Candida ogatae sp. nov., an anamorphic member of the Kuraishia clade.
Three methanol-assimilating yeast strains representing a hitherto undescribed species were isolated from rotten wood and freshwater samples collected in Hungary. Analysis of the D1/D2 large subunit rRNA gene sequences placed the strains in the Kuraishia clade; however, no ascospore formation was observed. These strains differ from Candida hungarica, the genetically most closely related recognized species, by four and five substitutions in D1/D2 and by >1% and 4% differences in the internal transcribed spacer and in the mitochondrial small subunit rRNA gene regions, respectively. Some phenotypic differences were also observed. Candida ogatae, a novel yeast species, is proposed to accommodate these isolates. The type culture is NCAIM Y.01845(T) (CBS 10924, NRRL Y-48474).


FAVR (Filtering and Annotation of Variants that are Rare): methods to facilitate the analysis of rare germline genetic variants from massively parallel sequencing datasets.
Characterising genetic diversity through the analysis of massively parallel sequencing (MPS) data offers enormous potential to significantly improve our understanding of the genetic basis for observed phenotypes, including predisposition to and progression of complex human disease. Great challenges remain in resolving genetic variants that are genuine from the millions of artefactual signals. FAVR is a suite of new methods designed to work with commonly used MPS analysis pipelines to assist in the resolution of some of the issues related to the analysis of the vast amount of resulting data, with a focus on relatively rare genetic variants. To the best of our knowledge, no equivalent method has previously been described. The most important and novel aspect of FAVR is the use of signatures in comparator sequence alignment files during variant filtering, and annotation of variants potentially shared between individuals. The FAVR methods use these signatures to facilitate filtering of (i) platform and/or mapping-specific artefacts, (ii) common genetic variants, and, where relevant, (iii) artefacts derived from imbalanced paired-end sequencing, as well as annotation of genetic variants based on evidence of co-occurrence in individuals. We applied conventional variant calling applied to whole-exome sequencing datasets, produced using both SOLiD and TruSeq chemistries, with or without downstream processing by FAVR methods. We demonstrate a 3-fold smaller rare single nucleotide variant shortlist with no detected reduction in sensitivity. This analysis included Sanger sequencing of rare variant signals not evident in dbSNP131, assessment of known variant signal preservation, and comparison of observed and expected rare variant numbers across a range of first cousin pairs. The principles described herein were applied in our recent publication identifying XRCC2 as a new breast cancer risk gene and have been made publically available as a suite of software tools. FAVR is a platform-agnostic suite of methods that significantly enhances the analysis of large volumes of sequencing data for the study of rare genetic variants and their influence on phenotypes.


Correlation of testicular volume, histology and LHRH test in adolescents with idiopathic varicocele.
Testicular growth failure as well as a well-documented histological damage have been reported in adolescents with varicocele. However, the proper management of this disorder in childhood is still debated due to the lack of seminal data and to the large number of adults with varicocele who have no fertility problems. In order to identify the subset of patients who are at risk of developing impaired testicular function and thus candidates for early varicocelectomy, we have studied 15 adolescents 10-17 years old with a grade II-III left-sided varicocele. Testicular volume and gonadotropin response to LHRH stimulation were evaluated preoperatively and compared to the histological findings from bilateral testicular biopsies. An abnormal left testicular histology was observed in 7 boys (46.6%) but only 3 of them (20%) showed severe testicular damage. The difference in size between the right and the left testis was never significant and was of no value in detecting subjects with abnormal histology. A good correlation was found between testicular injury and an increased response of both gonadotropins to the administration of LHRH. Decreased testicular volume does not seem to be a reliable predictor of early testicular damage. The assessment of gonadotropin response to LHRH may provide an additional objective means of evaluating adolescents with varicocele.


Binding of the integrin Mac-1 (CD11b/CD18) to the third immunoglobulin-like domain of ICAM-1 (CD54) and its regulation by glycosylation.
Both the integrins LFA-1 and Mac-1 bind to ICAM-1, an immunoglobulin superfamily member. Previously, we localized the binding sites of LFA-1 and the major group of human rhinoviruses to the first NH2-terminal immunoglobulin-like domain of ICAM-1. Here, we show that the binding site on ICAM-1 for Mac-1 is unexpectedly distinct from that for LFA-1 and maps to the third NH2-terminal immunoglobulin-like domain. These findings provide a function for the tandem duplication of immunoglobulin-like domains in ICAM-1 and have implications for other immunoglobulin superfamily members. Mutations at two sites in the third domain that destroy consensus sequences for N-linked glycosylation enhance binding to purified Mac-1. Agents that interfere with carbohydrate processing provide evidence that the size of the N-linked oligosaccharide side chains on ICAM-1 affects binding to Mac-1 but not to LFA-1. Thus, we suggest that the extent of glycosylation on ICAM-1 may regulate adhesion to LFA-1 or Mac-1 in vivo.


Relationship between human development and disappearance of unusually large von Willebrand factor multimers from plasma.
von Willebrand factor (vWF) multimers were examined in fetal, umbilical cord, and neonatal platelet-poor plasma (PPP) specimens. Sixty-five of 65 (100%) fetal PPP samples aged less than 35 weeks and seven of ten (70%) fetal samples aged greater than 35 weeks had unusually large vWF (ULvWF) multimers. Thirty of 46 (65%) cord PPP samples from neonates ranging in gestational age from 34 to 41 weeks had ULvWF. There was no significant relationship between either gestational age at time of delivery or birth weight and likelihood of finding ULvWF multimers in cord PPP samples. No maternal PPP sample contained ULvWF multimers. Serial heelstick samples from 16 preterm and term neonates were analyzed for 8 weeks. ULvWF multimers disappeared from the PPP of ten of the neonates during this time. The PPP of four neonates had vWF patterns similar to those in normal adult PPP throughout the sampling period. The ULvWF multimeric forms of fetal and neonatal PPP samples were similar to those constitutively released from endothelial cells. They were not as slowly migrating in a very porous 0.5% agarose gel system as the ULvWF multimers released from Weibel-Palade bodies in response to the calcium ionophore A23187. A vWF protomer was present in 97% of fetal samples, 83% of cord blood specimens, and 11% of neonatal heelstick samples, but was not found in any maternal sample. These results indicate that control mechanisms operative in older children and adults to prevent circulation of ULvWF multimers and vWF protomeric forms are normally acquired late in uterine life or during the neonatal period. ULvWF multimers, which are normal components of fetal, most cord, and some neonatal plasma samples, may contribute to in utero and postnatal hemostasis.


Isomerization of beta-HCH in aqueous solution.
beta-HCH was shaken with distilled water at 25 +/- 1 degree C for different time intervals and filtered. The ether extract of the filtrate on analysis by gas liquid chromatography showed the presence of four isomers, alpha, beta, gamma and delta, besides other compounds, thereby indicating isomerization of the beta-isomer in aqueous solution. Further, the ether extract, when dried and dissolved in acetone, showed high toxicity to mosquito larvae, flour beetle larvae and house-flies thereby indicating conversion of the inert beta-isomer to more toxic forms corroborating GLC analysis. Acetone solution of pure beta-HCH was found nontoxic against the insects tested.


Specificity of SARS-CoV-2 Real-Time PCR Improved by Deep Learning Analysis.
Real-time PCR (RT-PCR) is widely used to diagnose human pathogens. RT-PCR data are traditionally analyzed by estimating the threshold cycle (CT ) at which the fluorescence signal produced by emission of a probe crosses a baseline level. Current models used to estimate the CT value are based on approximations that do not adequately account for the stochastic variations of the fluorescence signal that is detected during RT-PCR. Less common deviations become more apparent as the sample size increases, as is the case in the current SARS-CoV-2 pandemic. In this work, we employ a method independent of CT value to interpret RT-PCR data. In this novel approach, we built and trained a deep learning model, qPCRdeepNet, to analyze the fluorescent readings obtained during RT-PCR. We describe how this model can be deployed as a quality assurance tool to monitor result interpretation in real time. The model's performance with the TaqPath COVID19 Combo Kit assay, widely used for SARS-CoV-2 detection, is described. This model can be applied broadly for the primary interpretation of RT-PCR assays and potentially replace the CT interpretive paradigm.


Carbohydrate restriction alters hepatic cholesterol metabolism in guinea pigs fed a hypercholesterolemic diet.
The current study was undertaken to evaluate the effect of carbohydrate restriction on hepatic cholesterol metabolism in guinea pigs fed a hypercholesterolemic diet. Hartley male guinea pigs (n = 10 per group) were fed 1 of 3 diets: a diet with a percent energy distribution of 42:23:35 carbohydrate:protein:fat and 0.04% cholesterol (control), a diet with the same macronutrient distribution but with 0.25% cholesterol (HChol), or a carbohydrate-restricted (CR) diet with a percent energy distribution of 11:30:59 carbohydrate:protein:fat and 0.25% cholesterol for 12 wk. There was more accumulation of hepatic cholesterol and triglycerides as well as lower 3-hydroxy-3-methyl glutaryl-CoA reductase messenger RNA abundance in guinea pigs fed the high-cholesterol diets (HChol and CR) (P < 0.01). Guinea pigs fed the CR diet had lower concentrations of hepatic total cholesterol and cholesteryl ester than those fed the HChol diet (P < 0.05). There was no diet effect on hepatic LDL receptor expression. Hepatic acyl CoA cholesteryl acyltransferase (ACAT) activity was lowest in guinea pigs fed the low-cholesterol diet (9.7 +/- 4.8 pmol.min(-1).mg(-1)), intermediate in those fed the CR diet (37.3 +/- 12.4 pmol.min(-1).mg protein(-1)), and highest in guinea pigs fed the HChol diet (55.9 +/- 11.2 pmol.min(-1).mg(-1)). ACAT activity was significantly correlated with hepatic cholesterol (r = 0.715; P < 0.01) and LDL cholesterol (r = 0.59; P < 0.01) for all dietary groups, suggesting a major role of this enzyme in hepatic cholesterol homeostasis and in lipoprotein concentrations. These results indicate that dietary cholesterol increases hepatic lipid accumulation and affects hepatic cholesterol homeostasis. Carbohydrate restriction in the presence of high cholesterol is associated with lower hepatic ACAT activity and an attenuation of hepatic cholesterol accumulation.


An overview of experimental designs in HPLC method development and validation.
Chemometric approaches have been increasingly viewed as precious complements to high performance liquid chromatographic practices, since a large number of variables can be simultaneously controlled to achieve the desired separations. Moreover, their applications may efficiently identify and optimize the significant factors to accomplish competent results through limited experimental trials. The present manuscript discusses usefulness of various chemometric approaches in high and ultra performance liquid chromatography for (i) methods development from dissolution studies and sample preparation to detection, considering the progressive substitution of traditional detectors with tandem mass spectrometry instruments and the importance of stability indicating assays (ii) method validation through screening and optimization designs. Choice of appropriate types of experimental designs so as to either screen the most influential factors or optimize the selected factors' combination and the mathematical models in chemometry have been briefly recalled and the advantages of chemometric approaches have been emphasized. The evolution of the design of experiments to the Quality by Design paradigm for method development has been reviewed and the Six Sigma practice as a quality indicator in chromatography has been explained. Chemometric applications and various strategies in chromatographic separations have been described.


Broadcast Propagation Time in SpaceFibre Networks with Various Types of Spatial Redundancy.
Various methods of spatial redundancy can be used in local networks based on the SpaceFibre standard for fault mitigation of network hardware and physical communication channels. Usually, a network developer chooses the method of spatial redundancy according to the number of failures that have to be mitigated, the time required for restoring the normal operation of the network, required overheads and hardware costs. The use of different spatial redundancy mechanisms can cause changes in the structure of the links between network nodes, in case of failure and subsequent mitigation. In turn, this may cause changes in the broadcast transmission paths and the temporal characteristics of their delivery from the source to the receivers. This article focuses on the change in the propagation time of broadcasts in SpaceFibre networks with spatial redundancy. Broadcast propagation rules significantly differ from data-packet propagation rules. Broadcast distribution time is very important for many applications, because broadcasts are generally used to send urgent messages, in particular for time synchronization. Various formal methods have been used to evaluate the propagation characteristics of the broadcast. A method for estimating broadcast propagation time along the shortest routes is proposed. In addition, we provide a formal method to estimate the number of failures, which occurred in the network during the broadcast propagation. This method is based on timed Petri nets; one of its features is the ability to calculate broadcast transmission delays. In addition, as an alternative solution, we propose a method for estimating delays based on time automata theory.


Two-Photon Excited Fluorescence Dynamics in Enzyme-Bound NADH: the Heterogeneity of Fluorescence Decay Times and Anisotropic Relaxation.
The dynamics of polarized fluorescence in NADH in alcohol dehydrogenase (ADH) in buffer solution has been studied using the TCSPC spectroscopy. A global fit procedure was used for determination of the fluorescence parameters from experiment. The interpretation of the results obtained was supported by ab initio calculations of the NADH structure. A theoretical model was developed describing the polarized fluorescence decay in ADH-NADH complexes that considered several interaction scenarios. A comparative analysis of the polarization-insensitive fluorescence decay using multiexponential fitting models has been carried out. As shown, the origin of a significant enhancement of the decay time in the ADH-NADH complex can be attributed to the decrease of nonradiative relaxation rates in the nicotinamide ring in the conditions of the apolar binding site environment. The existence of a single decay time in the ADH-NADH complex in comparison with two decay times observed in free NADH was attributed to a single NADH unfolded conformation in the ADH binding site. Comparison of the experimental data with the theoretical model suggested the existence of an anisotropic relaxation time of about 1 ns that is related with the rotation of fluorescence transition dipole moment due to the rearrangement of the excited state NADH nuclear configuration.


Macrophages produce IL-33 by activating MAPK signaling pathway during RSV infection.
It has been reported that RSV infection can enhance IL-33 production in lung macrophages. However, little is known about specific signaling pathways for activation of macrophages during RSV infection. In the present study, by using real-time RT-PCR as well as western blot assay, it became clear that RSV infection can enhance not only the expression of mRNAs for MAPK molecules (including p38, JNK1/2, and ERK1/2), but also the levels of MAPK proteins in lung macrophages as well as RAW264.7 cells. Furthermore, infection with RSV resulted in an increased level of phosphorylated MAPK proteins in RAW264.7 cells, suggesting that MAPK signaling pathway may participate in the process of RSV-induced IL-33 secretion by macrophages. In fact, the elevated production of IL-33 in RAW264.7 was attenuated significantly by pretreatment of the cells with special MAPK inhibitor before RSV infection, further confirming the function of MAPKs pathway in RSV-induced IL-33 production in macrophages. In contrast, the expression of NF-κB mRNA as well as the production of NF-κB protein in lung macrophages and RAW264.7 cells was not enhanced markedly after RSV infection. Moreover, RSV infection failed to induce the phosphorylation of NF-κB in RAW264.7 cells, suggesting that NF-κB signaling pathway may be not involved in RSV-induced IL-33 production in macrophages. Conclusion, these results indicate that RSV-induced production of IL-33 in macrophages is dependent on the activation of MAPK signaling pathway.


In vitro permeation study of a mucoadhesive drug delivery system for controlled delivery of nonoxynol-9.
A carbopol 934P-based drug delivery system (AmDDS) was developed to achieve the dual-controlled delivery of Nonoxynol-9 (N-9), a spermicidal agent, and EDTA, a potentiator of spermicidal activity. This gel-type system made intimate contact with vaginal mucosa and maintained an effective drug concentration within the vagina for a prolonged period of time. An existing mathematical model, based on a unilayer diffusion membrane, was applied for describing permeation of N-9 through vaginal mucosa. Vaginal permeation of N-9 from AmDDS was negligible over the initial 5 hr, then increased in a Q versus t pattern, but remained low. A relationship was observed between the permeation rate (Js) of drug through vaginal mucosa and the release flux (Q/t1/2) from AmDDS, which showed that the permeation rate (Js) increased with increased square of the release flux [(Q/t1/2)2] in a hyperbolic manner. This result indicated that release of N-9 from AmDDS is still the rate-limiting step at doses within the tested range. EDTA, at concentration of up to 0.32%, did not change the permeation rate of N-9 through vaginal mucosa. It is thus concluded that the developed AmDDS can control the intravaginal delivery of N-9 as well as its permeation through vaginal mucosa.


Clinical implications of food-drug interactions with small-molecule kinase inhibitors.
During the past two decades, small-molecule kinase inhibitors have proven to be valuable in the treatment of solid and haematological tumours. However, because of their oral administration, the intrapatient and interpatient exposure to small-molecule kinase inhibitors (SMKIs) is highly variable and is affected by many factors, such as concomitant use of food and herbs. Food-drug interactions are capable of altering the systemic bioavailability and pharmacokinetics of these drugs. The most important mechanisms underlying food-drug interactions are gastrointestinal drug absorption and hepatic metabolism through cytochrome P450 isoenzymes. As food-drug interactions can lead to therapy failure or severe toxicity, knowledge of these interactions is essential. This Review provides a comprehensive overview of published studies involving food-drug interactions and herb-drug interactions for all registered SMKIs up to Oct 1, 2019. We critically discuss US Food and Drug Administration (FDA) and European Medicines Agency (EMA) guidelines concerning food-drug interactions and offer clear recommendations for their management in clinical practice.


Improving the Toric Intraocular Lens Calculation by Considering Posterior Corneal Astigmatism and Surgically-induced Corneal Astigmatism.
To investigate the effect of surgically induced corneal astigmatism (SICA) and total corneal astigmatism (TCA) estimation on the anterior corneal astigmatism (ACA)-based toric intraocular lens (IOL) calculation. Data from preoperative and postoperative corneal astigmatism, postoperative visual acuities, and refractive outcomes were collected. The incision was superior in with the rule anterior corneal astigmatism (WTRA) eyes and temporal in against the rule anterior corneal astigmatism eyes. The following five methods of calculating the toric IOL were compared: (1) ACA only and estimated SICA; (2) ACA with a fixed posterior corneal astigmatism (PCA) and estimated SICA; (3) ACA with a fixed PCA value and actual SICA; (4) and (5) TCA derived from the regression equations of ACA and actual SICA. The residual astigmatism was simulated. The Alpins method was used to analyze the astigmatism. Sixty eyes from 46 patients were enrolled. Thirty eyes had WTRA and the other thirty had against the rule anterior corneal astigmatism. The vector and arithmetic means of the difference vector decreased when the information regarding the actual SICA and PCA was added to the calculation (from 0.59 diopters [D] @ 87.5° to 0.15 D @ 48.5°, and from 0.95 ± 0.53 to 0.71 ± 0.63 D, respectively; p < 0.001). The mean difference vector across the whole sample was lowest using model 5. The correction index was significantly closest to 1.0 in the WTRA group. Researchers may improve the accuracy of post-implantation predictions by calculating toric IOL using exact SICA and TCA, with consideration of the PCA derived from the regression equation of ACA.


Relationship between prosthesis-patient mismatch and pro-brain natriuretic peptides after aortic valve replacement.
It has been shown previously that elevated plasma levels of B-type natriuretic peptide (BNP) and its N-terminal fragment (NT-pro-BNP) are related to the degree and progression of native aortic valve disease. In addition, NT-pro-BNP levels have been shown to decrease after successful aortic valve replacement (AVR). The presence of a valve prosthesis-patient mismatch (PPM) may affect the beneficial effects of AVR, however. The study aim was to investigate the relationship between PPM and NT-pro-BNP plasma levels late after AVR. A series of consecutive patients (42 males, 31 females; mean age 66 +/- 13 years) who had undergone isolated AVR between May 2004 and July 2007 was enrolled into the study. Patients with preoperative moderate to severe mitral regurgitation, coronary artery disease, left ventricular (LV) dysfunction (ejection fraction <45%) and serum creatinine >150 mmol/l were excluded. PPM was defined severe as an indexed effective orifice area (EOAi) < or = 0.65 cm2/m2, or moderate when the EOAi was 0.66-0.85 cm2/m2. Plasma NT-pro-BNP levels and echocardiographic assessments were performed in all patients during routine follow up after surgery. The patients received either a biological (n = 42) or mechanical (n = 31) prosthesis. Among the patients, 21 had no PPM, 27 moderate PPM, and 25 severe PPM. At a median follow up of 18 months, the mean NT-pro-BNP plasma level was 532 pg/ml (95% CI: 393.1-671.6), and the mean LV mass index (LVMI) 120 +/- 4 g/m2, the LVEF 60 +/- 1%, the peak aortic prosthesis gradient 28 +/- 2 mmHg, and the EOAi 0.74 +/- 0.02 cm2/m2. Multivariate statistical analysis showed that NT-pro-BNP level correlated with age (beta = 0.57, p<0.0001), LVMI (beta = 0.32, p = 0.02), NYHA class (beta = 0.50, p = 0.003) and EOAi (beta = -0.38, p = 0.02). The study results showed that NT-pro-BNP levels were independently related to PPM late after isolated AVR in patients with preserved LV function. However, further investigations are required to confirm these findings and to identify their clinical implications.


Propagation of action potential activity in a predefined microtunnel neural network.
A polydimethylsiloxane microtunnel device with two wells is aligned and attached on top of a multi-electrode array. Neurons are grown first in one well and allow the propagation of axons through the tunnels into a second well. After 10 days, cells are plated in the second well, with much lower likelihood of extending axons back to the first well, with the intent of creating unidirectional connectivity between populations of neurons in the two wells. Here we report electrophysiological evidence that supports the hypothesis that the dominant information flow is in the desired direction. This was done by measuring the propagation speed and direction of individual action potentials, with the result that 84% of the spikes propagated in the desired direction. Further, we recorded globally synchronized burst activity on each of the electrodes, identified the timing of the first spike on each electrode, recorded locally synchronized burst activity which is found only in the second well and does not propagate back to the first well and concluded that this measure of burst propagation supports the hypothesis of a unidirectionally connected network. Two hypotheses are discussed for the mechanism underlying the activity pattern of the particular neural networks.


A novel immunomodulatory molecularly targeted strategy for refractory Hodgkin's lymphoma.
Although Hodgkin's lymphoma (HL) was one of the first human cancers to be cured by chemotherapy, no new agents other than brentuximab vedotin (Adcetris®, CD 30 directed antibody drug conjugate) have received US Food and Drug Administration (FDA) approval for HL since 1977. Subsets of young adult patients with HL continue to relapse, even after stem cell transplantation, warranting new approaches. Against this background, we report a dramatic response in a young patient with advanced HL refractory to the standard treatment who responded to the combination of a pan-histone deacetylase inhibitor (vorinostat, suberoylanilide hydroxamic acid, SAHA) and mammalian target of rapamycin (mTOR) inhibitor therapy (sirolimus,rapamume). In-depth immunohistochemical and morphoproteomic analyses of this exceptional responder to targeted therapy have yielded potential insights into the biology of advanced HL. The PI3K/AKT/mTOR pathway is a commonly activated pathway in multiple tumor types including HL. The patient was treated using therapy based on mechanistic in vitro data demonstrating that combined histone deacetylase (HDAC) and mTOR inhibition act together on this pathway, resulting in inhibition of reciprocal feedback networks, leading to better anti-proliferative activity. The in vivo response signature from this patient's tissue sample sheds light on immune dysregulation in HL. We describe the response signature achieved from targeting immune dysregulation in addition to targeting the key oncogenic PI3K/AKT/mTOR pathway. We also expand on the role of rapamycin analogs in oncology. This study supports a role for an immune-type pathogenesis that is amenable to immune modulating targeted therapy in refractory HL. We report an exceptional responder to molecularly targeted and immune modulator therapy in advanced Hodgkin's lymphoma. The morphoproteomic/morphometric findings in this "unusual responder" patient's relapsed HL that correlate best, as a response signature with the subsequent clinical remission following rapamycin (sirolimus) and vorinostat (SAHA) therapies, center on an immune dysregulation involving an imbalance between effector and functional T regulatory cells in addition to targeting the mTOR pathway. This underscores the need for an approach illustrated in our study--namely of focusing on pathogenetic mechanisms and combinatorial therapies that target both the pathogenesis and adaptive responses to contemplated therapies.


Propranolol blocks the unconditioned and conditioned hyperactive effects of methamphetamine in CD-1 mice.
The present experiment examined the contribution of the β-adrenergic receptor system in mediating the unconditioned (i.e. pharmacological) and conditioned (i.e. learned) hyperactive effects of methamphetamine. To this end, mice underwent an 8-day conditioning procedure involving two different, alternating session types (chamber and home-cage days). On chamber days (1, 3, 5, and 7), mice were injected (intraperitoneally) with vehicle (dH 2 O) or propranolol (16 or 32 mg/kg) and were injected (subcutaneously) 30 min (min) later by either vehicle (saline; unpaired) or methamphetamine (1.0 mg/kg; paired). On home-cage days (2, 4, 6, and 8), mice were injected (subcutaneously) with either vehicle (saline; paired) or methamphetamine (1.0 mg/kg; unpaired) in their home cages. The test day for conditioned hyperactivity occurred 48 h after the last chamber day. Propranolol dose-dependently blocked the unconditioned and conditioned hyperactive effects of methamphetamine, implicating a role for the β-adrenergic system in mediating these effects of methamphetamine.


Enhanced magnetotransport in nanopatterned manganite nanowires.
We have combined optical and focused ion beam lithographies to produce large aspect-ratio (length-to-width >300) single-crystal nanowires of La2/3Ca1/3MnO3 that preserve their functional properties. Remarkably, an enhanced magnetoresistance value of 34% in an applied magnetic field of 0.1 T in the narrowest 150 nm nanowire is obtained. The strain release at the edges together with a destabilization of the insulating regions is proposed to account for this behavior. This opens new strategies to implement these structures in functional spintronic devices.


A Novel MS-Cleavable Azo Cross-Linker for Peptide Structure Analysis by Free Radical Initiated Peptide Sequencing (FRIPS).
The chemical cross-linking/mass spectrometry (MS) approach is a growing research field in structural proteomics that allows gaining insights into protein conformations. It relies on creating distance constraints between cross-linked amino acid side chains that can further be used to derive protein structures. Currently, the most urgent task for designing novel cross-linking principles is an unambiguous and automated assignment of the created cross-linked products. Here, we introduce the homobifunctional, amine-reactive, and water soluble cross-linker azobisimidoester (ABI) as a prototype of a novel class of cross-linkers. The ABI-linker possesses an innovative modular scaffold combining the benefits of collisional activation lability with open shell chemistry. This MS-cleavable cross-linker can be efficiently operated via free radical initiated peptide sequencing (FRIPS) in positive ionization mode. Our proof-of-principle study challenges the gas phase behavior of the ABI-linker for the three amino acids, lysine, leucine, and isoleucine, as well as the model peptide thymopentin. The isomeric amino acids leucine and isoleucine could be discriminated by their characteristic side chain fragments. Collisional activation experiments were conducted via positive electrospray ionization (ESI) on two Orbitrap mass spectrometers. The ABI-mediated formation of odd electron product ions in MS/MS and MS3 experiments was evaluated and compared with a previously described azo-based cross-linker. All cross-linked products were amenable to automated analysis by the MeroX software, underlining the future potential of the ABI-linker for structural proteomics studies. Graphical Abstract .


The combined orthodontic and restorative treatment for patients with malocclusion and dentition defects: A randomized controlled trial.
To explore the effects of a combined orthodontic and restorative approach on chewing, swallowing, and language function in patients with malocclusion and dental defects. A total of 112 patients with malocclusion and dentition defects admitted to the Lianyungang Hospital of Traditional Chinese Medicine from June 2019 to June 2022 were prospectively selected. The patients were divided into study and control groups using a simple random number table method, with 56 patients in each group. The control group received routine restoration, whereas the study group received a combination of orthodontic and restorative treatments. The chewing function, swallowing and language function, and gingival periodontal condition before and after treatment in both groups were compared using t-test or Wilcoxon test, while treatment satisfaction were compared using chi-square test or Fisher exact test. After treatment, maximum area frame bite force/max movie force in both groups increased compared to before treatment, while occlusion time, bite force distrbution balance, and standard deviation hue decreased compared to before treatment. Moreover, maximum area frame bite force/max movie force in the study group was higher than that in the control group, whereas occlusion time, bite force distrbution balance, and standard deviation hue were lower than those in the control group (P < .05). The swallowing and language function scores of the study group were higher than those of the control group (P < .05). After treatment, the bleeding index, plaque index, and probing depth of both groups decreased compared to before treatment, and the study group was lower than the control group (P < .05). The treatment satisfaction of the study group (94.64%) was higher than that of the control group (82.14%) (P < .05). Adopting a combined orthodontic and restorative approach to intervene in patients with malocclusion and dentition defects is beneficial for improving their periodontal condition, effectively restoring chewing, swallowing, and language functions, and achieving high patient satisfaction.


Necessary condition for frequency synchronization in network structures.
We present the necessary condition for complete frequency synchronization of phase-coupled oscillators in network structures. The surface area of a set of sites is defined as the number of links between the sites within the set and those outside the set. The necessary condition is that the surface area of any set of cN (0 < c < 1) oscillators in the N-oscillator system must exceed square root of N in the limit N --> infinity. We also provide the necessary condition for macroscopic frequency synchronization. Thus, we identify networks in which one or both of the above mentioned types of synchronization do not occur.


Phase 3 CLEAR study in patients with advanced renal cell carcinoma: outcomes in subgroups for the lenvatinib-plus-pembrolizumab and sunitinib arms.
The phase 3 CLEAR study demonstrated that lenvatinib plus pembrolizumab significantly improved efficacy versus sunitinib as first-line treatment for patients with advanced renal cell carcinoma (RCC). Prognostic features including presence and/or site of baseline metastases, prior nephrectomy, and sarcomatoid features have been associated with disease and treatment success. This subsequent analysis explores outcomes in patients with or without specific prognostic features. In CLEAR, patients with clear cell RCC were randomly assigned (1:1:1) to receive either lenvatinib (20 mg/day) plus pembrolizumab (200 mg every 3 weeks), lenvatinib (18 mg/day) plus everolimus (5 mg/day), or sunitinib alone (50 mg/day, 4 weeks on, 2 weeks off). In this report, progression-free survival (PFS), overall survival (OS), and objective response rate (ORR) were all assessed in the lenvatinib-plus-pembrolizumab and the sunitinib arms, based on baseline features: lung metastases, bone metastases, liver metastases, prior nephrectomy, and sarcomatoid histology. In all the assessed subgroups, median PFS was longer with lenvatinib-plus-pembrolizumab than with sunitinib treatment, notably among patients with baseline bone metastases (HR 0.33, 95% CI 0.21-0.52) and patients with sarcomatoid features (HR 0.39, 95% CI 0.18-0.84). Median OS favored lenvatinib plus pembrolizumab over sunitinib irrespective of metastatic lesions at baseline, prior nephrectomy, and sarcomatoid features. Of interest, among patients with baseline bone metastases the HR for survival was 0.50 (95% CI 0.30-0.83) and among patients with sarcomatoid features the HR for survival was 0.91 (95% CI 0.32-2.58); though for many groups, median OS was not reached. ORR also favored lenvatinib plus pembrolizumab over sunitinib across all subgroups; similarly, complete responses also followed this pattern. Efficacy outcomes improved following treatment with lenvatinib-plus-pembrolizumab versus sunitinib in patients with RCC-irrespective of the presence or absence of baseline lung metastases, baseline bone metastases, baseline liver metastases, prior nephrectomy, or sarcomatoid features. These findings corroborate those of the primary CLEAR study analysis in the overall population and support lenvatinib plus pembrolizumab as a standard of care in 1L treatment for patients with advanced RCC. ClinicalTrials.gov, identifier NCT02811861.


[Dimensions of the upper airways and its impact on the dentoalveolar system development].
Due to the high mobility and variability of bone structures and soft tissues surrounding the upper respiratory tract, the exact boundaries for measuring and normalizing the size of the respiratory tract have not yet been determined. Studies have determined the relationship between the narrowing of the upper jaw and a decrease in the transverse dimensions of the airways, as well as a positive effect in changing the size of the airways after orthodontic treatment and/or orthognathic surgery. Nevertheless, the results of research in this area may differ greatly from different specialists, which indicates that the topic is poorly studied and it is necessary to continue and expand the range of scientific works to assess the state of the upper respiratory tract and their relationship with the orthodontic status.


Association between systemic immune-inflammatory markers and the risk of moyamoya disease: a case-control study.
Background:Systemic immune-inflammatory markers such as neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), lymphocyte-to-monocyte ratio (LMR) and systemic immune-inflammatory index (SII) are associated with the prognosis of many cardiovascular and neoplastic diseases. Moyamoya disease (MMD) is associated with inflammation, but the relationship between systemic immune-inflammatory markers between MMD is unclear. The aim of our study was to analyse the association between systemic immune-inflammatory markers and the risk of MMD and its subtypes.Methods:We consecutively recruited 360 patients with MMD and 89 healthy control subjects in a case-control study to calculate and analyse the association of systemic immune-inflammatory markers with the risk of MMD and its subtypes.Results:The risk of MMD increased with higher levels of NLR (OR 1.237, 95% CI [1.008, 1.520], p = .042). When NLR and SII were assessed as quartile-spaced subgroups, the third quartile grouping of NLR and SII had a higher risk of MMD than the first quartile grouping (NLR: OR 3.206, 95% CI [1.271, 8.088], p = .014; SII: OR 3.074,95% CI [1.232,7.672], p = .016). When NLR was combined with SII, the highest subgroup had a higher risk of MMD than the lowest subgroup (OR2.643, 95% CI [1.340, 5.212], p = .005). The risk of subtypes also increased with higher levels of NLR and SII. The association between the levels of NLR and SII with the staging of the Suzuki stage follows an inverted U-shape. The highest levels of NLR and SII were found in patients with MMD at Suzuki stages 3-4.Conclusion:The risk of MMD increases with elevated systemic immune-inflammatory markers. This study analysed the association of systemic immune-inflammatory markers with the risk of developing MMD and its subtypes, and identified novel inflammatory markers for MMD.


An arthroscopic hip documentation form.
Hip arthroscopy is becoming increasingly popular. A simple, precise, and practical means of recording arthroscopic findings will be useful for diagnostic, research, and audit purposes. Basic principles of cartography exist to produce two-dimensional paper representations of our spherical planet. We used the same principles to produce a two-dimensional map of the acetabulum and femoral head. The resulting hip diagram shows the acetabulum as viewed from the side and the femoral head as viewed from above. The ligamentum teres is attached to the medial margin of the head. The head-neck junction and part of the femoral neck is shown at the opposite margin of the ligamentum teres. The hip documentation form is simple, precise, and accurate. We use it to record our findings at hip arthroscopy, which we have used to assist us in our practice.


[Clinical applications of magnetic resonance spectroscopy].
In this overview the physics of magnetic resonance spectroscopy (MRS) are briefly discussed. The biochemical pathways which can be investigated by MRS in physiological and pathological states are presented. The potential clinical applications of this method to the musculoskeletal system, heart, brain, liver and kidney are examined.


Defining needs-based urban health planning areas is feasible and desirable: a population-based approach in Toronto, Ontario.
Reporting health data for large urban areas presents numerous challenges. In the case of Toronto, Ontario, amalgamation in 1998 merged six census subdivisions into one mega-city, resulting in the disappearance of standard reporting units. A population-based approach was used to define new health planning areas. Census tracts were used as building blocks and combined according to residential income homogeneity, respecting natural and man-made boundaries, forward sortation areas and the City of Toronto's community neighbourhoods whenever possible. Correlations and maps were used to establish area boundaries. The city was divided into 5 major planning areas which were further subdivided creating 15 minor areas. Both major and minor areas showed significant differences in population characteristics, health status and health service utilization. This commentary demonstrates the feasibility and describes the outcomes of one method for establishing planning and reporting areas in large urban centres. Next steps include the further generation of health data for these areas, comparisons with other Canadian urban areas, and application of these methods to recently announced Ontario Local Health Integration Networks. These areas can be used for planning and evaluating health service delivery, comparison with other Canadian urban areas and ongoing monitoring of and advocacy for equity in health.


Expression of somatostatin receptor and effects of somatostatin analog on pancreatic endocrine tumors.
Somatostatin analogs have been administered to patients with pancreatic endocrine tumors in an attempt to inhibit hormone hypersecretion and prevent tumor growth. It is speculated that their efficacy is correlated with the expression of specific subtypes of somatostatin receptors. The aim of this study was to immunohistochemically evaluate the expression of somatostatin receptor subtypes in human pancreatic endocrine tumors, and to determine whether the expression of these subtypes is correlated with the effectiveness of the somatostatin analogs. Somatostatin receptor subtypes 1, 2, and 3 (sst 1, 2, and 3) were immunohistochemically investigated in seven pancreatic endocrine tumors: four insulinomas, one VIPoma, and two nonfunctioning tumors associated with multiple endocrine neoplasia type I, using paraffin sections. Three of the four patients with insulinoma were given an octreotide injection. Cells were homogeneously stained in the tumor region. More than 85% of the specimens expressed sst 1, 2, and 3. There was no difference among the immunohistochemical stainings of somatostatin receptor subtypes according to most tumor characteristics; however, the expression of sst 2 was extremely positive, and the expression of sst 3 was moderately positive in the specimen from a patient in whom the octreotide injection had proven very effective. These findings indicate that the efficacy of octreotide may be correlated with the density of sst 2 and 3 in an immunohistological study using paraffin sections.


Oculo-otological Manifestations in a Case of Granulomatosis with Polyangiitis.
A 44 year old lady presented with acute onset of loss of vision in the right eye and cough with mucopurulent expectoration for two months. Ophthalmic examination revealed central retinal artery occlusion (CRAO). Chest radiograph showed multiple cavitatory nodules with fluid levels. Sputum was negative for AFB and ANCA was strongly positive suggestive of a diagnosis of Granulomatosis with Polyangiitis (GPA). Within the next few weeks the patient had rapid detenoration due to left eye CRAO, progressive bilateral hearing loss, facial palsy and retro orbital mass. The aggressive disease responded well to steroids and cyclophosphamide.


Emerging Role of Glioma Stem Cells in Mechanisms of Therapy Resistance.
Since their discovery at the beginning of this millennium, glioma stem cells (GSCs) have sparked extensive research and an energetic scientific debate about their contribution to glioblastoma (GBM) initiation, progression, relapse, and resistance. Different molecular subtypes of GBM coexist within the same tumor, and they display differential sensitivity to chemotherapy. GSCs contribute to tumor heterogeneity and recapitulate pathway alterations described for the three GBM subtypes found in patients. GSCs show a high degree of plasticity, allowing for interconversion between different molecular GBM subtypes, with distinct proliferative potential, and different degrees of self-renewal and differentiation. This high degree of plasticity permits adaptation to the environmental changes introduced by chemo- and radiation therapy. Evidence from mouse models indicates that GSCs repopulate brain tumors after therapeutic intervention, and due to GSC plasticity, they reconstitute heterogeneity in recurrent tumors. GSCs are also inherently resilient to standard-of-care therapy, and mechanisms of resistance include enhanced DNA damage repair, MGMT promoter demethylation, autophagy, impaired induction of apoptosis, metabolic adaptation, chemoresistance, and immune evasion. The remarkable oncogenic properties of GSCs have inspired considerable interest in better understanding GSC biology and functions, as they might represent attractive targets to advance the currently limited therapeutic options for GBM patients. This has raised expectations for the development of novel targeted therapeutic approaches, including targeting GSC plasticity, chimeric antigen receptor T (CAR T) cells, and oncolytic viruses. In this review, we focus on the role of GSCs as drivers of GBM and therapy resistance, and we discuss how insights into GSC biology and plasticity might advance GSC-directed curative approaches.


Incubation of premise plumbing water samples on Buffered Charcoal Yeast Extract agar at elevated temperature and pH selects for Legionella pneumophila.
Worldwide, over 90% of the notified cases of Legionnaires' disease are caused by Legionella pneumophila. However, the standard culture medium for the detection of Legionella in environmental water samples, Buffered Charcoal Yeast Extract (BCYE) agar of pH 6.9 ± 0.4 with or without antimicrobial agents incubated at 36 ± 1 °C, supports the growth of a large diversity of Legionella species. BCYE agar of elevated pH or/and incubation at elevated temperature gave strongly reduced recoveries of most of 26 L. non-pneumophila spp. tested, but not of L. pneumophila. BCYE agar of pH 7.3 ± 0.1, incubated at 40 ± 0.5 °C (BCYE pH 7.3/40 °C) was tested for selective enumeration of L. pneumophila. Of the L. non-pneumophila spp. tested, only L. adelaidensis and L. londiniensis multiplied under these conditions. The colony counts on BCYE pH 7.3/40 °C of a L. pneumophila serogroup 1 strain cultured in tap water did not differ significantly from those on BCYE pH 6.9/36 °C when directly plated and after membrane filtration and showed repeatability's of 13-14%. By using membrane filtration L. pneumophila was detected in 58 (54%) of 107 Legionella-positive water samples from premise plumbing systems under one or both of these culture conditions. The L. pneumophila colony counts (log-transformed) on BCYE pH 7.3/40 °C were strongly related (r2 = 0.87) to those on BCYE pH 6.9/36 °C, but differed significantly (p < 0.05) by a mean of - 0.12 ± 0.30 logs. L. non-pneumophila spp. were detected only on BCYE pH 6.9/36 °C in 49 (46%) of the samples. Hence, BCYE pH 7.3/40 °C can facilitate the enumeration of L. pneumophila and their isolation from premise plumbing systems with culturable L. non-pneumophila spp., some of which, e.g. L. anisa, can be present in high numbers.


Mangiferin Attenuates Myocardial Ischemia-Reperfusion Injury via MAPK/Nrf-2/HO-1/NF-κB In Vitro and In Vivo.
The aim of this study was to investigate the cardioprotective effect of mangiferin (MAF) in vitro and in vivo. Oxidative stress and inflammatory injury were detected in coronary artery ligation in rats and also in hypoxia-reoxygenation- (H/R-) induced H9c2 cells. MAF inhibited myocardial oxidative stress and proinflammatory cytokines in rats with coronary artery occlusion. The ST segment of MAF treatment groups also resumed. Triphenyltetrazolium chloride (TTC) staining and pathological analysis showed that MAF could significantly reduce myocardial injury. In vitro data showed that MAF could improve hypoxia/reoxygenation- (H/R-) induced H9c2 cell activity. In addition, MAF could significantly reduce oxidative stress and inflammatory pathway protein expression in H/R-induced H9c2 cells. This study has clarified the protective effects of MAF on myocardial injury and also confirmed that oxidative stress and inflammation were involved in the myocardial ischemia-reperfusion injury (I/R) model.


Interest of adjusting urine cannabinoids to creatinine level to monitor cannabis cessation therapy in heavy smokers with psychiatric disorders.
Up to 25% of hospitalized patients in a psychiatric department exhibit troubles linked to cannabis use. Weaning patients with psychiatric disorders off drugs of abuse requires specific care to improve their clinical outcome. The present study aims to develop a predictive model of urinary excretion of creatinine-normalized cannabinoids (UCNC ) and to determine UCNC thresholds corresponding to the widely used cut-offs of 20 ng/mL and 50 ng/mL for cannabinoids. One hundred thirty-two patients with 452 urine samples were included between 2013 and 2017. Urinary cannabinoids and UCNC elimination curves were computed for each patient. Using a mono-exponential mixed effect model with 88 samples from 26 subjects exhibiting at least 3 decreasing UCNC in a row, the average calculated elimination rate constant was 0.0108 ± 0.0026 h-1 , corresponding to a mean elimination half-life of 64 ± 12 hours. The use of UCNC is of particular interest because of a high inter- and intra-individual variability of urinary creatinine concentration (from 0.06 to 3.81 mg/mL). Moreover, UCNC allows for the detection of diluted or concentrated urine specimens that may lead to false positive (FP) or false negative (FN) results. Receiver operator characteristic (ROC) curves were used to assess UCNC thresholds of 32.4 and 124.7 ng/mg that provide a strong discrimination between positive and negative samples for cannabinoids cut-offs of 20 and 50 ng/mL respectively. The developed model and the defined UCNC thresholds allowed for the accurate prediction of the time needed to reach a negative UCNC result that could be used by clinicians to optimize clinical care.


Stem Xylem Characterization for Vitis Drought Tolerance.
Together with stomatal conductance and root conductivity, the stem water reserve and transport systems could be regulatory mechanisms able to participate in the regulation of the plant water status. Lianas, such as Vitis spp., minimize the trunk support role, and stems have evolved to improve their ability in water transport. In this work, stems of 10 different Vitis species were studied in relation to their expected drought tolerance using reflectance spectroscopy. Spectra were measured before (T0) and after coloration with Sudan IV dye. The T0 spectral signature showed characteristic species features. The partial least squares (PLS) regression and the self-organizing map (SOM) neural network analysis were able to predict the expected drought tolerance score; thus, reflectance spectroscopy was demonstrated to be a useful technique for drought tolerance phenotyping. These methods could be applied for the preliminary selection of new rootstocks/cultivars. Wood composition variation appeared to be correlated with the water stress susceptibility. To clarify this relationship, the attention was focused on the wood hydrophobicity. Sudan IV is a microscopy dye traditionally used to underline suberin, waxes, and, in general, hydrophobic substances. Differences between rough and colored spectra evidenced the absorption band of Sudan IV with a maximum at 539 nm. The coloration intensity was used to develop a hydrophobicity index. The obtained values were correlated with the expected drought tolerance score. Therefore, hydrophobic compounds seem to play an important role in water use efficiency, and an hydrophobic barrier in the xylem tissue appears to be a protective mechanism against water stress.


Synthesis and tunable reactivity of N-heterocyclic germylene.
Modifying the β-diketimine ligand LH 1 (LH=[ArN=C(Me)-CH=C(Me)-NHAr], Ar=2,6-iPr(2)C(6)H(3)) through replacement of the proton in 3-position by a benzyl group (Bz) leads to the new (Bz)LH ligand 2, which could be isolated in 77% yield. According to (1) H NMR spectroscopy, 2 is a mixture of the bis(imino) form [(ArN=C(Me)](2)CH(Bz) 2a and its tautomer [ArN=C(Me)-C(Bz)=C(Me)NHAr] 2b. Nevertheless, lithiation of the mixture of 2a and 2b affords solely the N-lithiated β-diketiminate [ArN=C(Me)-C(Bz)=C(Me)-NLiAr], (Bz)LLi 3. The latter reacts readily with GeCl(2)· dioxane to form the chlorogermylene (Bz)LGeCl 4, which serves as a precursor for a new zwitterionic germylene by dehydrochlorination with LiN(SiMe(3))(2). This reaction leads to the zwitterionic germylene (Bz)L'Ge: 5 ((Bz)L'=ArNC(=CH(2))C(Bz)=C(Me)NAr) which could be isolated in 83% yield. The benzyl group has a distinct influence on the reactivity of zwitterionic 5 in comparison to its benzyl-free analogue, as shown by the reaction of 5 with phenylacetylene, which yields solely the 1,4-addition product 6, that is, the alkynyl germylene (Bz)LGeCCPh. Compounds 2-6 have been fully characterized by multinuclear NMR spectroscopy, mass spectrometry, elemental analyses, and single-crystal X-ray diffraction analyses.


Alpha-smooth muscle actin in pathological human disc nucleus pulposus cells in vivo and in vitro.
That a contractile actin isoform has been found in cells of other cartilage tissues in healing and disease states prompted this investigation of the presence of alpha-smooth muscle actin (alpha-SMA) in pathological human intervertebral disc tissue. The presence of this isoform has been reported in human intervertebral disc specimens obtained at autopsy from subjects for whom there were no reported symptoms. An objective of this study was to evaluate the cell density and percentage of alpha-SMA-containing cells in pathological nucleus pulposus tissue obtained from lumbar disc surgery from 17 patients. Additionally, explants of nucleus pulposus material were cultured to determine how alpha-SMA expression changed with time in vitro. Seventy-six 5-mm diameter explants (approximately 2 mm thick) pooled from six lumbar surgeries were cultured for 1, 2, 4, or 6 weeks. Microtomed sections of paraffin-embedded specimens were stained with hematoxylin and eosin or a monoclonal antibody to alpha-SMA. Histologically, cells were categorized as to alpha-SMA phenotype (positive or negative), and the areal cell density was determined. The evaluation of the cultured nucleus pulposus explants also included documentation of the percentage of cells that were round or elongated and the percentage of the cells that were part of a group (group: >/= 2 cells). Every nucleus pulposus section exhibited the presence of alpha-SMA-containing cells, which accounted for approximately 24 percent of the cells in vivo. In vivo, the cell density was significantly higher in older individuals (p = 0.02). The average time for cell outgrowth from the explants was 8.6 days. Approximately 10-15 percent of the cells in the explants stained positive for alpha-SMA. The time in culture had no significant effect on any of the outcome measures except the percentage of alpha-SMA-containing cells that were round (p = 0.008), with values decreasing through 4 weeks and then slightly rising at 6 weeks. The role of alpha-SMA in intervertebral disc pathology warrants further investigation.


The School Health Innovative Programs: integrating school health and managed care in San Diego.
Managed care organizations (MCOs) are being recruited to support school health services delivered in school clinics. Schools without clinics already provide numerous health services and could provide more if they had support from managed care organizations. This article describes the first two years of a San Diego-based collaborative consisting of MCOs, school districts, and other health care agencies. By establishing trust, developing overriding principles, and creating an interagency communication infrastructure, this collaborative has encouraged shared management of many student health issues. Because the agreements apply to all schools, programs can reduce high rates of absenteeism district-wide and avoid unnecessary doctor appointments for common health problems. These collaborative agreements are designed to be financially self-sustaining. However, data collection, the logistics of obtaining parental consent, and getting health professionals to communicate with each other in new ways remain to be significant challenges.


WOLBACHIA INFECTION IN DROSOPHILA SIMULANS: DOES THE FEMALE HOST BEAR A PHYSIOLOGICAL COST?
Fitness traits of three Drosophila simulans strains infected by endocellular bacteria belonging to the genus Wolbachia have been compared with those of replicate stocks previously cured from the infection by an antibiotic treatment. The traits measured were development time, egg-to-adult viability, egg hatch, productivity, fecundity, and the number of functional ovarioles. Individuals of the first strain were bi-infected by two Wolbachia variants, wHa and wNo. The second strain was infected by wHa, the third one by wNo. The Wolbachia studied here cause cytoplasmic incompatibility (CI), a high embryonic mortality (70% to > 90%) when an infected male is crossed with an uninfected female. Three generations after antibiotic treatment, we observed in all strains a significant drop in productivity in the cured stocks. This drop was not due to antibiotic toxicity and was associated with the loss of the Wolbachia. However the effect had disappeared in two of the three strains five generations after treatment, and could not be found in the third strain in a third measurement carried out 14 generations after treatment. The temporary nature of the productivity difference indicates that Wolbachia do not enhance productivity in infected strains. On the other hand, in all traits measured, our results show the absence of any negative effects of the Wolbachia on their host. This could be explained when considering Wolbachia evolution, as maternally transmitted parasites bear a strong selective pressure not to harm their female host. However, CI would allow the bacteria to be maintained even when harming the female. The apparent absence of deleterious effects caused by these Wolbachia might result from a trade-off, where a relatively low bacteria density would advantage the Wolbachia by suppressing any deleterious effects on the female host, at the cost of a weaker maternal transmission rate of the infection.


Fast start-up of a pilot-scale deammonification sequencing batch reactor from an activated sludge inoculum.
Deammonification involves the combined application of aerobic and anoxic ammonium-oxidizing bacteria (AerAOB & AnAOB) and allows to treat wastewaters with a high ammonium concentration in a sustainable and cost-efficient way. So far, it could take more than one year to start up the process, even with the addition of AnAOB enriched inocula. In contrast, we started up a deammonifying reactor for the treatment of sludge digestate in less than four months without any AnAOB enriched inoculum. In a single sequencing batch reactor (SBR) of 3 m(3), nitritation and anammox were performed without nitrite accumulation. Larger biomass aggregates (>1.0 mm) had a typical reddish colour, but FISH also showed that small aggregates (<0.25 mm) contained a considerable amount of AnAOB. The AerAOB were related to Nitrosomonas halophila, N. eutropha and N. halophila, and the AnAOB to "Candidatus Kuenenia & Brocadia", as shown by FISH. Our results show that the deammonification inoculum does not play an important role, and that the AnAOB can quickly develop under the proper aerational conditions. Nitrogen was removed stably at high nitrogen loading rates (740 mg N/L/d) and removal efficiency (90%).


Effects of combined parenchymal and vascular injury on platelet aggregation in pial arterioles of living mice: evidence for release of aggregate-inhibiting materials.
Platelet aggregation was produced in pial arterioles by exposing them to appropriately filtered light from a mercury lamp, following intravascular injection of sodium fluorescein. The dye acted as a target for the radiant energy and initiated a sequence of events resulting in the platelet aggregation. The aggregates adhered to the vessel in which they first appeared. When a microneedle punctured the brain adjacent to a subsequently irradiated arteriole, platelet aggregation was inhibited, even though the arteriole was not touched by the microneedle. Inhibition was manifested by prolongation of the exposure time required for the light-dye stimulus to initiate an adherent aggregate and by lengthening the time required for the aggregate to grow and totally block erythrocyte flow in the affected vessel. It is suggested that a material(s) diffuses from the zone of brain puncture, to and through the arteriolar wall, with a resultant inhibition of aggregation. It is noted that the inhibiting effect is reduced as the distance between puncture and wall increases.


Open Access Fees: A Barrier to Scholarly Activity Among Neurology Trainees.
The open access publishing model provides readers of all backgrounds access to articles free of charge. To cover the costs of open access, however, many journals now charge substantial article processing fees. This has inadvertently created yet another barrier for trainees to engage in scholarly activity. Herein, we describe the issue, review the literature, and provide suggestions for addressing this barrier with the focus on the neurology specialty.


Paenibacillus sp. TS12 glucosylceramidase: kinetic studies of a novel sub-family of family 3 glycosidases and identification of the catalytic residues.
GCase (glucosylceramidase) from Paenibacillus sp. TS12, a family 3 glycosidase, hydrolyses the beta-glycosidic linkage of glucosylceramide with retention of anomeric configuration via a two-step, double-displacement mechanism. Two carboxyl residues are essential for catalysis, one functioning as a nucleophile and the other as a general acid/base catalyst. p-nitrophenyl beta-D-glucopyranoside [K(m)=0.27+/-0.02 mM and kcat/K(m)=(2.1+/-0.2)x10(6) M(-1) x s(-1)] and 2,4-dinitrophenyl beta-D-glucopyranoside [K(m)=0.16+/-0.02 mM and k(cat)/K(m)=(2.9+/-0.4)x10(6) M(-1) x s(-1)] were used for continuous assay of the enzyme. The dependence of kcat (and kcat/K(m)) on pH revealed a dependence on a group of pK(a)< or =7.8 in the enzyme-substrate complex which must be protonated for catalysis. Incubation of GCase with 2,4-dinitrophenyl 2-deoxy-2-fluoro-beta-D-glucopyranoside caused time-dependent inactivation (K(i)=2.4+/-0.7 mM and k(i)=0.59+/-0.05 min(-1)) due to the accumulation of a trapped glycosyl-enzyme intermediate. Electrospray ionization MS analysis of the peptic digest of this complex showed that the enzyme was covalently labelled by the reagent at Asp-223, consistent with its role as nucleophile. A mutant modified at this residue (D223G) showed substantially reduced activity compared with the wild type (>10(4)), but this activity could be partially restored by addition of formate as an external nucleophile. Kinetic analysis of the mutant E411A indicated that Glu-411 serves as the general acid/base catalytic residue since this mutant was pH-independent and since considerable GCase activity was restored upon addition of azide to E411A, along with formation of a glycosyl azide product.


The impact of dehydroepiandrosterone in poor ovarian responders on assisted reproduction technology treatment.
One in six couples worldwide will experience at least one infertility problem during their reproductive years. Between 5.6% and 35.1% of women will exhibit poor ovarian response. A variety of methods have been applied to improve ovarian response, including dehydroepiandrosterone. In the ovaries, dehydroepiandrosterone promotes follicular development and granulosa cell proliferation by increasing intraovarian androgen concentrations while simultaneously enhancing the level of follicular insulin-like growth factor-1, which promotes folliculogenesis. Dehydroepiandrosterone supplementation may improve in vitro fertilization outcomes and ovarian response in patients with poor ovarian response. However, a few questions still loom over the effectiveness of dehydroepiandrosterone.


Contributions to the history of psychology: XXXIV. Enrique O. Aragon, a pioneer in Mexican psychology.
Enrique O. Aragón (1880-1942) was a Mexican physician responsible, in great measure, for the development of academic psychology in Mexico in the first decades of the century. He wrote the first book devoted solely to psychology in Mexico, and his founding of the first psychological Wundtian-style laboratory at the National University of Mexico was instrumental in the development of sufficient academic interest in the discipline to lead to the establishment of a university program in psychology as a profession.


Clearance of drugs for multiple myeloma therapy during in vitro high-cutoff hemodialysis.
Current chemotherapy for multiple myeloma is based on bortezomib (BOR), dexamethasone (DEX), and thalidomide (THA). The purpose of the present study was to examine their clearance during high-cutoff (HCO) hemodialysis and to accordingly apply the results to the dialytic removal of protein-bound substances in general. During in vitro hemodialysis with human blood (blood, dialysate, and ultrafiltration flow rates 250, 500 and 5 mL/min, respectively) comparing a highly permeable HCO dialyzer (Theralite, 2.1 m(2) ) to a high-flux dialyzer (PFX; 2.1 m(2) ), ultrafiltered volume was replaced by saline containing 30 g/L urea. After recirculation for equilibration, BOR was injected, and arterial and venous samples were drawn after 10, 11, and 12 min to measure the plasma clearance (K) of both urea and BOR. The same procedure was performed with THA and DEX. By mathematical simulation, the influence of varying plasma albumin concentrations (CHSA ) on the protein-bound drug fraction (PBF) and K was assessed. Plasma K values of HCO and PFX for THA, BOR, and DEX were about 40% (80 ± 7 vs. 65 ± 6 mL/min; P < 0.05), 70% (40 ± 8 vs. 33 ± 4 mL/min; P < 0.05), and 65% (47 ± 11 vs. 38 ± 7 mL/min; P < 0.05), respectively-lower (P < 0.0001) compared with urea (125 ± 7 vs. 122 ± 5 mL/min). K was highest (P < 0.0001) for THA. K was negatively correlated with CHSA (THA, r(2) = 0.58, P < 0.001; BOR, r(2) = 0.24, P < 0.05; DEX, r(2) = 0.22, P < 0.05). CHSA continually decreased (P < 0.05) over time only with HCO, resulting in lower calculated PBF. Compared with BOR and DEX (minimum 72 and 56%, respectively), the PBF of THA (37%) was significantly lower (P < 0.001). A mathematical simulation based on the K values of urea and the drugs reliably estimated PBF (r(2) = 0.886, P < 0.001). Drugs for multiple myeloma therapy are significantly removed with both HCO and PFX, with important implications for the dosing and timing of administration, particularly in patients with cast nephropathy receiving extended dialysis. If the Kurea of a dialyzer and the PBF of any given drug are known, Kdrug can be reliably estimated by mathematical simulation.


Antibody-Dependent Cellular Phagocytosis of HIV-1-Infected Cells Is Efficiently Triggered by IgA Targeting HIV-1 Envelope Subunit gp41.
Antibodies mediate a broad array of non-neutralizing Fc-mediated functions against HIV-1 including antibody-dependent cellular cytotoxicity (ADCC) and antibody-dependent cellular phagocytosis (ADCP). Accordingly, ADCC and ADCP induced by anti-HIV envelope gp120 IgG have been correlated to the limited success of the HIV-1 phase III vaccine trial RV144. It remains elusive whether ADCP can also be triggered by IgA, the isotype predominant at mucosal surfaces through which HIV-1 is mainly transmitted. Yet, we have previously shown that the HIV envelope subunit gp41-specific broadly neutralizing antibody 2F5 under the IgA isotype (2F5-IgA) triggers ADCC and cooperates with 2F5-IgG to increase HIV-1-infected cell lysis. Here, we now demonstrate that 2F5-IgA, more efficiently than 2F5-IgG, induces ADCP not only of gp41-coated beads but also of primary HIV-1-infected cells in a FcαRI-dependent manner. Both primary monocytes and neutrophils can act as effector cells of 2F5-IgA-mediated ADCP, although with different kinetics with faster neutrophil phagocytosis. However, unlike for ADCC, 2F5-IgA and 2F5-IgG do not cooperate to increase ADCP. Altogether, our results reveal that gp41-specific IgA mediate the efficient phagocytosis of HIV-1-infected cells. Inducing such ADCC and ADCP-prone IgA response by vaccination in addition to anti-HIV envelope IgG, might increase the protection against HIV acquisition at mucosal level.


An improved enzyme-linked immunosorbent assay for whole-cell determination of methanogens in samples from anaerobic reactors.
An enzyme-linked immunosorbent assay was developed for the detection of whole cells of methanogens in samples from anaerobic continuously stirred tank digesters treating slurries of solid waste. The assay was found to allow for quantitative analysis of the most important groups of methanogens in samples from anaerobic digesters in a reproducible manner. Polyclonal antisera against eight strains of methanogens were employed in the test. The specificities of the antisera were increased by adsorption with cross-reacting cells. The reproducibility of the assay depended on the use of high-quality microtiter plates and the addition of dilute hydrochloric acid to the samples. In an experiment on different digester samples, the test demonstrated a unique pattern of different methanogenic strains present in each sample. The limited preparatory work required for the assay and the simple assay design make the test well suited for routine analysis of large numbers of samples and thus for process surveillance during operation of biogas digesters.


Comparing the Duration of Spinal Anesthesia Induced With Bupivacaine and a Bupivacaince-Lidocaine Combination in Trans-Urethral Resection of the Prostate (TURP).
Spinal anesthesia is a safe anesthetic mode for transurethral prostate resection (TUPR). There are several studies assessing the effect of bupivacaine, lonely or accompanied by other drugs, on short duration operations. However, there is controversy regarding the exact combination. The aim of the study was to compare the effects of spinal anesthesia with bupivacaine and low dose lidocaine with bupivacaine alone on postoperative pain in those undergoing transurethral resection of prostate (TURP). This was a randomized clinical trial performed in Shiraz university of medical sciences during one year. Eighty men scheduled for TURP were randomly assigned to receive spinal anesthesia with 1.5 mL bupivacaine 0.6% and 0.6 mL Lidocaine 1% or spinal anesthesia with 1.5 mL bupivacaine 0.5% in combination with 0.6 mL normal saline. The primary endpoint was the time lag between induction of spinal anesthesia and reaching the highest spinal block level. We also recorded the duration of spinal block declining to L1 level, operation duration and the admission duration. Both study groups were comparable regarding the baseline characteristics. We did not find any difference between the two study groups regarding the duration of anesthetic block reaching the maximum level (P = 0.433) and duration of decreasing it to L1 (P = 0.189). The course of postoperative recovery and duration of hospital admission were also comparable between the groups (P = 0.661). Lidocaine does not have additive effects on duration and quality of spinal anesthesia with bupivacaine in those undergoing TURP.


Cyclin A degradation by primate cytomegalovirus protein pUL21a counters its innate restriction of virus replication.
Cyclin A is critical for cellular DNA synthesis and S phase progression of the cell cycle. Human cytomegalovirus (HCMV) can reduce cyclin A levels and block cellular DNA synthesis, and cyclin A overexpression can repress HCMV replication. This interaction has only been previously observed in HCMV as murine CMV does not downregulate cyclin A, and the responsible viral factor has not been identified. We previously reported that the HCMV protein pUL21a disrupted the anaphase-promoting complex (APC), but a point mutant abrogating this activity did not phenocopy a UL21a-deficient virus, suggesting that pUL21a has an additional function. Here we identified a conserved arginine-x-leucine (RxL) cyclin-binding domain within pUL21a, which allowed pUL21a to interact with cyclin A and target it for proteasome degradation. Homologous pUL21a proteins from both chimpanzee and rhesus CMVs also contained the RxL domain and similarly degraded cyclin A, indicating that this function is conserved in primate CMVs. The RxL point mutation disabled the virus' ability to block cellular DNA synthesis and resulted in a growth defect similar to pUL21a-deficient virus. Importantly, knockdown of cyclin A rescued growth of UL21a-deficient virus. Together, these data show that during evolution, the pUL21a family proteins of primate CMVs have acquired a cyclin-binding domain that targets cyclin A for degradation, thus neutralizing its restriction on virus replication. Finally, the combined proteasome-dependent degradation of pUL21a and its cellular targets suggests that pUL21a may act as a novel suicide protein, targeting its protein cargos for destruction.


Identification of a novel interaction between the Ca(2+)-binding protein S100A11 and the Ca(2+)- and phospholipid-binding protein annexin A6.
S100A11 is a member of the S100 family of EF-hand Ca(2+)-binding proteins, which is expressed in smooth muscle and other tissues. Ca(2+) binding to S100A11 induces a conformational change that exposes a hydrophobic surface for interaction with target proteins. Affinity chromatography with immobilized S100A11 was used to isolate a 70-kDa protein from smooth muscle that bound to S100A11 in a Ca(2+)-dependent manner and was identified by mass spectrometry as annexin A6. Direct Ca(2+)-dependent interaction between S100A11 and annexin A6 was confirmed by affinity chromatography of the purified bacterially expressed proteins, by gel overlay of annexin A6 with purified S100A11, by chemical cross-linking, and by coprecipitation of S100A11 with annexin A6 bound to liposomes. The expression of S100A11 and annexin A6 in the same cell type was verified by RT-PCR and immunocytochemistry of isolated vascular smooth muscle cells. The site of binding of S100A11 on annexin A6 was investigated by partial tryptic digestion and deletion mutagenesis. The unique NH(2) terminal head region of annexin A6 was not required for S100A11 binding, but binding sites were identified in both NH(2)- and COOH-terminal halves of the molecule. We hypothesize that an agonist-induced increase in cytosolic free [Ca(2+)] leads to formation of a complex of S100A11 and annexin A6, which forms a physical connection between the plasma membrane and the cytoskeleton, or plays a role in the formation of signaling complexes at the level of the sarcolemma.


Role of 18 F-FDG PET-CT in Urethral Malignant Melanoma and Analysis of the UK Guidelines on Ano-uro-genital Melanomas.
Urethral melanomas are a rare subtype of noncutaneous melanomas. The disease has a tendency to have skip lesions and early metastases as compared with cutaneous melanomas. The role of fluorine-18 fluorodeoxyglucose ( 18 F-FDG) positron emission tomography computed tomography (PET-CT) is well established in cases of cutaneous melanomas and is recommended by the National Comprehensive Cancer Network (NCCN) for stage IIB to IV cancer. There are no established guidelines on the management of noncutaneous melanomas; however, a recently published United Kingdom national guideline aims to streamline the management of ano-uro-genital melanomas. The guideline describes a very limited role in the use of 18 F-FDG PET-CT in this case scenario. The tendency to skip lesions, early metastases, involvement of brain parenchyma, and finally the usage of anti-PD-1 medications are key areas where 18 F-FDG PET-CT has shown superiority over CT scan. With this case report, we aim to highlight the strength of 18 F-FDG PET-CT in the management of urethral melanomas, which can be extrapolated to other ano-uro-genital melanomas.


A 17-Year Experience in Hand and Digit Replantation at an Academic Center.
of replantation surgery following upper extremity traumatic amputation are extensively described in the literature, with success rates varying from 57 to 100 percent. The purpose of this study was to evaluate replantation success rate at a Level I trauma center over a 17-year period and to assess definable factors contributing to these results. A retrospective review of all digit and hand replantations at a Level I trauma center was performed using CPT codes from 2001 through 2018. Descriptive analyses, Mann-Whitney test, Kruskal-Wallis test, and logistic regressions were used. Significance was defined as p ≤ 0.05. Analysis consisted of 76 patients with 101 amputated parts (93 digits and eight hands). Fifty-six single digit amputations (30 percent success rate), 37 multidigit injuries (22 percent digit success rate), and eight hand amputations (50 percent success rate) were attempted. The overall success rate was 25 of 76 patients (33 percent) and 29 of 101 parts (29 percent). The most common mechanism of injury was laceration (n = 56), followed by crush (n = 30), and avulsion (n = 11), with repair of laceration-type injuries having the greatest success rate (36 percent). The authors report a lower success rate of hand and digit replantation than previously described in the literature. Whole hand and thumb replantations resulted in the highest survival rate in our series. Laceration mechanism showed a higher success rate than crush or avulsion-type injuries. The authors' modest results highlight the importance of effective internal auditing of low-volume replantation centers such as their own. Quality improvement measures are proposed for higher future success in replantation surgery at the authors' institution. Therapeutic, IV.


Corrective lens use and refractive error among United States Air Force aircrew.
Corrective lens use by military aviators is an important consideration in the design of head-mounted equipment. The United States Air Force (USAF) has periodically monitored lens use by aviators; however, it has been over a decade since the last study. We provide an update on the prevalence of corrective lenses and refractive error among USAF aircrew based on eyeglass orders processed through the Spectacle Request Transmission System (SRTS). Currently, 41% of active duty USAF pilots and 54% of other aircrew require corrective lenses to perform flight duties. Refractive errors are characterized by low to moderate levels of myopia with a mean spherical equivalent power of -1.01 diopters (D) for pilots and -1.68 D for others. Contact lenses, and more recently refractive surgery, reduce the number of aircrew that must rely on spectacles when flying; however, spectacle compatibility remains an important consideration in the cockpit.


Midterm results of bovine jugular vein conduit for right ventricular outflow tract reconstruction.
To evaluate the midterm results of Contegra conduit. The retrospective study comprised patient record at Aga Khan University Hospital, Karachi, of conduits implanted between May 2007 and June 2012. Data collection was made from the clinical notes and from serial echocardiograms by a single cardiologist. The last followup echocardiography was done at the time of data collection in June 2012. SPSS 19 was used for statistical analysis. A total of 18 conduits had been implanted (16-22 mm) during the study period. Median age at the time of surgery was 9 years (range: 2.5-16 years). Early mortality was seen in 3 (16.66%) patients, but none was Contegra related. Of the remaining 15 patients, 2 (13.33%) with a diagnosis of Pulmonary Atresia-Ventricular Septal Defect with hypoplastic peripheral Pulmonary Arteries (PA), developed severe distal pressure gradient (50 mmHg) across Contegra over a median period of 18 months (range: 12-24 months), with resultant severe regurgitation and needed percutaneous intervention. There was no thrombosis, calcification, anuerysmal dilation or late deaths. At midterm followup, Contegra conduit was associated with low re-intervention rates with satisfactory haemodynamic results. However, long-term durability must be determined for this conduit, especially in patients with Pulmonary Atresia-Ventricular Septal Defect with hypoplastic peripheral Pulmonary Arteries.


Exploring the Role of Carbonate in the Formation of an Organomanganese Tetramer.
The formation of metal-oxygen clusters is an important chemical transformation in biology and catalysis. For example, the biosynthesis of the oxygen-evolving complex in the enzyme photosystem II is a complicated stepwise process that assembles a catalytically active cluster. Herein we describe the role that carbonato ligands have in the formation of the known tetrameric complex [Mn(CO)3(μ3-OH)]4 (1). Complex 1 is synthesized in one step via the treatment of Mn2(CO)10 with excess Me3NO·2H2O. Alternatively, when anhydrous Me3NO is used, an OH-free synthetic intermediate (2) with carbonato ligands is produced. Complex 2 produces carbon dioxide, Me3NO·2H2O, and 1 when treated with water. Labeling studies reveal that the μ3-OH ligands in 1 are derived from the water and possibly the carbonato ligands in 2.


The influence of gastro-jejunal anastomosis on gastric carcinogenesis in rats.
The effect of reflux of the duodenal contents on the development of gastric stump carcinoma induced in male rats was studied. Two gastro-jejunal anastomoses were made in the resected stomach of 28 rats and about half of the rats were also given a single dose of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). Well-differentiated adenocarcinomas developed in the resected stomachs with and without MNNG administration at incidences of 40% in the former and 23% in the latter. All the carcinomas were localized in the vicinity of the gastro-jejunal anastomosis, at which the proximal jejunal segment was drained. Several mucosal changes were found predominantly in the fundic mucosa surrounding the anastomosis, i.e., ulcer, foveolar hyperplasia, intestinal metaplasia and atypical hyperplasia. On the other hand, there was little mucosal change surrounding the gastro-jejunal anastomosis of the distal jejunal segment. These findings suggest a direct correlation between the exposure of mucosa of the anastomotic region to the duodenal contents and the development of adenocarcinoma.


In vivo release and turnover of secreted platelet antiheparin proteins in rhesus monkey (Macaca mulatta).
Human and rhesus monkey platelets secrete at least two antiheparin proteins: platelet factor 4 (PF4) and low affinity platelet factor 4 (LA-PF4). Neither of these proteins showed species-related antigenic differences. As determined by radioimmunoassay, the levels of PF4 and LA-PF4 antigen per 10(9) monkey platelets amounted to 10.7 and 20.3 microgram, respectively. One milliliter of monkey plasma prepared from blood collected into an anticoagulant composed of EDTA, prostaglandin E1, and theophylline solution contained 22.4 ng LA-PF4 and 8.0 ng PF4. Concentrations of these two platelet-specific proteins in monkeys closely resembled levels found in human platelets and plasma. Infusion of prostacyclin (PGI2) (100 or 300 ng/kg/min) into monkeys for 15 min resulted in a significant decrease of plasma levels of LA-PF4 antigen and of PF4 by 40%--60% (p < 0.0001). This decrease was related to the inhibitory effect of PGI2 on the secretion of platelets stimulated by a catheter or by venipuncture. Longer infusion of PGI2 did not produce further significant change. The supernate obtained after aggregation of human platelets stimulated by thrombin was injected into monkeys receiving PGI2 infusion. The disappearance of LA-PF4 antigen in monkey plasma followed a biphasic exponential curve with half-lives for the fast and slow components of 8.4 and 63 min. PF4 disappeared faster but followed the same pattern (half-lives for the fast and slow component of 2.1 and 70 min). Analysis of the experimental data suggests that the low levels of secreted platelet proteins in monkey plasma are related to their minimal in vivo release and to their rapid clearance.


Recognition of septanose carbohydrates by concanavalin A.
The ability of the jack bean lectin concanavalin A (ConA) to bind seven membered ring (septanose) monosaccharides has been investigated by isothermal titration calorimetry (ITC) and saturation transfer difference (STD) NMR spectroscopy.


CRISPR/Cas9-Mediated CCR5 Ablation in Human Hematopoietic Stem/Progenitor Cells Confers HIV-1 Resistance In Vivo.
Transplantation of hematopoietic stem cells (HSCs) with a naturally occurring CCR5 mutation confers a loss of detectable HIV-1 in the patient, making ablation of the CCR5 gene in HSCs an ideal therapy for an HIV-1 cure. Although CCR5 disruption has been attempted in CD4+ T cells and hematopoietic stem/progenitor cells (HSPCs), efficient gene editing with high specificity and long-term therapeutic potential remains a major challenge for clinical translation. Here, we established a CRISPR/Cas9 gene editing system in human CD34+ HSPCs and achieved efficient CCR5 ablation evaluated in long-term reconstituted NOD/Prkdcscid/IL-2Rγnull mice. The CCR5 disruption efficiency in our system remained robust in secondary transplanted repopulating hematopoietic cells. More importantly, an HIV-1 resistance effect was observed as indicated by significant reduction of virus titration and enrichment of human CD4+ T cells. Hence, we successfully established a CRISPR/Cas9 mediated CCR5 ablating system in long-term HSCs, which confers HIV-1 resistance in vivo. Our study provides evidence for translating CCR5 gene-edited HSC transplantation for an HIV cure to the clinic.


Residential buffer, perceived neighborhood, and individual activity space: New refinements in the definition of exposure areas - The RECORD Cohort Study.
Neighborhood effects on health have been widely investigated; yet the definition of neighborhoods is usually arbitrary. This study analyses how disparities in environmental exposure according to urbanicity vary when considering a home-centered network-buffer, the perceived residential neighborhood, or the activity space. Exposures to the density of destinations and proportion of green space were compared for three spatial definitions of exposure areas, overall and stratified by urbanicity of the residence. Environmental exposure levels and gradients by urbanicity were found to vary depending on the spatial definition of the exposure area.


Chemical constituents from Licania cruegeriana and their cardiovascular and antiplatelet effects.
Three new lupane-type triterpenoids: 6β,30-dihydroxybetulinic acid glucopyranosyl ester (4), 6β,30-dihydroxybetulinic acid (5) and 6β-hydroxybetulinic acid (6), were isolated from Licania cruegeriana Urb. along with six known compounds. Their structures were elucidated on the basis of spectroscopic methods, including IR, ESIMS, 1D- and 2D-NMR experiments, as well as by comparison of their spectral data with those of related compounds. All compounds were evaluated in vivo for their effects on the mean arterial blood pressure (MABP) and heart rate (HR) of spontaneously hypertensive rats (SHR) and also in vitro for their capacity to inhibit the human platelet aggregation. None of the isolated flavonoids 1-3 showed cardiovascular effects on SHR and among the isolated triterpenoids 4-9 only 5 and 6 produced a significant reduction in MABP (60.1% and 17.2%, respectively) and an elevation in HR (11.0% and 41.2%, respectively). Compounds 3, 4, 5 and 6 were able to inhibit human platelet aggregation induced by ADP, collagen and arachidonic acid with different selectivity profiles.


Effects of repeated once daily dosing of three intranasal corticosteroids on basal and dynamic measures of hypothalamic-pituitary-adrenal-axis activity.
Intranasal corticosteroids are regarded as the first-line treatment for allergic rhinitis, but few studies have directly compared their systemic effects. We sought to compare the hypothalamic-pituitary-adrenal (HPA)-axis suppression with three intranasal corticosteroids in terms of basal and dynamic adrenocortical activity. Sixteen healthy volunteers (mean age, 30.7 years) were studied in a single-blind, randomized, four-way crossover study comparing placebo with 200 microg/day fluticasone propionate (FP), 220 microg/day triamcinolone acetonide (TAA), and 336 microg/day beclomethasone dipropionate (BDP). After 4 days of treatment, an overnight urine collection was taken for cortisol and creatinine excretion starting at 10 PM (14 hours after the fourth dose), and blood was taken for serum cortisol at 8 AM (24 hours after the fourth dose) and after stimulation with adrenocorticotrophic hormone (ACTH) (0.5 microg). For overnight urinary cortisol excretion compared with placebo (20.8 nmol), there was a significant (p < 0.05) degree of suppression with FP (11.8 nmol) but not with TAA (16.0 nmol) or BDP (16.5 nmol). In terms of fold difference (95% CI for difference) from placebo, this amounted to 1.75-fold (1.01 to 3.03) for FP (43% suppression), 1.30-fold (0.75 to 2.25) for TAA (23% suppression), and 1.26-fold (0.73 to 2.18) for BDP (21% suppression). There was also a trend towards suppression of overnight urinary cortisol/creatinine excretion, but this was not statistically significant (placebo, 5.2 nmol/mmol; TAA, 5.0 nmol/mmol; BDP, 4.3 nmol/mmol; and FP, 4.3 nmol/mmol). Values for serum cortisol before and after ACTH stimulation showed no significant suppression. Suppression of overnight urinary cortisol occurred with intranasal FP (43%), TAA (23%), and BDP (21%), although this was only statistically significant with FP. None of the drugs were associated with blunting of the response to ACTH stimulation. Further studies are indicated to establish whether the systemic effects of inhaled and intranasal corticosteroids are additive.


Inverse association between serum creatinine and mortality in acute kidney injury.
Sepsis is a leading precipitant of Acute Kidney Injury (AKI) in intensive care unit (ICU) patients, and is associated with a high mortality rate. We aimed to evaluate the risk factors for dialysis and mortality in a cohort of AKI patients of predominantly septic etiology. Adult patients from an ICU for whom nephrology consultation was requested were included. End-stage chronic renal failure and kidney transplant patients were excluded. 114 patients were followed. Most had sepsis (84%), AKIN stage 3 (69%) and oliguria (62%) at first consultation. Dialysis was performed in 66% and overall mortality was 70%. Median serum creatinine in survivors and non-survivors was 3.95 mg/dl (2.63 - 5.28) and 2.75 mg/dl (1.81 - 3.69), respectively. In the multivariable models, oliguria and serum urea were positively associated with dialysis; otherwise, a lower serum creatinine at first consultation was independently associated with higher mortality. In a cohort of septic AKI, oliguria and serum urea were the main indications for dialysis. We also described an inverse association between serum creatinine and mortality. Potential explanations for this finding include: delay in diagnosis, fluid overload with hemodilution of serum creatinine or poor nutritional status. This finding may also help to explain the low discriminative power of general severity scores - that assign higher risks to higher creatinine levels - in septic AKI patients.


Procedure incidence and in-hospital complication rates of bariatric surgery in the United States.
Complication rates for bariatric surgery have been reported primarily from academic centers with specialized programs. The rates may not reflect those occurring in the community. The National Hospital Discharge Survey (NHDS) database maintained by the Center for Disease Control (CDC) was queried to determine the national incidence and complication rate for bariatric surgery as performed in the United States. The number of bariatric procedures rapidly increased from 6,868 in 1996 to 45,473 in 2001, with most of the increase attributable to a very large rise in the annual number of Roux-en-Y gastric bypasses performed. The in-hospital complication rate was 9.6% and 8.6% of patients has a length of stay exceeding 7 days. Cholecystectomies were performed concurrently in 28% of cases and were not associated with increases in complication rates or longer hospital stays. For those undergoing surgery, the most common preoperative comorbid conditions were hypertension (34%), arthritis (27%), GERD (22%), sleep apnea (22%), and diabetes (18%). The rate at which bariatric procedures are being performed is rapidly increasing, resulting in the need to establish practice standards. In-house complication rates derived from a cross section of US practices compare favorably with those reported from specialized centers. Based on these nationally representative data, the expected in-house clinically significant complication rate for bariatric operations is approximately 10%. As that is the average complication rate observed nationally, it serves as the benchmark to which bariatric surgery programs can compare themselves.


Performance Characteristics of the Whole-Body Discovery IQ PET/CT System.
The aim of this study was to assess the physical performance of a new PET/CT system, the Discovery IQ with 5-ring detector blocks. Methods: Performance was measured using the National Electrical Manufacturers Association NU2-2012 methodology. Image quality was extended by accounting for different acquisition parameters (lesion-to-background ratios [8:1, 4:1, and 2:1] and acquisition times) and reconstruction algorithms (VUE-point HD [VPHD], VPHD with point-spread-function modeling [VPHD-S], and Q.Clear). Tomographic reconstruction was also assessed using a Jaszczak phantom. Additionally, 30 patient lesions were analyzed to account for differences in lesion volume and SUV quantification between reconstruction algorithms. Results: Spatial resolution ranged from 4.2 mm at 1 cm to 8.5 mm at 20 cm. Sensitivity measured at the center and at 10 cm was 22.8 and 20.4 kps/kBq, respectively. The noise-equivalent counting rate peak was 124 kcps at 9.1 kBq/cm3 The scatter fraction was 36.2%. The accuracy of correction for count losses and randoms was 3.9%. In the image quality test, contrast recovery for VPHD, VPHD-S, and Q.Clear ranged from 18%, 18%, and 13%, respectively (hot contrast; 10-mm sphere diameter; ratio, 2:1), to 68%, 67%, and 81%, respectively (cold contrast; 37-mm sphere diameter; ratio, 8:1). Background variability ranged from 3.4%, 3.0%, and 2.1%, respectively (ratio, 2:1), to 5.5%, 4.8%, and 3.7%, respectively (ratio, 8:1). On Q.Clear reconstruction, the decrease in the penalty term (β) increased the contrast recovery coefficients and background variability. With the Jaszczak phantom, image quality increased overall when a reconstruction algorithm modeling the point-spread function was used, and use of Q.Clear increased the signal-to-noise ratio. Lesions analyzed using VPHD-S and Q.Clear had an SUVmean of 6.5 ± 3 and 7 ± 3, respectively (P < 0.01), and an SUVmax of 11 ± 4.8 and 12 ± 4, respectively (P < 0.01). No significant difference in mean lesion volume was found between algorithms. Conclusion: Among the various Discovery bismuth germanium oxide-based PET/CT scanners, the IQ with 5-ring detector blocks has the highest overall performance, with improved sensitivity and counting rate performance. Q.Clear reconstruction improves the PET image quality, with higher recovery coefficients and lower background variability.