Mass drug administration and the global control of schistosomiasis: successes, limitations and clinical outcomes.
Preventive chemotherapy is advocated for the global control and elimination of schistosomiasis. Despite the well known short-term benefits of treating patients for schistosomiasis, the impact of mass drug administration (MDA) campaigns to control the disease in the long term remains unresolved. Many studies have advocated the success of MDA programs in order to attract donor funds for elimination efforts but such successes are often short-lived given the drug does not alter the life cycle of the organism or prevent reinfection. Within a matter of months to years after halting treatment, the prevalence, intensity of infection and morbidity of disease return to baseline levels. Other mitigating factors contribute to the failings of MDA campaigns namely: poverty, poor drug coverage, poor drug compliance, and, in the case of Asiatic schistosomiasis, zoonotic transmission. Genetic and innate and acquired immunologic mechanisms complicate the epidemiologic picture of schistosomiasis globally, and may contribute indirectly to MDA shortcomings. The possibility of drug resistance is an ever present concern because of the sole reliance on one drug, praziquantel. Preventive chemotherapy is advocated for the global control and elimination of schistosomiasis. The short-term benefits of MDA campaigns are well documented but the long-term benefits are questionable.


Mineral oil lipogranulomata in liver and spleen. A study of 465 autopsies.
Lipogranulomata (LG) are commonly seen in human tissues but the location and significance of these lesions are incompletely documented. In a study of 465 autopsies LG were found in 48% of livers and 46% of spleens. Hepatic LG were more common in portal tracts than adjacent to the terminal hepatic venules. There was a close correlation between hepatic LG in liver and in spleen. The incidence and severity of hepatic LG were higher in older adults, especially men. There was no evidence that LG caused hepatic dysfunction or portal hypertension. The absence of correlation with hepatic steatosis supports the suggestion that LG are secondary to mineral oil deposition rather than to steatosis.


Four biochemical tests for identification of probable enteroinvasive Escherichia coli strains.
Enteroinvasive Escherichia coli (EIEC) share important features with Shigella spp., but EIEC strains are difficult to identify because their biochemical reactions are variable, and Sereny tests or other biological and molecular assays are expensive or hard to perform. The aim of this work was to detect probable enteroinvasive E. coli strains by using four biochemical tests, in children under 5 years of age with and without acute diarrhea. 330 strains of E. coli isolated from children with diarrhea, and 660 strains from children without diarrhea were studied. All strains were tested with the following tests: mucus , lysine and ornithine decarboxylase and motility. The strains which were negative to the four tests were tested by Sereny assay. Twelve strains (3.6%) isolated from children with diarrhea were negative to the tests proposed; eleven were lactose positive and only one was lactose negative. Three strains (0.5%) from children without diarrhea were negative to the tests proposed and were lactose positive. All the 15 strains (100%) were positive in Sereny assay. We recommend the use of these four biochemical tests for initial detection of EIEC strains, because their cost is very low and it is feasible carry out them in small diagnostic laboratories.


[Hospital outbreak of salmonellosis with secondary cases].
Food poisoning by Salmonella in hospitals and other health care centers is not uncommon and occasionally results in high mortality rates. We report an outbreak of food poisoning that took place in June 1987 in a hospital. A staff relay was primarily affected, with an attack rate of 59.3%. The mean incubation period was 31.2 hours. The epidemiological evidence pointed to a rice with milk served as a dessert as the transmitting vehicle, although the contamination source could not be identified. Subsequently, 11 days after the start of the outbreak, 9 cases were identified among the patients cared for at the center, which were considered as secondary cases. The causative organism was Salmonella (serovariety enteritidis, phage type A). All the involved individuals had a favorable outcome. The transmission of the mentioned organism within the hospital milieu has not been previously reported. In food poisoning developing in health care centers, the exposed individuals should be extremely careful and absolutely conscious of the need for complying with hygienic rules. In our area, we recommend sick leave while symptoms persist or the culture of feces is positive.


Modification of the silver staining technique to detect lipopolysaccharide in polyacrylamide gels.
A silver staining method used routinely for detecting bacterial lipopolysaccharide (LPS) in sodium dodecyl sulfate-polyacrylamide gels (C. Tsai and E. Frasch, Anal. Biochem. 119:115-119, 1982) appeared to be inappropriate for visualizing certain LPS preparations. It did not stain S-form fractions of polyagglutinable Pseudomonas aeruginosa LPS or several partly deacylated (alkali-treated) S-form LPS after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. However, these LPS preparations could be detected by anti-LPS sera after electroblotting onto nitrocellulose, thereby confirming their integrity and presence in the polyacrylamide gel. This is because LPS fractions containing a low number of fatty acids are washed out of the gel during the initial fixing step (40% ethanol-4% acetic acid, overnight). By omitting this fixing step, which was originally developed for detecting proteins, and by increasing the LPS oxidation time (from 5 to 20 min), we restored the ability to detect LPS fractions that otherwise would not be stained. These modifications did not affect the detection of other S- and R-form LPSs. Thus, differences in the number of fatty acids present in polyagglutinable P. aeruginosa LPS may result in a selective loss of fatty acid-deficient S-form LPS in these apparent R-form LPS preparations. This modified procedure provides a fast, simple, and sensitive way to analyze LPS in polyacrylamide gels despite the number of acyl groups present.


Focused ultrasound for safe and effective release of brain tumor biomarkers into the peripheral circulation.
The development of noninvasive approaches for brain tumor diagnosis and monitoring continues to be a major medical challenge. Although blood-based liquid biopsy has received considerable attention in various cancers, limited progress has been made for brain tumors, at least partly due to the hindrance of tumor biomarker release into the peripheral circulation by the blood-brain barrier. Focused ultrasound (FUS) combined with microbubbles induced BBB disruption has been established as a promising technique for noninvasive and localized brain drug delivery. Building on this established technique, we propose to develop FUS-enabled liquid biopsy technique (FUS-LBx) to enhance the release of brain tumor biomarkers (e.g., DNA, RNA, and proteins) into the circulation. The objective of this study was to demonstrate that FUS-LBx could sufficiently increase plasma levels of brain tumor biomarkers without causing hemorrhage in the brain. Mice with orthotopic implantation of enhanced green fluorescent protein (eGFP)-transfected murine glioma cells were treated using magnetic resonance (MR)-guided FUS system in the presence of systemically injected microbubbles at three peak negative pressure levels (0.59, 1.29, and 1.58 MPa). Plasma eGFP mRNA levels were quantified with the quantitative polymerase chain reaction (qPCR). Contrast-enhanced MR images were acquired before and after the FUS sonication. FUS at 0.59 MPa resulted in an increased plasma eGFP mRNA level, comparable to those at higher acoustic pressures (1.29 MPa and 1.58 MPa). Microhemorrhage density associated with FUS at 0.59 MPa was significantly lower than that at higher acoustic pressures and not significantly different from the control group. MRI analysis revealed that post-sonication intratumoral and peritumoral hyperenhancement had strong correlations with the level of FUS-induced biomarker release and the extent of hemorrhage. This study suggests that FUS-LBx could be a safe and effective brain-tumor biomarker release technique, and MRI could be used to develop image-guided FUS-LBx.


Food, youth and the Mediterranean diet in Spain. Development of KIDMED, Mediterranean Diet Quality Index in children and adolescents.
To evaluate dietary habits in Spanish children and adolescents based on a Mediterranean Diet Quality Index tool, which considers certain principles sustaining and challenging traditional healthy Mediterranean dietary patterns. Observational population-based cross-sectional study. A 16-item Mediterranean Diet Quality Index was included in data gathered for the EnKid study (in which two 24-hour recalls, a quantitative 169-item food-frequency questionnaire and a general questionnaire about socio-economic, demographic and lifestyle items were administered). Spain. In total, 3850 children and youths aged 2-24 years residing in Spain. Of the sample, 4.2% showed very low KIDMED index results, 49.4% had intermediate values and 46.4% had high index results. Important geographical differences were seen, with subjects from the Northeast showing the most favourable outcomes (52% with elevated scores vs. 37.5% of those from the North). Lower percentages of high diet quality were observed in low socio-economic groups, compared with middle and upper income cohorts (42.8%, 47.6% and 54.9%, respectively). Large cities had more positive results and only slight variations were seen for gender and age. The KIDMED index, the first to evaluate the adequacy of Mediterranean dietary patterns in children and youth, confirms that this collective is undergoing important changes, which makes them a priority target for nutrition interventions. Results challenge certain commonly perceived notions tied to income level, population size and diet quality.


The Assessment of Reliability and Validity of Persian Version of the Endometriosis Health Profile (EHP-30).
The Endometriosis Health Profile-30 (EHP-30) is a disease-specific questionnaire to measure the health-related quality of life in patients with endometriosis. The aim of this study was to evaluate the validity and reliability of the Persian version of Endometriosis Health Profile (EHP-30) in women with endometriosis referring to three Gynecology Clinics in Tehran, Iran. One hundred women (20 to 50 years old) with surgically confirmed endometriosis recruited from three outpatient Gynecology Clinics affiliated to the Iran University of Medical Sciences. All 100 patients were asked to complete EHP-30 questionnaire while referring to the Clinics. The findings were analyzed using descriptive statistics, internal reliability consistency, construct validity (using short form-36, which had already been validated in Iran), factor analysis (with principle component analysis method), and item total correlation to assess the validity and reliability of the questionnaire. The internal consistency reliability of the questionnaire was high (Cronbach's α ranged between 0.80 and 0.93 for core, and 0.78 and 0.90 for modular parts). All items were loaded on their own factors except item 17 (feeling aggressive or violent) and item 18 (feeling unwell), which were loaded on pain and social support domains, respectively. Construct validity of EHP-30, established by using SF-36, indicates good correlations in several similar scales of these two questionnaires. The findings of the study demonstrate that Persian version of EHP-30 is a valid and reliable measure to assess the quality of life in women with endometriosis.


Source identification of heavy metals in peri-urban agricultural soils of southeast China: An integrated approach.
Intensive human activities, in particular agricultural and industrial production have led to heavy metal accumulation in the peri-urban agricultural soils of China threatening soil environmental quality and agricultural product security. A combination of spatial analysis (SA), Pb isotope ratio analysis (IRA), input fluxes analysis (IFA), and positive matrix factorization (PMF) model was successfully used to assess the status and sources of heavy metals in typical peri-urban agricultural soils from a rapidly developing region of China. Mean concentrations of Cd, As, Hg, Pb, Cu, Zn and Cr in surface soils (0-20 cm) were 0.31, 11.2, 0.08, 35.6, 44.8, 119.0 and 97.0 mg kg-1, respectively, exceeding the local background levels except for Hg. Spatial distribution of heavy metals revealed that agricultural activities have significant influence on heavy metal accumulation in the surface soils. Isotope ratio analysis suggested that fertilization along with atmospheric deposition were the major sources of heavy metal accumulation in the soils. Based on the PMF model, the relative contribution rates of the heavy metals due to fertilizer application, atmospheric deposition, industrial emission, and soil parent materials were 30.8%, 33.0%, 25.4% and 10.8%, respectively, demonstrating that anthropogenic activities had significantly higher contribution than natural sources. This study provides a reliable and robust approach for heavy metals source apportionment in this particular peri-urban area with a clear potential for future application in other regions.


Seasonal changes in VO2max among Division 1A collegiate women soccer players.
Aerobic capacity and body composition were measured at 3 time points over a 1-year period in 26 Division 1A women soccer players from Texas A&M University, in order to determine whether there were seasonal changes in these parameters. Subjects were tested in December, immediately following a 4-month competitive season; in April, following 15 weeks of strength and conditioning; and immediately prior to the start of the regular season in August, following a 12-week summer strength and conditioning program. A periodized strength and conditioning program design was incorporated in order to optimize anaerobic and oxidative capacity immediately prior to the regular competitive season. Significant differences in VO2max were measured between August (49.24 +/- 4.38 ml x kg(-1) x min(-1)) and December (44.87 +/- 4.61 ml x kg(-1) x min(-1)). No significant changes in aerobic capacity were found between April (47.43 +/- 4.01 ml x kg(-1) x min(-1)) and August (49.64 +/- 5.25 ml x kg(-1) x min(-1)). Significant increases in body fat were measured between August (15.71 +/- 2.92%) and December (18.78 +/- 2.79%), before and after the competitive season, respectively. No significant changes in body fat were found between April (16.24 +/- 2.95%) and August (15.71 +/- 2.92%). The results of this study suggest that decreases in muscle mass over the course of a regular competitive season contribute to decreases in aerobic capacity in collegiate women soccer players. Although it is unknown whether this decrease in muscle mass is the result of inadequate training or a normal adaptation to the physiological demands imposed by soccer, the results of the current study suggest that resistance training volume should be maintained during the competitive season, in order to maintain preseason levels of muscle mass.


Adenosine induces histamine release from human bronchoalveolar lavage mast cells.
Previous studies have shown that in vitro adenosine enhances histamine release from activated human lung mast cells obtained by enzymic dispersion of lung parenchyma. However, adenosine alone has no effect on histamine release from these cells. Given the evidence for direct activation of mast cells after endobronchial challenge with adenosine and previous studies indicating that mast cells obtained at bronchoalveolar lavage are a better model for asthma studies than those obtained by enzymic dispersion of lung tissue, the histamine-releasing effect of adenosine was examined on lavage mast cells. Bronchoalveolar lavage fluid was obtained from patients attending hospital for routine bronchoscopy (n=54). Lavage cells were challenged with adenosine or adenosine receptor agonists (20 min, 37 degrees C) and histamine release determined using an automated fluorometric assay. Endogenous adenosine levels were also measured in lavage fluid (n=9) via an HPLC method. Adenosine alone caused histamine release from lavage mast cells in 37 of 54 patients with a maximal histamine release of 20.56+/-2.52% (range 5.2-61%). The adenosine receptor agonists (R)-N6-(2-phenylisopropyl)adenosine, 5'-N-ethylcarboxamidoadenosine and CGS21680 also induced histamine release from lavage mast cells. Preincubation of lavage mast cells with the adenosine receptor antagonist xanthine amine congener caused significant inhibition of the response to adenosine (P=0.007). There was an inverse correlation between endogenous adenosine levels in the lavage fluid and the maximal response to in vitro adenosine challenge of the lavage cells. The findings of the present study indicate a means by which adenosine challenge of the airways can induce bronchoconstriction and support a role for adenosine in the pathophysiology of asthma. The results also suggest that cells obtained from bronchoalveolar lavage fluid may provide the ideal model for the testing of novel, adenosine receptor, targeted therapies for asthma.


Late respiratory infection after lung transplantation.
Aiming to improve outcome of lung transplantation (LTx) patients, we reviewed risk factors and treatment practices for the LTx recipients who experienced respiratory infection in the late post-LTx period (>1 month after LTx). We analyzed the clinical data of 48 recipients and donors from 61 LTx, who experienced late respiratory infections. Late respiratory infections were classified according to the etiology, time of occurrence, and frequency of donor-to-host transmission or colonization of the recipient prior to transplantation. During the period of observation, 42 episodes of respiratory infections occurred. The organisms most frequently involved were gram (-) bacteria: Acinetobacter baumannii (n=13, 31.0%), Pseudomonas aeruginosa (n=7, 16.7%), and Klebsiella pneumoniae (n=4, 10.0%). Among the 42 episodes recorded, 14 occurred in the late post-LTx period. These were bacterial (n=6, 42.9%), fungal (n=2, 14.3%), viral (n=4, 28.5%), and mycobacterial (n=2, 14.3%) infections. Of 6 bacterial infections, 2 were from multidrug-resistant (MDR) A. baumannii and one from each of MDR P. aeruginosa, extended spectrum β-lactamase (+) K. pneumoniae, methicillin-resistant Staphylococcus aureus and Streptococcus pneumoniae. Infection-related death occurred in 6 of the 14 episodes (43%). Although the frequency of respiratory infection decreased sharply in the late post-LTx period, respiratory infection was still a major cause of mortality. Gram (-) MDR bacteria were the agents most commonly identified in these infections.


Nanotopography as a trigger for the microscale, autogenous and passive lysis of erythrocytes.
Microscale devices are increasingly being developed for diagnostic analysis although conventional lysis as an initial step presents limitations due to its scale or complexity. Here, we detail the physical response of erythrocytes to the surface nanoarchitecture of black Si (bSi) and foreshadow their potential in microanalysis. The physical interaction brought about by the spatial convergence of the two topologies: (a) the nanopillar array present on the bSi and (b) the erythrocyte cytoskeleton present on the red blood cells (RBCs), provides spontaneous stress-induced cell deformation, rupture and passive lysis within an elapsed time of ∼3 min from immobilisation to rupture and without external chemical or mechanical intervention. The mechano-responsive bSi surface provides highly active yet autogenous RBC lysis and a prospect as a front-end platform technology in evolving micro-fluidic platforms for cellular analyses.


Cutting edge: a role for the adaptor protein LAT in human NK cell-mediated cytotoxicity.
Stimulation of NK cell-mediated cytotoxicity involves the coupling of proximal Src and Syk family protein tyrosine kinases to downstream effectors. However, the mechanisms linking these second messenger pathways are incompletely understood. Here, we describe a key role for the LAT (p36) adaptor protein in human NK cell activation. LAT is tyrosine phosphorylated upon stimulation of NK cells through FcgammaRIII receptors and following direct contact with NK-sensitive target cells. This NK stimulation induces the association of LAT with several phosphotyrosine-containing proteins. In addition to the biochemical evidence showing LAT involvement in NK cell activation, a genetic model shows that LAT is required for FcR-dependent phosphorylation of phospholipase C-gamma. Furthermore, overexpression of LAT in NK cells leads to increased Ab-dependent cell-mediated cytotoxicity and "natural cytotoxicity," thus demonstrating a functional role for LAT in NK cells. These data suggest that LAT is an important adaptor protein for the regulation of human NK cell-mediated cytotoxicity.


Quantitative Glycopeptide Changes in Rat Sperm During Epididymal Transit.
Mammalian spermatozoa acquire fertilizing potential as they undergo a series of changes during epididymal transit. One major facet of such is the alterations in the sperm glycome. Modifications of the sialic acid content within glycan moieties are known to regulate epitope presentation and cellular adhesion and signaling, all of which may be critical for sperm to successfully reach and fertilize the egg. To date, there is paucity of information regarding the sialic acid changes that occur on spermatozoa during epididymal transit. Therefore, the aim of this study was to identify N-linked sialylated glycoproteins in rat epididymal sperm and investigate whether they are regulated during epididymal transit. Sialylated glycopeptides from caput, corpus, and cauda spermatozoa were enriched using titanium dioxide beads. Bound N-linked glycopeptides were released by enzymatic deglycosylation using PNGase F and then analyzed by liquid chromatography tandem-mass spectrometry. A total of 92 unique N-linked sialylated glycopeptides were identified from 65 different proteins. These included members of the disintegrin and metalloproteinase domain-containing protein family (ADAM), Basigin, and Testis-expressed protein 101 (TEX101). Remarkably, label-free quantification showed that more than half of these peptides (48/92) were regulated during epididymal transit. Of interest, the protein TEX101 exhibited PNGase F-resistant deglycosylation under the conditions used in this study. The results from this study showed that changes in the N-linked sialoglycoprotein profile is a major hallmark of sperm maturation in rats.


Influence of an Al2O3 surface coating on the response of polymeric waveguide sensors.
The responses of a polymer ridge waveguide Young interferometer with and without a bilayer of Al2O3/TiO2, fabricated by atomic layer deposition, are studied and compared when applied as an aqueous chemical sensor. The phase shift of the guided mode, as a result of the change in refractive index of the cover medium, is monitored. The results indicate that the over-coating affects the linearity of the sensor response. The effect of concentration on the linearity of the sensor response is investigated by applying different concentrations of water-ethanol solution. Although the performance of the sensor is improved by the additional layers, the study reveals a non-monotonic behavior of the device. We show that it comes mainly from the adsorption of ethanol molecules on the surface of the films. Such an understanding of the platform is crucial for sensing of analytes involving polar molecules.


IGPRED: Combination of convolutional neural and graph convolutional networks for protein secondary structure prediction.
There is a close relationship between the tertiary structure and the function of a protein. One of the important steps to determine the tertiary structure is protein secondary structure prediction (PSSP). For this reason, predicting secondary structure with higher accuracy will give valuable information about the tertiary structure. Recently, deep learning techniques have obtained promising improvements in several machine learning applications including PSSP. In this article, a novel deep learning model, based on convolutional neural network and graph convolutional network is proposed. PSIBLAST PSSM, HHMAKE PSSM, physico-chemical properties of amino acids are combined with structural profiles to generate a rich feature set. Furthermore, the hyper-parameters of the proposed network are optimized using Bayesian optimization. The proposed model IGPRED obtained 89.19%, 86.34%, 87.87%, 85.76%, and 86.54% Q3 accuracies for CullPDB, EVAset, CASP10, CASP11, and CASP12 datasets, respectively.


Insulin resistance and its treatment by thiazolidinediones.
Insulin resistance is a change in physiologic regulation such that a fixed dose of insulin causes less of an effect on glucose metabolism than occurs in normal individuals. The normal compensatory response to insulin resistance is an increase in insulin secretion that results in hyperinsulinemia. If the hyperinsulinemia is sufficient to overcome the insulin resistance, glucose regulation remains normal; if not, type 2 diabetes ensues. Associated with insulin resistance, however, is a cluster of other metabolic abnormalities involving body fat distribution, lipid metabolism, thrombosis and fibrinolysis, blood pressure regulation, and endothelial cell function. This cluster of abnormalities is referred to as the insulin resistance syndrome or the metabolic syndrome. It is causally related not only to the development of type 2 diabetes but also to cardiovascular disease. A major unresolved issue is whether there is a single underlying cause of this syndrome and, if so, what might it be? Several promising hypotheses have been proposed. There are some data to support the hypothesis that fetal malnutrition imprints on metabolic regulatory processes that, in later adult life, predispose to the development of the insulin resistance syndrome. Visceral obesity also has been a candidate for the cause of the syndrome. Whatever mechanism is ultimately found to be responsible, it will undoubtedly have both genetic and environmental components. Among the biochemical mediators that are likely to be responsible for the interference with insulin's effects on intermediary metabolism are free fatty acids and other products from adipose tissue. Recent data suggest that the substances stimulate serine phosphorylation of molecules involved in the initial steps of insulin action, thereby blocking the ability of these molecules to be tyrosine phosphorylated and initiate the subsequent steps of the insulin action cascade. The thiazolidinediones are a new class of agents that have been developed to treat type 2 diabetic patients. These drugs act as peroxisome proliferator-activated receptor gamma (PPARgamma) agonists. Following their binding to the receptor, the heterodimer molecule that contains the binding site is activated. The activated complex binds to the response elements of specific genes that regulate molecules that effect insulin action and lipid metabolism. These genes are either activated or inhibited. Specifically, the thiazolidinediones improve insulin action and decrease insulin resistance. The exact mechanism by which these agents decrease insulin resistance is not clear but they do decrease the elevated free fatty acid levels present in insulin-resistant patients and they appear to change the body distribution of adipose tissue. Treatment of insulin-resistant type 2 diabetic patients with thiazolidinediones not only improves glycemic control and decreases insulin resistance, it also improves many of the abnormalities that are part of the insulin resistance syndrome.


Targeted irradiation of shoot apical meristem of Arabidopsis embryos induces long-distance bystander/abscopal effects.
Bystander effects induced by low-dose ionizing radiation have been shown to occur widely in many cell types and may have a significant impact on radiation risk assessment. Although the region of radiation damage is known to be much greater than the initial target volume irradiated, it remains to be seen whether this response is limited to the specific organ irradiated, spans a limited region of the body, or even covers the whole body of the target. To determine whether long-distance bystander/abscopal effects exist in whole organisms and to clarify the problem of intercellular communication, in the present study a specific cell group, the shoot apical meristem in Arabidopsis embryo, was irradiated with a defined number of protons and examined for root development postirradiation. The results showed that after direct damage to the shoot apical meristem from ion traversals, root hair differentiation, primary root elongation and lateral root initiation were all inhibited significantly in postembryonic development, suggesting that radiation-induced long-distance bystander/abscopal responses might exist in the whole organism. To further scrutinize the mechanism(s) underlying these inhibitory effects, a DR5-GUS transgenic Arabidopsis was used. The results showed that accumulation of the reporter GUS gene transcript in irradiated shoot apical meristem embryos decreased in the postembryonic development. Treatment with either 2,4-dichlorophenoxyacetic acid, a synthetic plant auxin, or DMSO, a effective reactive oxygen species (ROS) scavenger, could rescue the reporter GUS enzyme accumulation and the length of primary root in irradiated shoot apical meristem embryos, indicating that ROS or probably the ROS related auxin and auxin-dependent transcription process may be involved in radiation-induced long-distance bystander/abscopal effects.


Recognition of PF4-VWF complexes by heparin-induced thrombocytopenia antibodies contributes to thrombus propagation.
Heparin-induced thrombocytopenia (HIT) is a prothrombotic disorder mediated by complexes between platelet factor 4 (PF4) and heparin or other polyanions, but the risk of thrombosis extends beyond exposure to heparin implicating other PF4 partners. We recently reported that peri-thrombus endothelium is targeted by HIT antibodies, but the binding site(s) has not been identified. We now show that PF4 binds at multiple discrete sites along the surface of extended strings of von Willebrand factor (VWF) released from the endothelium following photochemical injury in an endothelialized microfluidic system under flow. The HIT-like monoclonal antibody KKO and HIT patient antibodies recognize PF4-VWF complexes, promoting platelet adhesion and enlargement of thrombi within the microfluidic channels. Platelet adhesion to the PF4-VWF-HIT antibody complexes is inhibited by antibodies that block FcγRIIA or the glycoprotein Ib-IX complex on platelets. Disruption of PF4-VWF-HIT antibody complexes by drugs that prevent or block VWF oligomerization attenuate thrombus formation in a murine model of HIT. Together, these studies demonstrate assembly of HIT immune complexes along VWF strings released by injured endothelium that might propagate the risk of thrombosis in HIT. Disruption of PF4-VWF complex formation may provide a new therapeutic approach to HIT.


Protease inhibitors and carcinogenesis: a review.
This brief review article deals with the subject of anticarcinogenic activity of protease inhibitors (PI). Three basic premises are made: (1) Although PI are prevalent constituents of dietary staples such as soy products, which have been epidemiologically associated with reduced cancer incidences at multiple target sites, they are unlikely to be the active anticarcinogenic entities. Cooked soy products, which are devoid of PI activity, are equally as effective at reducing cancer development as raw soy products. Isoflavones are likely to represent major chemopreventive agents in soy, although other constituents may well contribute. (2) Although supplementation of diets with PI (natural or synthetic), or direct topical administration, results in lower cancer incidences in many experimental models in vivo, this effect appears to be indirect. Dietary PI are, in general, poorly absorbed from the GI tract, and never reach target organs in any measurable quantity. The most attractive hypothesis is that dietary PI could induce synthesis and distribution of endogenous PI (acute-phase reactants), which have widespread effects on cell growth and behavior. Effects of topical administration of PI also encompass prominent anti-inflammatory effects. (3) A spectrum of PI inhibit in vitro transformation induced by a variety of carcinogenic agents. Their effects can be grouped into three basic categories, affecting: (a) signal transduction pathways; (b) DNA repair processes; and (c) nuclear proteases. I suggest that the nuclear multicatalytic protease activity, in particular the chymotrypsin-like activity, represents an important cellular target for which considerable anecdotal support can be garnered.


Pharmacological perturbation of cholinergic systems: applications to the study of pathophysiological mechanisms in affective disorders.
Clinical, behavioral, physiological, biochemical and receptor binding parameters are useful as dependent measures in studies into the pathophysiology of affective disorders. Pharmacological perturbation of cholinergic mechanisms and modification of cholinergic-monoaminergic interaction mimics aspects of the neurobiology of affective disorders. The effects of these pharmacological manipulations can be quantitatively assessed by measuring their impact on variables in each of these classes. These deviations are easily and safely produced by several classes of drugs which directly or indirectly modify the function of central muscarinic cholinergic networks. Methods for inducing these changes in cholinergic systems and their application to clinical and basic research in the field of affective disorders are highlighted.


Cyclic adenosine 3':5'-monophosphate phosphodiesterase and its role in learning in Drosophila.
Drosophila carrying the X-linked mutation dunce (dnc) showed poor learning in a negative reinforcement olfactory conditioning paradigm (Dudai, Y., Y.-N. Jan, D. Byers, W.G. Quinn, and S. Benzer (1976) Proc. Natl. Acad. Sci. U.S.A. 73: 1684-1688). More recently, dnc flies were shown to have reduced activity for one of two cAMP phosphodiesterases (PDEs) present in normal flies, PDE II, whereas PDE form I was unaffected (Byers, D., R. L. Davis, and J. A. Kiger, Jr. (1981) Nature 289: 79-81). A micro-assay technique is described that allows the separate measurement of PDE I and PDE II in crude extracts, based on specific inhibition of PDE I [3H]cAMP hydrolysis by cGMP. Using this technique, PDE II is shown to occur normally at high specific activity in the nervous system, consistent with the hypothesis that this enzyme plays a role in neuronal function. Reduced PDE II activity correlates with poor learning in dnc flies at three developmental stages (first and third instar larva and adult), as well as in response to genetic modification of dnc gene activity. Biochemical and genetic experiments fail to reveal any abnormal regulation of PDE II in dnc. The specific activity of PDE II is shown to correlate in a one to one fashion with the level of normal dnc gene (dnc+) activity at five different doses of dnc+. These results support the hypothesis that PDE II represents the primary product of the dnc gene, indicating a role for this enzyme in Drosophila learning.


The occurrence of polycystic echinococcosis in Suriname.
The occurrence of polycystic echinococcosis (PE) caused by Echinococcus vogeli in Suriname is reviewed. Six of the eight known cases, including the first (which was incorrectly diagnosed as cystic hydatid disease caused by E. granulosus) in 1982, are described. The distribution in Suriname of the natural hosts of E. vogeli--the bush dog (Speothus venaticus) and the paca (Agouti paca)--is discussed. Characteristics of the four species recognized in the genus Echinococcus are compared, and all known cases of PE in 11 countries in the Neotropics are summarized. Finally, it is stressed, that, in Suriname, PE should be considered during the differential diagnosis of all subjects from rural areas who present with abdominal pain and palpable, intraperitoneal masses.


Skin vascular resistance and skin perfusion pressure as predictors of healing of ischemic lesion of the lower limb: influences of diabetes mellitus, hypertension, and age.
An isotope washout technique that requires inexpensive equipment and is well tolerated by patients has been developed to measure skin perfusion pressure (SPP) and skin vascular resistance (SVR). The SPP is a measure of the severity of peripheral vascular disease and the SVR is an indicator of microangiopathy associated with diabetes mellitus and hypertension. This test has been applied to 87 patients who had ischemic lesions of the lower limb. Fourty-four patients had lesions that healed with local treatment while the other 43 required major surgery. The presence of diabetes, hypertension, and old age reduced the likelihood of healing. Similarly if the SPP less than 40 mm Hg, only 18% of patients' lesions healed. If the SPP lay between 40 and 50 mm Hg and the SVR less than 1000 U, 67% of lesions healed. However, if the SPP was between 40 and 50 mm Hg and the SVR greater than 1000 U, no lesions healed. If the SPP greater than 50 mm Hg and SVR less than 1000 U, 83% of lesions healed, where as if the SVR greater than 1000 U, healing occurred in only 23% of lesions. This test can be used to predict the likelihood of healing of ischemic lesions of the lower leg, thus rationalizing their management. The importance of microangiopathy is also demonstrated.


The interface engineering and structure design of an alloying-type metal foil anode for lithium ion batteries: a review.
An alloying-type metal foil serves as an integrated anode that is distinct from the prevalent powder-casting production of lithium ion batteries (LIBs) and emerging lithium metal batteries (LMBs), and also its energy density and processing technology can be profoundly developed. However, besides their apparent intriguing advantages of a high specific capacity, electrical conductivity, and the ease of formation, metal foil anodes suffer from slow lithiation kinetics, a trade-off between specific capacity and cycle life, and a low initial Coulombic efficiency (ICE) owing to their multi-scaled structural geometry, huge volume change, and induced interfacial issues during the alloying process. In this review, we attempt to present a comprehensive overview on the recent research progress with respect to alloying-type metal foil anodes toward high-energy-density and low-cost LIBs. The failure mechanism of metal foil anodes during lithiation/delithiation and existing challenges are also summarized. Subsequently, the structural design and interface engineering strategies that have witnessed significant achievements are highlighted, which can promote the practical development of LIBs, including artificial SEI, alloying, structural design, and grain refinement. Furthermore, scientific perspectives are proposed to further improve the overall performance and decouple the complex mechanisms in terms of interdisciplinary fields of electrochemistry, metallic materials science, mechanics, and interfacial science, demonstrating that metal foil anode-based LIBs require more research efforts.


Insights into the interaction between influenza virus and pneumococcus.
Bacterial infections following influenza are an important cause of morbidity and mortality worldwide. Based on the historical importance of pneumonia as a cause of death during pandemic influenza, the increasingly likely possibility that highly pathogenic avian influenza viruses will trigger the next worldwide pandemic underscores the need to understand the multiple mechanisms underlying the interaction between influenza virus and bacterial pathogens such as Streptococcus pneumoniae. There is ample evidence to support the historical view that influenza virus alters the lungs in a way that predisposes to adherence, invasion, and induction of disease by pneumococcus. Access to receptors is a key factor and may be facilitated by the virus through epithelial damage, by exposure or up-regulation of receptors, or by provoking the epithelial regeneration response to cytotoxic damage. More recent data indicate that alteration of the immune response by diminishing the ability of the host to clear pneumococcus or by amplification of the inflammatory cascade is another key factor. Identification and exploration of the underlying mechanisms responsible for this synergism will provide targets for prevention and treatment using drugs and vaccines.


A correlative study of ophthalmoscopy and fluorescein angiography in systemic hypertension.
This study correlates the fundus signs with the severity and signs of hypertension and evaluates the role of fluorescein angiography in detecting changes in the retinal and choroidal capillary bed in hypertension and defines its advantages over direct ophthalmoscopy. 37 hypertensives belonging to all grades of hypertension were studied. A thorough physical examination, hypertension work up, direct ophthalmoscopy and fluorescein angiography was done in all cases. A significant association was found between the presence of marked arteriolar narrowing and the presence of severe hypertension, left ventricular hypertrophy (LVH) and cardiomegaly. Patients having definite arterio-venous crossing changes and exudative retinopathy had a higher incidence of LVH and cardiomegaly. Renal functions and neurological signs in hypertension showed no correlation with the fundus signs. Capillary bed and choriodal abnormalities could be better studied on fluorescein angiography. Hard exudates were not visualized on fluorescein angiography. There was total resolution of exudative phenomenon on treatment.


Decreased cardiac output in humans during laparoscopic antireflux surgery: direct measurements.
In a porcine model, we demonstrated that laparoscopic Nissen fundoplication causes a significant drop in cardiac output (30%) because it exposes both the peritoneal cavity and the mediastinum to CO(2) under pressure. To determine if this occurs in humans, we examined cardiovascular physiology during laparoscopic Nissen fundoplication. Because of invasiveness required in this pilot trial, only six patients were studied. The arterial blood pressure (via radial arterial catheter) and the pulmonary artery diastolic pressure and cardiac index (via pulmonary artery thermodilution catheter) were measured at seven points in time during each laparoscopic Nissen fundoplication. The systolic blood pressure decreased in all patients, and the cardiac index decreased in all but one patient. The exception was a patient with Huntington disease, in whom the cardiac output did not decrease. In four of the five patients, the cardiac output was lowest during hiatal dissection, and in the fifth, it was lowest after reverse Trendelenburg positioning. No significant change in the pulmonary artery diastolic pressure was noted. All patients received adequate intravenous fluid replacement (average, 58 +/- 16 mL/kg) to support blood pressure. In one patient, with a particularly large paraesophageal hernia, profound hypotension (40/25 mm Hg) developed during the mediastinal phase of the procedure, and this patient required alpha-adrenergic support followed by laparotomy to eliminate a surgical cause (none found). Although it is a tremendous advance for patients, laparoscopic Nissen fundoplication can be associated with a significant reduction in cardiac output and blood pressure. Surgeons and anesthesiologists must be alert to changes reflecting these decreases during procedures, which violate both the peritoneal cavity and the mediastinum. We propose careful hemodynamic monitoring during these procedures, especially in patients with coronary artery disease or significant left ventricular dysfunction.


Egg cortical granule N-acetylglucosaminidase is required for the mouse zona block to polyspermy.
The mammalian egg must be fertilized by only one sperm to prevent polyploidy. In most mammals studied to date, the primary block to polyspermy occurs at the zona pellucida, the mammalian egg coat, after exocytosis of the contents of the cortical granules into the perivitelline space. The exudate acts on the zona, causing it to lose its ability to bind sperm and to be penetrated by sperm previously bound to the zona. However, the cortical granule components responsible for the zona block have not been identified. Studies described herein demonstrate that N-acetylglucosaminidase is localized in cortical granules and is responsible for the loss in sperm-binding activity leading to the zona block to polyspermy. Before fertilization, sperm initially bind to the zona by an interaction between sperm surface GalTase and terminal N-acetylglucosamine residues on specific oligosaccharides of the zona glycoprotein ZP3 (Miller, D. J., M. B. Macek, and B. D. Shur. 1992. Nature (Lond.). 357:589-593). These GalTase-binding sites are lost from ZP3 after fertilization, an effect that can be duplicated by N-acetylglucosaminidase treatment. Therefore, N-acetylglucosaminidase, or a related glycosidase, may be present in cortical granules and be responsible for ZP3's loss of sperm-binding activity at fertilization. Of eight glycosidases assayed in exudates of ionophore-activated eggs, N-acetylglucosaminidase was 10-fold higher than any other activity. The enzyme was localized to cortical granules using immunoelectron microscopy. Approximately 70 or 90% of the enzyme was released from cortical granules after ionophore activation or in vivo fertilization, respectively. The isoform of N-acetylglucosaminidase found in cortical granules was identified as beta-hexosaminidase B, the beta, beta homodimer. Inhibition of N-acetylglucosaminidase released from activated eggs, with either competitive inhibitors or with specific antibodies, resulted in polyspermic binding to the zona pellucida. Another glycosidase inhibitor or nonimmune antibodies had no effect on sperm binding to activated eggs. Therefore, egg cortical granule N-acetylglucosaminidase is released at fertilization, where it inactivates the sperm GalTase-binding site, accounting for the block in sperm binding to the zona pellucida.


Socioeconomic status, obesity, individual behaviors, diabetes, and risk for frozen shoulder: A Mendelian randomization study.
There are few studies on risk factors for frozen shoulder, and even fewer Mendelian randomization (MR) studies on frozen shoulder. Therefore, we conducted a two-sample MR study to explore whether socioeconomic status (years of schooling, average total household income before tax), obesity (body mass index and waist circumference), individual behaviors (smoking initiation, alcohol intake frequency, coffee intake, nonoily fish intake, tea intake, beef intake, bread intake, cheese intake, oily fish intake, and fresh fruit intake), and diabetes (type 1 and type 2 diabetes) are associated with frozen shoulder. The exposure datasets and the outcome dataset were extracted from the MRC Integrative Epidemiology Unit at the University of Bristol Open genome-wide association studies project (https://gwas.mrcieu.ac.uk/). We conducted MR analyses using the inverse variance weighted (primary method), MR-Egger, and weighted median methods and conducted heterogeneity and pleiotropy analyses. Type 1 diabetes (OR: 1.103; 95% CI: 1.053-1.156; P = .0000410) was associated with an increased risk of frozen shoulder. Cheese intake (OR: 0.490; 95% CI: 0.267-0.899; P = .0213), non-oily fish intake (OR: 0.0993; 95% CI: 0.0220-0.448; P = .00267), years of schooling (OR: 0.453; 95% CI: 0.349-0.588; P = .00000000277), and average total household income before tax (OR: 0.434; 95% CI: 0.253-0.743; P = .00236) were discovered as protective factors. No horizontal pleiotropy was found in all analyzes we performed (P > .05). Our study indicated that type 1 diabetes was a risk factor for frozen shoulder while cheese intake, non-oily fish intake, years of schooling, and average total household income before tax were considered as protective factors for frozen shoulder.


Staff Perceptions of Improving Emergency Care for Children: A Qualitative Study.
The objective of this study was to identify staff perceptions of a service improvement for pediatric emergency care at a university teaching hospital. Semistructured qualitative interviews of stakeholders were conducted, and grounded theory approach was used for analysis. Forty-one interviews were conducted with physicians, nurses, managers, and health care workers. Major themes emerging from the analysis included the physical space of and flow within the pediatric emergency department (ED), impact of technology, staffing in the ED, the effects of frontline pediatricians and emergency physicians managing children in the ED, and the need for and expectations of a pediatric emergency medicine (PEM) consultant. Human interactions among health care providers, leadership, and teaching are considered as equally important as providing the appropriate environment and qualified professionals for improving care for children in the ED. Appointment of a PEM consultant was suggested to provide leadership and education to manage relationships and implement changes. Subsequent to the study, the model of care for PEM was changed, the pediatric care delivery became more integrated with the main ED, and two PEM consultants were appointed to the institution.


Plasma testosterone levels in the male rhesus: influences of sexual and social stimuli.
Four adult male rhesus monkeys were provided access individually to a group of receptive females. Each male assumed dominance and engaged in frequent copulations. Plasma testosterone levels increased two- to threefold during this period. Next, each male was subjected to sudden and decisive defeat by a large all-male group, and plasma testosterone fell following this experience. Two males were later reintroduced to the females, and plasma testosterone rose rapidly to the previous elevated levels.


Effect of Shortened Light-Curing Modes on Bulk-Fill Resin Composites.
Shortened light curing does not affect volumetric polymerization shrinkage or cohesive tensile strength but negatively affects the shear bond strength of some bulk-fill resin composites. When performing shortened light curing, clinicians should be aware of the light output of their light-curing units. Purpose: To evaluate volumetric polymerization shrinkage (VPS), shear bond strength (SBS) to dentin, and cohesive tensile strength (CTS) of bulk-fill resin composites (BFRCs) light activated by different modes.Methods and Materials: Six groups were evaluated: Tetric EvoCeram bulk fill + high mode (10 seconds; TEC H10), Tetric EvoFlow bulk fill + high mode (TEF H10), experimental bulk fill + high mode (TEE H10), Tetric EvoCeram bulk fill + turbo mode (five seconds; TEC T5), Tetric EvoFlow bulk fill + turbo mode (TEF T5), and experimental bulk fill + turbo mode (TEE T5). Bluephase Style 20i and Adhese Universal Vivapen were used for all groups. All BFRC samples were built up on human molar bur-prepared occlusal cavities. VPS% and location were evaluated through micro-computed tomography. SBS and CTS tests were performed 24 hours after storage or after 5000 thermal cycles; fracture mode was analyzed for SBS.Results: Both TEC H10 and TEE H10 presented lower VPS% than TEF H10. However, no significant differences were observed with the turbo-curing mode. No differences were observed for the same BFRC within curing modes. Occlusal shrinkage was mostly observed. Regarding SBS, thermal cycling (TC) affected all groups. Without TC, all groups showed higher SBS values for high mode than turbo mode, while with TC, only TEC showed decreased SBS from high mode to turbo modes; modes of fracture were predominantly adhesive. For CTS, TC affected all groups except TEE H10. In general, no differences were observed between groups when comparing the curing modes.Conclusions: Increased light output with a shortened curing time did not jeopardize the VPS and SBS properties of the BFRCs, although a decreased SBS was observed in some groups. TEE generally showed similar or improved values for the tested properties in a shortened light-curing time. The VPS was mostly affected by the materials tested, whereas the SBS was affected by the materials, curing modes, and TC. The CTS was not affected by the curing modes.


Oct4 and Hypoxia Dual-Regulated Oncolytic Adenovirus Armed with shRNA-Targeting Dendritic Cell Immunoreceptor Exerts Potent Antitumor Activity against Bladder Cancer.
Immunotherapy has emerged as a promising modality for cancer treatment. Dendritic cell immunoreceptor (DCIR), a C-type lectin receptor, is expressed mainly by dendritic cells (DCs) and mediates inhibitory intracellular signaling. Inhibition of DCIR activation may enhance antitumor activity. DCIR is encoded by CLEC4A in humans and by Clec4a2 in mice. Gene gun-mediated delivery of short hairpin RNA (shRNA) targeting Clec4a2 into mice bearing bladder tumors reduces DCIR expression in DCs, inhibiting tumor growth and inducing CD8+ T cell immune responses. Various oncolytic adenoviruses have been developed in clinical trials. Previously, we have developed Ad.LCY, an oncolytic adenovirus regulated by Oct4 and hypoxia, and demonstrated its antitumor efficacy. Here, we generated a Clec4a2 shRNA-expressing oncolytic adenovirus derived from Ad.LCY, designated Ad.shDCIR, aimed at inducing more robust antitumor immune responses. Our results show that treatment with Ad.shDCIR reduced Clec4a expression in DCs in cell culture. Furthermore, Ad.shDCIR exerted cytolytic effects solely on MBT-2 bladder cancer cells but not on normal NIH 3T3 mouse fibroblasts, confirming the tumor selectivity of Ad.shDCIR. Compared to Ad.LCY, Ad.shDCIR induced higher cytotoxic T lymphocyte (CTL) activity in MBT-2 tumor-bearing immunocompetent mice. In addition, Ad.shDCIR and Ad.LCY exhibited similar antitumor effects on inhibiting tumor growth. Notably, Ad.shDCIR was superior to Ad.LCY in prolonging the survival of tumor-bearing mice. In conclusion, Ad.shDCIR may be further explored as a combination therapy of virotherapy and immunotherapy for bladder cancer and likely other types of cancer.


Antibody-dependent cell cytotoxicity against Trypanosoma cruzi is only mediated by protective antibodies.
The antibody-dependent cell cytotoxicity (ADCC) to Trypanosoma cruzi blood forms (Btry) using non-adherent spleen cells is only mediated by sera from chronic chagasic patients or mice. Both display 'lytic antibodies' (LA), which are immunoglobulins directed against epitopes only present in living BTry, and 'conventional serology antibodies' (CSA), which are responsible for the positive diagnostic tests in Chagas disease. Sera from mice immunized with different T. cruzi antigens or from treated patients (displaying only CSA but not LA) are unable to mediate ADCC. These data confirm the central role played by LA in the host resistance against T. cruzi. Moreover, they probably explain why most immunizing agents used as vaccines in Chagas' disease and which elicit CSA but not LA, do not display significant protection against T. cruzi. We also demonstrate that trypsinization of BTry increases significantly the rate of parasite destruction by ADCC, suggesting that enzyme sensitive membrane components may help BTry to evade from this immune effector mechanism.


Enhanced remediation of carbon tetrachloride by Fe(II)-Fe(III) systems in the presence of copper ions.
The effect of Cu(II) ion on the dechlorination of carbon tetrachloride (CT) by Fe(II) associated with various iron oxides was investigated. Iron oxides including goethite, hematite, ferrihydrite and magnetite were selected as the model compounds. CT was dechlorinated to chloroform (CF) by 3 mM Fe(II) in iron oxide suspensions at pH 7.2. The dechlorination followed pseudo first-order kinetics and the pseudo first-order rate constants (k(obs)) were 0.048 h(-1), 0.0836 h(-1), 0.0609 h(-1) and 0.0144 h(-1) in goethite-, hematite-, ferrihydrite- and magnetite-amended systems, respectively. Addition of Cu(II) into systems increased the k(obs) for CT dechlorination significantly. A 3- to 120-fold increase in k(obs) relative to the systems without Cu(II) was observed when 0.5 mM Cu(II) was added to the Fe(II)-Fe(III) suspensions. The pH of the system is an important factor controlling the dechlorination rate of CT. The increase in concentrations of Fe(II) and iron oxides also enhanced the dechlorination efficiency and rate of CT. Moreover, a linear relationship between the k(obs) and Cu(II) concentration ranging between 0 and 0.4 mM was observed. Results obtained demonstrate the feasibility of using surface-bound iron species with Cu(III) for the detoxification of chlorinated solvents in the contaminated aquifers.


Progress in cell based assays for botulinum neurotoxin detection.
Botulinum neurotoxins (BoNTs) are the most potent human toxins known and the causative agent of botulism, and are widely used as valuable pharmaceuticals. The BoNTs are modular proteins consisting of a heavy chain and a light chain linked by a disulfide bond. Intoxication of neuronal cells by BoNTs is a multi-step process including specific cell binding, endocytosis, conformational change in the endosome, translocation of the enzymatic light chain into the cells cytosol, and SNARE target cleavage. The quantitative and reliable potency determination of fully functional BoNTs produced as active pharmaceutical ingredient (API) requires an assay that considers all steps in the intoxication pathway. The in vivo mouse bioassay has for years been the 'gold standard' assay used for this purpose, but it requires the use of large numbers of mice and thus causes associated costs and ethical concerns. Cell-based assays are currently the only in vitro alternative that detect fully functional BoNTs in a single assay and have been utilized for years for research purposes. Within the last 5 years, several cell-based BoNT detection assays have been developed that are able to quantitatively determine BoNT potency with similar or greater sensitivity than the mouse bioassay. These assays now offer an alternative method for BoNT potency determination. Such quantitative and reliable BoNT potency determination is a crucial step in basic research, in the development of pharmaceutical BoNTs, and in the quantitative detection of neutralizing antibodies.


Characterization of (4-hydroxy-3-nitrophenyl)acetyl (NP)-specific germinal center B cells and antigen-binding B220- cells after primary NP challenge in mice.
Previous studies examining the primary germinal center (GC) response to SRBC in mice demonstrated a steady ratio of IgM(+) to isotype-switched GC B cells and a persistent population of GC B cells with a founder phenotype. These characteristics held true at the inductive, plateau, and dissociative phases of the GC response, suggesting a steady-state environment. To test whether these characteristics apply to the primary response of other T cell-dependent Ags, the present study examined the GC response after challenge with (4-hydroxy-3-nitrophenyl)acetyl (NP) in C57BL/6 mice. Multiparameter flow cytometric analysis was used to assess the phenotype of splenic NP-reactive cells at multiple time points after immunization. Results of these studies demonstrated the characteristics of the SRBC-induced GC reaction to be fully maintained in the NP response. In particular, there was a steady ratio of nonswitched to switched B cells, with the majority of NP-reactive GC B cells displaying IgM. In addition, a substantial frequency of B220(-) NP-binding cells was observed in the spleen at later time points after NP challenge. Although these cells were IgE(+), they were found to express both kappa and lambda L chains and display the high-affinity IgE Fc (FcepsilonRI) receptor, suggesting that this population is not of B cell origin. Adoptive transfer studies further demonstrated the B220(-) NP-binding subset to be derived from the myeloid lineage.


[Flow cytometry analysis of retinoblastoma and its clinical significance].
Flow cytometry (FCM) is a new technic that determines with precision the cellular DNA content and its distribution in various phases of the cell cycle. Using FCM, we analysed paraffin embedded tissue masses of retinoblastoma from 20 patients with the results that the rate of occurrence of DNA heteroploid was 35%; the occurrence rate of heteroploid in the glaucoma stage and the extraocular stage was higher than that in the intraocular stage (P less than 0.02); the postoperative survival rate of the diploid and near-diploid tumors was obviously higher than that of the heteroploid (P = 0.01); the cellular proportion in the G2M stage of the heteroploid and the index of cell proliferation was obviously higher than that of the diploid and near-diploid (P less than 0.05). This technique is of value in estimating the prognosis of retinoblastoma.


The crosstalk between ferroptosis and autophagy in cancer.
In order to better understand the interplay between ferroptosis and autophagy, enhance the interpretation of the crosstalk between these two forms of regulated cell death, develop the effective pharmacological mechanisms for cancer treatment, discover novel biomarkers for better diagnostic, and envisage the future hotspots of the research on ferroptosis and autophagy, we harnessed bibliometric tools to study the articles published from 2012 to 2022 on the relationship between ferroptosis and autophagy. Web of Science Core Collection (WOSCC) database was used to conduct a comprehensive search and analysis of articles in this field from January 1, 2012, to September 1, 2022. The Citespace 6.1.R2 software and VOS viewer 6.1.8 software were utilized to analyze the overall structure of the network, network clusters, links between clusters, key nodes or pivot points, and pathways. A total of 756 articles associated with the crosstalk between ferroptosis and autophagy were published in 512 journals by 4183 authors in 980 organizations from 55 countries or regions. The distribution of countries and organizations was demonstrated using CiteSpace and VOS viewer. The top three countries with the most articles were China (n = 511), United States (n = 166), and Germany (n = 37). The most productive institutions were Guangzhou Medical University and Central South University (n = 42), but their centralities were relatively low, which values were respective 0.04 and 0.03. Kang and Tang published the most articles related to ferroptosis and autophagy (n = 49), followed by Jiao Liu (n = 22), Guido Kroemer (n = 20), and Daniel Klionsky (n = 12). Published studies on ferroptosis and asthma have the most cited counts. The top three keywords with the highest frequencies were autophagy (n = 283), cell death (n = 243), and oxidative stress (n = 165). Our results provide insights into the development of recognition related to the crosstalk between ferroptosis and autophagy, and the current molecular crosslinked mechanisms in the context of common signal transduction pathways or affecting cellular environment to induce the adaptive stress response and to activate the particular form of regulated cell death (RCD), and the development of cancer treatment based on novel targets and signaling regulatory networks provided by ferroptosis and autophagy.


Neural Differentiation of Human Pluripotent Stem Cells for Nontherapeutic Applications: Toxicology, Pharmacology, and In Vitro Disease Modeling.
Human pluripotent stem cells (hPSCs) derived from either blastocyst stage embryos (hESCs) or reprogrammed somatic cells (iPSCs) can provide an abundant source of human neuronal lineages that were previously sourced from human cadavers, abortuses, and discarded surgical waste. In addition to the well-known potential therapeutic application of these cells in regenerative medicine, these are also various promising nontherapeutic applications in toxicological and pharmacological screening of neuroactive compounds, as well as for in vitro modeling of neurodegenerative and neurodevelopmental disorders. Compared to alternative research models based on laboratory animals and immortalized cancer-derived human neural cell lines, neuronal cells differentiated from hPSCs possess the advantages of species specificity together with genetic and physiological normality, which could more closely recapitulate in vivo conditions within the human central nervous system. This review critically examines the various potential nontherapeutic applications of hPSC-derived neuronal lineages and gives a brief overview of differentiation protocols utilized to generate these cells from hESCs and iPSCs.


Evaluation of the alopecia areata patients on tofacitinib treatment during the COVID-19 pandemic.
Tofacitinib is a Janus Kinase 3 inhibitor that is used in the treatment of alopecia areata. We recommended our alopecia areata patients to discontinue their tofacitinib treatment during the COVID-19 pandemic for an average of 80 days. We aimed to evaluate the drug use and the SARS-CoV-2 infection status of alopecia areata patients; and the relationships of recurrence to age, gender, treatment duration, and tofacitinib discontinuation. One-hundred and ninety-one (61.4%) patients were off the drug and 120 (38.6%) were on therapy during the pandemic. The relationship between drug discontinuation due to the COVID-19 pandemic and recurrence was statistically significant (P < .001). Statistically significant relationships of age (P = .013) and treatment duration (P < .001) to recurrence were also found. The change in the SALT score differed between the patients on therapy and off therapy during the pandemic (P < .001). A significant negative correlation was found between the change in the SALT score and treatment duration: the spearman correlation test P = .018. We concluded that the patients may continue to the tofacitinib therapy during the rest of the COVID-19 pandemic if the benefit outweighed the risk.


[Primary cerebral non-Hodgkin lymphoma in non-immunocompromised subjects].
Cerebral non Hodgkin's lymphomas (NHL) of immunocompetent patients are rare disease (1% of all NHL), but their incidence has been steadily increasing in the last 20 years. Most cerebral NHL are diffuse large cell NHL with a B phenotype. These tumors are most often localized in the central nervous system, the eye or meninges; other systemic tumor sites are rare, either at initial diagnosis and relapse. The overall survival of primary cerebral lymphoma (PCL) is poor in the published retrospective series with a median survival in the range of 12 to 16 months and a 5 year survival between 5 et 20%. PCL are however curable brain tumors, and the outcome of these patients has probably improved in the past 10 years, using a multidisciplinary therapeutic approach. Complete resection of the tumor dose not improve the outcome of these patients. The treatment of PCL relies on combinations of chemotherapy and radiotherapy, but the type of regimen and the dose to the brain are still a matter of debate, in particular because of late neurological sequelae. Multicentric prospective clinical trials in France and Europe will provide answers to these questions in the next years.


The solution structure of an N-terminally truncated version of the yeast CDC24p PB1 domain shows a different beta-sheet topology.
Phox and Bem1 (PB1) domains mediate protein-protein interactions via the formation of homo- or hetero-dimers. The C-terminal PB1 domain of yeast cell division cycle 24 (CDC24p), a guanine-nucleotide exchange factor involved in cell polarity establishment, is known to interact with the PB1 domain occurring in bud emergence MSB1 interacting 1 (BEM1p) during the regulation of the yeast budding process via its OPR/PC/AID (OPCA) motif. Here, we present the structure of an N-terminally truncated version of the Sc CDC24p PB1 domain. It shows a different topology of the beta-sheet than the long form. However, the C-terminal part of the structure shows the conserved PB1 domain features including the OPCA motif with a slight rearrangement of helix alpha1. Residues which are important for the heterodimerization with BEM1p are structurally preserved.


Field measurements of isoprene emission from trees in response to temperature and light.
The atmospheric hydrocarbon budget is important for predicting ozone episodes and the effects of pollution mitigation strategies. Isoprene emission from plants is an important part of the atmospheric hydrocarbon budget. We measured isoprene emission capacity at the bottom, middle, and top of the canopies of a white oak (Quercus alba L.) tree and a red oak (Quercus rubra L.) tree growing adjacent to a tower in the Duke University Forest. Leaves at the top of the white oak tree canopy had a three- to fivefold greater capacity for emitting isoprene than leaves at the bottom of the tree canopy. Isoprene emission rate increased with increasing temperature up to about 42 degrees C. We conclude that leaves at the top of the white oak tree canopy had higher isoprene emission rates because they were exposed to more sunlight, reduced water availability, and higher temperature than leaves at the bottom of the canopy. Between 35 and 40 degrees C, white oak photosynthesis and stomatal conductance declined, whereas red oak (Quercus rubra) photosynthesis and stomatal conductance increased over this range. Red oak had lower rates of isoprene emission than white oak, perhaps reflecting the higher stomatal conductance that would keep leaves cool. The concentration of isoprene inside the leaf was estimated with a simplified form of the equation used to estimate CO(2) inside leaves.


Predictors of components of expressed emotion in major caregivers of Turkish patients with schizophrenia.
Expressed emotion (EE) is a concept reflecting the emotional atmosphere of the home environment. Specific components of EE, namely criticism, hostility and emotional over-involvement, have been found to be important predictors of relapse for schizophrenic patients. The main aim of this study was to examine the predictive power of patient and caregiver characteristics and caregivers' perceptions of frequency, coping, distress/discomfort, control of symptom behaviours by the patient, and attributions on locus of causality for the development of the illness on two components of EE (criticism/hostility and emotional over-involvement) in a sample of major caregivers of Turkish schizophrenic patients. Seventy-two caregivers of schizophrenic patients were administered a set of questionnaires tapping socio-demographic and illness-related variables, the family questionnaire tapping perceived frequency, distress/discomfort, coping and control of symptom behaviours, causal attributions for illness and, finally, the Expressed Emotion Scale in the hospital setting. The results showed that caregivers' perceptions of coping with specific symptom behaviours decreased criticism/hostility (C/H), whereas perceptions of higher frequency of symptom behaviours increased C/H. For emotional over-involvement (EOI), the number of individuals living in the household, being the mother, father or the spouse, perceptions of coping with symptom behaviours and reported distress/discomfort about symptom behaviours were significant predictors. Caregivers' perceptions of their ability to cope with symptom behaviours and their reported distress due to these behaviours are important variables related to components of EE and need to be targets in intervention studies. The cultural and clinical implications of the results for the management of schizophrenia and for support for the caregivers are discussed.


A retrospective comparison of inpatients with mixed and pure depression.
Some authors advocate a broadening of the narrow concept of mixed episodes in the direction of mania leading to the concept of mixed mania, and in the direction of depression leading to the concept of mixed depression. The latter has been little investigated so far. In the present article, we retrospectively compare 49 patients with pure depression with 51 patients with mixed depression in terms of socio-demographic and clinical variables in order to contribute to the validation of the distinction between mixed and pure depression. Supporting this distinction, we observed that mixed depressive patients more frequently had past histories of bipolar disorder and alcohol abuse and had longer durations of hospital stay. These last two points remain significant even when we control for the effect of the association with bipolarity.


Emergency postexposure vaccination with vesicular stomatitis virus-vectored Ebola vaccine after needlestick.
Safe and effective vaccines and drugs are needed for the prevention and treatment of Ebola virus disease, including following a potentially high-risk exposure such as a needlestick. To assess response to postexposure vaccination in a health care worker who was exposed to the Ebola virus. Case report of a physician who experienced a needlestick while working in an Ebola treatment unit in Sierra Leone on September 26, 2014. Medical evacuation to the United States was rapidly initiated. Given the concern about potentially lethal Ebola virus disease, the patient was offered, and provided his consent for, postexposure vaccination with an experimental vaccine available through an emergency Investigational New Drug application. He was vaccinated on September 28, 2014. The vaccine used was VSVΔG-ZEBOV, a replicating, attenuated, recombinant vesicular stomatitis virus (serotype Indiana) whose surface glycoprotein gene was replaced by the Zaire Ebola virus glycoprotein gene. This vaccine has entered a clinical trial for the prevention of Ebola in West Africa. The vaccine was administered 43 hours after the needlestick occurred. Fever and moderate to severe symptoms developed 12 hours after vaccination and diminished over 3 to 4 days. The real-time reverse transcription polymerase chain reaction results were transiently positive for vesicular stomatitis virus nucleoprotein gene and Ebola virus glycoprotein gene (both included in the vaccine) but consistently negative for Ebola virus nucleoprotein gene (not in the vaccine). Early postvaccination cytokine secretion and T lymphocyte and plasmablast activation were detected. Subsequently, Ebola virus glycoprotein-specific antibodies and T cells became detectable, but antibodies against Ebola viral matrix protein 40 (not in the vaccine) were not detected. It is unknown if VSVΔG-ZEBOV is safe or effective for postexposure vaccination in humans who have experienced a high-risk occupational exposure to the Ebola virus, such as a needlestick. In this patient, postexposure vaccination with VSVΔG-ZEBOV induced a self-limited febrile syndrome that was associated with transient detection of the recombinant vesicular stomatitis vaccine virus in blood. Strong innate and Ebola-specific adaptive immune responses were detected after vaccination. The clinical syndrome and laboratory evidence were consistent with vaccination response, and no evidence of Ebola virus infection was detected.


New service lines for hospital emergency departments.
One way for a hospital to increase revenue, improve the delivery of health care, and boost patient satisfaction is to develop new services as an adjunct to its emergency department. New service lines that treat emergency department patients quickly and efficiently and eliminate negative experiences patients often associate with emergency department visits can offer significant benefits to a hospital as well as its patients.


SdiA sensing of acyl-homoserine lactones by enterohemorrhagic E. coli (EHEC) serotype O157:H7 in the bovine rumen.
Chemical communication mediates signaling between cells. Bacteria also engage in chemical signaling, termed quorum sensing (QS), to coordinate population-wide behavior. The bacterial pathogen enterohemorrhagic E. coli (EHEC), responsible for outbreaks of bloody diarrhea worldwide, exploits QS to promote expression of virulence factors in humans. Although EHEC is a human pathogen, it is a member of the gastrointestinal (GI) flora in cattle, the main reservoir for this bacterium. EHEC cattle colonization requires SdiA, a QS transcription factor that uses acyl-homoserine lactones (AHLs), for proper folding and function. EHEC harbors SdiA, but does not produce AHLs, consequently having to sense AHLs produced by other bacterial species. We recently showed that SdiA is necessary for efficient EHEC passage through the bovine GI tract, and show that AHLs are prominent within cattle rumen, but absent from the other sections of the GI tract. EHEC utilizes the locus of enterocyte effacement (LEE) to colonize the recto-anal junction of cattle, and the glutamate decarboxylase (gad) system to colonize cows. Transcription of the LEE genes is decreased by rumen AHLs through SdiA, while transcription of the gad acid resistant system is increased. It would be expensive for EHEC to express the LEE genes in the rumen where they are not necessary. However, in preparation for the acidic distal stomachs the EHEC gad is activated in the rumen. Hence AHL signaling through SdiA aids EHEC in gauging these environments, and modulates gene expression towards adaptation to a commensal life-style in cattle.1 Inasmuch as EHEC is largely prevalent in cattle herds, interference with SdiA-mediated QS inhibition of cattle colonization could be an attractive approach to diminish contamination of food products due to cattle shedding of this pathogen.


A proposed framework for developing quality assessment tools.
Assessment of the quality of included studies is an essential component of any systematic review. A formal quality assessment is facilitated by using a structured tool. There are currently no guidelines available for researchers wanting to develop a new quality assessment tool. This paper provides a framework for developing quality assessment tools based on our experiences of developing a variety of quality assessment tools for studies of differing designs over the last 14 years. We have also drawn on experience from the work of the EQUATOR Network in producing guidance for developing reporting guidelines. We do not recommend a single 'best' approach. Instead, we provide a general framework with suggestions as to how the different stages can be approached. Our proposed framework is based around three key stages: initial steps, tool development and dissemination. We recommend that anyone who would like to develop a new quality assessment tool follow the stages outlined in this paper. We hope that our proposed framework will increase the number of tools developed using robust methods.


Management of odontoid fractures in the elderly.
Odontoid fractures are frequent in patients over 70 years of age, and in patients over 80 years of age they form the majority of spinal fractures. In a retrospective analysis of 23 geriatric (> 70 years) patients with a fracture of the odontoid, we compared some of the clinical features to a contemporary series of patients younger than 70 years of age. Whereas in the younger patients high-energy trauma accounted for the majority of the fractures, low-energy falls were the underlying cause in 90% of the odontoid fractures in the elderly. In contrast to the younger age group, in elderly patients predominantly type II fractures (95%) were identified. Anterior and posterior displacement were recorded with equal frequency on the first postinjury radiograph in the younger age group, whereas in geriatric patients displacement was mainly posterior. The number of associated injuries was significantly higher in younger patients. There was no difference in the occurrence of neurological deficits (13%) between the two age groups, and neurological compromise was mainly related to posterior dislocation of the odontoid in both groups. The overall complication rate was significantly higher in elderly patients (52.2% vs 32.7%), with an associated in-hospital mortality of 34.8%. Loss of reduction and non-union after non-operative treatment, a complicated postoperative course and complications due to associated injuries accounted primarily for this high complication rate. Elderly patients with a fracture of the odontoid are a high-risk group with a high morbidity and mortality rate. An aggressive diagnostic approach to detect unstable fractures and application of a halo device or early primary internal stabilisation of these fractures is recommended.


Functional analysis and treatment of problem behavior exhibited by children with fragile X syndrome.
The efficacy of function-based interventions for the treatment of severe problem behavior exhibited by individuals with intellectual and developmental disabilities (IDD) is well established. However, few studies have reported on behavioral interventions in fragile X syndrome (FXS) specifically. The present study is a consecutive case-series analysis that reports on functional analysis and treatment of problem behavior of nine children with FXS. Assessment findings were consistent with previous research indicating that among individuals with FXS, problem behavior is more commonly maintained by escape from demands and access to tangible items, relative to the broader population of individuals with IDD. Functional analysis-based behavioral interventions resulted in a mean reduction in problem behavior of 95.2% across the nine participants. Additionally, generalization of treatment effects from controlled clinical settings to home, school, and community was demonstrated. The current findings suggest that function-based behavioral interventions shown to be effective with the broader population of individuals with IDD are also effective for individuals with FXS. Our results in combination with those of previous studies describing functional analysis outcomes provide additional evidence for a unique functional behavioral phenotype for severe problem behavior in individuals with FXS. Implications of study findings for early intervention and prevention of problem behavior in children with FXS are discussed.


Expression of NK1 receptor at the protein and mRNA level in the porcine female reproductive system.
The presence and distribution of substance P (SP) receptor NK1 was studied in the ovary, the oviduct and the uterus (uterine horn and cervix) of the domestic pig using the methods of molecular biology (RT-PCR and immunoblot) and immunohistochemistry. The expression of NK1 receptor at mRNA level was confirmed with RT-PCR in all the studied parts of the porcine female reproductive system by the presence of 525 bp PCR product and at the protein level by the detection of 46 kDa protein band in immunoblot. Immunohistochemical staining revealed the cellular distribution of NK1 receptor protein. In the ovary NKI receptor was present in the wall of arterial blood vessels, as well as in ovarian follicles of different stages of development. In the tubular organs the NK1 receptor immunohistochemical stainings were observed in the wall of the arterial blood vessels, in the muscular membrane, as well as in the mucosal epithelium. The study confirmed the presence of NK1 receptor in the tissues of the porcine female reproductive tract which clearly points to the possibility that SP can influence porcine ovary, oviduct and uterus.


Controlling Gut Microbiota by Twendee X® May Contribute to Dementia Prevention.
The human gut microbiota (GM) is a complex and dynamic ecosystem that hosts trillions of commensal and potentially pathogenic microorganisms. It is crucial in protecting humans from pathogens and in maintaining immune and metabolic homeostasis. Numerous studies have demonstrated that GM has a significant impact on health and disease, including Alzheimer's disease (AD). AD is a progressive neurodegenerative disorder characterized by impaired short-term memory and cognitive deficits. Patients with AD have been reported to exhibit abnormalities in GM density and species composition. Oxidative stress (OS) has been implicated in the onset and progression of AD; however, the relationship between OS and gut microbiota in AD onset and progression is not clear. Twendee X® (TwX), an oral supplement consisting of eight active ingredients, has been shown to prevent dementia in mild cognitive impairment (MCI) in humans and substantially improve cognitive impairment in mouse models of AD. This positive effect is achieved through the potency of the combined antioxidants that regulate OS; therefore, similar results cannot be achieved by a single antioxidant ingredient. To examine the impact of long-term OS elevation, as seen in AD on the body and GM, we examined GM alterations during the initial OS elevation using a two-week OS loading rat model, and examined the effects of TwX on OS and GM. Furthermore, using a questionnaire survey and fecal samples, we analyzed the impact of TwX on healthy individuals' gut bacteria and the associated effect on their quality of life (QOL). TwX was found to increase the number of bacteria species and their diversity in GM, as well as butyrate-producing bacteria, which tend to be reduced in AD patients. Additionally, TwX improved defecation condition and QOL. The gut bacteria function as part of the homeostatic function during OS elevation, and the prophylactic administration of TwX strengthened this function. The results suggest that the preventative effect of TwX on dementia may involve the GM, in addition to the other previously demonstrated effects.


Mechanical bowel preparation for colorectal surgery.
Nosocomial infections are the most frequent complications of surgical patients. Most surgical site infections (SSI) are acquired intraoperatively and arise from the flora of the patient's skin, gastrointestinal tract, or mucous membranes. Although preoperative mechanical cleansing of the bowel is considered by many surgeons a cornerstone of modern elective colorectal surgery and, in association with antibiotic prophylaxis, a fundamental component of an intestinal antisepsis program, many surgeons do not perform preoperative mechanical preparation routinely. Review of the pertinent literature. Some recent randomized trials and a Cochrane review found no proof that mechanical bowel preparation reduces the risk of complications after elective colorectal surgery. Indeed, there is some evidence that this intervention is associated with a higher rate of anastomotic leakage and wound complications. The dogma that mechanical bowel preparation is necessary before elective colorectal surgery may need to be reconsidered. On the other hand, such preparation decreases operating time by improving bowel handling during construction of the anastomosis. Moreover, it is helpful when intestinal palpation will be necessary for identification of a lesion.


On the molecular organization, diversity and functions of desmosomal proteins.
After a brief review of epithelial cell junctions, the authors describe a series of observations and experiments directed toward elucidating the molecular organization and functions of the proteins of desmosomes (maculae adherentes). Their immunofluorescence observations reveal differences between the proteins of junctional complex and other desmosomes of the same cells. Quantitative immuno-localization studies are described using protein A-colloidal gold markers and a series of monospecific, polyclonal antibodies directed against cytokeratins and four desmosomal protein families. It is shown that desmoplakin III (about 81 kDa) is restricted to the desmosomal cytoplasmic plaque, which possesses outer and inner dense zones. Desmoplakins I/II (about 240 kDa and 210 kDa) extend across the same range and beyond into the intermediate filament-rich cytoplasm. Two glycoprotein families, desmoglein I (DGI; about 150 kDa) and desmoglein II (DGII; about 97-118 kDa), extend from the desmosomal midline (or beyond) across the desmoglea (intercellular space), through the plasma membrane and across both layers of the plaque, terminating near its cytoplasmic border. Cytokeratins do not extend into the desmosome. With new procedures utilizing guanidine HCl for preparing and fractionating desmosomal proteins, DGII has been purified to homogeneity. DGII, DGI and a third protein are all shown to bind Ca2+, which is known to promote desmosome assembly.


The false-positive to false-negative ratio in epidemiologic studies.
The ratio of false-positive to false-negative findings (FP:FN ratio) is an informative metric that warrants further evaluation. The FP:FN ratio varies greatly across different epidemiologic areas. In genetic epidemiology, it has varied from very high values (possibly even >100:1) for associations reported in candidate-gene studies to very low values (1:100 or lower) for associations with genome-wide significance. The substantial reduction over time in the FP:FN ratio in human genome epidemiology has corresponded to the routine adoption of stringent inferential criteria and comprehensive, agnostic reporting of all analyses. Most traditional fields of epidemiologic research more closely follow the practices of past candidate gene epidemiology, and thus have high FP:FN ratios. Further, FP and FN results do not necessarily entail the same consequences, and their relative importance may vary in different settings. This ultimately has implications for what is the acceptable FP:FN ratio and for how the results of published epidemiologic studies should be presented and interpreted.


Common treatment goals of antipsychotics: acute treatment.
When a patient with an acute exacerbation of schizophrenia is admitted into the hospital, the target symptoms include pathologic excitement/agitation and exacerbated psychotic symptoms. The goal of hospitalization becomes attenuation of these symptoms to a level compatible with safe discharge. The mainstay of stabilization is antipsychotic treatment. A risk/benefit analysis of the conventional versus the newer antipsychotics favors the use of the newer agents as first-line drugs. These newer antipsychotic agents represent the first significant advance in the pharmacologic treatment of schizophrenia in the past four decades. They are at least as effective as conventional agents and are clearly superior from a safety perspective. Because of short inpatient stays, the challenge for clinicians is to provide an adequate treatment period without aggressively escalating the dose.


Jatropha curcas hemagglutinin is similar to a 2S albumin allergen from the same source and has unique sugar affinities.
We have previously reported the purification and preliminary X-ray characterization of a hemagglutinin from the seeds of Jatropha curcas and, with the detailed sequencing information available now, we find that it is similar to a 2S albumin allergen isolated from the same source. Through a search of Jatropha genome database (http://www.kazusa.or.jp/jatropha/), we map it to the sequence id JcCA0234191 (now referred to as Jcr4S00619.70 in the new version, release 4.5) which has a conserved alpha amylase inhibitor/seed storage protein domain found in the 2S albumin allergens. The putative sequence of the small and large chains of the protein is assigned and the total mass of the two subunits matches with the intact mass 10 kDa determined through MALDI. The protein retains hemagglutination activity between pH 6-9 and up to 60 °C on heat treatment and its hemagglutination activity is inhibited by sialic acid and fetuin. Bioinformatics studies show that the isolated protein sequence clusters in close association with a 2S albumin from Ricinus communis in phylogeny analysis and has a conservation of the characteristic four disulfide linkage pattern. Hemagglutinins and lectins are known to have allergenic effects through their interaction with immunoglobulin E and histamine release and earlier studies have shown that this interaction can be inhibited by lectin-specific sugars. We hope this report bridges the plant allergens and hemagglutinins further for exploring possible mediation of allergenic activity through sialic acid and complex sugar interactions and generates further interest in the area.


Formation of non-infective herpesvirus particles in cultured cells treated with human interferon.
Treatment of HeLa cells with human lymphoblastoid interferon [HuIFN-alpha(Ly)] induced an antiviral state that rendered the cells refractory to infection by herpes simplex virus type 1 (HSV-1), and the yield of infectious HSV-1 was reduced by 98%. Analysis of the mechanism of the anti-herpes action of IFN indicated that no gross inhibition of viral protein synthesis took place and late proteins appeared, although the synthesis of some of them was partially inhibited. No differences were apparent between the glycoproteins or phosphoproteins synthesized in control and IFN-treated HSV-1-infected HeLa cells. No inhibition of the formation of new virus particles from IFN-treated cells was evident as assessed by electron microscopy and biochemical analyses, although their infectivity was drastically reduced. These virions exhibited some differences in their content of a few proteins as determined by PAGE. Studies on the second infection cycle with virions obtained from IFN-treated cells suggested that they attached and penetrated HeLa cells but that their development was impaired, since no viral protein synthesis was observed during the second infection. Altogether these results do not support the idea that the molecular mechanism of action of human IFN against HSV-1 is mediated via the 2'-5' A system, or by the inactivation of initiation factors. Rather, they focus the problem of the anti-herpes action of IFN at the level of formation of defective virions.


Immediate, full weightbearing cast treatment of acute Achilles tendon ruptures: a long-term follow-up study.
Thirty-two patients with 33 ruptures were available for comprehensive evaluation, which included a questionnaire, physical examination, and manual as well as objective (Kin-Com) strength testing. Data excluding Kin-Com strength testing were available in an additional seven patients with seven ruptures; hence, the data are based on these 39 patients with 40 ruptures. Five patients with eight ruptures were excluded from strength testing but were included in our rerupture data, which totaled 44 patients with 48 ruptures. All patients were treated using a standard protocol by a single surgeon. All patients were treated within 11 days of injury, and the average follow-up was 55 months (range, 24-133). Ninety-five percent (38/39) of patients were completely satisfied with their treatment. The American Orthopaedic Foot and Ankle Society (AOFAS) ankle hindfoot scores were >80 in 97.4% of patients, and the rerupture rate was 6.25% (3/48). Calf circumference and ankle range of motion were not statistically different between the two legs. The injured leg tended to be slightly weaker than the uninjured leg, but the difference was not significant for most of the quantitative strength measurements. This study provides data for the Achilles rupture patient who asks, "Is there a treatment option other than surgery?" In addition, the standardized protocol can be utilized to obtain successful outcomes in those who choose nonoperative treatment or are poor surgical candidates.


The prognostic effect of immunoscore in patients with clear cell renal cell carcinoma: preliminary results.
This study aimed to evaluate the density of CD8+ and CD3+ tumor-infiltrating lymphocytes (TILs) and determine whether the immunoscore has any prognostic effect on the oncological outcomes in patients with clear cell renal cell carcinoma (RCC). A total of 129 patients diagnosed with clear cell RCC following radical or partial nephrectomy between 2009 and 2014 were retrospectively analyzed. Both tumor core (CT) and the invasive margin of nephrectomy specimens were assessed. The specimens were immunostained for anti-CD8+ and anti-CD3+ TILs. The patients were divided into three groups (favorable, intermediate, and poor risk) according to immunoscore levels. In the multivariate analysis, a favorable immunoscore (I3-4) was associated with prolonged disease-free survival (DFS), progression-free survival (PFS), and overall survival (OS) (HR 2.652, 2.848, and 2.933, respectively; all p < 0.001). The lower Fuhrman grade and pathological tumor-node-metastasis (TNM) stage had better DFS, PFS, and OS, whereas prolonged PFS was associated with a higher density of CD8+ CT (HR 1.602, 95% CI 0.934-3.470; p = 0.014). The shorter DFS, PFS, and OS were observed in the group with poor immunoscore (I0-1) at the early TNM stage of RCC (p < 0.001). In the metastatic subgroup analysis, the immunoscore showed better estimation than the International Metastatic RCC Database Consortium model and the Memorial Sloan-Kettering Cancer Center risk model for progression and OS (p < 0.001). The additional contributions of immunoscore to TNM stage, Fuhrman grade, and the WHO/ISUP 2016 grade for estimating oncological outcomes were found in ROC analysis. According to our preliminary results, immunoscore can be a promising prediction tool in clear cell RCC for postoperative oncological outcomes following nephrectomy.


Rapid preparation of procyanidins B2 and C1 from Granny Smith apples by using low pressure column chromatography and identification of their oligomeric procyanidins.
Research in the field of procyanidins is always hindered by the lack of procyanidin standards, and the preparation of procyanidins, especially in large scale, remains difficult and time-consuming. Commercial sources of procyanidin standards are scarce. In this study, a rapid preparation method of procyanidins by using low-pressure column chromatography was developed. Procyanidins in Granny Smith apples were extracted with boiled water and purified on an ADS-17 macroporous resin column to obtain a Granny Smith apple procyanidin extract (GSE). GSE was fractionated according to its degree of polymerization on a Toyopearl TSK HW-40s column. Procyanidins B2 (epicatechin-(4beta-8)-epicatechin) and C1 (epicatechin-(4beta-8)-epicatechin-(4beta-8)-epicatechin) were prepared without HPLC separation. Oligomeric procyanidins from Granny Smith apples were also identified by liquid chromatography-electrospray ionization-mass spectrometry.