Effects of metoprolol therapy on cardiac gap junction remodelling and conduction in human chronic atrial fibrillation.
We investigated the influence of metoprolol on gap junction proteins connexin43 (Cx43) and connexin40 (Cx40) in atrial tissue from patients with/without atrial fibrillation (AF). Left atrial tissue samples from 160 patients with AF or sinus rhythm (SR) with or without metoprolol (mean daily dose: 65.2 ± 9.1 mg·day⁻¹) were analysed for Cx43 and Cx40 by Western blot and immunohistology. Transverse and longitudinal conduction velocities were determined by 64 multi-electrode mapping. Both Cx43 and Cx40 expression were significantly increased in patients with AF versus SR. Cx43-expression in AF was significantly higher in patients receiving metoprolol, while Cx40 expression was unaffected by metoprolol treatment. In AF, the ratio of lateral/polar expression of Cx43 and Cx40 was enhanced due to increased expression at the sides of the cells (lateral) and a loss at the cell poles. This AF-induced increase in lateral/polar expression of Cx43, but not of Cx40, was significantly antagonized by metoprolol treatment. Functionally, in AF patients, transverse conduction velocity in atrial samples was significantly enhanced and this change was also significantly antagonized by metoprolol. AF induced enhanced lateral expression of Cx43 and Cx40 together with enhanced transverse conduction velocity in left atrial tissue. Alterations in localization of Cx43 and conduction changes were both antagonized by metoprolol, showing that pharmacological modulation of gap junction remodelling seems, in principle, possible. This finding may open new approaches to the development of anti-arrythmic drugs.


Sentinel Lymph Node Biopsy and Completion Lymph Node Dissection for Melanoma.
This review critically evaluates recent trials which have challenged the practice of completion lymph node dissection (CLND) for melanoma patients diagnosed with regional metastasis by positive sentinel lymph node biopsy (SLNB). Two trials in the last 2 years, DeCOG-SLT and MSLT-II, found no significant differences in melanoma-specific survival between patients, whether they received immediate CLND or observation after positive SLNB, despite decreases in nodal recurrence achieved by dissection. These trials together disfavor routine CLND in most patients after positive SLNB. However, their conclusions are limited by study populations which overall harbored a lower burden of SLN disease. Special attention needs to be given to patients who do have higher risk disease, with SLN tumor burdens exceeding 1 mm in diameter, for whom CLND may remain both prognostic and therapeutic. Current guidelines thus recommend either CLND or careful observation after positive SLNB after appropriate risk stratification of patients. While a decline in CLND is inevitable, treatment of stage III melanoma is witnessing the concurrent rise of effective adjuvant therapies. PD-1 inhibitors such as nivolumab, or combination BRAF/MEK inhibitors for V600E or K mutant melanoma, which were previously available to only trial patients with completely resected stage III disease, are now approved for use in patients with positive SLNB alone. Providers are better equipped than ever to treat clinically occult, regional metastatic disease with SLNB followed by adjuvant therapy for most patients, but should take steps to avoid undertreatment of high-risk patients who may proceed to disease relapse or progression.


A test of hypothesis on familial correlations.
When familial data are analysed, the model usually employed assumes independence of family observations and constancy of interclass and intraclass correlations. A statistic for testing the validity of the assumptions of family independence and constant interclass and intraclass correlations is developed. The test statistic is for constant family size; it has an asymptotic chi-square distribution. An example to illustrate the theory is given using Frets's data on head lengths. A recommendation is made on how to apply the test to the general case of varying sizes. Another recommendation is made on models to be tried once the null hypothesis is rejected.


Fighting malaria with engineered symbiotic bacteria from vector mosquitoes.
The most vulnerable stages of Plasmodium development occur in the lumen of the mosquito midgut, a compartment shared with symbiotic bacteria. Here, we describe a strategy that uses symbiotic bacteria to deliver antimalaria effector molecules to the midgut lumen, thus rendering host mosquitoes refractory to malaria infection. The Escherichia coli hemolysin A secretion system was used to promote the secretion of a variety of anti-Plasmodium effector proteins by Pantoea agglomerans, a common mosquito symbiotic bacterium. These engineered P. agglomerans strains inhibited development of the human malaria parasite Plasmodium falciparum and rodent malaria parasite Plasmodium berghei by up to 98%. Significantly, the proportion of mosquitoes carrying parasites (prevalence) decreased by up to 84% for two of the effector molecules, scorpine, a potent antiplasmodial peptide and (EPIP)(4), four copies of Plasmodium enolase-plasminogen interaction peptide that prevents plasminogen binding to the ookinete surface. We demonstrate the use of an engineered symbiotic bacterium to interfere with the development of P. falciparum in the mosquito. These findings provide the foundation for the use of genetically modified symbiotic bacteria as a powerful tool to combat malaria.


Modulation of the oxidative stress and inflammatory cytokine response by thymoquinone in the collagen induced arthritis in Wistar rats.
Thymoquinone (TQ) is the major active compound derived from Nigella sativa. Our aim of this work was to evaluate the antioxidant and antiarthritic activity of TQ in Wistar rat by collagen induced arthritis (CIA). TQ was administered at a dose of 5mgkg(-1) body weight once daily for 21days. The effects of treatment in the rats were assessed by biochemical (articular elastase, MPO, LPO, GSH, catalase, SOD and NO), inflammatory mediators (IL-1β, IL-6, TNF-α, IL-10, IFN-γ and PGE(2)) and histological studies in joints. TQ was effective in bringing significant changes on all the parameters (articular elastase, MPO, LPO, GSH, catalase, SOD and NO) studied. Oral administration of TQ resulted in significantly reduced the levels of pro-inflammatory mediators (IL-1β, IL-6, TNF-α, IFN-γ and PGE(2)) and increased level of IL-10. The protective effects of TQ against RA were also evident from the decrease in arthritis scoring and bone histology. In conclusion, the fact that TQ abolished a number of factors known to be involved in RA pathogenesis indicates that the administration of thymoquinone may have potential value in the treatment of inflammatory disease.


The discriminative stimulus effects of dopamine D2- and D3-preferring agonists in rats.
Previous research has found the stimulus effects of dopamine D2- and D3-preferring agonists difficult to distinguish in drug discrimination studies. Antagonism studies suggest that the stimulus effects of both types of agonists may be mediated primarily through D2 receptors. The current study was designed to further assess the receptors mediating the stimulus effects of these agonists and to attempt to train rats to discriminate directly between D2- and D3-preferring dopamine agonists. Four groups of eight rats were trained to discriminate either 0.1 mg/kg of the D3-preferring agonist pramipexole from saline, 1.0 mg/kg of the D2-preferring agonist sumanirole from saline, 0.1 mg/kg pramipexole from either saline or 1.0 mg/kg sumanirole, or 1.0 mg/kg sumanirole from either saline or 0.1 mg/kg pramipexole. Three of eight rats in the 0.1 mg/kg pramipexole vs. 1.0 mg/kg sumanirole or saline failed to meet the training criteria, and the discrimination in this group was tenuous. The D2-preferring antagonist L-741,626 at 1.0 mg/kg was more effective at shifting to the right the pramipexole dose-response curve in pramipexole-trained rats, while 32 mg/kg of the selective D3 antagonist PG01037 had little effect. Quinpirole and 7-OH-DPAT fully or partially substituted for both pramipexole and sumanirole in each group tested, while cocaine did not substitute in any group. Antagonist data along with the pattern of training and substitution data suggested that D2 receptor activation is primarily responsible for the stimulus effects of both sumanirole and pramipexole with D3 receptor activation playing little or no role.


Organ Procurement Organization and transplant center effects on cadaver renal transplant outcomes.
1. A majority of transplant centers had significant improvements in cadaver donor kidney graft survival rates since 1991. A 10% or greater increase in one-year graft survival rates was reported by 36 of 128 large transplant centers. 2. When transplants were grouped according to the organ procurement agency that procured the kidney, large variations in one-year graft survival rates among 65 OPOs were noted. The "OPO effect" was of a magnitude similar to that of the "center effect" when analyzed separately. 3. Fourteen OPOs with the lowest graft survival rates procured a significantly higher fraction of kidneys from older, hypertensive or Black donors and donors whose deaths were not from motor vehicle accidents. Transplants were more often given to Black recipients, and kidneys were less frequently shared compared with 15 OPOs with the highest graft survival rates. 4. Significantly higher proportions of non-Black donors and recipients, patients with no activity limitation at the time of listing, patients working fulltime at the time of transplantation, and patients on dialysis less than one year were more frequently transplanted at 21 centers with excellent one-year graft survival rates compared with centers in groups with poorer results. 5. Increased numbers of mismatched HLA-B,DR antigens were detrimental to graft survival at centers with good or fair one-year graft survival rates, but had a minimal effect at centers with excellent results. 6. The OPO effect on graft survival rates was significantly associated with the center effect. Kidneys procured by OPOs associated with low graft survival rates and transplanted at centers with low graft survival rates resulted in the worst graft outcome. Interestingly, kidneys procured by OPOs associated with high graft survival rates, but transplanted by centers with low graft survival rates had better outcomes than kidneys from OPOs with lower graft survival rates. 7. Based on univariate analyses, the center and OPO effects ranked third and fifth among 16 significant factors. However, after adjusting for these 16 potentially confounding variables using a multivariate Cox regression model, the differences between the best and worst center and OPO groups were the fourth and eleventh most detrimental risk factors on graft outcome, respectively.


Anti-inflammatory Activity of 1-docosanoyl Cafferate Isolated from Rhus verniciflua in LPS-stimulated BV2 Microglial Cells.
Although various derivatives of caffeic acid have been reported to possess a wide variety of biological activities such as protection of neuronal cells against excitotoxicity, the biological activity of 1-docosanoyl cafferate (DC) has not been examined. The objective of the present study was to evaluate the anti-inflammatory effects of DC, isolated from the stem bark of Rhus verniciflua, on lipopolysaccharide (LPS)-stimulated BV2 microglial cells. Pretreatment of cells with DC significantly attenuated LPS-induced NO production, and mRNA and protein expression of iNOS in a concentration-dependent manner. DC also significantly suppressed LPS-induced release of cytokines such as TNF-α and IL-1β . Consistent with the decrease in cytokine release, DC dose-dependently and significantly attenuated LPS-induced mRNA expression of these cytokines. Furthermore, DC significantly suppressed LPS-induced degradation of IKB, which retains NF-kB in the cytoplasm. Therefore, nuclear translocation of NF-kB induced by LPS stimulation was significantly suppressed with DC pretreatment. Taken together, the present study suggests that DC exerts its anti-inflammatory activity through the suppression of NF-kB translocation to the nucleus.


Five essential oils from aromatic plants of Cameroon: their antibacterial activity and ability to permeabilize the cytoplasmic membrane of Listeria innocua examined by flow cytometry.
To investigate the antibacterial effect of five essential oils (EO) extracted from aromatic plants (Cymbopogon citratus, Ocimumbasilicum, Ocimum gratissimum, Thymus vulgaris and Zingiber officinale) of Cameroon against strains of Listeria monocytogenes, L. innocua and Staphylococcus aureus. The ability of selected EO to permeabilize the cytoplasmic membrane of L. innocua was also examined. The antibacterial activity of the EO determined by the agar diffusion method showed that T. vulgaris had the highest activity followed by O. gratissimum and C. citratus. Lowest activity was recorded from Z. officinale and O. basilicum. Significant differences in sensitivity between strains of Listeria and S. aureus were observed. Flow cytometry of L. innocua stained with carboxy-fluorescein diacetate showed that the fluorescence intensity of cells exposed to EO decreased faster than nonexposed cells, indicating that EO permeabilized the cytoplasmic membrane with the leakage of carboxy-fluorescein. Almost all the EO tested showed antibacterial activity to a different extent. The antibacterial effect was due to permeabilization of the cytoplasmic membrane. This study has identified the preservative potential of the EO examined. The use of sensitive method, such as flow cytometry, is advantageous for quick generation of data on the antibacterial effect of EO.


Surfactant effects on the kinetics of Michael-type addition reaction in reverse emulsion polymeric systems.
To ensure patients' safety during vascular surgery involving polymer injection, it is important to ensure a high degree of control over the gelation rate of the materials to prevent the formation of uncontrolled emboli from shear or dispersion. Through manipulation of factors affecting the kinetics of in situ forming polymeric systems, patients' safety can be maximized. The rate of Michael-Type Addition is known to be affected by the pH of the accompanying solution, as thiols will become nucleophilic thiolates in an aqueous environment. This work evaluates the effect of the addition of 2 and 4 wt % of several common surfactants on the gelation rate of reverse emulsion polymeric systems as a method for augmenting control of in vivo Michael-type addition kinetics. This work shows that both Tween-20 (HLB 16.7) and Pluronic type surfactants (HLB reverse similar 6.7) stabilize our emulsion, with gelation rates being significantly different in the presence of Pluronic type surfactants (p < 0.05). This work has demonstrated the ability of surfactants to promote faster gelation at lower pH by maximizing emulsion stability and reducing cohesion of smaller bubbles, thereby maximizing surface area between the initiator and the bulk phases.


Involvement of Toll-like receptors on Helicobacter pylori-induced immunity.
Dendritic cells (DCs) play a major role in the innate immune response since they recognize a broad repertoire of PAMPs mainly via Toll-like receptors (TLRs). During Helicobacter pylori (H. pylori) infection, TLRs have been shown to be important to control cytokine response particularly in murine DCs. In the present study we analyzed the effect of blocking TLRs on human DCs. Co-incubation of human DCs with H. pylori resulted in the release of the pro-inflammatory cytokines IL-12p70, IL-6 and IL-10. Release of IL-12p70 and IL-10 was predominantly influenced when TLR4 signaling was blocked by adding specific antibodies, suggesting a strong influence on subsequent T cell responses through TLR4 activation on DCs. Co-incubation of H. pylori-primed DC with allogeneic CD4+ T cells resulted in the production of IFN-γ and IL-17A as well as the expression of Foxp3, validating a mixed Th1/Th17 and Treg response in vitro. Neutralization of TLR4 during H. pylori infection resulted in significantly decreased amounts of IL-17A and IFN-γ and reduced levels of Foxp3-expressing and IL-10-secreting T cells. Our findings suggest that DC cytokine secretion induced upon TLR4-mediated recognition of H. pylori influences inflammatory and regulatory T cell responses, which might facilitate the chronic bacterial persistence.


The NSLS 100 element solid state array detector.
X-ray absorption studies of dilute samples require fluorescence detection techniques. Since signal-to-noise ratios are governed by the ratio of fluorescent to scattered photons counted by a detector, solid state detectors which can discriminate between fluorescence and scattered photons have become the instruments of choice for trace element measurements. Commercially available 13 element Ge array detectors permitting total count rates < 500000 counts per second are now in routine use. Since X-ray absorption beamlines at high brightness synchrotron sources can already illuminate most dilute samples with enough flux to saturate the current generation of solid state detectors, the development of next-generation instruments with significantly higher total count rates is essential. We present the design and current status of the 100 element Si array detector being developed in a collaboration between the NSLS and the Instrumentation Division at Brookhaven National Laboratory. The detecting array consists of a 10×10 matrix of 4 mm×4 mm elements laid out on a single piece of ultrahigh purity silicon mounted at the front end of a liquid nitrogen dewar assembly. A matrix of charge sensitive integrating preamplifiers feed signals to an array of shaping amplifiers, single channel analyzers, and scalers. An electronic switch, delay amplifier, linear gate, digital scope, peak sensing A/D converter, and histogramining memory module provide for complete diagnostics and channel calibration. The entire instrument is controlled by a LabView 2 application on a MacII ci; the software also provides full control over beamline hardware and performs the data collection.


Organisational and individual characteristics associated with glycaemic control among patients with type 2 diabetes: cross-sectional study in China.
There is a high prevalence of poor glycaemic control among patients with type 2 diabetes (T2DM) in China. This study aimed to explore both organisational and individual characteristics associated with glycaemic control among patients with T2DM. Cross-sectional survey. Shandong Province, China. The participants were 2166 patients with T2DM and 337 healthcare providers from 36 urban communities and 36 rural villages in Shandong Province. Multistage stratified sampling procedures were used to measure demand-side individual demographic, clinical and self-management characteristics, and supply-side organisational characteristics, and the status of glycaemic control. Multilevel logistic regression analysis was performed to assess key determinants of glycaemic control. Only 42.8% of the patients with T2DM achieved good glycaemic control. Age, income, hypertension and self-efficacy were significantly positive predictors of optimal glycaemic control, while duration of diabetes, antidiabetic drugs and monitoring of blood glucose were significantly negative predictors of that. Private VCs (OR=0.48, 95% CI 0.29 to 0.82, p<0.01) and lack of healthcare providers (OR=0.69, 95% CI 0.53 to 0.89, p<0.01; OR=0.71, 95% CI 0.52 to 0.98, p<0.05) were significantly negative predictors of optimal glycaemic control, while diabetes knowledge level of healthcare providers (OR=1.36, 95% CI 1.02 to 1.83, p<0.05; OR=1.45, 95% CI 1.00 to 2.10, p<0.05) and kinds of antidiabetic drugs (OR=1.37, 95% CI 0.97 to 1.93, p<0.1; OR=1.46, 95% CI 1.07 to 2.00, p<0.05) were significantly positive predictors of that. Glycaemic control was suboptimal among patients with T2DM in China. The determinants of failing to achieve good glycaemic control included both organisational and individual characteristics. Potential interventions that target patients, providers and the healthcare organisations should be taken to improve the glycaemic control and health outcome among patients with T2DM.


Ethanol-sedative hypnotic interactions: in vitro studies on Na-K-activated adenosine triphosphatase.
The combined in vitro effects of ethanol wih methaqualone, phenobarbital, pyrazole or disulfiram were studied using rat brain microsomal NA-K-ATPase. Methaqualone and phenobarbital, either alone or in combination with ethanol, showed a dose-dependent inhibition of Na-K-ATPase activity. Kinetic studies showed that the inhibitory effects were competitive with the respect to K+ and uncompetitive with respect to MgATP for phenobarbital either alone or in combination with ethanol. Methaqualone showed a noncompetitive type of inhibition with respect to K+, whereas a mixed type of inhibition was seen in combinations with ethanol. Furthermore, methaqualone either alone or in combination with ethanol also showed an uncompetitive type of inhibition with respect to MgATP. The dose-dependent inhibition of brain Na-K-ATPase activity by disulfiram was independent of K+ and additive in combination with ethanol. Pyrazole showed no effect on brain Na-K-ATPase activity. The inhibitions by phenobarbital and methaqualone, either alone or in combination with ethanol, were significantly greater at low K+ (1 mM) and high Na+ (100-200 mM). A greater inhibition by these drugs was also obtained when Na+ is low (5 mM) and K+ is high (15 mM). The inhibition by these drugs appears to depend on K+ and Na/K ratio in the reaction medium. The combined effects of ethanol and the other CNS depressants on brains Na-K-ATPase may be important to their interaction toxicities.


The human nucleolar protein FTSJ3 associates with NIP7 and functions in pre-rRNA processing.
NIP7 is one of the many trans-acting factors required for eukaryotic ribosome biogenesis, which interacts with nascent pre-ribosomal particles and dissociates as they complete maturation and are exported to the cytoplasm. By using conditional knockdown, we have shown previously that yeast Nip7p is required primarily for 60S subunit synthesis while human NIP7 is involved in the biogenesis of 40S subunit. This raised the possibility that human NIP7 interacts with a different set of proteins as compared to the yeast protein. By using the yeast two-hybrid system we identified FTSJ3, a putative ortholog of yeast Spb1p, as a human NIP7-interacting protein. A functional association between NIP7 and FTSJ3 is further supported by colocalization and coimmunoprecipitation analyses. Conditional knockdown revealed that depletion of FTSJ3 affects cell proliferation and causes pre-rRNA processing defects. The major pre-rRNA processing defect involves accumulation of the 34S pre-rRNA encompassing from site A' to site 2b. Accumulation of this pre-rRNA indicates that processing of sites A0, 1 and 2 are slower in cells depleted of FTSJ3 and implicates FTSJ3 in the pathway leading to 18S rRNA maturation as observed previously for NIP7. The results presented in this work indicate a close functional interaction between NIP7 and FTSJ3 during pre-rRNA processing and show that FTSJ3 participates in ribosome synthesis in human cells.


Vitamin D3 affects differentiation, maturation, and function of human monocyte-derived dendritic cells.
We studied the effects of 1alpha,25-dihydroxyvitamin D3 (1alpha, 25-(OH)2D3) on differentiation, maturation, and functions of dendritic cells (DC) differentiated from human monocytes in vitro in the presence of GM-CSF and IL-4 for 7 days. Recovery and morphology were not affected by 1alpha,25-(OH)2D3 up to 100 nM. DC differentiated in the presence of 10 nM 1alpha,25-(OH)2D3 (D3-DC) showed a marked decrease in the expression of CD1a, while CD14 remained elevated. Mannose receptor and CD32 were significantly increased, and this correlated with an enhancement of endocytic activity. Costimulatory molecules such as CD40 and CD86 were slightly decreased or nonsignificantly affected (CD80 and MHC II). However, after induction of DC maturation with LPS or incubation with CD40 ligand-transfected cells, D3-DC showed marginal increases in MHC I, MHC II, CD80, CD86, CD40, and CD83. The accessory cell function of D3-DC in classical MLR was also inhibited. Moreover, allogeneic T cells stimulated with D3-DC were poor responders in a second MLR to untreated DC from the same or an unrelated donor, thus indicating the onset of a nonspecific hyporesponsivity. In conclusion, our data suggest that 1alpha,25-(OH)2D3 may modulate the immune system, acting at the very first step of the immune response through the inhibition of DC differentiation and maturation into potent APC.


African neurosurgery part II: current state and future prospects.
A survey conducted among African neurosurgeons shows that there are now 500 neurosurgeons in Africa; that is, one neurosurgeon for 1,350,000 inhabitants, and 70,000 km2. The distribution of these neurosurgeons shows a striking regional disparity: North Africa has 354 neurosurgeons for 119 million inhabitants; that is, one neurosurgeon for 338,000 inhabitants; and South Africa has 65 neurosurgeons for 40 million inhabitants; that is, one neurosurgeon for 620,000 inhabitants. Between these two areas where neurosurgery is developing quite well, we have the majority of African countries with a scant density of neurosurgeons (81 neurosurgeons for 515 million inhabitants; that is, one neurosurgeon for 6,368,000 inhabitants). The Panafrican Association of Neurological Sciences (PAANS) brings together African neurosurgeons. This continental African association represents African neurosurgeons in the World Federation of Neurosurgical Societies (WFNS). In addition to this continental association, there are national societies of neurosciences. However, there are only six societies of neurosurgery. Two systems of training exist in Africa: (1) local training, and (2) training abroad. These two systems have unequal quality and specific difficulties that are pointed out. Among the optimistic elements that make us believe in the development of neurosurgery in Africa, are the existence of a quite good level of neurosurgery at the two extremities of the continent (North Africa and South Africa), the development of neurosciences in African universities, and the increasing interest that the international community bears to Africa in the last years. However, the real factor of optimism is the African neurosurgeons who should promote neurosurgery in their continent, at the level of their own countries by developing information and health education, setting their specialty in the education syllabus and health planning, and settling into active and performing societies. At the continental and international level, African neurosurgeons should institutionalize inter-African cooperation, expedite their continental association (PAANS), and further exchanges with the other continents through the WFNS. The latter, together with other associations such as the European Association of Neurosurgical Societies (EANS) could provide help to the development of neurosurgery in Africa as far as training, exchanges, research, and organization are concerned.


Ellagic acid induced p53/p21 expression, G1 arrest and apoptosis in human bladder cancer T24 cells.
It is well known that dietary phenolic compounds can elicit vital cellular responses such as cytotoxicity, cell cycle arrest and apoptosis by activating a cascade of molecular events. Ellagic acid is one of these phenolic compounds, but the exact mechanism of its action is still unclear. The objective of this study was to investigate ellagic acid-induced cell cycle arrest and apoptosis in T24 human bladder cancer cells in vitro. Assays were performed to determine cell viability, cell cycle arrest, apoptosis, caspases-3 activity and gene expression, measured by flow cytometric assay, polymerase chain reaction (PCR) and determination of caspase-3 activity. Ellagic acid significantly reduced the viable cells, induced G0/G1-phase arrest of the cell cycle and apoptosis. Ellagic acid also increased p53 and p21 and decreased CDK2 gene expression, that may lead to the G0/G1 arrest of T24 cells. Ellagic acid also promoted caspase-3 activity after exposure for 1, 3, 6, 12 and 24 h, which led to induction of apoptosis. Furthermore, the ellagic acid-induced apoptosis on T24 cells was blocked by the broad-spectrum caspase inhibitor (z-VAD-fmk).


Sensing and tuning microfiber chirality with nematic chirogyral effect.
Microfibers with their elongated shape and translation symmetry can act as important components in various soft materials, notably for their mechanics on the microscopic level. Here we demonstrate the mechanical response of a micro-object to imposed chirality, in this case, the tilt of disclination rings in an achiral nematic medium caused by the chiral surface anchoring on an immersed microfiber. This coupling between chirality and mechanical response, used to demonstrate sensing of chirality of electrospun cellulose microfibers, is revealed in the optical micrographs due to anisotropy in the elastic response of the host medium. We provide an analytical explanation of the chirogyral effect supported with numerical simulations and perform an experiment to test the effect of the cell confinement and fiber size. We controllably twist the microfibers and demonstrate the response of the nematic medium. More generally the demonstrated study provides means for experimental discrimination of surface properties and allows mechanical control over the shape of disclination rings.


Growth of Saccharomyces cerevisiae is controlled by its limited respiratory capacity: Formulation and verification of a hypothesis.
A novel mechanistic model for the growth of baker's yeast on glucoseis presented. It is based on the fact that glucose degradation proceeds via two pathways under conditions of aerobic ethanol formation. Part is metabolized oxidatively and part reductively, with ethanol being the end product of reductive energy metabolism. The corresponding metabolic state is designated oxidoreductive. Ethanol can be used oxidatively only. Maximum rates of oxidative glucose and ethanol degradation are governed by the respiratory capacity of the cells. The model is formulated by using the stoichiometric growth equations for pure oxidative and reductive (fermentative) glucose and ethanol metabolism. Together with the experimentally determinable yield coefficients (Y(X/S)) for the respective metabolic pathways, the resulting equation system is sufficiently determined. The superiority of the presented model over hitherto published ones is based on two essential novelities. (1) The model was developed on experimentally easily accessible parameters only. (2) For the modeling of aerobic ethanol formation, the substrate flow was split into two simultaneously operating (i.e., in parallel) metabolic pathways that exhibit different but constant energy-generating efficiencies (respiration and fermentation) and consequently different and constant biomass yields (Y(X/S)). The model allows the prediction of experimental data without parameter adaption in a biologically dubious manner.


Real-Time Kinetic Probes Support Monothiol Glutaredoxins As Intermediate Carriers in Fe-S Cluster Biosynthetic Pathways.
Iron-sulfur (Fe-S) clusters are protein cofactors that are required for many essential cellular functions. Fe-S clusters are synthesized and inserted into target proteins by an elaborate biosynthetic process. The insensitivity of most Fe-S assembly and transfer assays requires high concentrations for components and places major limits on reaction complexity. Recently, fluorophore labels were shown to be effective at reporting cluster content for Fe-S proteins. Here, the incorporation of this labeling approach allowed the design and interrogation of complex Fe-S cluster biosynthetic reactions that mimic in vivo conditions. A bacterial Fe-S assembly complex, composed of the cysteine desulfurase IscS and scaffold protein IscU, was used to generate [2Fe-2S] clusters for transfer to mixtures of putative intermediate carrier and acceptor proteins. The focus of this study was to test whether the monothiol glutaredoxin, Grx4, functions as an obligate [2Fe-2S] carrier protein in the Fe-S cluster distribution network. Interestingly, [2Fe-2S] clusters generated by the IscS-IscU complex transferred to Grx4 at rates comparable to previous assays using uncomplexed IscU as a cluster source in chaperone-assisted transfer reactions. Further, we provide evidence that [2Fe-2S]-Grx4 delivers clusters to multiple classes of Fe-S targets via direct ligand exchange in a process that is both dynamic and reversible. Global fits of cluster transfer kinetics support a model in which Grx4 outcompetes terminal target proteins for IscU-bound [2Fe-2S] clusters and functions as an intermediate cluster carrier. Overall, these studies demonstrate the power of chemically conjugated fluorophore reporters for unraveling mechanistic details of biological metal cofactor assembly and distribution networks.


Sex-Specific Effects of Apolipoprotein ε4 Allele on Mortality in Very Old and Centenarian Japanese Men.
The apolipoprotein E4 (APOE ε4) allele has attracted attention as an age-related genetic factor, both in neurology and gerontology. To understand the effects of the APOE ε4 allele on mortality in elderly individuals, we combined Japanese prospective cohort studies comprising 535 very old individuals (85-99 years of age) and 930 centenarians (over 100 years of age) and analyzed the association between mortality rates and candidate factors, including the APOE ε4 allele. APOE genotyping revealed an inverse correlation between the APOE ε4 allele carrier rate and age. Additionally, APOE ε4 allele carrier rate in centenarian men was significantly lower than that in centenarian women. The association analysis between APOE ε4 allele carriers and all-cause mortality indicated that APOE ε4 carriers showed significantly higher mortality rates than the APOE ε4 noncarriers among men in the very old group. Further analysis using Cox proportional hazard models indicated that cause-specific mortalities, including pneumonia and severe dementia, were associated with APOE ε4 carriers. These findings indicate that the APOE ε4 allele shows phenotypic male-specific adverse effects in the very old, which would explain the high mortality rate observed in this group, resulting in a low APOE ε4 allele carrier rate in centenarian men.


Isolation and characterization of chondroitin sulfates from sturgeon (Acipenser sinensis) and their effects on growth of fibroblasts.
Chondroitin sulfate (CS) is a glycosaminoglycan that composed of hexosamine (D-galactosamine) and hexuronic acid (D-glucuronic acid) unit arranged in an alternating unbranched sequence. CS is an essential component of the extracellular matrix (ECM) of connective tissue. It is mainly covalently attached to core proteins in the form of proteoglycans so that it exhibits specific interactions with proteins for cell growth, differentiation, division and migration. In this study, CSs were purified from the cartilage and backbone of sturgeon (Acipenser sinensis). To characterize their biochemical properties, we performed disaccharide compositional analysis after chondroitinase ABC digestion, high performance size exclusion chromatography (HPSEC) and (1)H-NMR spectroscopy. We also investigated the effects of CSs on fibroblast proliferation and adhesion to determine whether wound healing was accelerated in vitro and proliferation of different mitogen-activated protein kinases (MAPK) signaling pathways was facilitated. The CS purified from sturgeon cartilage was primarily composed of 4-sulfated CS (88.8%) and sturgeon backbone CS contains more than 60% 6-sulfated CS. The average molecular weights of CSs obtained from sturgeon cartilage and backbone were found to be 8 and 43 kDa, respectively. Our results showed that both CSs are able to increase cell adhesion, induce proliferation and migration on fibroblasts and may accelerate wound healing by inducing MAPK signaling pathways.


Conjunctival transcriptomics in ocular mucous membrane pemphigoid.
Ocular Mucous Membrane Pemphigoid (OcMMP) is an orphan disease characterized by chronic autoimmune-driven conjunctival inflammation leading to progressive scarring, debilitating symptoms, and blinding sequelae. This feasibility study aims to demonstrate conjunctival genetic transcriptomic analyses as a putative tool for interrogation of pathogenic signaling pathways in OcMMP. Conjunctival RNA profiling using the NanoString nCounter Human Fibrosis panel was undertaken on RNA extracted from conjunctival swabs obtained from 6 MMP patients (8 eyes; 4 M/2F; median age 78 [range 64-84] years); and 8 age-matched control participants (15 eyes; 3 M/5F; median age 69.5 [range 69-88] years). Data from 770 genes were analyzed with ROSALIND HyperScale architecture and stratified according to the level of clinically visible bulbar conjunctival inflammation. Normalization, fold-changes (≥+1.5-fold or ≤ -1.5-fold) and p-values adjustment (<0.05) using the Benjamini-Hochberg method were calculated. 93 differentially expressed genes (DEGs) were observed between OcMMP versus controls of which 48 were upregulated, and 45 downregulated. The top 4 upregulated DEGs represented fibrosis (COL3A1, COL1A1, FN1 and THBS1) while the key under-expressed genes (SCIN, HMGS2, XCL1/2) were indicative of ocular surface failure (goblet cell loss, keratinization, vulnerability to secondary infections). Forty-four pathways had a global significance score ≥2, the most significant being those related to extracellular matrix (ECM) remodeling, synthesis, and degradation. These pathways were accentuated in eyes with visible inflammation. NanoString methodology acquired via a simple conjunctival swab identifies profibrotic genes in OcMMP group and differentiates inflamed eyes. Longitudinal sampling and following investigative intervention will further mechanistic insight and development of novel biomarkers to monitor disease progression.


Visual tracking synchronization as a metric for concussion screening.
Our goal was to determine whether performance variability during predictive visual tracking can provide a screening measure for mild traumatic brain injury (mTBI). Seventeen subjects with chronic postconcussive syndrome and 9 healthy control subjects were included in this study. Eye movements were recorded with video-oculography as the subject visually tracked a target that moved through a circular trajectory. We compared the variability of gaze positional errors relative to the target with the microstructural integrity of white matter tracts as measured by the fractional anisotropy (FA) parameter of diffusion tensor imaging. Gaze error variability was significantly correlated with the mean FA values of the right anterior corona radiata (ACR) and the left superior cerebellar peduncle, tracts that support spatial processing and sustenance of attention, and the genu of the corpus callosum. Because the ACR and the genu are among the most frequently damaged white matter tracts in mTBI, the correlations imply that gaze error variability during visual tracking may provide a useful screening tool for mTBI. Gaze error variability was also significantly correlated with attention and working memory measures in neurocognitive testing; thus, measurement of visual tracking performance is promising as a fast and practical screening tool for mTBI.


Metastatic melanoma: a regional review and future directions.
. The incidence of malignant melanoma has risen steadily over recent decades. NCI data from 2005-2007 have suggested that 1.93% of individuals born today in the US will develop melanoma at some stage. Approximately 15% of patients with MM either present with metastatic disease or develop metastases during the course of their illness. Unfortunately, metastatic MM remains a challenge with limited treatment options, and median overall survival is 6-9 months. We reviewed our data for the treatment of metastatic MM over a period of four years. Data from all patients with metastatic MM treated with systemic therapy without clinical trials from 2006 to 2009 were reviewed. Response rate was determined as per RECIST criteria. Sixty four patients were treated with one or more lines of cytotoxic therapy. Median age was 62 years (range, 23-82) with 53% males. Primary site of the disease was the skin in 75%, mucosal in 12.5%, ocular in 9.4% and nodal with an occult primary in 3.1%. Visceral metastases were present in 75% of patients at the start of treatment, including pulmonary (39.6%) and hepatic (34.4%). All patients were screened for brain metastases, which were present in 26.5% of patients. ECOG performance status was 0 in 7.8%, 1 in 68.7%, 2 in 9.4% and undocumented in the remaining 14%. Patients without brain metastases received single agent DTIC as first line; those with brain metastases received temozolomide. Response rate was 7% for DTIC and 28% for temozolomide, with median progression-free survival of 2.4 and 3.2 months, respectively. Seven patients who received DTIC are alive on follow-up, 2 have ongoing stable disease post-DTIC at 41 months and 18 months. Second line therapy with vinblastine was given to 21 patients (32%), with a response rate of 9.5% and median progression-free survival of 3.4 months. Median overall survival from initiation of therapy was 7.7 months for DTIC and 3.6 months for patients with brain metastases receiving temozolomide. A performance status of 2 was associated with shorter median overall survival (2.0 months). . Our results are comparable to published data. Malignant melanoma is a disease with rising incidence and limited treatment options. These patients are best treated in the context of clinical trials as new targeted therapies are promising as future strategies.


Male reproductive organs are at risk from environmental hazards.
Male reproductive disorders that are of interest from an environmental point of view include sexual dysfunction, infertility, cryptorchidism, hypospadias and testicular cancer. Several reports suggest declining sperm counts and increase of these reproductive disorders in some areas during some time periods past 50 years. Except for testicular cancer this evidence is circumstantial and needs cautious interpretation. However, the male germ line is one of the most sensitive tissues to the damaging effects of ionizing radiation, radiant heat and a number of known toxicants. So far occupational hazards are the best documented risk factors for impaired male reproductive function and include physical exposures (radiant heat, ionizing radiation, high frequency electromagnetic radiation), chemical exposures (some solvents as carbon disulfide and ethylene glycol ethers, some pesticides as dibromochloropropane, ethylendibromide and DDT/DDE, some heavy metals as inorganic lead and mercury) and work processes such as metal welding. Improved working conditions in affluent countries have dramatically decreased known hazardous workplace exposures, but millions of workers in less affluent countries are at risk from reproductive toxicants. New data show that environmental low-level exposure to biopersistent pollutants in the diet may pose a risk to people in all parts of the world. For other toxicants the evidence is only suggestive and further evaluation is needed before conclusions can be drawn. Whether compounds as phthalates, bisphenol A and boron that are present in a large number of industrial and consumer products entails a risk remains to be established. The same applies to psychosocial stressors and use of mobile phones. Finally, there are data indicating a particular vulnerability of the fetal testis to toxicants-for instance maternal tobacco smoking. Time has come where male reproductive toxicity should be addressed form entirely new angles including exposures very early in life.


Genetic services for familial cancer patients: a follow-up survey of National Cancer Institute Cancer Centers.
Anecdotal reports suggest that the volume of services offered to individuals concerned with hereditary cancer risk has increased substantially in recent years. As a follow-up to our 1993 survey, we sought to determine how the scope and volume of genetic services has changed between 1993 and 2002. We surveyed the 61 National Cancer Institute-designated cancer centers in operation in 2002 using an updated version of the questionnaire from 1993. Analysis included frequencies and summary statistics. The majority of cancer centers responding (46 of 56 centers; 82.1%) provided some genetic services for evaluation of familial cancer, which is a higher proportion than in 1993 (50%; P < .01). Almost all centers (42 of 46 centers; 91.3%) provided services not only to cancer patients and their families, but also to individuals concerned with risk, which is a change (P = .01) from 1993, when 64.7% of centers offered such services. In addition, increases have been found for most other measures of services rendered for familial genetic services. As public awareness of cancer susceptibility genes has grown markedly in recent years, the demand has also grown for genetic services to assess familial cancer risk. Major deleterious genetic mutations are rare, and much of the current research in genetic variation focuses on higher prevalence variants that carry lower risks. This may suggest that testing for mutations will move from genetics clinics to primary care and specialty practices. Thus, it is unclear whether the scope and volume of cancer center genetics services will continue to grow as rapidly as they have over the last decade.


Subcellular localization of selectively permeable aquaporins in the male germ line of a marine teleost reveals spatial redistribution in activated spermatozoa.
In oviparous vertebrates such as the marine teleost gilthead seabream, water and fluid homeostasis associated with testicular physiology and the external activation of spermatozoa is potentially mediated by multiple aquaporins. To test this hypothesis, we isolated five novel members of the aquaporin superfamily from gilthead seabream and developed paralog-specific antibodies to localize the cellular sites of protein expression in the male reproductive tract. Together with phylogenetic classification, functional characterization of four of the newly isolated paralogs, Aqp0a, -7, -8b, and -9b, demonstrated that they were water permeable, while Aqp8b was also permeable to urea, and Aqp7 and -9b were permeable to glycerol and urea. Immunolocalization experiments indicated that up to seven paralogous aquaporins are differentially expressed in the seabream testis: Aqp0a and -9b in Sertoli and Leydig cells, respectively; Aqp1ab, -7, and -10b from spermatogonia to spermatozoa; and Aqp1aa and -8b in spermatids and sperm. In the efferent duct, only Aqp10b was found in the luminal epithelium. Ejaculated spermatozoa showed a segregated spatial distribution of five aquaporins: Aqp1aa and -7 in the entire flagellum or the head, respectively, and Aqp1ab, -8b, and -10b both in the head and the anterior tail. The combination of immunofluorescence microscopy and biochemical fractionation of spermatozoa indicated that Aqp10b and phosphorylated Aqp1ab are rapidly translocated to the head plasma membrane upon activation, whereas Aqp8b accumulates in the mitochondrion of the spermatozoa. In contrast, Aqp1aa and -7 remained unchanged. These data reveal that aquaporin expression in the teleost testis shares conserved features of the mammalian system, and they suggest that the piscine channels may play different roles in water and solute transport during spermatogenesis, sperm maturation and nutrition, and the initiation and maintenance of sperm motility.


Brain size, head size and behaviour of a passerine bird.
A recent increase in comparative studies of the ecological and evolutionary consequences of brain size in birds and primates in particular have suggested that cognitive abilities constitute a central link. Surprisingly, there are hardly any intraspecific studies investigating how individuals differing in brain size behave, how such individuals are distributed and how brain size is related to life history and fitness components. Brain mass of the barn swallow Hirundo rustica was strongly predicted by external head volume, explaining 99.5% of the variance, allowing for repeatable estimates of head volume as a reflection of brain size. Repeatability of head volume within and between years was high, suggesting that measurement errors were small. In a 2 years study of 501 individual adult barn swallows, I showed that head volume differed between sexes and age classes, with yearlings having smaller and more variable heads than older individuals, and females having smaller and more variable heads than males. Large head volume was not a consequence of large body size, which was a poor predictor of head volume. Birds with large heads arrived early from spring migration, independent of sex and age, indicating that migratory performance may have an important cognitive component. Head volume significantly predicted capture date and recapture probability, suggesting that head volume is related to learning ability, although morphological traits such as wing length, aspect ratio and wing area were unimportant predictors. Intensity of defence of offspring increased with head volume in females, but not in males. Barn swallows with large heads aggregated in large colonies, suggesting that individuals with large heads were more common in socially complex environments. These results suggest that brain size is currently under natural and sexual selection, and that micro-evolutionary processes affecting brain size can be studied under field conditions.


Mood disorders and parity - a clue to the aetiology of the postpartum trigger.
Episodes of postpartum psychosis have been associated with first pregnancies in women with bipolar I disorder. It is unclear, however, if the effect extends to episodes at other times in relation to childbirth and to women with other mood disorders such as major depression and bipolar II disorder. This primiparity effect, which is also seen in other pregnancy related conditions such as pre-eclampsia, is a potentially important clue to the aetiology of childbirth related mood episodes. Participants were interviewed and case notes reviewed. Best-estimate diagnoses were made according to DSM-IV criteria. Data on the occurrence of episodes in pregnancy and the postpartum were available on 3345 full term deliveries from 1667 participants, 934 with bipolar I disorder (BD-I), 278 with bipolar II disorder (BD-II) and 455 with recurrent major depression (RMD). Onsets of psychosis/mania within 6 weeks of childbirth were overrepresented in primiparae (p=0.007) with BD-I. Although primiparity was not associated with perinatal bipolar depression, there was an association with the onset of depression within 6 weeks in women with RMD (p=0.035). Whilst women experiencing a postpartum episode were less likely to go on to have further children, this did not account for the association with primiparity. Data were collected retrospectively. Information on pharmacological treatment was not available. Primiparity is associated not only with postpartum psychosis/mania in BD-I, but also with postpartum depression in RMD. Psychosocial factors and biological differences between first and subsequent pregnancies may play a role and are candidates for examination in further studies.


Limb Hypothermia for Preventing Paclitaxel-Induced Peripheral Neuropathy in Breast Cancer Patients: A Pilot Study.
Peripheral neuropathy (PN) due to paclitaxel is a common dose-limiting toxicity with no effective prevention or treatment. We hypothesize that continuous-flow limb hypothermia can reduce paclitaxel-induced PN. An internally controlled pilot trial was conducted to investigate the neuroprotective effect of continuous-flow limb hypothermia in breast cancer patients receiving weekly paclitaxel. Patients underwent limb hypothermia of one limb for a duration of 3 h with every paclitaxel infusion, with the contralateral limb used as control. PN was primarily assessed using nerve conduction studies (NCSs) before the start of chemotherapy, and after 1, 3, and 6 months. Skin temperature and tolerability to hypothermia were monitored using validated scores. Twenty patients underwent a total of 218 cycles of continuous-flow limb hypothermia at a coolant temperature of 22°C. Continuous-flow limb hypothermia achieved mean skin temperature reduction of 1.5 ± 0.7°C and was well tolerated, with no premature termination of cooling due to intolerance. Grade 3 PN occurred in 2 patients (10%), grade 2 in 2 (10%), and grade 1 in 12 (60%). Significant correlation was observed between amount of skin cooling and motor nerve amplitude preservation at 6 months (p < 0.0005). Sensory velocity and amplitude in the cooled limbs were less preserved than in the control limbs, but the difference did not attain statistical significance. One patient with a history of diabetes mellitus had significant preservation of compound muscle action potential in the cooled limb on NCS analysis. This study suggests that continuous limb hypothermia accompanying paclitaxel infusion may reduce paclitaxel-induced PN and have therapeutic potential in select patients and warrants further investigation. The method is safe and well tolerated.


Glasgow Prognostic Score predicts prognosis of intrahepatic cholangiocarcinoma.
High Glasgow Prognostic Score (GPS) has been associated with poor prognosis in patients with lung, ovarian, colorectal and renal cancer, as well as hepatocellular carcinoma. The aim of this study was to investigate the prognostic value of GPS in patients with intrahepatic cholangiocarcinoma (ICC) undergoing partial hepatectomy. A total of 72 patients with pathologically confirmed ICC were classified according to their GPS scores assigned based on the preoperative levels of C-reactive protein (CRP) and albumin. Their clinicopathological data were retrospectively assessed using univariate and multivariate analysis to determine their association with overall survival and recurrence. High GPS scores in ICC patients were associated with preoperative levels of CRP (P<0.001) and albumin (P<0.001), frequency of ascites accumulation (P=0.035), lymph node metastasis (P=0.002) and tumour size (P=0.005). On univariate analysis, preoperative levels of CRP (P<0.001), albumin (P=0.016) and carbohydrate antigen 19-9 (P=0.038), hepatitis B virus (HBV) positivity (P=0.009), occurrence of lymph node metastasis (P=0.001), Child-Pugh class B (P=0.013) and high tumour-node-metastasis (TNM) stage (P=0.002) were found to be associated with the 1- and 3-year overall survival. Multivariate analysis suggested that GPS score (HR=2.037, 95% CI: 1.092-3.799, P=0.025), TNM classification (HR=2.000, 95% CI: 1.188-3.367, P=0.009) and HBV positivity (HR=0.559 95% CI: 0.328-0.953, P=0.032) were independently associated with patient survival. High GPS scores also predicted ICC recurrence. In conclusion, our results demonstrated that GPS may serve as an independent marker of prognosis in patients with ICC following partial hepatectomy.


Clinical Outcomes Associated with Achromobacter Species Infection in Patients with Cystic Fibrosis.
Achromobacter species are increasingly identified in individuals with cystic fibrosis (CF), but the clinical outcomes in these patients remain poorly understood. We aimed to determine the association of Achromobacter infection on clinical outcomes in pediatric and adult patients with CF. A cohort study of pediatric and adult patients with CF was conducted from 1997 to 2014 in Toronto, Ontario, Canada. Achromobacter spp. infection was categorized as no history of infection, intermittent infection, and chronic infection (two or more positive cultures in the preceding 12 months). Cox models were used to estimate risk of death or transplantation. Mixed-effects models were used to assess odds of pulmonary exacerbations and effect on lung function (FEV1%) by Achromobacter spp. A total of 1,103 patients were followed-up over the course of 18 years; 88 patients (7.3%) had one or more culture for Achromobacter species. Chronic Achromobacter infection was associated with a greater risk of death or transplantation compared with in patients with no history of infection (adjusted hazard ratio, 2.03; 95% confidence interval, 1.05-3.95; P = 0.036). Pulmonary exacerbations were more common in patients with chronic infection, but after adjusting for confounding factors, the effect was no longer significant. The chronic group had lower FEV1%, but it did not worsen after developing chronic infection. Patients with CF and chronic Achromobacter infection are at increased risk of death or transplantation.


Molecular characterization of rabbit scavenger receptor class B types I and II: portal to central vein gradient of expression in the liver.
To further elucidate the role of scavenger receptor class B type I (SR-BI) in reverse cholesterol transport and in atherogenesis, we performed studies in the rabbit, an animal model displaying a lipoprotein profile similar to that of human, expressing cholesteryl ester transfer protein in plasma and having been demonstrated to be susceptible to atherosclerosis. In this report, we describe for the first time the isolation and characterization of rabbit cDNA fragments encoding SR-BI and scavenger receptor class B type II (SR-BII). Development of an isoform-specific Taqman Real Time PCR system and generation of isoform-specific polyclonal antibodies allowed us to measure SR-BI and SR-BII expression in various rabbit organs on mRNA and protein levels, respectively. We found the highest expression of SR-BI in adrenal gland, liver, and proximal intestine; lesser expression was found in appendix and spleen. Immunohistochemical staining of frozen sections showed SR-BI expression in the cortex but not in the medulla of adrenal gland. An increasing portal to central vein gradient of expression was found within the hepatic lobule. As shown in this report, identification and characterization of SR-BI expression in the rabbit affords a powerful tool to elucidate the role of SR-BI in cholesterol homeostasis and atherogenesis in human.


Cryptococcus neoformans population diversity and clinical outcomes of HIV-associated cryptococcal meningitis patients in Zimbabwe.
HIV and cryptococcal meningitis co-infection is a major public health problem in most developing countries. Cryptococcus neoformans sensu stricto is responsible for the majority of HIV-associated cryptococcosis cases in sub-Saharan Africa. Despite the available information, little is known about cryptococcal population diversity and its association with clinical outcomes in patients with HIV-associated cryptococcal meningitis in sub-Saharan Africa. In a prospective cohort, we investigated the prevalence and clinical outcome of Cryptococcusneoformans sensu stricto meningitis among HIV-infected patients in Harare, Zimbabwe, and compared the genotypic diversity of the isolates with those collected from other parts of Africa. Molecular typing was done using amplified fragment length polymorphism genotyping and microsatellite typing. The majority of patients with HIV-associated Cryptococcusneoformans sensu stricto meningitis in this cohort were males (n=33/55; 60.0 %). The predominant Cryptococcus neoformans sensu stricto genotype among the Zimbabwean isolates was genotype AFLP1/VNI (n=40; 72.7 %), followed by AFLP1A/VNB/VNII (n=8; 14.6 %), and AFLP1B/VNII was the least isolated (n=7; 12.7 %). Most of the isolates were mating-type α (n=51; 92.7 %), and only four (7.3 %) were mating-type a. Overall in-hospital mortality was 55.6 % (n=30), and no difference between infecting genotype and clinical outcome of patient (P=0.73) or CD4+ counts (P=0.79) was observed. Zimbabwean Cryptococcusneoformans sensu stricto genotypes demonstrated a high level of genetic diversity by microsatellite typing, and 51 genotypes within the main molecular types AFLP1/VNI, AFLP1A/VNB/VNII and AFLP1B/VNII were identified. This study demonstrates that Cryptococcusneoformans sensu stricto in Zimbabwe has a high level of genetic diversity when compared to other regional isolates.


Decelerating burst and complex repetitive discharges in the striated muscle of the urethral sphincter, associated with urinary retention in women.
A type of electromyographic activity, formerly referred to as "pseudomyotonia", can be recorded from the striated muscle of the urethral sphincter using a concentric needle electrode. There are two components to this activity, complex repetitive discharges and decelerating bursts. The latter usually dominate recordings and sound very like myotonic discharges. Analysis of these discharges indicates that they are a form of "bizarre repetitive discharge", and as such, result from ephaptic spread of excitation between muscle fibres rather than from excitation arising in the terminal branches of the motor axon. Profuse activity of this type has been found in 15 women with symptoms of urethral dysfunction, including 11 with urinary retention. It is suggested that this activity is associated with a failure of urethral sphincter relaxation.


Bensingtonia rectispora sp. nov. and Bensingtonia bomiensis sp. nov., ballistoconidium-forming yeast species from Tibetan plant leaves.
Five yeast strains isolated from plant leaves collected in south-east Tibet formed cream to brownish colonies and produced asymmetrical ballistoconidia and CoQ-9 as the major ubiquinone. Sequence analysis of the 26S rRNA D1/D2 domain and the internal transcribed spacer region indicated that these strains represented two novel species of the genus Bensingtonia. The names Bensingtonia rectispora sp. nov. (type strain XZ 4C5(T) = CGMCC 2.02635(T) = CBS 10710(T)) and Bensingtonia bomiensis sp. nov. (type strain XZ 33D1(T) = CGMCC 2.02670(T) = CBS 10713(T)) are proposed for the two novel species, which are phylogenetically closely related to Bensingtonia naganoensis, Bensingtonia pseudonaganoensis and the type species of the genus, Bensingtonia ciliata.


Three-mode factor analysis by means of Candecomp/Parafac.
A three-mode covariance matrix contains covariances of N observations (e.g., subject scores) on J variables for K different occasions or conditions. We model such an JK×JK covariance matrix as the sum of a (common) covariance matrix having Candecomp/Parafac form, and a diagonal matrix of unique variances. The Candecomp/Parafac form is a generalization of the two-mode case under the assumption of parallel factors. We estimate the unique variances by Minimum Rank Factor Analysis. The factors can be chosen oblique or orthogonal. Our approach yields a model that is easy to estimate and easy to interpret. Moreover, the unique variances, the factor covariance matrix, and the communalities are guaranteed to be proper, a percentage of explained common variance can be obtained for each variable-condition combination, and the estimated model is rotationally unique under mild conditions. We apply our model to several datasets in the literature, and demonstrate our estimation procedure in a simulation study.


Predictors for persistent neuropathic pain--a Delphi survey.
Chronic neuropathic pain has a major effect on quality of life. In order to prevent neuropathic pain from becoming chronic and improve neuropathic pain care, it is important to identify predictors associated with the persistence of neuropathic pain. To identify potential predictors associated with the persistence of neuropathic pain. A 2-round Delphi study. University Medical Center and Pain Management Research Center. A 2-round Delphi study was conducted among 17 experts in the field of neuropathic pain. Selection of the panel was based on the citation index ranking for neuropathic pain-related research and/or membership in the neuropathic pain special interest group of the International Association for the Study of Pain (IASP), complemented with experts with demonstrated field knowledge.Potential predictors were categorized according to the International Classification of Functioning, Disability and Health model. Participants were asked to identify important predictors, suggest new predictors, and grade the importance on a 0-10 scale. For the second round, predictors were considered important if the median score was ≥ 7 and the interquartile range (IQR) ≤ 3. In the first round, 20 predictors were selected and 58 were added by the experts (patient characteristics [15], environmental factors [25], functions & structure [4], participation & health related quality of life [14]). In the second round, 12 predictors were considered important (patient characteristics [4; e.g., depression, pain catastrophizing], environmental factors [surgery as treatment for neuropathic pain], functions & structure [6; e.g., allodynia, duration of the complaints], participation & trait anxiety/depression as a part of health related quality of life). Presence of depression and pain catastrophizing were considered the most important predictors for chronic neuropathic pain (median ≥ 8; IQR ≤ 2). The study design did not include plenary discussion among the experts. The meaning of the individual topics used in this study could have been subject to interpretation bias. Overall, psychological factors and factors related to sensory disturbances were considered important predictors for persistence of neuropathic pain. Activity related factors and previously received paramedical and alternative treatment were considered to be less important. The list of possible predictors obtained by this study may serve as a basis for development of a clinical prediction rule for chronic neuropathic pain.


Near infrared spectroscopy monitoring in the pediatric cardiac catheterization laboratory.
Near-infrared spectroscopy (NIRS) is a noninvasive method used to evaluate tissue oxygenation. We evaluated the relationship between cerebral and renal NIRS parameters during transcatheter intervention and adverse events in the catheterization room. Between January 1 and May 31, 2012, 123 of 163 pediatric patients undergoing cardiac catheterization were followed by NIRS. All were monitored by electrocardiography, noninvasive blood pressure measurement, pulse oxymetry, initial and final blood lactate level measurement. The number of interventional procedures was 73 (59%). During the procedures, 39 patients experienced a total of 41 adverse events: 18 (19.5%) had desaturation, 10 (8.1%) arrhythmia, three (2.4%) had respiratory difficulty, six (4.8%) had a situation calling for cardiopulmonary resuscitation, three (2.4%) had anemia necessitating transfusion, and one (0.8%) had a cyanotic spell. Cranial NIRS values worsened in 12 (9.8%) and renal measurements worsened in 13 (12.5%) patients. The sensitivity and specificity of a 9% impairment of cranial values were 90 and 61%, respectively, while the corresponding calculations for a 21% fall in renal measurements were 54% sensitivity and 90% specificity. When arrhythmia developed, NIRS values fell simultaneously, while the development of a desaturation problem was heralded by NIRS falling 10-15 s earlier than changes in pulse oxymetry; on improving saturation, NIRS returned to earlier values 10-15 s before pulse oxymetry readings. NIRS monitoring may provide an early warning with regard to complications likely to develop during a procedure. A fall of 9% in cranial NIRS values, or of 21% in renal measurements, should raise clinician awareness.


Characterization of ebastine, hydroxyebastine, and carebastine metabolism by human liver microsomes and expressed cytochrome P450 enzymes: major roles for CYP2J2 and CYP3A.
Ebastine undergoes extensive metabolism to form desalkylebastine and hydroxyebastine. Hydroxyebastine is subsequently metabolized to carebastine. Although CYP3A4 and CYP2J2 have been implicated in ebastine N-dealkylation and hydroxylation, the enzyme catalyzing the subsequent metabolic steps (conversion of hydroxyebastine to desalkylebastine and carebastine) have not been identified. Therefore, we used human liver microsomes (HLMs) and expressed cytochromes P450 (P450s) to characterize the metabolism of ebastine and that of its metabolites, hydroxyebastine and carebastine. In HLMs, ebastine was metabolized to desalkyl-, hydroxy-, and carebastine; hydroxyebastine to desalkyl- and carebastine; and carebastine to desalkylebastine. Of the 11 cDNA-expressed P450s, CYP3A4 was the main enzyme catalyzing the N-dealkylation of ebastine, hydroxyebastine, and carebastine to desalkylebastine [intrinsic clearance (CL(int)) = 0.44, 1.05, and 0.16 microl/min/pmol P450, respectively]. Ebastine and hydroxyebastine were also dealkylated to desalkylebastine to some extent by CYP3A5. Ebastine hydroxylation to hydroxyebastine is mainly mediated by CYP2J2 (0.45 microl/min/pmol P450; 22.5- and 7.5-fold higher than that for CYP3A4 and CYP3A5, respectively), whereas CYP2J2 and CYP3A4 contributed to the formation of carebastine from hydroxyebastine. These findings were supported by chemical inhibition and kinetic analysis studies in human liver microsomes. The CL(int) of hydroxyebastine was much higher than that of ebastine and carebastine, and carebastine was metabolically more stable than ebastine and hydroxyebastine. In conclusion, our data for the first time, to our knowledge, suggest that both CYP2J2 and CYP3A play important roles in ebastine sequential metabolism: dealkylation of ebastine and its metabolites is mainly catalyzed by CYP3A4, whereas the hydroxylation reactions are preferentially catalyzed by CYP2J2. The present data will be very useful to understand the pharmacokinetics and drug interaction of ebastine in vivo.


Comparison of tuber proteomes of potato varieties, landraces, and genetically modified lines.
Crop improvement by genetic modification remains controversial, one of the major issues being the potential for unintended effects. Comparative safety assessment includes targeted analysis of key nutrients and antinutritional factors, but broader scale-profiling or "omics" methods could increase the chances of detecting unintended effects. Comparative assessment should consider the extent of natural variation and not simply compare genetically modified (GM) lines and parental controls. In this study, potato (Solanum tuberosum) proteome diversity has been assessed using a range of diverse non-GM germplasm. In addition, a selection of GM potato lines was compared to assess the potential for unintended differences in protein profiles. Clear qualitative and quantitative differences were found in the protein patterns of the varieties and landraces examined, with 1,077 of 1,111 protein spots analyzed showing statistically significant differences. The diploid species Solanum phureja could be clearly differentiated from tetraploid (Solanum tuberosum) genotypes. Many of the proteins apparently contributing to genotype differentiation are involved in disease and defense responses, the glycolytic pathway, and sugar metabolism or protein targeting/storage. Only nine proteins out of 730 showed significant differences between GM lines and their controls. There was much less variation between GM lines and their non-GM controls compared with that found between different varieties and landraces. A number of proteins were identified by mass spectrometry and added to a potato tuber two-dimensional protein map.


Helicobacter pylori Vacuolating Toxin and Gastric Cancer.
Helicobacter pylori VacA is a channel-forming toxin unrelated to other known bacterial toxins. Most H. pylori strains contain a vacA gene, but there is marked variation among strains in VacA toxin activity. This variation is attributable to strain-specific variations in VacA amino acid sequences, as well as variations in the levels of VacA transcription and secretion. In this review, we discuss epidemiologic studies showing an association between specific vacA allelic types and gastric cancer, as well as studies that have used animal models to investigate VacA activities relevant to gastric cancer. We also discuss the mechanisms by which VacA-induced cellular alterations may contribute to the pathogenesis of gastric cancer.


Monoclonal antibody Y13-259 recognizes an epitope of the p21 ras molecule not directly involved in the GTP-binding activity of the protein.
The p21 products of ras proto-oncogenes are GTP-binding proteins with associated GTPase activity. Recent studies have indicated that ras p21 may be required for the initiation of normal cell DNA synthesis, since microinjection of a monoclonal antibody, Y13-259, blocks serum stimulation of DNA synthesis in quiescent cell cultures (L. S. Mulcahy, M.R. Smith, and D. W. Stacey, Nature [London] 313:241-243, 1985). We localized the structural domain within the p21 molecule recognized by the Y13-259 monoclonal antibody. By analysis of a series of bacterially expressed p21 deletion mutants, the monoclonal antibody was found to interact with a region between positions 70 and 89 in the p21 amino acid sequence. By comparison of the coding sequences of different p21 proteins recognized by this monoclonal antibody, a highly conserved amino acid region between positions 70 and 81 was found to be the most likely site for the epitope detected by the Y13-259 antibody. This monoclonal antibody was further shown not to interfere directly with in vitro biochemical functions of the molecule, including GTP binding, GTPase, and autokinase activities.


PD-1H/VISTA mediates immune evasion in acute myeloid leukemia.
Acute myeloid leukemia (AML) presents a pressing medical need in that it is largely resistant to standard chemotherapy as well as modern therapeutics such as targeted therapy and immunotherapy, including anti-PD therapy. We demonstrate that Programmed Death-1 Homolog (PD-1H), an immune co-inhibitory molecule is highly expressed in blasts from the bone marrow of AML patients, while normal myeloid cell subsets and T cells have the expression of PD-1H. In studies employing syngeneic and humanized AML mouse models, overexpression of PD-1H promoted the growth of AML cells, mainly by evading T cell-mediated immune responses. Importantly, ablation of AML cell surface PD-1H by antibody blockade or genetic targeting significantly inhibited AML progression by promoting T cell activity. In addition, the genetic deletion of PD-1H from host normal myeloid cells inhibited AML progression as well and the combination of PD-1H blockade with PD-1 blockade conferred a synergistic anti-leukemia effect. Our findings provide the basis for PD-1H as an attractive therapeutic target to treat human AML.


Teratogenic effects of vigabatrin in TO mouse fetuses.
Vigabatrin (VGB) is a relatively recently introduced antiepileptic drug that enhances the brain levels of gamma aminobutyric acid (GABA). Few data on its teratogenic effects appear to have been reported. Our objective was to determine if VGB was teratogenic in the TO mouse. Single doses of 300-600 mg/kg of VGB dissolved in saline were administered intraperitoneally (IP) to groups of TO mice on one of gestation days (GD) 7-12. The controls were saline treated or untreated. No maternal toxic effects were observed in the 300 or 450 mg/kg groups, and the 600 mg/kg dose was totally lethal to the mothers. Fetuses were collected on GD 18. Both 300 and 450 mg/kg doses induced a consistently significant intrauterine growth retardation irrespective of the developmental stage at administration. VGB did not augment the spontaneous incidence of neural tube defects characteristic of this strain, but accelerated destruction of the brain in spontaneous exencephalic embryos. Mandibular and maxillary hypoplasia, arched palate, cleft palate (two cases), limb defects (one case), and exomphalos were observed in the malformed fetuses. The high incidence of exomphalos appears to be a unique result of VGB treatment. Alizarin red-S/alcian blue-stained, skeletons revealed hypoplasia of mid facial bones, stage-dependent increase in the frequency of cervical and lumbar ribs, rib fusion, and sternal and vertebral malformations in the drug-treated fetuses. Middle and distal phalanges of the forepaw and mid phalanges and tarsals of the hindpaw failed to ossify in a significant number of experimental fetuses. Homeotic shift in terms of presacral vertebral number and a high incidence of lumbar and cervical ribs in the treated group are suggestive of treatment-related alterations in gene expression. In view of the paucity of human and animal data on the reproductive toxicologic effects of VGB, the results of the present study assume particular importance and suggest that VGB should be used in pregnancy with extreme caution.


Sub-nm resolution cavity enhanced microspectrometer.
A novel on-chip spectrometer device using combined functionalities of a micro-ring resonator and a planar diffraction grating is proposed. We investigate the performance of this architecture by implementing it in a silicon-on-insulator platform. We experimentally demonstrate such a device with 100 channels, 0.1 nm channel spacing and a channel crosstalk less than -10 dB. The entire device occupies an area of less than 2 mm(2). Based on our initial results we envision that this device enables the possibility of the realization of low-cost and high-resolution ultra-compact spectroscopy.


Inclusion of the glucocorticoid receptor in a hypothalamic pituitary adrenal axis model reveals bistability.
The body's primary stress management system is the hypothalamic pituitary adrenal (HPA) axis. The HPA axis responds to physical and mental challenge to maintain homeostasis in part by controlling the body's cortisol level. Dysregulation of the HPA axis is implicated in numerous stress-related diseases. We developed a structured model of the HPA axis that includes the glucocorticoid receptor (GR). This model incorporates nonlinear kinetics of pituitary GR synthesis. The nonlinear effect arises from the fact that GR homodimerizes after cortisol activation and induces its own synthesis in the pituitary. This homodimerization makes possible two stable steady states (low and high) and one unstable state of cortisol production resulting in bistability of the HPA axis. In this model, low GR concentration represents the normal steady state, and high GR concentration represents a dysregulated steady state. A short stress in the normal steady state produces a small perturbation in the GR concentration that quickly returns to normal levels. Long, repeated stress produces persistent and high GR concentration that does not return to baseline forcing the HPA axis to an alternate steady state. One consequence of increased steady state GR is reduced steady state cortisol, which has been observed in some stress related disorders such as Chronic Fatigue Syndrome (CFS). Inclusion of pituitary GR expression resulted in a biologically plausible model of HPA axis bistability and hypocortisolism. High GR concentration enhanced cortisol negative feedback on the hypothalamus and forced the HPA axis into an alternative, low cortisol state. This model can be used to explore mechanisms underlying disorders of the HPA axis.


Antimicrobial Susceptibility Testing for Polymyxins: Challenges, Issues, and Recommendations.
Polymyxins, including polymyxin B and polymyxin E (colistin), are now increasingly being used worldwide to treat patients with multidrug-resistant (MDR) Gram-negative bacterial infections. This necessitates that laboratories employ an accurate and reliable method for the routine performance of polymyxin susceptibility testing. A number of reasons have accounted for the difficulties with susceptibility testing for the polymyxins, including their multicomponent composition, poor diffusion in the agar medium, adsorption to microtiter plates, the lack of a reliable susceptibility test, the lack of a specific breakpoint from professional organizations, the synergistic effect of polysorbate 80, and the development of heteroresistance. This minireview discusses such problems that impact the results of currently available susceptibility testing methods. We also provide emerging concepts on mechanisms of polymyxin resistance, including chromosomally and plasmid-mediated mcr-related resistance. Broad-range investigations on such critical issues in relation to polymyxins can be beneficial for the implementation of effective treatment against MDR Gram-negative bacterial infections.


Blood transfusion determines postoperative morbidity in pediatric cardiac surgery applying a comprehensive blood-sparing approach.
Recently we suggested a comprehensive blood-sparing approach in pediatric cardiac surgery that resulted in no transfusion in 71 infants (25%), postoperative transfusion only in 68 (24%), and intraoperative transfusion in 149 (52%). We analyzed the effects of transfusion on postoperative morbidity and mortality in the same cohort of patients. The effect of transfusion on the length of mechanical ventilation and intensive care unit stay was assessed using Kaplan-Meier curves. To assess whether transfusion independently determined the length of mechanical ventilation and length of intensive care unit stay, a multivariate model was applied. Additionally, in the subgroup of transfused infants, the effect of the applied volume of packed red blood cells was assessed. The median length of mechanical ventilation was 11 hours (interquartile range, 9-18 hours), 33 hours (interquartile range, 18-80 hours), and 93 hours (interquartile range, 34-161 hours) in the no transfusion, postoperative transfusion only, and intraoperative transfusion groups, respectively (P < .00001). The corresponding median lengths of intensive care unit stay were 1 day (interquartile range, 1-2 days), 3.5 days (interquartile range, 2-5 days), and 8 days (interquartile range, 3-9 days; P < .00001). The multivariate hazard ratio for early extubation was 0.24 (95% confidence interval, 0.16-0.35) and 0.37 (95% confidence interval, 0.25-0.55) for the intraoperative transfusion and postoperative transfusion only groups, respectively (P < .00001). In addition, the cardiopulmonary time, body weight, need for reoperation, and hemoglobin during cardiopulmonary bypass affected the length of mechanical ventilation. Similar results were obtained for the length of intensive care unit stay. In the subgroup of transfused infants, the volume of packed red blood cells also independently affected both the length of mechanical ventilation and the length of intensive care unit stay. The incidence and volume of blood transfusion markedly affects postoperative morbidity in pediatric cardiac surgery. These results, although obtained by retrospective analysis, might stimulate attending physicians to establish stringent blood-sparing approaches in their institutions.


Regulatory T cells: customizing for the clinic.
Immune-suppressive cellular immunotherapy requires large numbers of antigen-specific regulatory T cells (T(reg) cells), lymphocytes that suppress certain immune responses. Together, three papers in this issue of Science Translational Medicine describe protocols for the ex vivo expansion of human T(reg) cells and assess the immune-suppressive function of ex vivo-manipulated T(reg) cells after transfer into humanized mouse disease models. Along with recent phase I clinical trial results, these new data provide a platform for clinical use of T(reg) cells as personalized therapeutic agents for the treatment of autoimmune diseases, graft-versus-host disease, and transplant rejection.


Triacylglycerols and Other Lipids Profiling of Hemp By-Products.
Hemp seed by-products, namely hemp cake (hemp meal) and hemp hulls were studied for their lipid content and composition. Total lipid content of hemp cake and hemp hulls was 13.1% and 17.5%, respectively. Oil extraction yields using hexane, on the other hand, were much lower in hemp cake (7.4%) and hemp hulls (12.1%). Oil derived from both hemp seeds and by-products were primarily composed of neutral lipids (>97.1%), mainly triacylglycerols (TAGs), determined by SPE and confirmed by NMR study. Linoleic acid was the major fatty acid present in oils derived from hemp by-products, covering almost 55%, followed by α-linolenic acid, covering around 18% of the total fatty acids. For the first time, 47 intact TAGs were identified in the hemp oils using UPLC-HRMS. Among them, TAGs with fatty acid acyl chain 18:3/18:2/18:2 and 18:3/18:2/18:1 were the major ones, followed by TAGs with fatty acid acyl chain of 18:3/18:3/18:2, 18:2/18:2/16:0, 18:2/18:2/18:1, 18:3/18:2.18:0, 18:2/18:2/18:0, 18:2/18:1/18:1 and 18:3/18:2:16:0. Besides TAGs, low levels of terpenes, carotenoids and cannabidiolic acid were also detected in the oils. Moreover, the oils extracted from hemp by-products possessed a dose-dependent DPPH radical scavenging property and their potencies were in a similar range compared to other vegetable oils.


Laser-assisted 3D bioprinting of exocrine pancreas spheroid models for cancer initiation study.
Pancreatic ductal adenocarcinoma (PDAC) is the most common malignancy of the pancreas. It has shown a poor prognosis and a rising incidence in the developed world. Other pathologies associated with this tissue include pancreatitis, a risk condition for pancreatic cancer. The onset of both pancreatitis and pancreatic cancer follows a common pattern: exocrine pancreatic acinar cells undergo a transdifferentiation to duct cells that triggers a 3D restructuration of the pancreatic tissue. However, the exact mechanism underlying this process remains partially undefined. Further understanding the cellular events leading to PDAC could open new avenues in the development of novel therapeutic approaches. Since current 2D cell culture models fail to mimic the tridimensional complexity of the pancreatic tissue, new in vitro models are urgently needed. Here, we generated 3D pancreatic cell spheroid arrays using laser-assisted bioprinting and characterized their phenotypic evolution over time through image analysis and phenotypic characterization. We show that these bioprinted spheroids, composed of both acinar and ductal cells, can replicate the initial stages of PDAC development. This bioprinted miniaturized spheroid-based array model should prove useful for the study of the internal and external factors that contribute to the formation of precursor PDAC lesions and to cancer progression, and may therefore shed light on future PDAC therapy strategies.


Fish intake and risk of chronic obstructive pulmonary disease in 2 large US cohorts.
Despite mechanistic data that linked fish and omega-3 (n-3) PUFAs with lower risk of chronic obstructive pulmonary disease (COPD), epidemiologic data remain scarce. Fish and n-3 PUFAs are an important component of the prudent dietary pattern that is thought to be protective in the onset of COPD. We examined the role of fish and PUFA intakes on risk of developing COPD while taking into account the overall dietary pattern. We investigated the objective in 120,175 women and men from the Nurses' Health Study and Health Professionals Follow-Up Study. Over the study period (1984-2000), there were 889 cases of newly diagnosed COPD. Cumulative average intakes of fish, eicosapentaenoic acid, docosahexaenoic acid, n-3 PUFAs, n-6 PUFAs, and the n-3:n-6 ratio were calculated from repeated food-frequency questionnaires. Because fish is a food group included in the prudent pattern, we derived a new prudent pattern without the contribution from fish, and we termed this pattern the "modified prudent" pattern. We performed multivariable Cox proportional hazards models. Before the dietary pattern was taken into account, and with 14 factors controlled for, we showed that more-frequent fish intake (≥4 servings/wk) was inversely associated with risk of COPD [adjusted pooled HR for the highest intake compared with the lowest intake (<1 serving/wk): 0.71; 95% CI: 0.54, 0.94]. After additional adjustment for the dietary pattern (modified prudent and Western patterns), the association was NS (0.84; 95% CI: 0.63, 1.13). No significant associations were shown between PUFA intakes and risk of COPD. Although COPD-prevention efforts should continue to focus on smoking cessation, these prospective findings support the importance of promoting a healthy diet in multi-interventional programs to prevent COPD instead of focusing on changes in an isolated food or nutrient.


[Ilio-pelvic lymphatic dissection in the rectal cancer].
The method of iliopelvic lymphodissection was used in 61 patients aged from 42 to 72 years having histologically verified rectum cancer. The results of treatment by this method have shown that in 83.6% of the patients the tumor spread corresponded to pT3-T4, the metastatic involvement of regional lymph nodes was found in 52.5% of the patients. The most justified reason for a radical surgery and lymphodissection was a combination of the endoscopic signs of a considerable extension of the tumor around the circumference of the rectum with the CT signs of enlargement of the regional lymph nodes or the ingrowth of the tumor in the pararectal fat. In cases with the lymphogenic metastasing the cumulative 5 year survival was 43.6 +/- 8.7%, in cases without it it was 55.3 +/- 11.8%. A thorough preoperative evaluation of the extension of the tumor process allows to optimize indications for iliopelvic lymphodissection.


Differential effects of divalent cations, cation chelators and an ionophore (A23187) on morphine and dibutyryl guanosine 3': 5'-cyclic monophosphate antinociception.
The effects of intraventricularly (i.c.v.) administered divalent cations, cation chelators and an ionophore (A23187) on antinociception produced by i.c.v. administration of morphine or dibutyryl guanosine 3':5'-cyclic monophosphate (db c-GMP) were quantitated in the mouse tail-flick procedure. Ca++ pretreatment produced a dose-related potentiation (> 10-fold) of db c-GMP and a dose-related antagonism (> 20-fold) of morphine antinociception. Mg++ pretreatment antagonized db c-GMP, whereas morphine antinociception was unaffected. Ba++ and Sr++ were observed to possess intrinsic antinociceptive activity. Administration of Ba++ or Sr++ had greater than additive effects on db c-GMP and morphine antinociception. EDTA pretreatment did not affect db c-GMP or morphine antinociception. Ethylene glycol bis(beta-aminoethyl ether)N,N-tetraacetic acid had no effect on db c-GMP but potentiated the morphine response. The ionophore A23187 had no effect on db c-GMP or morphine in the tail-flick test. However, A23187 potentiated the effect of high doses of Ca++ on db c-GMP and increased the antagonistic effect of a low dose of Ca++ on morphine antinociception. The results provide further evidence that the mechanism of db c-GMP antinociception is different from that of morphine.


Conditional inference trees: a method for predicting intimate partner violence.
A number of different methodologies have been employed to investigate the complex relationship between psychological and physical aggression. Herein, a method of unbiased recursive partitioning (conditional inference trees) was applied to a longitudinal sample to identify cutoffs of psychological aggression at baseline that differentiate between individuals who do and do not perpetrate physical aggression at follow-up. The algorithm categorized men into low- and high-risk groups, and women into mild-, moderate-, or high-risk categories of perpetration. Couples responded anonymously to a self-report measure of psychological and physical aggression (CTS2) at baseline and a 12-month follow-up. Sensitivity analyses for predicting physical aggression reached as high as 59% for women and 60% for men.


Analysis of the impact of COVID-19 on collisions, fatalities and injuries using time series forecasting: The case of Greece.
The current study aims to investigate the impact of the COVID-19 pandemic on road traffic collisions, fatalities, and injuries using time series analyses. To that aim, a database containing road collisions, fatalities, and slight injuries data from Greece were derived from the Hellenic Statistical Authority (HSA) and covered a ten-year timeframe (from January 2010 to August 2020. The chosen time period contained normal operations, as well as the period of the first COVID-19-induced lockdown period in Greece. Three different Seasonal Autoregressive Integrated Moving Average (SARIMA) time series models were implemented in order to compare the observed measurements to forecasted values that were intended to depict assumed conditions; namely, without the appearance of the COVID-19 pandemic. Modelling results revealed that the total number of road collisions, fatalities, and slightly injured were decreased, mainly due to the sharp traffic volume decrease. However, the percentage reduction of the collision variables and traffic volume were found to be disproportionate, which probably indicates that more collisions occurred with regard to the prevailing traffic volume. An additional finding is that fatalities and slightly injured rates were significantly increased during the lockdown period and the subsequent month. Overall, it can be concluded that a worse performance was identified in terms of road safety. Since subsequent waves of COVID-19 cases and other pandemics may reappear in the future, the outcomes of the current study may be exploited for the improvement of road safety from local authorities and policymakers.


Recent advances in cellulose and chitosan based membranes for water purification: A concise review.
Recently membrane technology has emerged as a new promising and pervasive technology due to its innate advantages over traditional technologies such as adsorption, distillation and extraction. In this article, some of the recent advances in developing polymeric composite membrane materials for water purification from natural polysaccharide based polymers namely cellulose derivatives and chitosan are concisely reviewed. The impact of human social, demographic and industrial evolution along with expansion through environment has significantly affected the quality of water by pollution with large quantities of pesticides, minerals, drugs or other residues. At the forefront of decontamination and purification techniques, we found the membrane materials from polymers as a potential alternative. In an attempt to reduce the number of technical polymers widely used in the preparation of membranes, many researchers have reported new solutions for desalination or retention of organic yeasts, based on bio renewable polymers like cellulose derivatives and chitosan. These realizations are presented and discussed in terms of the most important parameters of membrane separation especially water flux and retention in this article.


Synchrony of spontaneous calcium activity in mouse neocortex before synaptogenesis.
Spontaneous calcium activity can be detected in embryonic mouse cortical slices as fluorescence intensity variations, in the presence of a fluorescent calcium indicator. Current methods to detect and quantify these variations depend heavily on experimenters whose judgement may interfere with measurement. In the present work, we developed new software called CalSignal for automatic detection and tracking of cellular bodies and quantification of spontaneous calcium activity on time-series of confocal fluorescence images. Analysis of 28 neocortical slices revealed that 21.0% of detected cells displayed peaks of fluorescence corresponding to spontaneous activity, with a mean frequency of one peak per 4 min. This activity was blocked in the absence of extracellular calcium but was not modified after depletion of calcium stores with thapsigargin or blockade of voltage-gated calcium channels with Ni2+. Further, statistical analysis of calcium activity revealed concomitant activation of distant cells in 24 slices, and the existence of a significant network of synchrony based on such coactivations in 17 slices out of 28. These networks enclosed 84.3% of active cells, scattered throughout the neocortical wall (mean distance between cellular bodies, 111.7 microm). Finally, it was possible to identify specific cells which were synchronously active with more neighbouring cells than others. The identity of these nodal cells remains to be investigated to fully comprehend the role of spontaneous calcium activity, before synaptogenesis, in shaping cortical neurogenesis.


Investigation of Ocular Tear Ferning in Controlled and Uncontrolled Diabetic Subjects.
To investigate the ocular tear film in controlled and uncontrolled diabetes mellitus (DM) subjects and the relationship between dry eye and diabetes. One hundred twenty-four diabetes adults (65 male and 59 female, with 62 controlled and 62 uncontrolled DM) were assessed by McMonnies dry eye questionnaire, tear break-up time (TBUT), phenol red thread (PRT) test. In addition, tear samples were collected from the lower tear meniscus of right eyes and dried onto glass slides under controlled room temperature (25°C) and humidity (40%). The glass slides were observed using a digital microscopy and the tear ferning (TF) patterns observed were graded based on Masmali TF grading scale. Patients with a HbA1c value below 6.5% were defined as "controlled DM" and those with a HbA1c value above 6.5% were defined as "uncontrolled DM." The Spearman's correlation coefficient was used to determine the relationship between various tests. Several subjects having dry eye varied depending on the test used. McMonnies questionnaire scores suggested that 28 subjects (23%) showed dry eye (score >14.5), of whom 23 subjects (82%) showed uncontrolled diabetes. The TF patterns showed that 44 subjects (35.5%) showed dry eyes (Grade ≥2), of whom 32 subjects (72.7%) showed uncontrolled diabetes. Phenol red thread test found that 46 subjects (37%) showed dry eye (wetting <10 mm), of whom 38 subjects (82.6%) showed uncontrolled diabetes. Tear break-up time found that 57 subjects (46%) showed dry eye (time <10 sec), of whom 45 subjects (78.9%) showed uncontrolled diabetes. In controlled diabetic subjects, TF grades showed medium negative correlations with both TBUT and PRT (-0.374 and -0.349, respectively). In uncontrolled subjects, TF grades have a strong negative correlation with TBUT (-0.539) and a medium negative correlation with PRT (-0.410). Dry eye disease is common in diabetic subjects, particularly in people with poor diabetic control. Uncontrolled blood glucose level seemed to be a risk factor for dry eye syndrome. Tear ferning test showed a correlation between diabetes and dry eye in diabetic subjects and can be used as a reliable test to evaluate the quality of the ocular tear film.