with a protein conformational change. This type of regulation is well characterized, and its mechanistic elucidation is an interdisciplinary field bordering on physics, chemistry and biology. Changes in lipid composition that alter bilayer physical properties (including cholesterol, polyunsaturated fatty acids, other lipid metabolites and amphiphiles) regulate a wide range of membrane proteins in a seemingly non-specific manner. The commonality of the changes in protein function suggests an underlying physical mechanism, and recent studies show that at least some of the changes are caused by altered bilayer physical properties. This advance is because of the introduction of new tools for studying lipid bilayer regulation of protein function. The present review provides an introduction to the regulation of membrane protein function by the bilayer physical properties. We further describe the use of gramicidin channels as molecular force probes for studying this mechanism, with a unique ability to discriminate between consequences of changes in monolayer curvature and bilayer elastic moduli.


[The hypersensitive respiratory system in childhood].
Aetiological factors and mechanisms of bronchial hyperreactivity are discussed. The hypothesis of increased permeability of the bronchial mucosa--caused by either endogenous or exogenous stimuli--as the most plausible explanation for the connection between hyperreactivity and childhood asthma bronchiale is emphasized. Finally, unspecific inhalative provocation tests for proof of bronchial hyperreactivity, its indications and contraindications as well as the criteria of judgement are mentioned.


Dry needling for the treatment of muscle spasticity in a patient with multiple sclerosis: a case report.
Spasticity is a common cause of disability in multiple sclerosis (MS), which can negatively affect the patient's walking and balance. To evaluate the immediate effect of dry needling (DN) on spasticity and mobility in a female with MS. In this case, a 38-year-old female with a 4-year history of MS was treated. The hamstring muscles (biceps femoris and semitendinosus) were needled for 1 minute in a single session. The main outcome measures were the Modified Modified Ashworth Scale (MMAS) to evaluate spasticity, the Timed 25-Foot Walk (T25FW) for the assessment of mobility and leg function performance, and stiffness as a biomechanical index of spasticity measured by a dynamometer. The assessments were done before and immediately after DN. The MMAS scores decreased in the hamstrings (1 to 0) and quadriceps (2 to 1). The mobility improved as the time for T25FW decreased from 16.30 to 9.29 seconds. The stiffness of hamstring decreased after treatment (0.451 to 0.312). One session of DN for the hamstring muscle decreased spasticity and improved mobility in this patient with MS. Further studies are suggested.


A prospective evaluation of positron emission tomography scanning, sentinel lymph node biopsy, and standard axillary dissection for axillary staging in patients with early stage breast cancer.
Positron emission tomography (PET) is a noninvasive imaging modality that can detect malignant lymph nodes. This study determined the sensitivity, specificity, predictive values, and likelihood ratios of PET scanning compared with standard axillary lymph node dissection (ALND) and sentinel lymph node biopsy (SLNB) in staging the axilla in women with early stage breast cancer. Women with clinical stage I or II breast cancer had whole body PET scanning before ALND and SLNB, in a prospective, blinded protocol. ALND were evaluated by standard hematoxylin and eosin (H&E) staining techniques, while sentinel nodes were also examined for micrometastatic disease. A total of 98 patients were recruited. PET compared with ALND demonstrated sensitivity of 0.40 (95% CI, 0.16, 0.68), specificity 0.97 (CI, 0.90, 0.99), positive likelihood ratio 14.4 (CI, 3.21, 64.5), positive predictive value 0.75 (CI, 0.35, 0.97), and false-negative rate of 0.60 (CI, 0.32, 0.84). Test properties were similar for PET compared with sentinel nodes positive by H&E staining. A few false-positive scans (0.028, CI, 0.003, 0.097) were seen. Multiple logistic regression analysis found that PET accuracy was better in patients with high grade and larger tumors. Increased size and number of positive nodes were also associated with a positive PET scan. The sensitivity of PET compared with ALND and SLNB was low, whereas PET scanning had high specificity and positive predictive values. The study suggests that PET scanning cannot replace histologic staging in early stage breast cancer. The low rate of false-positive findings suggests that PET can identify women who can forego SLNB and require full axillary dissection.


Comparative Pharmacokinetics of High and Low Doses of the Herbicide Propanil in Mice.
We have documented that the herbicide propanil is immunotoxic in mice, and our in vitro tissue culture experiments largely recapitulate the in vivo studies. Laboratory studies on environmental contaminants are the most meaningful when these studies are conducted using concentrations that approximate levels in the environment. Many techniques to measure the distribution and pharmacokinetics (PK) on compounds rely on techniques, such as liquid scintillation counting (LSC) of radio-labeled starting compound, that require concentrations higher than environmental levels. The aim of this study was to compare tissue PK after exposure to propanil concentrations more relevant to levels of exposure to agricultural workers and the general population to concentrations previously reported for laboratory studies. To this end, we conducted a study to measure propanil distribution in three immune organs, using ultrasensitive accelerator mass spectrometry (AMS). We used two doses: the lower dose modeled levels expected in the environment or long-term occupational exposure to low doses, while the higher dose was to model the effects of an accidental exposure. Our results showed that the distribution and PK profiles from these two different concentrations was markedly different. The profile of the high dose (concentration) exposure was indicative of saturation of the detoxifying capability of the animal. In contrast, at the lower environmentally relevant concentration, in vivo concentrations of propanil in spleen, liver, and blood dropped to a very low level by 720 min. In conclusion, these studies highlight the differences in PK of propanil at these two doses, which suggests that the toxicity of this chemical should be re-investigated to obtain better data on toxic effects at doses relevant for humans.


Management of non-urgent paediatric emergency department attendances by GPs: a retrospective observational study.
Non-urgent emergency department (ED) attendances are common among children. Primary care management may not only be more clinically appropriate, but may also improve patient experience and be more cost-effective. To determine the impact on admissions, waiting times, antibiotic prescribing, and treatment costs of integrating a GP into a paediatric ED. Retrospective cohort study explored non-urgent ED presentations in a paediatric ED in north-west England. From 1 October 2015 to 30 September 2017, a GP was situated in the ED from 2.00 pm until 10.00 pm, 7 days a week. All children triaged as 'green' using the Manchester Triage System (non-urgent) were considered to be 'GP appropriate'. In cases of GP non-availability, children considered non-urgent were managed by ED staff. Clinical and operational outcomes, as well as the healthcare costs of children managed by GPs and ED staff across the same timeframe over a 2-year period were compared. Of 115 000 children attending the ED over the study period, a complete set of data were available for 13 099 categorised as 'GP appropriate'; of these, 8404 (64.2%) were managed by GPs and 4695 (35.8%) by ED staff. Median duration of ED stay was 39 min (interquartile range [IQR] 16-108 min) in the GP group and 165 min (IQR 104-222 min) in the ED group (P<0.001). Children in the GP group were less likely to be admitted as inpatients (odds ratio [OR] 0.16; 95% confidence interval [CI] = 0.13 to 0.20) and less likely to wait >4 hours before being admitted or discharged (OR 0.11; 95% CI = 0.08 to 0.13), but were more likely to receive antibiotics (OR 1.42; 95% CI = 1.27 to 1.58). Treatment costs were 18.4% lower in the group managed by the GP (P<0.0001). Given the rising demand for children's emergency services, GP in ED care models may improve the management of non-urgent ED presentations. However, further research that incorporates causative study designs is required.


Safety and efficacy of cardiopoietic stem cells in the treatment of post-infarction left-ventricular dysfunction - From cardioprotection to functional repair in a translational pig infarction model.
To date, clinical success of cardiac cell-therapies remains limited. To enhance the cardioreparative properties of stem cells, the concept of lineage-specification through cardiopoietic-guidance has been recently suggested. However, so far, only results from murine studies and from a clinical pilot-trial in chronic heart-failure (CHF) are available, while systematic evidence of its therapeutic-efficacy is still lacking. Importantly, also no data from large animals or for other indications are available. Therefore, we here investigate the therapeutic-efficacy of human cardiopoietic stem cells in the treatment of post-infarction LV-dysfunction using a translational pig-model. Using growth-factor priming, lineage-specification of human bone-marrow derived MSCs was achieved to generate cardiopoietic stem cells according to GMP-compliant protocols. Thereafter, pigs with post-infarction LV-dysfunction (sub-acute phase;1-month) were randomized to either receive transcatheter NOGA 3D electromechanical-mapping guided intramyocardial transplantation of cardiopoietic cells or saline (control). After 30days, cardiac MRI (cMRI) was performed for functional evaluation and in-vivo cell-tracking. This approach was coupled with a comprehensive post-mortem cell-fate and mode-of-repair analysis. Cardiopoietic cell therapy was safe and ejection-fraction was significantly higher when compared to controls (p = 0.012). It further prevented maladaptive LV-remodeling and revealed a significantly lower relative and total infarct-size (p = 0.043 and p = 0.012). As in-vivo tracking and post-mortem analysis displayed only limited intramyocardial cardiopoietic cell-integration, the significant induction of neo-angiogenesis (∼40% higher; p = 0.003) and recruitment of endogenous progenitors (∼2.5x higher; p = 0.008) to the infarct border-zone appeared to be the major modes-of-repair. This is the first report using a pre-clinical large animal-model to demonstrate the safety and efficacy of cardiopoietic stem cells for the treatment of post-infarction LV-dysfunction to prevent negative LV-remodeling and subsequent CHF. It further provides insight into post-delivery cardiopoietic cell-fate and suggests the mechanisms of cardiopoietic cell-induced cardiac-repair. The adoption of GMP-/GLP-compliant methodologies may accelerate the translation into a phase-I clinical-trial in patients with post-ischemic LV-dysfunction broadening the current indication of this interesting cell-type.


Pulmonary thromboembolism in cats: 29 cases (1987-1997).
To determine clinical signs, clinicopathologic abnormalities, radiographic findings, histologic abnormalities, and predisposing factors or diseases in cats with pulmonary thromboembolism (PTE). Retrospective study. 29 cats in which PTE was confirmed at necropsy. Information on signalment, body weight, history, results of physical examination, results of CBC and serum biochemical analyses, whether PTE was suspected prior to death, type of indwelling venous catheter and duration of venous catheterization, results of thoracic radiography, and whether cats had any concurrent diseases was obtained from medical records. PTE was identified in cats of various ages (median, 8.7 years), weights (median, 4.1 kg [9 lb]), and breeds. The most common owner-reported problems included lethargy (17 cats), anorexia (14), weight loss (10), and difficulty breathing (8); physical abnormalities included lethargy (21), tachypnea or dyspnea (16), and dehydration (13). Clinicopathologic abnormalities reflected concurrent or underlying diseases. Common radiographic abnormalities included pulmonary vessel abnormalities (11), pleural effusion (8), and peripheral noncircumscribed consolidations (6). Underlying or predisposing conditions, including cardiac disease (12), neoplasia (10), corticosteroid administration (8), disseminated intravascular coagulation (5), protein-losing nephropathy (4) or enteropathy (4), immune-mediated hemolytic anemia (2), and sepsis (2) were identified in all cats. Results suggest that PTE can develop in cats of any age, sex, or breed. Because PTE is a serious, potentially life-threatening disease, it should be suspected in cats with thoracic radiographic changes suggestive of uneven distribution of blood flow between lung lobes, especially in cats that have predisposing factors or diseases.


Immunological detection of Chinese hamster ovary cells expressing a multidrug resistance phenotype.
A monoclonal antibody (IgG1) has been prepared that specifically detects Chinese hamster ovary cells expressing a multidrug-resistant phenotype. This antibody recognizes the membrane P-glycoprotein (Mr 170,000) associated with drug resistance as determined by enzyme-linked immunosorbent assay with purified P-glycoprotein and by Western blot analysis of cell extracts from drug-resistant and drug-sensitive cells. By immunofluorescence methods, the antibody also reacts strongly with viable and ether:ethanol-fixed resistant cells but does not react with the parent drug-sensitive cell line. Thus, this antibody can bind with live cells allowing discrimination by immunohistochemistry between drug-resistant and drug-sensitive Chinese hamster ovary cells.


[Effect of natural light during "white nights" and melatonin on circadian rhythm of total antioxidant capacity of human saliva].
The results of our previous experiments indicate that total antioxidant capacity (TAC) of human saliva has a distinct circadian rhythm with maximum in the night and minimum in the daytime. The decrease of amplitude and other parameters of circadian rhythm of TAC of saliva in summer is marked. The aim of the present study was to evaluate an effect of light at night and exogenous melatonin (MT) on this parameter. There were 6 participants in experiments, at which 28 circadian rhythms have been analysed. The method of coulometric titration was used for measuring TAC of saliva. Light at night significantly decreases amplitude (P<0.001) and exogenous MT increases the level of AUC (P<0.05) and mesor (P<0.05). MT in vitro has a highest antioxidant activity among water soluble antioxidants tested (MT, quercetin, ascorbat, mitophen, white lignin, rutin). Pharmacological quantities of MT (0.1%) increases TAC of saliva in vitro as high as three times. The results of calculations indicate that specific TAC of MT is not enough to explain increase in TAC of saliva in vivo conditions. Consequently, MT stimulates biosynthesis and/or redistribution of low-molecular weight antioxidants in the human organism.


Oral Development and Pathology.
The mouth is integral in the development of feeding, the initiation of digestion, and for speech and socialization. Signs of systemic disease and nutritional deficiencies often manifest in the mouth, and poor oral health can exacerbate many systemic conditions. This review addresses the fetal development of the mouth, major anomalies, common minor physical findings, and pathologic conditions and their management. Pediatric practitioners have historically been poorly trained in diagnosis and management of oral conditions, so this article provides an overview of oral embryology and pathology, with a focus on hard and soft tissue disease identification, triage, and management. For primary prevention to be effective, pediatric providers must be knowledgeable about the process of dental caries, prevention of the disease, and available interventions, including fluoride. The embryology and anatomy of the oral cavity are complex, and the mouth is crucial to many physiologic processes. Pediatric primary care providers are uniquely positioned to prevent, identify, and triage dental caries, the most common chronic disease of childhood.


Consequences of warming and resource quality on the stoichiometry and nutrient cycling of a stream shredder.
As a result of climate change, streams are warming and their runoff has been decreasing in most temperate areas. These changes can affect consumers directly by increasing their metabolic rates and modifying their physiology and indirectly by changing the quality of the resources on which organisms depend. In this study, a common stream detritivore (Echinogammarus berilloni Catta) was reared at two temperatures (15 and 20°C) and fed Populus nigra L. leaves that had been conditioned either in an intermittent or permanent reach to evaluate the effects of resource quality and increased temperatures on detritivore performance, stoichiometry and nutrient cycling. The lower quality (i.e., lower protein, soluble carbohydrates and higher C:P and N:P ratios) of leaves conditioned in pools resulted in compensatory feeding and lower nutrient retention capacity by E. berilloni. This effect was especially marked for phosphorus, which was unexpected based on predictions of ecological stoichiometry. When individuals were fed pool-conditioned leaves at warmer temperatures, their growth rates were higher, but consumers exhibited less efficient assimilation and higher mortality. Furthermore, the shifts to lower C:P ratios and higher lipid concentrations in shredder body tissues suggest that structural molecules such as phospholipids are preserved over other energetic C-rich macromolecules such as carbohydrates. These effects on consumer physiology and metabolism were further translated into feces and excreta nutrient ratios. Overall, our results show that the effects of reduced leaf quality on detritivore nutrient retention were more severe at higher temperatures because the shredders were not able to offset their increased metabolism with increased consumption or more efficient digestion when fed pool-conditioned leaves. Consequently, the synergistic effects of impaired food quality and increased temperatures might not only affect the physiology and survival of detritivores but also extend to other trophic compartments through detritivore-mediated nutrient cycling.


DETECTION OF HUMAN HERPESVIRUS-8 ANTIGEN IN HIV-INFECTED PATIENTS IN EAST JAVA, INDONESIA.
Co-infection of human herpesvirus-8 (HHV-8) in HIV-positive people might cause Kaposi's sarcoma. Early detection of HHV-8 may prevent the onset of clinical manifestations. In Indonesia, detection of HHV-8 antigen in HIV-positive patients has yet to be reported. The intention of this research was to examine the presence of HHV-8 antigen in HIV-positive patients in East Java, Indonesia. 103 serum samples were collected from HIV-positive patients in Surabaya and Tulungagung, East Java, Indonesia. Serums were then tested for the presence of HHV-8 antigen by using sandwich ELISA. Human Herpesvirus-8 antigen was detected in 15 samples (14.5%). The presence of HHV-8 infection in HIV-positive patients did not present differently in males and females and among different age groups. Human immunodeficiency virus-positive serum samples were collected from 23 homosexual men, 25 intravenous drug users (IVDUs) and 52 heterosexuals. In the male homosexual group, HHV-8 antigen was detected in 21.7% (5/23) of the samples, while in the intravenous drug user group (IVDUs), 16% (4/25) of the samples were found to have HHV-8 antigen. This research found the presence of HHV-8 antigen in 14.5% of patients in East Java, Indonesia. It is recommended that patients with a positive result should receive further examination to detect any clinical manifestations related to HHV-8 infection, especially in the form of Kaposi's sarcoma lesions, so that the illness can be appropriately managed.


A comparison of recombinant human thyrotropin and thyroid hormone withdrawal for the detection of thyroid remnant or cancer.
Recombinant human TSH has been developed to facilitate monitoring for thyroid carcinoma recurrence or persistence without the attendant morbidity of hypothyroidism seen after thyroid hormone withdrawal. The objectives of this study were to compare the effect of administered recombinant human TSH with thyroid hormone withdrawal on the results of radioiodine whole body scanning (WBS) and serum thyroglobulin (Tg) levels. Two hundred and twenty-nine adult patients with differentiated thyroid cancer requiring radioiodine WBS were studied. Radioiodine WBS and serum Tg measurements were performed after administration of recombinant human TSH and again after thyroid hormone withdrawal in each patient. Radioiodine whole body scans were concordant between the recombinant TSH-stimulated and thyroid hormone withdrawal phases in 195 of 220 (89%) patients. Of the discordant scans, 8 (4%) had superior scans after recombinant human TSH administration, and 17 (8%) had superior scans after thyroid hormone withdrawal (P = 0.108). Based on a serum Tg level of 2 ng/mL or more, thyroid tissue or cancer was detected during thyroid hormone therapy in 22%, after recombinant human TSH stimulation in 52%, and after thyroid hormone withdrawal in 56% of patients with disease or tissue limited to the thyroid bed and in 80%, 100%, and 100% of patients, respectively, with metastatic disease. A combination of radioiodine WBS and serum Tg after recombinant human TSH stimulation detected thyroid tissue or cancer in 93% of patients with disease or tissue limited to the thyroid bed and 100% of patients with metastatic disease. In conclusion, recombinant human TSH administration is a safe and effective means of stimulating radioiodine uptake and serum Tg levels in patients undergoing evaluation for thyroid cancer persistence and recurrence.


Bundle to reduce unplanned extubation in a neonatal intensive care unit.
Objective: To evaluate the effectiveness of a bundle to reduce unplanned extubations (UE) in ventilated newborn infants (NB) and to verify the factors associated to UE.Method: Intervention study with a historical control group in a university hospital neonatal intensive care unit (NICU) between June 2014-May 2015 (Period I) and September 2015-August 2016 (Period II). All ventilated NB were included except those with facial malformations. The bundle (new tracheal tube fixation model, team training, identification of NB at risk of UE, and debriefing after UE episodes) was implemented between Periods I and II. Rates of UE/100 NB ventilated-day were compared between periods for the entire sample and according to the cause: accidental or by medical indication. Factors associated to the first UE episode of each NB were studied by logistic regression.Results: A total of 231 intubations were performed in 120 infants in Period I (gestational age 33.6 ± 4.7 W; birth weight: 2020 ± 929 g) and 212 intubations in 131 infants in Period II (34.2 ± 4.7 W; 2080 ± 997 g). UE occurred in 19.9% and 14.6% of the NB, in Periods I and II, respectively. Accidental extubation and change of the tube by medical indication were observed in 58.7% and 41.3% of UE in Period I and in 51.6% and 48.4% in Period II. Higher birth weight, lower SNAPPE-II score, and daytime period were associated with a lower chance of UE in all newborns.Conclusion: The bundle did not reduce the UE in NB ventilated in NICU but continued control of UE rates is crucial for improved care, especially for immature and critically neonates.What is new about the paper? The study presented a strategy for assessing the causes of unplanned extubations in a Neonatal Intensive Care Unit, considering not only the accidental extubations, but aldo the medical ordered extubations, which contributes to the definition of actions for the reduction of unplanned extubations in the NICU setting.


In silico and in vivo anti-angiogenic validation on ethanolic extract of Curcuma longa and curcumin compound in hepatocellular carcinoma through mitogen activated protein kinase expression in male and female wistar rats.
Hepatocellular carcinoma (HCC) is the most frequent primary malignancy of the liver. The aim of this study is to evaluate the comparative in silico and in vivo ameliorative potential of the ethanolic extract of Curcuma longa (EECL) in male and female Wistar rats administered N-nitrosodiethylamine-induced hepatocellular carcinoma. The MAPK compound was obtained from a protein data bank (PDB ID: 7AUV) for molecular docking. One hundred and twenty Wistar rats, were randomly selected into twelve groups (n = 5): Group A received regular diets as a basal control; groups B to G were administered 100 mg/kg NDEA twice in two weeks; while groups C to E received 200 mg/kg, 400 mg/kg, and 600 mg/kg of EECL; group F was treated with 200 mg/kg pure curcumin; and group G received 100 mg/kg Sylibon-140. Group H received only 200 mg/kg pure curcumin, and group I received 200 mg/kg of dimethylsulfoxide (DMSO). Groups J, K, and L received 200 mg/kg, 400 mg/kg and 600 mg/kg of EECL. MAPK and AFP mRNA in Wistar rats administered NDEA were upregulated as compared to EECL groups. In conclusion, the in silico and in vitro study validates the mitigating role of ethanolic extract of Curcuma longa and pure curcumin.


Formation of mitoxantrone adducts in human tumor cells: potentiation by AN-9 and DNA methylation.
The ability of mitoxantrone to form DNA adducts was investigated in a series of human tumor cell lines consisting of human cervical cancer (HeLa), human breast cancer (MCF-7), and human neuroblastoma (IMR-32) cells. The mitoxantrone-resistant human promyelocytic leukemia cell line HL60/MX2 was also compared to the parental cell line HL60 in terms of adduct formation in cellular DNA, RNA, and protein. DNA adduct formation detected using [14C]mitoxantrone as a single agent occurred at very low levels but addition of the formaldehyde-releasing prodrug AN-9 (pivaloyloxymethyl butyrate) increased adduct formation considerably in all cell lines tested. Adduct formation increased when increasing ratios of AN-9 were used, and were observed at maximal levels when AN-9 addition was 4 h after the addition of mitoxantrone. However, low levels of adducts were observed when AN-9 addition was 16 h prior to mitoxantrone. The ability of [14C]mitoxantrone to form adducts with DNA, RNA, and protein was assessed in HL60 cells, and DNA was found to be the major substrate for adduct formation. RNA was also shown to be a good substrate while protein adduct levels were consistently very low. In mitoxantrone-resistant HL60/MX2 cells, DNA adduct levels were approximately fourfold lower. To establish the influence of DNA methylation on the ability of mitoxantrone to form adducts in cells, decitabine was used to reduce DNA methylation levels in cells prior to mitoxantrone treatment. This was clearly shown to influence adduct formation, with increasing decitabine levels leading to a decrease in the level of adducts observed in both IMR-32 and MCF-7 cell lines. Collectively, these results suggest that two major factors that influence the extent of mitoxantrone adduct formation in cells are the availability of formaldehyde and the extent of genomic DNA methylation.


Claudin-5 Redistribution Induced by Inflammation Leads to Anti-VEGF-Resistant Diabetic Macular Edema.
Approximately 40% of patients with diabetic macular edema (DME) are resistant to anti-vascular endothelial growth factor (VEGF) therapy (rDME). Here, we demonstrate that significant correlations between inflammatory cytokines and VEGF, as observed in naive DME, are lost in patients with rDME. VEGF overexpression in the mouse retina caused delayed inflammatory cytokine upregulation, monocyte/macrophage infiltration (CD11b+ Ly6C+ CCR2+ cells), macrophage/microglia activation (CD11b+ CD80+ cells), and blood-retinal barrier disruption due to claudin-5 redistribution, which did not recover with VEGF blockade alone. Phosphorylated protein analysis of VEGF-overexpressed retinas revealed rho-associated coiled-coil-containing protein kinase (ROCK) activation. Administration of ripasudil, a selective ROCK inhibitor, attenuated retinal inflammation and claudin-5 redistribution. Ripasudil also contributed to the stability of claudin-5 expression by both transcriptional enhancement and degradation suppression in inflammatory cytokine-stimulated endothelium. Notably, the anti-VEGF agent and the ROCK inhibitor were synergic in suppressing cytokine upregulation, monocyte/macrophage infiltration, macrophage/microglia activation, and claudin-5 redistribution. Furthermore, in vitro analysis confirmed that claudin-5 redistribution depends on ROCK2 but not on ROCK1. This synergistic effect was also confirmed in human rDME cases. Our results suggest that ROCK-mediated claudin-5 redistribution by inflammation is a key mechanism in the anti-VEGF resistance of DME.


Genomic models of short-term exposure accurately predict long-term chemical carcinogenicity and identify putative mechanisms of action.
Despite an overall decrease in incidence of and mortality from cancer, about 40% of Americans will be diagnosed with the disease in their lifetime, and around 20% will die of it. Current approaches to test carcinogenic chemicals adopt the 2-year rodent bioassay, which is costly and time-consuming. As a result, fewer than 2% of the chemicals on the market have actually been tested. However, evidence accumulated to date suggests that gene expression profiles from model organisms exposed to chemical compounds reflect underlying mechanisms of action, and that these toxicogenomic models could be used in the prediction of chemical carcinogenicity. In this study, we used a rat-based microarray dataset from the NTP DrugMatrix Database to test the ability of toxicogenomics to model carcinogenicity. We analyzed 1,221 gene-expression profiles obtained from rats treated with 127 well-characterized compounds, including genotoxic and non-genotoxic carcinogens. We built a classifier that predicts a chemical's carcinogenic potential with an AUC of 0.78, and validated it on an independent dataset from the Japanese Toxicogenomics Project consisting of 2,065 profiles from 72 compounds. Finally, we identified differentially expressed genes associated with chemical carcinogenesis, and developed novel data-driven approaches for the molecular characterization of the response to chemical stressors. Here, we validate a toxicogenomic approach to predict carcinogenicity and provide strong evidence that, with a larger set of compounds, we should be able to improve the sensitivity and specificity of the predictions. We found that the prediction of carcinogenicity is tissue-dependent and that the results also confirm and expand upon previous studies implicating DNA damage, the peroxisome proliferator-activated receptor, the aryl hydrocarbon receptor, and regenerative pathology in the response to carcinogen exposure.


[Rescue maneuver for entrapped stent in the coronary tree].
We describe two patients in whom stents dislodged from the unexpanded balloon during angioplasty and remained trapped in the coronary tree. In one patient the stent was located in the curvature of the circumflex artery and, in the other patient, it was detected in the distal left main artery. Retrieval of the undeployed stents was accomplished in the first case after advancing a low profile balloon through the stent and, in the second case, after placing a second guidewire exterior to the stent. In both cases, the balloon was inflated distally and the stents were dislodged after pulling the balloon back towards the guiding catheter and then withdrawing the whole system.


Schistosomiasis and soil-transmitted helminthiasis preventive chemotherapy: Adverse events in children from 2 to 15 years in Bengo province, Angola.
Preventive chemotherapy campaigns with praziquantel and albendazole are being implemented in Angola, as a high priority public health intervention. However, there are no published data regarding adverse events associated with these medications. In this context, we analysed adverse events due to co-administration of praziquantel and albendazole in endemic areas of schistosomiasis and soil-transmitted helminths in Bengo, Angola. In the context of a targeted drug administration, between December 2012 and September 2013, we conducted two surveys after co-administrating single oral doses of praziquantel and albendazole tablets to children 2 to 15 years of age. About 24 hours after each treatment, participants answered a questionnaire about adverse events. At baseline, 605 children (55.0% male; mean age: 9.7 years) were treated; 460 were interviewed and 257 (55.9%) reported at least one adverse event, 62.3% (160/257) of children being infected with schistosoma haematobium. After six months of treatment, among 339 children surveyed, 184 (54.3%) reported adverse events, with 49.5% (91/184) of infected children. Adverse events were most common in preschool-aged children, with no significant difference between genders. The most frequent adverse events in the two surveys were abdominal pain (18.5%, 25.7%), headache (20.9%, 23.0%) and dizziness (15.7%, 19.8%). Children aged 12 to 15 years (adjusted OR = 0.40, p = 0.040) and those with mixed infection (adjusted OR = 0.04, p = 0.011) had lower odds of adverse events. After the second treatment, those with heavy infection (adjusted OR = 2.72, p = 0.018) and aged 9-11 years (adjusted OR = 2.01, p = 0.049) had significantly fewer adverse events. About 2.0% of children experienced severe adverse events. This study adds evidence that preventive chemotherapy for schistosomiasis and soil-transmitted helminths control is safe, but cases of adverse events are expected. Standardized methodologies to discriminate drug-related adverse events from the clinical manifestations of the infections are needed.


Cosmetic rhinoplasty using the external approach.
Although the external rhinoplasty remains controversial it is difficult to understand why. The columella scar is of little significance since this becomes nearly invisible if the closure is carefully performed. There is a relative postoperative prolongation of nasal tip swelling in comparison with the closed technique, however this also becomes negligible with time. On the other hand, in our experience the open approach will achieve better understanding of the patient's individual anatomy and thus lead to a more predictable result through increased exposure and precision tailoring. The external technique facilitates the application of the great variety of tip refinements that have been developed over the years to allow facial plastic surgeons to get consistently excellent results in rhinoplasty.


Diagnostic accuracy of CT scan-based criteria compared with surgical exploration for the analysis of cervical fusion and nonunion.
Computed tomography (CT) scans are accepted as the imaging standard of reference to define union after anterior cervical discectomy and fusion (ACDF). However, ideal CT criteria to diagnose union have not been identified or validated. The objective of this study was to analyze the diagnostic value of 9 CT-based criteria and identify the ideal criteria among them to assess cervical fusion after ACDF using surgical exploration as the standard of reference. The authors performed a retrospective radiographic study of a single surgeon's prospective assessment of osseous fusion during cervical revision surgery by analyzing complete radiographic data in 44 patients who underwent anterior cervical revision surgery due to symptomatic suspected nonunion or adjacent level disease. All patients received standard preoperative CT scans, which were assessed by an independent radiologist to evaluate 9 diagnostic criteria for osseous union. During revision surgery, scar tissue was removed and manual segmental translation tests were performed. Nonunion was defined by visualized motion at the treated ACDF level. In total, 44 patients were included in the study (30 men; patient age 54 ± 6 years, BMI 28 ± 5 kg/m2). For analysis of fusion, 75 cervical levels were explored, of which 61 levels (81%) showed intraoperative movement indicating nonunion. Statistical analysis showed that of the 9 parameters used to diagnose bone union, "bridging bone on ≥ 3 CT slices" yielded the highest sensitivity (100%) and specificity (58%). Multivariate analysis revealed that prediction accuracy was not increased if several criteria were combined to determine fusion. The authors found that the best indicator of bone union was the item bridging bone on ≥ 3 CT slices. Combining the scoring of more than one criterion did not increase the diagnostic accuracy.


Bacilysin from Bacillus amyloliquefaciens FZB42 has specific bactericidal activity against harmful algal bloom species.
Harmful algal blooms, caused by massive and exceptional overgrowth of microalgae and cyanobacteria, are a serious environmental problem worldwide : In the present study, we looked for Bacillus strains with sufficiently strong anticyanobacterial activity to be used as biocontrol agents. Among 24 strains, Bacillus amyloliquefaciens FZB42 showed the strongest bactericidal activity against Microcystis aeruginosa, with a kill rate of 98.78%. The synthesis of the anticyanobacterial substance did not depend on Sfp, an enzyme that catalyzes a necessary processing step in the nonribosomal synthesis of lipopeptides and polyketides, but was associated with the aro gene cluster that is involved in the synthesis of the sfp-independent antibiotic bacilysin. Disruption of bacB, the gene in the cluster responsible for synthesizing bacilysin, or supplementation with the antagonist N-acetylglucosamine abolished the inhibitory effect, but this was restored when bacilysin synthesis was complemented. Bacilysin caused apparent changes in the algal cell wall and cell organelle membranes, and this resulted in cell lysis. Meanwhile, there was downregulated expression of glmS, psbA1, mcyB, and ftsZ-genes involved in peptidoglycan synthesis, photosynthesis, microcystin synthesis, and cell division, respectively. In addition, bacilysin suppressed the growth of other harmful algal species. In summary, bacilysin produced by B. amyloliquefaciens FZB42 has anticyanobacterial activity and thus could be developed as a biocontrol agent to mitigate the effects of harmful algal blooms.


Bell's palsy during pregnancy: is it associated with adverse perinatal outcome?
To determine whether an association exists between Bell's palsy during pregnancy and adverse perinatal outcomes. A retrospective study comparing all singleton pregnancies of patients with and without Bell's palsy was conducted. Multiple logistic regression model was performed to control for confounders. Out of 242,216 deliveries, 0.017% (n = 42) were diagnosed with Bell's palsy during pregnancy. Risk factors for Bell's palsy were chronic hypertension (9.5% vs. 1.5%, P < .001) and maternal obesity (7.1% vs. 0.8%, P < .001). Patients with Bell's palsy during pregnancy had higher rates of severe preeclampsia (9.5% vs. 1.1%, P < .001) and Cesarean deliveries (31.0% vs. 13.3%, P = .001) compared to the comparison group. In contrast, no significant association was documented between Bell's palsy and adverse perinatal outcomes such as low Apgar scores (<7) at 5 minutes (4.8% vs. 3.1%; P = .524) and perinatal mortality (2.4% vs. 1.4%; P = .57). Using multivariable analysis, controlling for confounders such as maternal age, fertility treatments, and ethnicity, Bell's palsy during pregnancy was significantly associated with obesity (odds ratio [OR] = 9.08 95% confidence interval [CI] 2.8-29.46; P < .001), chronic hypertension (OR = 6.69 95% CI 2.38-18.76; P < .001), and severe preeclampsia (OR = 9.46 95% CI 3.37-26.53; P < .001). Chronic hypertension and obesity are independent risk factors for Bell's palsy. Bell's palsy during pregnancy is significantly associated with severe preeclampsia. Nevertheless, no significant association exists between Bell's palsy and adverse perinatal outcomes.


[Medical aspects of diving in otorhinolaryngology. I. Barotrauma and decompression sickness].
Recreational scuba diving has become immensely popular in recent years and is no longer restricted to individualists or adventurers. During a dive, the human body with its gas-filled cavities is exposed to an increased ambient pressure. In the present review article, aspects of diving and hyperbaric medicine related to the otolaryngology field are presented. The basics of physics, physiology and pathophysiology that are essential to understand the effects of an increased and varying ambient pressure are reviewed. Barotrauma of the outer, middle and inner ear, the paranasal sinuses, face, teeth and larynx are explained and classified in those during the compression and decompression phase. This is followed by a discussion of inner ear decompression sickness. The present article will provide a background and foundation for both, an adequate treatment of these diseases and a critical and responsible health education of the diver.


Detecting archaic introgression using an unadmixed outgroup.
Human populations outside of Africa have experienced at least two bouts of introgression from archaic humans, from Neanderthals and Denisovans. In Papuans there is prior evidence of both these introgressions. Here we present a new approach to detect segments of individual genomes of archaic origin without using an archaic reference genome. The approach is based on a hidden Markov model that identifies genomic regions with a high density of single nucleotide variants (SNVs) not seen in unadmixed populations. We show using simulations that this provides a powerful approach to identifying segments of archaic introgression with a low rate of false detection, given data from a suitable outgroup population is available, without the archaic introgression but containing a majority of the variation that arose since initial separation from the archaic lineage. Furthermore our approach is able to infer admixture proportions and the times both of admixture and of initial divergence between the human and archaic populations. We apply the model to detect archaic introgression in 89 Papuans and show how the identified segments can be assigned to likely Neanderthal or Denisovan origin. We report more Denisovan admixture than previous studies and find a shift in size distribution of fragments of Neanderthal and Denisovan origin that is compatible with a difference in admixture time. Furthermore, we identify small amounts of Denisova ancestry in South East Asians and South Asians.


Pneumoretroperitoneum in two patients with Clostridium perfringens necrotizing pancreatitis.
Pancreatic gas gangrene is an uncommon and often fatal complication of acute pancreatitis, due to the sporulating anaerobe Clostridium perfringens. C. perfringens is a normal constituent of colonic flora, but infects the pancreas by either transmural spread from the colon or via the biliary tree. Only three reported cases in the world literature describe acute pancreatitis with pneumoretroperitoneum and clostridial infection. Two separate cases, at the same institution, of acute pancreatitis complicated by C. perfringens were analyzed. The records of patients were reviewed for admission history, laboratory and radiology results, intensive care support, surgical intervention, and outcome. Retroperitoneal air was visualized early in the clinical course of both patients by computed tomography. Early surgical debridement, drainage, parental antibiotics, and reexploration resulted in an uncomplicated recovery. Early computed tomography in patients with suspected necrotizing pancreatitis contributes to early intervention and may advantageously enhance survival.


Shabbir Syndrome: Case Report of a Rare Disease.
To report a case of Laryngeal Onycho Cutaneous Syndrome in a 10 year old child presenting with an ocular surface mass. A 10 year old boy presented in the out-patient department with a painless mass in left eye. There was history of airway obstruction requiring tracheotomy in early childhood and recurrent skin ulcerations. Aslit lamp examination revealed fleshy mass arising from the supero-nasalconjunctiva of left eye. Systemic examination revealed ulcerative lesions on cheeks and ears with dystrophic nails in hands and feet. The ocular surface lesion was treated successfully with topical steroid drops. Steroids may be useful in treating early lesion of ocular surface in LOCS. The importance of regular follow must be emphasized for timely management of frequent granulation tissue outgrowth in bid to avoid development of severe symblepharon and blindness.


Interactions of Candida albicans yeast cells, germ tubes and hyphae with human polymorphonuclear leucocytes in vitro.
Suspensions of Candida albicans yeast cells, germ tubes and hyphae with biomass standardized by ATP measurement were compared for their relative susceptibilities to phagocytosis and intracellular killing by human polymorphonuclear leucocytes. All three forms were ingested to a similar extent, but significantly fewer yeast cells were killed intracellularly after ingestion than were filamentous forms of the fungus. Ketoconazole pretreatment significantly enhanced the susceptibility of hyphae, but not of germ tubes, to phagocytosis and intracellular killing. The opsonic requirements of the yeasts and filamentous forms for efficient phagocytosis and killing differed.


[Current diagnosis and treatment of pancreatic cancer].
Adenocarcinoma of the pancreas (pancreatic cancer) is the most frequent tumor entity in the pancreas. While the results of surgical therapy of pancreatic cancer were disappointing in the past due to high perioperative mortality rates, resection of pancreatic cancer nowadays represents the standard treatment for non-metastatic cancer with a mortality rate below 5%. This decrease in perioperative mortality of the Whipple operation is inversely correlated to the case load of the hospital and the responsible surgeon, and is mainly related to improvements in the intensive care management, the surgical technique and patient selection. In particular, the perioperative use of octreotide resulted in a significant decrease in the rate of pancreatic fistula. Furthermore, modern staging examinations such as diagnostic laparoscopy, PET, or endoscopic ultrasound resulted in improved patient selection. In addition, the long-term results of the surgical treatment of pancreatic cancer has been improved by adjuvant and neoadjuvant chemotherapy in the past 10 years. Similar progress has been made in the palliative treatment of metastatic or locally advanced cancer. Nowadays, endoscopic procedures can replace surgical palliation of obstructive jaundice in most cases and sometimes even gastric outlet obstruction. Moreover, systemic chemotherapy using gemcitabine-based protocols has resulted in a significant prolongation of survival. However, further progress in the treatment of pancreatic cancer can only be achieved by an interdisciplinary management of this disease.


Movement pattern of Spartocera dentiventris (Berg) (Hemiptera: Coreidae) in an experimental crop of Nicotiana tabacum L. (Solanaceae).
In the present work we investigated the movement pattern of Spartocera (= Corecoris) dentiventris (Berg) (Hemiptera: Coreidae) adults through the mark-release-recapture method in an experimental tobacco crop (Nicotiana tabacum L., Solanaceae) in Porto Alegre, RS, Brazil. We marked 217 specimens from August 1999 to April 2000. Females moved 2.94 +/- 0.198 times and males 1.46 +/- 0.171 times among host plants. The number of movements per capture was 0.53 +/- 0.03 for females and 0.54 +/- 0.06 for males. Newly recruited adults took 12.75 +/- 0.919 days to leave their original plant, females moved 9.87 +/- 0.711 m and males 9.16 +/- 1.29 m in this first movement. We estimate that females move an average of 26.22 m and males 13.89 m during their permanence in the area.


Blister-like Aneurysms in Atypical Locations: A Single-Center Experience and Comprehensive Literature Review.
Blister-like aneurysms (BLAs) were originally described to arise typically along the nonbranching segment of the dorsal wall of the internal carotid artery (ICA); however, BLAs located in areas other than the dorsal ICA have been described more recently. We present a case series of "atypical" BLAs and a systematic review of the literature on this subject. We conducted a literature search using the key word "blister-like aneurysm." Studies reporting BLAs in locations other than the dorsal ICA wall were selected. Clinical presentation, treatment modality, complications, and outcomes (modified Rankin Scale for neurologic outcomes and Roy scale for radiologic outcomes) were extracted from each study. We also reviewed our single-institution experience with atypical BLAs and analyzed the topography and outcomes of all the atypical BLAs according to each specific treatment modality. Atypical BLAs were observed in the anterior communicating, middle cerebral, basilar, posterior cerebral, anterior cerebral, and posterior inferior cerebellar arteries. Surgery was the treatment in 65% of patients, an endovascular approach was used in 30%, and a combined approach was used in 5%. A good outcome (modified Rankin Scale 0-1-2) was experienced by 88% and 55% of the patients in the endovascular and surgical groups, respectively. There were 4 deaths, 2 in the endovascular group and 2 in the surgical group. Endovascular treatment of BLAs seems to be associated with reduced morbidity and mortality and to provide a better outcome compared with surgical approaches. Further prospective studies are needed to confirm these results. It is important for clinicians to remember that BLAs may also occur in sites other than the typical ICA location.


Novel nociceptin analogues.
A series of new nociceptin analogues containing cysteine was tested for their nociceptive effects in tail-flick test on mice after icv injection. The cysteines were introduced in order to get irreversibly binding analogues based on the assumption that the cysteines in the ligand can interact with the cysteines from the receptor to form an S-S bridge. In vivo tests revealed that Cys1-nociceptin (1-13)-NH2 (Cys1-NC) is an antagonist, whereas Cys7-NC is an agonist. Gly1[Phe(p-NO2)]4-NC was less active indicating that the antagonist properties of Cys1-NC are associated with the presence of the sulfhydryl group of cysteine. The analogues D-Cys2 and Cys3 were also almost inactive.


Safety and efficacy of vaccination against hepatitis B in patients with rheumatoid arthritis.
Hepatitis B infection and vaccination against it have been implicated in the potential triggering or flare of some autoimmune diseases, including rheumatoid arthritis (RA). However, the safety of hepatitis B vaccination in patients with pre-existing RA is not known. To assess the safety and antibody response of immunisation with a recombinant DNA hepatitis B vaccine in patients with RA. The study comprised 44 patients with RA, of whom 22 received three doses (the second and third dose being given after one and six months) of a recombinant DNA hepatitis B vaccine (study group) and 22 did not receive the vaccine (control group). Both groups had comparable proportions of women and similar mean age (51 years). Clinical assessment before and two and seven months after the first immunisation included evaluation of daytime pain with a 10 cm visual analogue scale, duration of morning stiffness, and number of tender and swollen joints. Erythrocyte sedimentation rate (ESR) and C reactive protein (CRP) were measured at each visit. Antibodies to hepatitis B surface antigen (HBsAg) were determined by a commercial enzyme linked immunosorbent assay (ELISA) test kit. Hepatitis B vaccination was not associated with an appreciable deterioration in any clinical or laboratory measure of disease. The measures of disease activity of the patients and controls during the study period did not differ significantly: p=0.76 for daytime pain, p=0.1 for morning stiffness, p=0.24 and p=0.3 for tender and swollen joints respectively, p=0.08 for CRP, and p=0.12 for ESR. Fifteen of the 22 patients responded to vaccination, with an antibody level against HBsAg of 10 IU/l after seven months. Lack of response was associated with older age and higher scores of daytime pain. Hepatitis B vaccination is safe in RA and produces antibodies in 68% of the patients.


Current status of transluminal angioplasty.
Although the published results of transluminal angioplasty have been satisfactory, its role in the practice of vascular surgery has not been well documented. Of 137 members of the Canadian Society for Vascular Surgery polled, 102 (74.5%) responded to a questionnaire drawn up to assess the current status of transluminal angioplasty in Canada. Of 1343 angioplasties reported, 64% were iliac, 21% femoral, 12% renal and 3% of other arteries. Claudication was the indication in 80%, limb salvage in 20%. In 15% angioplasty was used as an adjunct to subsequent arterial surgery. The lesion and results of treatment were assessed by clinical, noninvasive and pressure-gradient evaluation in most hospitals. Complications varied widely, reflecting the large number of physicians reporting. Most angioplasties were performed by vascular radiologists, and joint decision-making by radiologist and surgeon was the most common approach. The use of angioplasty increased in hospitals carrying out the most intensive evaluation, suggesting that it should have an expanded role.


CD83 orchestrates immunity toward self and non-self in dendritic cells.
Dendritic cells (DCs) are crucial to balance protective immunity and autoimmune inflammatory processes. Expression of CD83 is a well-established marker for mature DCs, although its physiological role is still not completely understood. Using a DC-specific CD83-conditional KO (CD83ΔDC) mouse, we provide new insights into the function of CD83 within this cell type. Interestingly, CD83-deficient DCs produced drastically increased IL-2 levels and displayed higher expression of the costimulatory molecules CD25 and OX40L, which causes superior induction of antigen-specific T cell responses and compromises Treg suppressive functions. This also directly translates into accelerated immune responses in vivo. Upon Salmonella typhimurium and Listeria monocytogenes infection, CD83ΔDC mice cleared both pathogens more efficiently, and CD83-deficient DCs expressed increased IL-12 levels after bacterial encounter. Using the experimental autoimmune encephalomyelitis model, autoimmune inflammation was dramatically aggravated in CD83ΔDC mice while resolution of inflammation was strongly reduced. This phenotype was associated with increased cell influx into the CNS accompanied by elevated Th17 cell numbers. Concomitantly, CD83ΔDC mice had reduced Treg numbers in peripheral lymphoid organs. In summary, we show that CD83 ablation on DCs results in enhanced immune responses by dysregulating tolerance mechanisms and thereby impairing resolution of inflammation, which also demonstrates high clinical relevance.


Training community-based paraprofessionals as behavior therapists with families of alcohol-abusing adolescents.
To combat the increase in chronic alcohol use among adolescents in a large metropolitan area, community-based paraprofessionals were trained in behavior therapy and treated these adolescents and their families in the home. Preliminary evaluation of the program revealed encouraging results. This training supplied structure for the paraprofessionals' interventions with families and provided them with skills they are continuing to use in their community work.


Coupling of I1-imidazoline receptors to diacylglyceride accumulation in PC12 rat pheochromocytoma cells.
The I1-subtype of imidazoline binding sites has been characterized concerning binding specificity and tissue localization, and several physiological functions have been ascribed to it. However, the signaling pathways coupled to this putative receptor are not known. Pheochromocytoma PC12 cells express I1-imidazoline binding sites in plasma membrane and lack alpha2-adrenergic receptors, which recognize many I1-imidazoline ligands. In this cellular model, diacylglycerol (DAG), a second messenger, is generated in response to the putative I1-imidazoline agonist moxonidine. Using radioflux with [3H]myristate and direct measurements of DAG mass, we showed a rapid and transient peak of DAG in undifferentiated PC12 cells within the first 1 min of agonist exposure. In PC12 cells treated with nerve growth factor to initiate differentiation, DAG accumulation at 15 sec was facilitated, and the increase in DAG mass persisted throughout 10 min of agonist treatment. Efaroxan, a putative I1-antagonist, attenuated the effect of moxonidine on DAG accumulation in nerve growth factor-treated cells, as did D609, an inhibitor of phosphatidylcholine-selective phospholipase C. Phospholipase D did not seem to be involved in generation of DAG in response to I1-receptor activation, nor was there accumulation of phosphatidic acid. These findings suggest coupling of I1-imidazoline receptors to a phospholipase C to generate DAG as a second messenger, a process regulated by neuronal differentiation and possibly participating in the physiological responses to I1-imidazoline receptor activation.


Risk Factors for Bites and Diseases Associated With Black-Legged Ticks: A Meta-Analysis.
The emergence and spread of Lyme disease and other infections associated with black-legged ticks is causing a public health crisis. No human vaccines are currently available, and both diagnosis and treatment are sometimes ineffectual, leading to advocacy for self-directed preventative measures. These recommendations are widely communicated to the public, but there is limited evidence for their efficacy. We undertook a systematic review and mixed-effects meta-regression analysis of factors purported to increase or decrease risk of black-legged tick bites and tick-borne disease. Published articles used in the study spanned the years 1984-2018. Variables associated with increased probability of tick-borne disease, with odds ratios significantly greater than 1, included deer abundance, high density of nymph-stage black-legged ticks, landscapes with interspersed herbaceous and forested habitat, low human population density, gardens, cat ownership, and race. Contrary to recommendations, use of landscape-related tick control measures, such as clearing brush, trimming branches, and having a dry barrier between lawn and woods, tended to increase risk. Pet ownership increased bite risk. Bite risk was highest for children aged 5 years or less, with a secondary peak in persons aged 50-70 years. Although some widely disseminated recommendations are supported by the research analyzed, others require further evaluation. Additional research is also needed to understand the mechanisms underlying significant relationships.


Bone health. New role for vitamin K?
To assess growing evidence that vitamin K (phylloquinone) plays an important role in bone health and, subsequently, in prevention of osteoporotic fractures. We searched MEDLINE from January 1972 to December 2002 using the key words vitamin K and bone health. We reviewed 30 articles that seemed relevant or had a human focus. All evidence can be categorized as level II. Evidence suggests that dietary phylloquinone intake of <100 microg daily might not be optimal for bone health. Low intake of vitamin K could contribute to osteoporosis and subsequent fracture due to the undercarboxylation of osteocalcin. Family physicians need to be aware of the importance of encouraging adequate vitamin K intake, particularly among institutionalized elderly people, to prevent increased bone resorption. Further study is needed to determine the exact role of vitamin K in bone metabolism, and methods of assessing vitamin K requirements need to be standardized.


Simple coumarins and analogues in medicinal chemistry: occurrence, synthesis and biological activity.
Coumarins, also known as benzopyrones, are present in remarkable amounts in plants, although their presence has also been detected in microorganisms and animal sources. The structural diversity found in this family of compounds led to the division into different categories, from simple coumarins to many other kinds of policyclic coumarins, such as furocoumarins and pyranocoumarins. Simple coumarins and analogues are a large class of compounds that have attracted their interest for a long time due to their biological activities: they have shown to be useful as antitumoural, anti-HIV agents and as CNS-active compounds. Furthermore, they have been reported to have multiple biological activities (anticoagulant, anti-inflammatory), although all these properties have not been evaluated systematically. In addition, their enzyme inhibition properties, antimicrobial and antioxidant activities are other foremost topics of this field of research. The present work is to survey the information published or abstracted from 1990 till 2003, which is mainly related to the occurrence, synthesis and biological importance of simple coumarins and some analogues, such as biscoumarins and triscoumarins. Data are also highlighted, concerning the development of new synthetic strategies that could help in drug design and in the work on SAR or QSAR.


The lateral thoracodorsal flap in breast reconstruction: a comparison between two plastic surgical centres.
In a retrospective study the results of breast reconstruction with 146 lateral thoracodorsal flaps operated on during the period 1991-94 at the Departments of Plastic Surgery at the Sahlgrenska University Hospital in Göteborg and the Stockholm Söder Hospital were evaluated. The median age of the 135 patients was 51 years (range 37-74). About two thirds of the patients were also operated on on the opposite side at the same time; and this was done more often in Stockholm, which partly explains the longer operating time in Stockholm. Perioperative bleeding (median 100 ml, range 25-400) was similar in both groups. Median postoperative bleeding, which was measured only in Stockholm, was 300 ml (range 30-1000). Seromas were seen only in Göteborg where postoperative drains were rarely used. The rate of early complications (infection and partial necrosis) were higher in Stockholm where more flaps were raised in irradiated tissue. We conclude that the results were similar in the two centres, and the thoracodorsal flap gives good results. The incidence of early morbidity could be reduced by excluding patients who had been irradiated, by inserting drains, and by giving antibiotic prophylaxis.