Long, abundantly expressed non-coding transcripts are altered in cancer.
Recent studies with tiling arrays have revealed more genomic transcription than previously anticipated. Whole new groups of non-coding transcripts (NCTs) have been detected. Some of these NCTs, including miRNAs, can regulate gene expression. To date, most known NCTs studied have been relatively short, but several important regulatory NCTs, including XIST, MALAT-1, BC1 and BC200, are considerably larger in length and represent a novel class of long, non-coding RNA species. Whole-genome tiling arrays were utilized to identify novel long NCTs across the entire human genome. Our results have identified a new group of long (>400 nt), abundantly expressed NCTs and have found that a subset of these are also highly evolutionarily conserved. In this report, we have begun to characterize 15 long, conserved NCTs. Quantitative real-time RT-PCR was used to analyze their expression in different normal human tissue and also in breast and ovarian cancers. We found altered expression of many of these NCTs in both cancer types. In addition, several of these NCTs have consistent mutations when sequences of normal samples were compared with a panel of cancer-derived cell lines. One NCT was found to be consistently mutated in a panel of endometrial cancers compared with matched normal blood. These NCTs were among the most abundantly expressed transcripts detected. There are probably many long, conserved NCTs, albeit with lower levels of expression. Although the function of these NCTs is currently unknown, our study indicates that they may play an important function in both normal cells and in cancer development.


High fat diet induces formation of spontaneous liposarcoma in mouse adipose tissue with overexpression of interleukin 22.
Interleukin 22 (IL-22) is a T-cell secreted cytokine that modulates inflammatory response in nonhematopoietic tissues such as epithelium and liver. The function of IL-22 in adipose tissue is currently unknown. We generated a transgenic mouse model with overexpression of IL-22 specifically in adipose tissue. The IL-22 transgenic mice had no apparent changes in obesity and insulin resistance after feeding with high fat diet (HFD). Unexpectedly, all the IL-22 transgenic mice fed with HFD for four months developed spontaneous tumors in epididymal adipose tissue. Histological analysis indicated that the tumors were well-differentiated liposarcomas with infiltration of inflammatory cells. IL-22 overexpression promotes production of inflammatory cytokines such as IL-1β and IL-10 and stimulates ERK phosphorylation in adipose tissue. Furthermore, IL-22 treatment in differentiated 3T3-L1 adipocytes could induce IL-1β and IL-10 expression, together with stimulation of ERK phosphorylation. Taken together, our study not only established a novel mouse model with spontaneous liposarcoma, but also revealed that IL-22 overexpression may collaborate with diet-induced obesity to impact on tumor development in mouse.


Comparative levels of CALLA/neutral endopeptidase on normal granulocytes, leukemic cells, and transfected COS-1 cells.
We discovered that the common acute lymphoblastic leukemia antigen, CALLA (CD10), was identical to human neutral endopeptidase 184.108.40.206 (NEP), a Zn-binding glycoprotein with an extracellular active site capable of hydrolyzing several biologically active peptides. In this study we compare the expression of CALLA/NEP in terms of antigenic density and enzymatic activity at the cell surface and of messenger RNA (mRNA) levels on granulocytes, leukemic cells, and CALLA-transfected COS-1 cells. Mature granulocytes, the only readily available source of normal human CALLA, express relatively low but constant levels of antigen, NEP activity (3.5 pmol/min/10(6) cells), and mRNA. The two major CALLA-mRNA species of 6.5 kb and 3.8 kb, observed to date in a variety of cells and tissues, were also found in four independent granulocyte preparations. With leukemia cell lines, a correlation was established between the density of CALLA antigen and the level of enzymatic activity (3.4 to 21.0 pmol/min/10(6) cells). This paper constitutes the first report of NEP activity on blast cells derived from patients with non-T acute lymphoblastic leukemia (ALL); the levels of activity were variable (1.5 to 35.9 pmol/min/10(6) cells for six cases) but correlated with the level of CALLA assessed by flow cytometry. Heterogeneous levels of expression of the CALLA-mRNA species were also observed in non-T ALL cases that correlated with the level of CALLA expression at the surface of these cells. Very high levels of NEP activity were achieved by transfecting COS-1 cells with pSV-CALLA; 20% of the transfected cells were CALLA+ and expressed 550 pmol/min/10(6) cells. Extracts prepared from COS-1 cells transfected with pSV-CALLA (carrying human NEP cDNA) and pSVENK19 (carrying rabbit NEP-cDNA), respectively, gave Michaelis constant (Km) values of 50 mumol/L and similar inhibition curves with thiorphan. Thus the recombinant proteins encoded by these two genes have similar enzymatic properties, confirming the high degree of their structural relatedness. The expression of high levels of CALLA/NEP on COS-1 cells should allow the use of this system to test the effects of specific mutations on activity and might lead to the understanding of the role of CALLA in the onset and/or progression of leukemia.


Critical thinking of psychology students: A within- and cross-cultural study using Rasch models.
The primary aim of this study was to conduct a cross-cultural comparison of the level of critical thinking of Danish and Spanish psychology students (N = 788) attending innately different bachelor-level courses; personality psychology and introductory statistics/research methods. Several instruments are available for the assessment of critical thinking (CTh), but most are, however, unsuitable for use in large surveys with many constructs, due to length or single-person administration formats. One brief and much-used scale is the CTh scale in the Motivated Strategies for Learning Questionnaire (MSLQ), used in this study. To ensure unbiased comparisons across countries and courses, our secondary aim was to investigate the content validity, as well as the psychometric properties, of the CTh scale to ensure the most accurate and unbiased comparison. For this purpose, Rasch and graphical loglinear Rasch models were used. After reducing the number of items due to content validity issues, the CTh scale fitted Rasch models within each national sample, and was measurement invariant relative to age group, gender, course and university. In the cross-cultural item analysis, two items functioned differentially relative to nationality, and the scale fitted a graphical loglinear Rasch model. CTh scores related to introductory statistics/research methods courses differed significantly from scores related to personality psychology, for Danish students only.


Use of general practitioners versus mental health professionals in six European countries: the decisive role of the organization of mental health-care systems.
To investigate patterns of use of general practitioners (GPs) and/or mental health professionals (MHPs) for mental health reasons, across six European countries, and the extent to which these patterns of use are associated with differences in mental health-care delivering systems. Data are based on the European Study of the Epidemiology of Mental Disorders (ESEMeD): a cross-sectional survey of a representative sample of 8,796 non-institutionalized adults from six European countries, conducted between 2001 and 2003 using computer-assisted interviews with the CIDI-3.0. Countries were classified into: MHP- for countries where access to medical professionals tends to predominate (Belgium, France, Italy), and MHP+ where access to non-medical MHPs predominates (Germany, Spain, The Netherlands). Among respondents consulting GPs and/or MHPs in the past year (n = 1,019), respondents from the MHP- group more often consulted GPs (68 vs. 55 % in MHP+ group), GPs and psychiatrists (23 vs. 14 %). People from the MHP+ group more often used MHPs only (45 vs. 32 %), GPs and non-medical MHPs (16 vs. 8 %). GPs from the MHP+ group were more inclined to refer patients to MHPs. Factors associated with use of GPs versus MHPs were: being over 49 years, not highly educated, lower income and suffering from mood or severe mental disorders. Differences in the use of GPs versus MHPs are markedly linked to individual as well as organizational factors. Interventions are needed, in countries fostering medical access, to reimburse sessions with non-medical MHPs and improve cooperation between professionals to obtain better practice in access to care.


Malt1 deficient mice develop osteoporosis independent of osteoclast-intrinsic effects of Malt1 deficiency.
This study tested the hypothesis that mucosa associated lymphoid tissue 1 (Malt1) deficiency causes osteoporosis in mice by increasing osteoclastogenesis and osteoclast activity. A patient with combined immunodeficiency (CID) caused by MALT1 deficiency had low bone mineral density resulting in multiple low impact fractures that was corrected by hematopoietic stem cell transplant (HSCT). We have reported that Malt1 deficient Mϕs, another myeloid cell type, are hyper-responsive to inflammatory stimuli. Our objectives were to determine whether Malt1 deficient mice develop an osteoporosis-like phenotype and whether it was caused by Malt1 deficiency in osteoclasts. We found that Malt1 deficient mice had low bone volume by 12 weeks of age, which was primarily associated with reduced trabecular bone. Malt1 protein is expressed and active in osteoclasts and is induced by receptor activator of NF-κB ligand (RANKL) in preosteoclasts. Malt1 deficiency did not impact osteoclast differentiation or activity in vitro. However, Malt1 deficient (Malt1-/- ) mice had more osteoclasts in vivo and had lower levels of serum osteoprotegerin (OPG), an endogenous inhibitor of osteoclastogenesis. Inhibition of Malt1 activity in Mϕs induced MCSF production, required for osteoclastogenesis, and decreased OPG production in response to inflammatory stimuli. In vitro, MCSF increased and OPG inhibited osteoclastogenesis, but effects were not enhanced in Malt1 deficient osteoclasts. These data support the hypothesis that Malt1 deficient mice develop an osteoporotic phenotype with increased osteoclastogenesis in vivo, but suggest that this is caused by inflammation rather than an effect of Malt1 deficiency in osteoclasts.


Microwave-assisted one-pot synthesis of N-succinimidyl-4[ ¹⁸F]fluorobenzoate ([¹⁸F]SFB).
Biomolecules, including peptides¹⁻⁹, proteins¹⁰⁻¹¹, and antibodies and their engineered fragments¹²⁻¹⁴, are gaining importance as both potential therapeutics and molecular imaging agents. Notably, when labeled with positron-emitting radioisotopes (e.g., Cu-64, Ga-68, or F-18), they can be used as probes for targeted imaging of many physiological and pathological processes.¹⁵⁻¹⁸ Therefore, significant effort has devoted to the synthesis and exploration of ¹⁸F-labeled biomolecules. Although there are elegant examples of the direct ¹⁸F-labeling of peptides,¹⁹⁻²² the harsh reaction conditions (i.e., organic solvent, extreme pH, high temperature) associated with direct radiofluorination are usually incompatible with fragile protein samples. To date, therefore, the incorporation of radiolabeled prosthetic groups into biomolecules remains the method of choice.²³(,)²⁴ N-Succinimidyl-4-[¹⁸F]fluorobenzoate ([¹⁸F]SFB),²⁵⁻³⁷ a Bolton-Hunter type reagent that reacts with the primary amino groups of biomolecules, is a very versatile prosthetic group for the ¹⁸F-labeling of a wide spectrum of biological entities, in terms of its evident in vivo stability and high radiolabeling yield. After labeling with [¹⁸F]SFB, the resulting [F]fluorobenzoylated biomolecules could be explored as potential PET tracers for in vivo imaging studies.¹ Most [¹⁸F]SFB radiosyntheses described in the current literatures require two or even three reactors and multiple purifications by using either solid phase extraction (SPE) or high-performance liquid chromatography (HPLC). Such lengthy processes hamper its routine production and widespread applications in the radiolabeling of biomolecules. Although several module-assisted [¹⁸F]SFB syntheses have been reported²⁹⁻³²,⁴¹⁻⁴² they are mainly based on complicated and lengthy procedures using costly commercially-available radiochemistry boxes (Table 1). Therefore, further simplification of the radiosynthesis of [¹⁸F]SFB using a low-cost setup would be very beneficial for its adaption to an automated process. Herein, we report a concise preparation of [¹⁸F]SFB, based on a simplified one-pot microwave-assisted synthesis (Figure 1). Our approach does not require purification between steps or any aqueous reagents. In addition, microwave irradiation, which has been used in the syntheses of several PET tracers,³⁸⁻⁴¹ can gives higher RCYs and better selectivity than the corresponding thermal reactions or they provide similar yields in shorter reaction times.³⁸Most importantly, when labeling biomolecules, the time saved could be diverted to subsequent bioconjugation or PET imaging step. ²⁸(,)⁴³The novelty of our improved [¹⁸F]SFB synthesis is two-fold: (1) the anhydrous deprotection strategy requires no purification of intermediate(s) between each step and (2) the microwave-assisted radiochemical transformations enable the rapid, reliable production of [¹⁸F]SFB.


The Pathogenesis of Human Cervical Epithelium Cells Induced by Interacting with Trichomonas vaginalis.
Trichomonas vaginalis is a protozoan parasite that occurs in the urogenital-vaginal tract and is the primary causative agent of trichomoniasis, a common sexually transmitted disease in humans. The aggregation of this protozoan tends to destroy epithelial cells and induce pathogenesis. This study cultured T. vaginalis and human cervical epithelial cells (Z172) under the same conditions in the experiments. Following co-culturing for ten hours, the protozoans became attached to Z172, such that the cells presented a round shape and underwent shrinkage. Time-lapse recording and flow cytometry on interacted Z172 revealed that 70% had been disrupted, 18% presented a necrosis-like morphology and 8% showed signs of apoptosis. Gene expression profiling revealed in the seven inflammatory Z172 genes as well as in T. vaginalis genes that code for adhesion proteins 65 and 65-1. These results suggest that cytopathogenic effects progress while Z172 is in contact with T. vaginalis, and the resulting morphological changes can be categorized as disruption.


Imagination and remembrance: what role should historical epidemiology play in a world bewitched by mathematical modelling of COVID-19 and other epidemics?
Although every emerging infectious disease occurs in a unique context, the behaviour of previous pandemics offers an insight into the medium- and long-term outcomes of the current threat. Where an informative historical analogue exists, epidemiologists and policymakers should consider how the insights of the past can inform current forecasts and responses.


Estimating human drug oral absorption kinetics from Caco-2 permeability using an absorption-disposition model: model development and evaluation and derivation of analytical solutions for k(a) and F(a).
Intestinal transcellular permeability (P(m)), measured across cell lines such as Caco-2 cells in vitro, is often used for assessing oral drug absorption potential in humans. However, the quantitative link between in vitro permeability and apparent in vivo absorption kinetics, based on drug appearance in plasma, is poorly understood. In the current study, a novel absorption-disposition kinetic model that links traditional pharmacokinetic and mass transfer models was developed. Analytical solutions of k(a) and F(a) were deduced, and using Caco-2 permeability, F(a) in humans was predicted for 51 structurally diverse compounds. Predicted F(a) values were similar to and correlated highly with their corresponding experimental values with an average error of 1.88 +/- 1.06% (-17 to 22%) and r2 = 0.934. Simulated concentration profiles for 17 of 18 drugs corresponded to observed plasma concentration profiles in healthy volunteers. The equilibrium solution for k(a) (k(a,eq)) was found to be a key determinant of F(a), whereas under sink conditions, k(a) is likely to be a determinant of plasma concentration kinetics. The current version of the model offers a quantitative approach for predicting human oral absorption kinetics from in vitro permeability. It also establishes, for the first time, a quantitative link between P(m) and k(a) and between k(a,eq) and F(a). This will facilitate better in vitro or in situ-in vivo correlations since it establishes a basis for incorporating permeability coefficients from the various experimental formats based on drug loss or appearance that are commonly used in the laboratory for permeability determination.


In situ cryo-electron tomography and subtomogram averaging of intraflagellar transport trains.
In situ cryo-electron tomography (cryo-ET) and subtomogram averaging are powerful tools, able to provide 3D structures of biological samples at sub-nanometer resolution, while preserving information about cellular context and higher-order assembly. Best results are typically achieved, when applied to highly repetitive structures, such as viruses. Other typical examples are protein complexes that decorate long stretches along ciliary microtubules at stereotypical and precise repeats, such as axonemal dyneins. For such cases, a plethora of subtomogram averaging protocols exist. In this chapter, we show how we use cryo-ET and subtomogram averaging to study the architecture of the intraflagellar transport (IFT) machinery, a more challenging target that appears only in low copy numbers per tomogram. In the IFT trains, repeating units of IFT adaptor proteins engage two oppositely directed molecular motors to quickly shuttle ciliary building blocks and other proteins to the tip of the cilium and/or back to the base. This dynamic and sporadic nature of IFT trains poses challenges for determining the localization or precise orientation of the particles to be averaged. Solutions to these problems are described in this chapter.


Capacitative calcium entry and TRPC channel proteins are expressed in rat distal pulmonary arterial smooth muscle.
Mammalian homologs of transient receptor potential (TRP) genes in Drosophila encode TRPC proteins, which make up cation channels that play several putative roles, including Ca2+ entry triggered by depletion of Ca2+ stores in endoplasmic reticulum (ER). This capacitative calcium entry (CCE) is thought to replenish Ca2+ stores and contribute to signaling in many tissues, including smooth muscle cells from main pulmonary artery (PASMCs); however, the roles of CCE and TRPC proteins in PASMCs from distal pulmonary arteries, which are thought to be the major site of pulmonary vasoreactivity, remain uncertain. As an initial test of the possibility that TRPC channels contribute to CCE and Ca2+ signaling in distal PASMCs, we measured [Ca2+]i by fura-2 fluorescence in primary cultures of myocytes isolated from rat intrapulmonary arteries (>4th generation). In cells perfused with Ca2+-free media containing cyclopiazonic acid (10 microM) and nifedipine (5 microM) to deplete ER Ca2+ stores and block voltage-dependent Ca2+ channels, restoration of extracellular Ca2+ (2.5 mM) caused marked increases in [Ca2+]i whereas MnCl2 (200 microM) quenched fura-2 fluorescence, indicating CCE. SKF-96365, LaCl3, and NiCl2, blocked CCE at concentrations that did not alter Ca2+ responses to 60 mM KCl (IC50 6.3, 40.4, and 191 microM, respectively). RT-PCR and Western blotting performed on RNA and protein isolated from distal intrapulmonary arteries and PASMCs revealed mRNA and protein expression for TRPC1, -4, and -6, but not TRPC2, -3, -5, or -7. Our results suggest that CCE through TRPC-encoded Ca2+ channels could contribute to Ca2+ signaling in myocytes from distal intrapulmonary arteries.


Effects of Caffeine on Behavioural and Cognitive Deficits in Rats.
There are many studies that have sought to find drug therapies to prevent harm arising from sepsis. Such studies have represented a progress in the support to septic patients and also in the development of new pharmacological alternatives. Our interest was to investigate the caffeine effect on sepsis behavioural and memory impairments. Male rats were anaesthetized and the surgery was made to allow exposure of the caecum, which was then squeezed to extrude a small amount of faeces from the perforation site, which was later placed back into the peritoneal cavity. This procedure, which served to generate experimental sepsis, is herein referred to as ceccum ligation and perforation (CLP). The caffeine (10 mg/kg) was administered by gavage route, once daily, during 7 or 14 consecutive days to investigate the effects of acute or subchronic caffeine treatment on long-term behavioural and cognitive deficits induced by CLP. On the last day, 1 hr after caffeine administration, the animals were submitted to open-field, elevated plus maze (EPM), forced swimming and step-down inhibitory avoidance tests. The results showed that caffeine increased the percentage of open arm entries and open arm time in the EPM test, and reduced the immobility time when compared to the sham-operated group. The caffeine also increased the latency in the inhibitory avoidance test platform. Our results demonstrated that the caffeine improved behavioural changes and improved the neurocognitive deficits of sepsis-surviving animals. It is possible that blockage of the adenosine receptors may be responsible for the results here observed.


The effect of the androstane lung cancer inhibitor content on the cell-selective toxicity of hydroxyapatite-chitosan-PLGA nanocomposites.
An androstane (17β-hydroxy-17α-picolyl-androst-5-en-3β-yl-acetate (derivative A)) cancer inhibitor was successfully captured in a carrier made of nano-sized hydroxyapatite (HAp) coated with chitosan-PLGA polymer blends (Ch-PLGA). In our previous studies, we demonstrated that it was convenient to use spherical HAp/Ch-PLGA carriers as vehicles to target the lungs following intravenous administration. In this study, we used emulsification and subsequent freeze-drying to load the spherical HAp/Ch-PLGA carriers with varying contents of the derivative A, in order to examine the selective toxicity towards cancerous/healthy lung cells. The XRD and FT-IR techniques confirmed the drug loading process, and the content of the poorly water soluble derivative A was estimated directly via the DSC technique. The particles were spherical in shape with the d50 distribution varying between 167 and 231 nm, whereas the content of the derivative A ranged from 6.5 to 19.3 wt%. Cell-selective cytotoxicity was examined simultaneously on two cell lines: human lung carcinoma (A549 ATCC CCL 185) and human lung fibroblasts (MRC-5 ATCC CCL 171). All particles exhibited nearly three times larger cytotoxicity towards cancer cells (A549) than towards healthy cells (MRC5), where the particles with the derivative A content of 6.5 wt% allowed for the viability of healthy cells >80%. Ninety-six hours after the treatment of cells with particles with different contents of derivative A (after incubation and recovery), recovery was faster in damaged healthy cells than in cancerous cells.


Coronary computed tomography angiography.
Significant limitations exist for traditional noninvasive cardiac imaging with regard to equivocal or indeterminate findings that result in repetitive testing or unnecessary referral to invasive coronary angiography (ICA). Recent hardware and software advances in multislice computed tomography angiography have achieved high spatial and temporal resolution to allow accurate noninvasive assessment of coronary arteries. This poses a paradigm shift in management of patients with suspected coronary artery disease (CAD). Multicenter studies showed that coronary computed tomography angiography (CCTA) has a very high diagnostic accuracy, and, in particular, a very high negative predictive value (>98%) in detecting coronary stenosis when compared with ICA. In addition to its diagnostic ability, recent evidence-based outcome data have also validated the value of CCTA in predicting cardiac events. Absence of CAD on CCTA conveys excellent prognosis, whereas increasing disease severity and extent are associated with worsening outcome. Furthermore, CCTA allows comprehensive assessment of coronary stenosis, plaque burden, left ventricular morphology, function, perfusion and viability. One concern with CCTA is the issue of ionizing radiation exposure. Recent technical progress allows dramatic reduction of radiation dose. The newest generation scanner is capable of producing CCTA of diagnostic quality with a dose of less than 1 mSv. A multisociety guideline for appropriate clinical indications for cardiac computed tomography was recently published. When used appropriately, CCTA has been established as a valid noninvasive imaging alternative to ICA in selected patients at low to intermediate risk of CAD.


Supramolecular Nanopumps with Chiral Recognition for Moving Organic Pollutants from Water.
Since organic pollutants in water resources have raised concerns on aquatic ecosystems and human health, mechanical machines such as a nanopump for rapid and efficient removal of pollutants from water with regeneration properties remains a challenge. Here, a pH-responsive artificial pump from left-handed porous tubules into right-handed solid fibers was presented by the self-assembly of bent-shaped aromatic amphiphiles. The bent-shaped amphiphile with a pH-sensitive segment was demonstrated in aromatic hexameric macrocycles, which could contract into dimeric disks. Such a switchable aromatic pore with superhydrophobicity was well-suited for an efficient removal and controlled release of organic pollutants from water through pulsating motion. The removal efficiency is found to be 78% for ethinyloestradiol and 82% for bisphenol. Additionally, the pumping accompanied by chiral inversion was endowed with a rapid removal and convenient regenerable ability. The inflation from right-handed solid fibers into left-handed tubules for efficient removal pollutants was remarkably promoted by (-)-acidic enantiomer of malic acid, whereas the contraction with full desorption of pollutants was incisively responsive to alkaline with (+)-conformation. The kinetically regulable porous device with a chiral recognition will provide a promising platform for the construction of rapid responsible machine for sewage treatment.


Metachronous/concomitant B-cell neoplasms with discordant light-chain or heavy-chain isotype restrictions: evidence of distinct B-cell neoplasms rather than clonal evolutions.
Metachronous/concomitant B-cell neoplasms with distinct morphology are usually considered clonally related. We retrospectively analyzed 4 cases of metachronous/concomitant B-cell neoplasms with discordant light-chain/heavy-chain restrictions. The primary diagnoses included chronic lymphocytic leukemia (CLL; n = 2), lymphoplasmacytic lymphoma (n = 1), and pediatric follicular lymphoma (FL; n = 1). The respective secondary diagnoses included diffuse large B-cell lymphoma (DLBCL; n = 2), plasmablastic myeloma, and pediatric FL. The secondary B-cell neoplasm occurred after the primary diagnosis in 3 cases, with the median interval of 120 months (range, 21-216), whereas the remaining 1 case had the 2 neoplasms (CLL/DLBCL) diagnosed concurrently. Histology suggested aggressive transformation in 3 cases and recurrence in 1 case (FL). Nonetheless, 3 cases showed discordant light-chain restrictions between the 2 B-cell neoplasms, whereas in the remaining case (lymphoplasmacytic lymphoma/plasmablastic myeloma), the 2 neoplasms shared κ light-chain restriction but expressed different heavy-chain isotypes (IgM versus IgA). The 2 CLL/DLBCL cases had polymerase chain reaction-based IGH/K gene rearrangement study and amplicon sequence analysis performed, which demonstrated distinct clonal amplicons between the 2 B-cell neoplasms in each case. Concomitant/metachronous B-cell neoplasms may be clonally unrelated, which can be confirmed by immunoglobulin isotype analysis and/or genotypic studies. We advocate analysis of clonal identities in large cell transformation or recurrent disease compared with primary indolent B-cell neoplasm because of a potential difference in prognosis between clonally related and unrelated secondary B-cell neoplasms.


Crystallophore: a versatile lanthanide complex for protein crystallography combining nucleating effects, phasing properties, and luminescence.
Macromolecular crystallography suffers from two major issues: getting well-diffracting crystals and solving the phase problem inherent to large macromolecules. Here, we describe the first example of a lanthanide complex family named "crystallophore" (Xo4), which contributes to tackling both bottlenecks. This terbium complex, Tb-Xo4, is an appealing agent for biocrystallography, combining the exceptional phasing power of the Tb(iii) heavy atom with powerful nucleating properties, providing ready-to-use crystals for structure determination. Furthermore, protein/Tb-Xo4 co-crystals can be easily detected and discriminated from other crystalline by-products using luminescence. We demonstrate the potential of this additive for the crystallisation and structure determination of eight proteins, two of whose structures were unknown.


Value of repeated measures of nerve conduction and quantitative sensory testing in a diabetic neuropathy trial.
Conduct of a large, multicenter trial of the aldose reductase inhibitor zenarestat provided data on the reproducibility of multiple electrophysiologic (nerve conduction studies, NCS) and quantitative sensory (QST) tests. Baseline and 12-month electrophysiologic data from approximately 1100 patients at multiple centers were available for analysis. Intersite variability contributed minimally to overall test variance. All NCS tests were highly reproducible. Cool thermal and vibration QST thresholds, as measured by CASE IV instrumentation, were also highly reproducible. Intersubject variance accounted for the majority of variance for all parameters measured. Repeating NCS and QST measures decreased sample sizes needed to show statistical significance. Consideration of these observations, particularly with regard to QST, should aid in the design of future clinical trials investigating neuropathy.


Effect of Sildenafil on Pulmonary Circulation and Cardiovascular Function in Near-Term Fetal Sheep During Hypoxemia.
Sildenafil is a potential new treatment for placental insufficiency in human pregnancies as it reduces the breakdown of vasodilator nitric oxide. Pulmonary vasodilatation is observed in normoxemic fetuses following sildenafil administration. Placental insufficiency often leads to fetal hypoxemia that can cause pulmonary vasoconstriction and fetal cardiac dysfunction as evidenced by reduced isovolumic myocardial velocities. We tested the hypotheses that sildenafil, when given directly to the hypoxemic fetus, reverses reactive pulmonary vasoconstriction, increases left ventricular cardiac output by increasing pulmonary venous return, and ameliorates hypoxemic myocardial dysfunction. We used an instrumented sheep model. Fetuses were made hypoxemic over a mean (standard deviation) duration of 41.3 (9.5) minutes and then given intravenous sildenafil or saline infusion. Volume blood flow through ductus arteriosus was measured with an ultrasonic transit-time flow probe. Fetal left and right ventricular outputs and lung volume blood flow were calculated, and ventricular function was examined using echocardiography. Lung volume blood flow decreased and the ductus arteriosus volume blood flow increased with hypoxemia. There was a significant reduction in left ventricular and combined cardiac outputs during hypoxemia in both groups. Hypoxemia led to a reduction in myocardial isovolumic velocities, increased ductus venosus pulsatility, and reduced left ventricular myocardial deformation. Direct administration of sildenafil to hypoxemic fetus did not reverse the redistribution of cardiac output. Furthermore, fetal cardiac systolic and diastolic dysfunction was observed during hypoxemia, which was not improved by fetal sildenafil treatment. In conclusion, sildenafil did not improve pulmonary blood flow or cardiac function in hypoxemic sheep fetuses.


Do Acupuncture Services Reduce Subsequent Utilization of Opioids and Surgical Interventions Compared to Noninvasive Therapies among Patients with Pain Conditions?
To compare prescribed opioid use and invasive surgical interventions between patients using acupuncture and those using non-steroidal anti-inflammatory drugs (NSAIDs)/physical therapy (PT). Retrospective observational study of administrative claims. Large commercial insurance plan. 52 346 each treated with either acupuncture or NSAIDs/PT. Users of acupuncture and NSAIDs/PT were identified from January 1, 2014, to December 31, 2017. The first date of each service was defined as the index date. Acupuncture patients were 1:1 propensity score matched to the NSAIDs/PT group on baseline characteristics. Outcomes included opioid use, subsequent invasive surgical procedures, healthcare utilization such as hospitalizations or emergency department (ED) visits, and costs. These were assessed in the 12-month period before index date (baseline) and 12-month period following index date (follow-up) using difference-in-difference (DID) analysis. Results for opioid use were stratified by those with and without baseline opioid use. The acupuncture group had fewer patients initiating opioids post-index both among those with (49.2% vs 56.5%, P < .001) and without (15.9% vs 22.6%, P < .001) baseline opioid use. There was a small increase in invasive surgical procedures with acupuncture (3.1% vs 2.8%, P = .006). A reduction in ED visits was observed with acupuncture (DID -4.6% for all-cause; -3.3% for pain-related, all P < .001). Acupuncture was associated with higher total medical and pharmacy costs (DID +$1331 per patient, P = .006). Acupuncture showed a modest effect in reducing opioid use and ED visits. More research on acupuncture's place in emergency care, pain relief, and comparison to other types of non-opioid treatment is needed.


Filgrastim and infliximab biosimilar uptake in Medicare Advantage compared with Traditional Medicare, 2016-2019.
Medicare Advantage (MA) and Traditional Medicare face different financing structures and incentives and may implement different strategies to encourage biosimilar uptake. Strategies used by health insurers can influence biosimilar uptake, which can in turn promote savings to insurers and patients. To compare filgrastim and infliximab biosimilar uptake between MA and Traditional Medicare from 2016 to 2019 and examine biosimilar uptake by different MA carriers and plan types (Health Maintenance Organization [HMO] or Preferred Provider Organization). We use a 2016-2019 nationally representative random 20% sample of the carrier (physician) and outpatient paid claims for Traditional Medicare data and final-action carrier and outpatient records for MA data. We compare quarterly biosimilar uptake from 2016 to 2019 for the first 2 drugs with biosimilar competition: (1) filgrastim, (Neupogen, originator), and biosimilars tbo-filgrastim (GRANIX) and filgrastim-sndz (ZARXIO), and (2) infliximab (Remicade, originator), and biosimilars infliximab-dyyb (Inflectra) and infliximab-abda (Renflexis). From their introduction, there was consistently greater uptake of filgrastim and infliximab biosimilars in MA compared with Traditional Medicare. By Q4 2019, filgrastim biosimilar uptake was 7.6 percentage points higher in MA (80.3%) than Traditional Medicare (72.7%). By Q4 2019, infliximab biosimilar uptake was 28.7% and 15.4% in MA and Traditional Medicare, respectively. Kaiser HMO plans were primarily responsible for the higher uptake of biosimilars in MA; in Q4 2019, filgrastim and infliximab biosimilar uptake was 98.8% and 78.8%, respectively. Our findings suggest that filgrastim and infliximab biosimilar uptake is greater in MA compared with Traditional Medicare, which is driven in part by particularly high uptake of biosimilars in MA Kaiser HMO plans. This highlights the need for future work to examine specific strategies and levers employed by MA Kaiser HMO plans and other insurers to increase biosimilar uptake, which can lead to cost savings for physician-administered drugs.


Determining calmodulin binding to metabotropic glutamate receptors with distinct protein-interaction methods.
mGluRs (metabotropic glutamate receptors) are G-protein-coupled receptors that modulate synaptic transmission. The eight mammalian mGluRs form three groups based on sequence and functional similarities: group I (1 and 5), group II (2 and 3) and group III (4, 6-8) mGluRs. In the present study, we used a Y2H (yeast two hybrid) screen to identify proteins that interact with the C-terminal intracellular tail of mGluR3. Prominent among the candidate receptor interacting proteins was calmodulin, a Ca(2+) sensor known to bind identifiable sequences in group I and III mGluRs. The Y2H method was used to investigate calmodulin binding to mGluRs but failed to confirm the documented interaction with group III mGluRs. Furthermore, subsequent biochemical analysis showed that calmodulin does not interact with group II mGluRs. This illustrates that certain Ca(2+)-dependent interactions are not recapitulated in yeast. Moreover, it highlights the necessity for supporting biochemical data to substantiate interactions identified with Y2H methods.


Mindfulness on-the-go: Effects of a mindfulness meditation app on work stress and well-being.
We investigated whether a mindfulness meditation program delivered via a smartphone application could improve psychological well-being, reduce job strain, and reduce ambulatory blood pressure during the workday. Participants were 238 healthy employees from two large United Kingdom companies that were randomized to a mindfulness meditation practice app or a wait-list control condition. The app offered 45 prerecorded 10- to 20-min guided audio meditations. Participants were asked to complete one meditation per day. Psychosocial measures and blood pressure throughout one working day were measured at baseline and eight weeks later; a follow-up survey was also emailed to participants 16 weeks after the intervention start. Usage data showed that during the 8-week intervention period, participants randomized to the intervention completed an average of 17 meditation sessions (range 0-45 sessions). The intervention group reported significant improvement in well-being, distress, job strain, and perceptions of workplace social support compared to the control group. In addition, the intervention group had a marginally significant decrease in self-measured workday systolic blood pressure from pre- to post-intervention. Sustained positive effects in the intervention group were found for well-being and job strain at the 16-week follow-up assessment. This trial suggests that short guided mindfulness meditations delivered via smartphone and practiced multiple times per week can improve outcomes related to work stress and well-being, with potentially lasting effects. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Retroperitoneal dissection in gynecologic surgery for benign disease.
In this study, the pelvic retroperitoneal anatomy was dissected during 140 consecutive exploratory laparotomies for benign gynecologic disease. In addition, 110 bilateral hypogastric artery ligations were done both to teach the technique and to attempt to determine whether such a procedure reduces blood loss in elective abdominal hysterectomy. Fifty-eight hysterectomies with hypogastric artery ligation were compared to an equal number done without bilateral hypogastric ligation. The data indicate that prophylactic hypogastric artery ligation during hysterectomy did decrease blood loss, though other variables may account for this, and that teaching retroperitoneal dissection to residents is paramount, teaching them a potentially life-saving procedure essential to gynecologic and obstetric training. Only one complication occurred in this series.


Research in nutritional supplements and nutraceuticals for health, physical activity, and performance: moving forward 1.
This Horizons is part of a series that identifies key, forward-thinking research questions and challenges that need to be addressed. Specifically, this Horizons paper discusses research in nutritional supplements and nutraceuticals for health, physical activity, and performance, and is the product of a discussion by an expert panel that took place in January 2018 prior to the Canadian Nutrition Society Thematic Conference "Advances in Sport Nutrition from Daily Living to High Performance Sport". The objective of this Horizons paper was to identify core considerations for future studies for this research area, and how scientists can be leaders in the field to ensure the best quality science is available for decision makers. It is strongly recommended that the various elements highlighted throughout this Horizons paper will increase the awareness of the significant before-, during-, and after-research due-diligence required to produce research of the highest quality. While it is recognized that many scientists will not be able to meet all of these aspects, it is nonetheless important to consider the points outlined and to recognize that those elements that are not met in studies may be significant limitations. Highlights Research questions that are hypothesis-driven are the strongest, and when combined with careful planning of the study, the result will often be of the best quality. Studies with a strong experimental design help discern between evidence-based findings and those that have not been substantiated.


[Bacterial resistance against antibiotics].
Antibiotic resistance has progressed over many decades and is increasingly problematic. This paper gives a short summary of antibiotic resistance and its biology. The authors have worked in this field for many years. References to major overviews and important work are given, but no systematic literature search has been done. Development of resistance is driven by positive selection of resistant clones of bacteria. There are multiple, often interlinked molecular mechanisms behind this resistance, and they all lead to a less effective interaction between antibiotics and their target. Many observations of antibiotic resistance phenomena and their development over the last decades indicate that the problem is substantial, persisting and increasing. It will probably have an important impact on many medical disciplines in the future. Work to counteract this development is needed in every medical field in order to halt and hopefully counteract resistance development as strongly as we can.


Pediatric carve outs. The use of disease-specific conditions as risk adjusters in capitated payment systems.
To address the issue of adverse selection in capitated payment systems by developing a list of disease-specific pediatric conditions (i.e., "carve outs") to be considered for separate reimbursement. A descriptive study using a large Medicaid database. With the use of fiscal year 1993 Washington State Medicaid cost data for 302,240 pediatric patients, a list of disease-specific carve outs was developed to meet the following criteria: high cost, low variability in cost, and association with a large proportion of medical spending. Six-hundred seventy-three patients (0.2%) in the database had annual costs totaling $25,000 or more. Ten percent of the cases accounted for approximately two thirds of spending, while the least expensive 70% of cases made up only 15% of the expenditures. Prematurity and complications of prematurity, neoplasms, congenital heart disease, organ transplantations, congenital anomalies, and respiratory problems were general categories of disease that met our criteria for a carve out. The association of a major surgical procedure with a diagnosis increased the predictive accuracy for high cost. Lists of disease-specific carve outs such as this one can be used by Medicaid plans and other insurers who are concerned about risk selection to identify conditions for separate reimbursement in capitated payment systems.


Life expectancy of HIV-positive adults: a review.
We review studies estimating life expectancy (LE), the average number of additional years a person can expect to live, for HIV-positive adults and show variation by sociodemographic factors and level of immunosuppression. The LE of patients starting antiretroviral therapy early in the course of HIV infection and attaining restoration of a normal CD4 count may approach that of the general population. Improvements in treatment and the associated increase in LE imply that increasing numbers of HIV-positive patients will live to older age. Late diagnosis and presentation to care and treatment not concordant with guidelines contribute to reducing LE.


"My Family Was Also Happy": Couples' Qualitative Reports of a Combined Behavioral Economics and Cognitive Behavioral Therapy Intervention to Reduce Alcohol Use and Intimate Partner Violence.
Improving intimate partner violence interventions requires understanding pathways to change among couples participating in these interventions. This article presents qualitative data from 18 males and 16 females who participated in a combined behavioral economics (contingency management) and cognitive behavioral therapy alcohol and violence reduction intervention trial in Bengaluru, India. Results confirmed several theorized pathways of change, as well as identified further mechanisms through which the intervention supported the change. These included the emotional impacts of incentives, perceived and actual accountability via breathalyzers and family involvement, and enhanced support gained through counseling skills. Findings reveal critical insights into intervention design for future implementation.


An Herbivore-Induced Plant Volatile From Saltcedar (Tamarix spp.) Is Repellent to Diorhabda carinulata (Coleoptera: Chrysomelidae).
The leaf beetle Diorhabda carinulata Desbrochers (Coleoptera: Chrysomelidae) was introduced into the United States in 1999 for classical biological control of the exotic woody invader saltcedar (Tamarix spp. L. [Caryophyllales: Tamaricaceae]). The recent southern expansion of the range of D. carinulata in the United States has precipitated conflict between proponents of biological control of Tamarix and those with concerns over habitat conservation for avian species. Several semiochemicals that mediate aggregations by this species have been reported, but no repellent compounds have been recorded thus far. We now report a repellent compound, 4-oxo-(E)-2-hexenal, induced by adult D. carinulata feeding on saltcedar foliage. Collection of headspace volatiles, gas chromatography mass spectrometry, and electroantennographic analyses identified 4-oxo-(E)-2-hexenal as an insect-induced compound that is antennally active. Behavioral and exposure assays were conducted to test for repellency and toxicity in adults and larvae. Headspace volatiles were also collected from adult males exposed to 4-oxo-(E)-2-hexenal to determine the impact exposure might have on the emission of the aggregation pheromone. 4-Oxo-(E)-2-hexenal elicited electrophysiological responses in adults of both sexes. Behavioral responses indicated repellency across multiple doses for reproductive D. carinulata adults but not in nonreproductive adults. Exposure assays indicated altered behaviors in first instar larvae and adults, but not in third instar larvae. Collection of headspace volatiles indicated that exposure to 4-oxo-(E)-2-hexenal did not alter emission of the D. carinulata aggregation pheromone by adult males. The continued development and field deployment of this repellent compound may provide a new tool for the management of D. carinulata.


The SPUR adherence profiling tool: preliminary results of algorithm development.
The SPUR (Social, Psychological, Usage, and Rational) Adherence Profiling Tool is a recently developed adaptive instrument for measuring key patient-level risk factors for adherence problems. This study describes the SPUR questionnaire's psychometric refinement and evaluation. Data were collected through an online survey among individuals with type 2 diabetes in the United States. 501 participants completed multiple questionnaires, including SPUR and several validated adherence measures. A Partial Credit Model (PCM) analysis was performed to evaluate the structure of the SPUR tool and verify the assumption of a single underlying latent variable reflecting adherence. Partial least-squares discriminant analyses (PLS-DA) were conducted to identify which hierarchically-defined items within each dimension needed to be answered by a given patient. Lastly, correlations were calculated between the latent trait of SPUR adherence and other patient-reported adherence measures. Of the 45 candidate SPUR items, 39 proved to fit well to the PCM confirming that SPUR responses reflected one underlying latent trait hypothesized as non-adherence. Correlations between the latent trait of the SPUR tool and other adherence measures were positive, statistically significant, and ranged from 0.32 to 0.48 (p-values < .0001). The person-item map showed that the items reflected well the range of adherence behaviors from perfect adherence to high levels of non-adherence. The PLS-DA results confirmed the relevance of using four meta-items as filters to open or close subsequent items from their corresponding SPUR dimensions. The SPUR tool represents a promising new adaptive instrument for measuring adherence accurately and efficiently using the digital behavioral diagnostic tool.


Condom access in San Antonio, Texas: the "three A's" and HIV/STI incidence.
Condoms are highly effective at reducing STI transmission. Access is a critical precursor to use. This socio-ecologic study operationalizes the concept of condom access by defining it in terms of availability, affordability, and affect, and explores the relationship between these and STI incidence. Three pairs of zip codes, each with similar demographics but different Chlamydia and HIV incidence rates, were identified. Supermarkets, convenience stores, and pharmacies (N = 102) were evaluated for the three A's. Nonparametric methods were used to compare the moderate-Chlamydia areas to the high-incidence areas. High-Chlamydia areas were significantly less likely to have 12-packs available compared to moderate-Chlamydia areas. Prices averaged over $1 per condom and did not vary by incidence. High-incidence areas were significantly less likely to allow patrons to select condoms unassisted. High-HIV areas placed more positively-connoted items near condoms. Improving these factors may improve condom acquisition and in turn reduce STI and HIV incidence rates.


Divergent patterns of colonization and immune response elicited from two intestinal Lactobacillus strains that display similar properties in vitro.
Lactobacilli derived from the endogenous flora of normal donors are being increasingly used as probiotics in functional foods and as vaccine carriers. However, a variety of studies done with distinct strains of lactobacilli has suggested heterogeneous and strain-specific effects. To dissect this heterogeneity at the immunological level, we selected two strains of lactobacilli that displayed similar properties in vitro and studied their impact on mucosal and systemic B-cell responses in monoxenic mice. Germfree mice were colonized with Lactobacillus johnsonii (NCC 533) or Lactobacillus paracasei (NCC 2461). Bacterial loads were monitored for 30 days in intestinal tissues, and mucosal and systemic B-cell responses were measured. Although both Lactobacillus strains displayed similar growth, survival, and adherence properties in vitro, they colonized the intestinal lumen and translocated into mucosal lymphoid organs at different densities. L. johnsonii colonized the intestine very efficiently at high levels, whereas the number of L. paracasei decreased rapidly and it colonized at low levels. We determined whether this difference in colonization correlated with an induction of different types of immune responses. We observed that colonization with either strain induced similar germinal center formation and immunoglobulin A-bearing lymphocytes in the mucosa, suggesting that both strains were able to activate mucosal B-cell responses. However, clear differences in patterns of immunoglobulins were observed between the two strains in the mucosa and in the periphery. Therefore, despite similar in vitro probiotic properties, distinct Lactobacillus strains may colonize the gut differently and generate divergent immune responses.


Gonococcal urethral stricture and watering-can perineum.
A total of sixteen patients with urethral stricture and/or perineal urinary fistulae (water-can perineum) complicating gonorrhoea were seen at the Special Treatment Clinic, University College Hospital, Ibadan, Nigeria. The patients were aged between 25 and 80 years, and the latent period between the time of original attack of gonococcal infection and the development of complications varied from 4 to 50 years. The rate of divorce or marital separation is high among these patients with late sequelae of gonorrhoea. The factors responsible for the present higher incidence of early and late complications of gonorrhoea among patients in Nigeria and other tropical countries compared with their counterparts in Europe and North American include: (a) Lack of medical facilities in most rural areas; (b) Inadequate treatment of veneral diseases, including the urban areas where self-medication is practised on a large scale by the general population; (c) Illiteracy and ignorance of venereal diseases. The cases of watering-can perineum reported here, and the subsequent chronic pyelonephritis and hypertension, reinforce the plea for early and energetic treatment of acute gonorrhoea in Africa as well as large-scale control measures by the health authorities.


Mindset × Context: Schools, Classrooms, and the Unequal Translation of Expectations into Math Achievement.
When do adolescents' dreams of promising journeys through high school translate into academic success? This monograph reports the results of a collaborative effort among sociologists and psychologists to systematically examine the role of schools and classrooms in disrupting or facilitating the link between adolescents' expectations for success in math and their subsequent progress in the early high school math curriculum. Our primary focus was on gendered patterns of socioeconomic inequality in math and how they are tethered to the school's peer culture and to students' perceptions of gender stereotyping in the classroom. To do this, this monograph advances Mindset × Context Theory. This orients research on educational equity to the reciprocal influence between students' psychological motivations and their school-based opportunities to enact those motivations. Mindset × Context Theory predicts that a student's mindset will be more strongly linked to developmental outcomes among groups of students who are at risk for poor outcomes, but only in a school or classroom context where there is sufficient need and support for the mindset. Our application of this theory centers on expectations for success in high school math as a foundational belief for students' math progress early in high school. We examine how this mindset varies across interpersonal and cultural dynamics in schools and classrooms. Following this perspective, we ask: 1. Which gender and socioeconomic identity groups showed the weakest or strongest links between expectations for success in math and progress through the math curriculum? 2. How did the school's peer culture shape the links between student expectations for success in math and math progress across gender and socioeconomic identity groups? 3. How did perceptions of classroom gender stereotyping shape the links between student expectations for success in math and math progress across gender and socioeconomic identity groups? We used nationally representative data from about 10,000 U.S. public school 9th graders in the National Study of Learning Mindsets (NSLM) collected in 2015-2016-the most recent, national, longitudinal study of adolescents' mindsets in U.S. public schools. The sample was representative with respect to a large number of observable characteristics, such as gender, race, ethnicity, English Language Learners (ELLs), free or reduced price lunch, poverty, food stamps, neighborhood income and labor market participation, and school curricular opportunities. This allowed for generalization to the U.S. public school population and for the systematic investigation of school- and classroom-level contextual factors. The NSLM's complete sampling of students within schools also allowed for a comparison of students from different gender and socioeconomic groups with the same expectations in the same educational contexts. To analyze these data, we used the Bayesian Causal Forest (BCF) algorithm, a best-in-class machine-learning method for discovering complex, replicable interaction effects. Chapter IV examined the interplay of expectations, gender, and socioeconomic status (SES; operationalized with maternal educational attainment). Adolescents' expectations for success in math were meaningful predictors of their early math progress, even when controlling for other psychological factors, prior achievement in math, and racial and ethnic identities. Boys from low-SES families were the most vulnerable identity group. They were over three times more likely to not make adequate progress in math from 9th to 10th grade relative to girls from high-SES families. Boys from low-SES families also benefited the most from their expectations for success in math. Overall, these results were consistent with Mindset × Context Theory's predictions. Chapters V and VI examined the moderating role of school-level and classroom-level factors in the patterns reported in Chapter IV. Expectations were least predictive of math progress in the highest-achieving schools and schools with the most academically oriented peer norms, that is, schools with the most formal and informal resources. School resources appeared to compensate for lower levels of expectations. Conversely, expectations most strongly predicted math progress in the low/medium-achieving schools with less academically oriented peers, especially for boys from low-SES families. This chapter aligns with aspects of Mindset × Context Theory. A context that was not already optimally supporting student success was where outcomes for vulnerable students depended the most on student expectations. Finally, perceptions of classroom stereotyping mattered. Perceptions of gender stereotyping predicted less progress in math, but expectations for success in math more strongly predicted progress in classrooms with high perceived stereotyping. Gender stereotyping interactions emerged for all sociodemographic groups except for boys from high-SES families. The findings across these three analytical chapters demonstrate the value of integrating psychological and sociological perspectives to capture multiple levels of schooling. It also drew on the contextual variability afforded by representative sampling and explored the interplay of lab-tested psychological processes (expectations) with field-developed levers of policy intervention (school contexts). This monograph also leverages developmental and ecological insights to identify which groups of students might profit from different efforts to improve educational equity, such as interventions to increase expectations for success in math, or school programs that improve the school or classroom cultures.


Genotype and Long-term Clinical Course of Bietti Crystalline Dystrophy in Korean and Japanese Patients.
To investigate the genotype and long-term clinical phenotype of patients with Bietti crystalline dystrophy (BCD) in Korea and Japan. Retrospective case series. We analyzed 62 patients with clinical features of BCD who harbor pathogenic biallelic CYP4V2 variants in their homozygote or compound heterozygote. Data were collected from patient charts, including age, best-corrected visual acuity (BCVA), Goldmann perimetry results, fundus photography, OCT findings, fundus autofluorescence results, and electroretinography findings. We compared the clinical course of the patients with homozygous c.802-8_810de117insGC [exon7del], the most common mutation in the East Asian population, with those of the patients with other genotypes. Best-corrected visual acuity, visual field (VF), and their changes during follow-up. The mean age at the first visit was 55.2 years, with a mean follow-up of 7.1 years. The mean BCVAs at the first and last visits were 0.28 logarithm of the minimum angle of resolution (logMAR) and 0.89 logMAR, respectively. In genetic testing, c.802-8_810de117insGC was detected in 86 of 124 alleles of the patients, and 36 patients were homozygous for this mutation. The age, BCVA, VF area, central foveal thickness, and abnormal hypoautofluorescent area at either the first or last visit were not different between the exon7del homozygotes and the others. The mean BCVA changes per year were 0.089 logMAR in the exon7del homozygotes and 0.089 logMAR in the others. An age- and gender-adjusted linear regression analysis showed no association between the exon7del homozygote status and the rate of vision loss. Characteristic crystalline deposits in the posterior pole were generally observed in younger patients and disappeared over time along with progressive retinochoroidal atrophy. Patients with BCD and a homozygote for c.802-8_810de117insGC accounted for more than 50% of this cohort of Korean and Japanese patients, and the clinical effect of this deleterious variant was not severe in the spectrum of CYP4V2 retinopathy.


SCF(Fbxo22)-KDM4A targets methylated p53 for degradation and regulates senescence.
Recent evidence has revealed that senescence induction requires fine-tuned activation of p53, however, mechanisms underlying the regulation of p53 activity during senescence have not as yet been clearly established. We demonstrate here that SCF(Fbxo22)-KDM4A is a senescence-associated E3 ligase targeting methylated p53 for degradation. We find that Fbxo22 is highly expressed in senescent cells in a p53-dependent manner, and that SCF(Fbxo22) ubiquitylated p53 and formed a complex with a lysine demethylase, KDM4A. Ectopic expression of a catalytic mutant of KDM4A stabilizes p53 and enhances p53 interaction with PHF20 in the presence of Fbxo22. SCF(Fbxo22)-KDM4A is required for the induction of p16 and senescence-associated secretory phenotypes during the late phase of senescence. Fbxo22(-/-) mice are almost half the size of Fbxo22(+/-) mice owing to the accumulation of p53. These results indicate that SCF(Fbxo22)-KDM4A is an E3 ubiquitin ligase that targets methylated p53 and regulates key senescent processes.


Comparison of PRAS II, RapID ANA, and API 20A systems for identification of anaerobic bacteria.
This study evaluated the PRAS II, RapID ANA, and API 20A systems for the identification of anaerobic bacteria. A total of 80 isolates (68 fresh clinical isolates and 12 stock cultures) were examined and included 25 Bacteriodes spp., 7 Fusobacterium spp., 12 Clostridium spp., 2 Veillonella spp., 16 gram-positive cocci, and 18 gram-positive nonsporeforming bacilli. All isolates were initially identified by the procedures outlined in Holdeman et al. (ed.), Anaerobe Laboratory Manual, Virginia Polytechnic Institute and State University, Blacksburg, Va., 1977; identifications from the PRAS II, RapID ANA, and API 20A systems were compared with these initial identifications. If no supplemental tests were required, the RapID ANA and API 20A systems had incubation times of 4 and 24 h, respectively; the PRAS II system generally required 2 to 5 days of incubation, depending on the growth rate of the isolate. PRAS II identified 74% correct to species level, 14% correct to genus only, and 6% incorrect; 6% could not be identified. PRAS II data were reevaluated according to a revised data base that was provided after completion of the study; PRAS II (revised) identified 82% correct to species, 12% correct to genus only, and 6% incorrect. RapID ANA identified 62% correct to the species level, 28% correct to genus only, and 10% incorrect. API 20A identified 71% correct to the species level, 10% correct to genus only, and 3% incorrect; 16% could not identified. The API 20A is a more established system for identification of anaerobic bacteria; PRAS II and RapID ANA appear to be promising new methods for the identification of anaerobic bacteria.


Localization in the fibrinogen gamma-chain of a new site that is involved in the acceleration of the tissue-type plasminogen activator-catalysed activation of plasminogen.
In previous publications [e.g. Voskuilen, Vermond, Veeneman, Van Boom, Klasen, Zegers & Nieuwenhuizen (1987) J. Biol. Chem. 262, 5944-5946] we have shown that fibrin(ogen) chain fragment A alpha-(148-160) contains a site that contributes to the acceleration of Glu-plasminogen activation by tissue-type plasminogen activator (t-PA). In contrast with fibrin, this peptide, however, does not enhance the rate of mini-plasminogen activation. Therefore, possibly more stimulatory sites than A alpha-(148-160) are present in fibrin. In the present investigation we have localized a possible second type of stimulatory site in the fibrin(ogen) molecule. A whole CNBr digest of fibrinogen was applied to a Bio-Gel P-2 column run in water, pH 4. Two peaks with stimulatory activity were observed, one at the void volume and one between the void volume and the total volume. The former contained the previously described stimulating fragment FCB-2 [which comprises A alpha-(148-160)]; the latter had not been observed before and was characterized further. The stimulating material in the low-M(r) fraction of the Bio-Gel P-2 column was precipitated at pH 8.3 in a virtually pure form. It has a high tryptophan content, and an M(r) of 6500 as assessed by SDS/PAGE. On reduction, a main band of M(r) 2500 is seen, plus a weakly staining band of M(r) 4000. These properties plus the amino acid sequence data identify the fragment as FCB-5. FCB-5 consists of two chains, i.e. gamma-(311-336) and gamma-(337-379), linked by a single disulphide bond between Cys-gamma-326 and Cys-gamma-339. Both these chains and the disulphide bond appear to be essential for rate enhancement. FCB-5 enhances the activation rates of Glu-, mini- and micro-plasminogen, with all five kringles, only kringle V and without kringles respectively. FCB-5 binds t-PA, but none of the plasminogen forms binds to FCB-5. This indicates that the rate enhancements induced by FCB-5 are due to an effect on t-PA.


Differences in Screening and Treatment for Antepartum Versus Postpartum Patients: Are Providers Implementing the Guidelines of Care for Perinatal Depression?
National guidelines recommend universal depression screening for perinatal patients and provide treatment recommendations to prevent adverse maternal/child health outcomes. However, providers rarely screen all patients, and most women with perinatal depression remain undertreated. This study investigated predictors of universal screening and guideline-congruent care for perinatal depression by obstetrician-gynecologists and examined differences in practices with pregnant and postpartum patients. A random, national sample of obstetric providers (n = 483) from a three-wave mailing completed the survey. Regression analysis showed that providers universally screened (53.04% vs. 82.40%; p < 0.001) and implemented guideline-congruent care (33.61% vs. 58.51%; p < 0.001) with pregnant patients at a lower rate than with postpartum patients. Predictors of antenatal universal screening included the following: external influences, including screening as a clinic priority (adjusted odds ratio [AOR] 1.85; 95% confidence intervals [CIs]: 1.42-2.40), and intrinsic factors related to familiarity with mental health providers. Predictors of universal postpartum screenings included the following: clinic priority (AOR 3.01; 95% CIs: 2.12-4.28), provider comfort with diagnosing depression (AOR 1.58; 95% CIs: 1.04-2.39), and provider belief of patients unlikely to disclose depression unprompted (AOR 0.40; 95% CIs: 0.24-0.68). The only predictor of guideline-congruent care with pregnant patients was provider motivation (AOR 1.32; 95% CIs: 1.09-1.61). Predictors of postpartum guideline-congruent care included the following: comfort with prescribing antidepressants (AOR 1.98; 95% CIs: 1.53-2.57) and provider belief that depression will not resolve on its own (AOR 1.33; 95% CIs: 1.07-1.65). This study displays differences in perinatal depression care for pregnant and postpartum patients, particularly disparities for pregnant patients. The findings highlight facilitating factors for translating guidelines into routine practices.


Mechanical punctate pain threshold is associated with headache frequency and phase in patients with migraine.
Previous studies regarding the quantitative sensory testing are inconsistent in migraine. We hypothesized that the quantitative sensory testing results were influenced by headache frequency or migraine phase. This study recruited chronic and episodic migraine patients as well as healthy controls. Participants underwent quantitative sensory testing, including heat, cold, and mechanical punctate pain thresholds at the supraorbital area (V1 dermatome) and the forearm (T1 dermatome). Prospective headache diaries were used for headache frequency and migraine phase when quantitative sensory testing was performed. Twenty-eight chronic migraine, 64 episodic migraine and 32 healthy controls completed the study. Significant higher mechanical punctate pain thresholds were found in episodic migraine but not chronic migraine when compared with healthy controls. The mechanical punctate pain thresholds decreased as headache frequency increased then nadired. In episodic migraine, mechanical punctate pain thresholds were highest (p < 0.05) in those in the interictal phase and declined when approaching the ictal phase in both V1 and T1 dermatomes. Linear regression analyses showed that in those with episodic migraine, headache frequency and phase were independently associated with mechanical punctate pain thresholds and accounted for 29.7% and 38.9% of the variance in V1 (p = 0.003) and T1 (p < 0.001) respectively. Of note, unlike mechanical punctate pain thresholds, our study did not demonstrate similar findings for heat pain thresholds and cold pain thresholds in migraine. Our study provides new insights into the dynamic changes of quantitative sensory testing, especially mechanical punctate pain thresholds in patients with migraine. Mechanical punctate pain thresholds vary depending on headache frequency and migraine phase, providing an explanation for the inconsistency across studies.


Contextualizing and assessing the social capital of seniors in congregate housing residences: study design and methods.
This article discusses the study design and methods used to contextualize and assess the social capital of seniors living in congregate housing residences in Calgary, Alberta. The project is being funded as a pilot project under the Institute of Aging, Canadian Institutes for Health Research. Working with seniors living in 5 congregate housing residencies in Calgary, the project uses a mixed method approach to develop grounded measures of the social capital of seniors. The project integrates both qualitative and quantitative methods in a 3-phase research design: 1) qualitative, 2) quantitative, and 3) qualitative. Phase 1 uses gender-specific focus groups; phase 2 involves the administration of individual surveys that include a social network module; and phase 3 uses anamolous-case interviews. Not only does the study design allow us to develop grounded measures of social capital but it also permits us to test how well the three methods work separately, and how well they fit together to achieve project goals. This article describes the selection of the study population, the multiple methods used in the research and a brief discussion of our conceptualization and measurement of social capital.


Potential functions of histone H3.3 lysine 56 acetylation in mammals.
H3K56 acetylation (H3K56Ac) was first identified in yeast and has recently been reported to play important roles in maintaining genomic stability, chromatin assembly, DNA replication, cell cycle progression and DNA repair. Although H3.1K56Ac has been relatively well studied, the function of H3.3K56Ac remains mostly unknown in mammals. In this study, we used H3.3K56Q and H3.3K56R mutants to study the possible function of H3.3K56 acetylation. The K-to-Q substitution mimics a constitutively acetylated lysine, while the K-to-R replacement mimics a constitutively unmodified lysine. We report that cell lines harbouring mutation of H3.3K56R exhibit increased cell death and dramatic morphology changes. Using a Tet-Off inducible system, we found an increased population of polyploid/aneuploid cells and decreased cell viability in H3.3K56R mutant cells. Consistent with these results, the H3.3K56R mutant had compromised H3.3 incorporation into several pericentric and centric heterochromatin regions we tested. Moreover, mass spectrometry analysis coupled with label-free quantification revealed that biological processes regulated by the H3.3-associating proteins, whose interaction with H3.3 was markedly increased by H3.3K56Q mutation but decreased by H3.3K56R mutation, include sister chromatid cohesion, mitotic nuclear division, and mitotic nuclear envelope disassembly. These results suggest that H3.3K56 acetylation is crucial for chromosome segregation and cell division in mammals.


Increased Urine IgM and IgG(2) Levels, Indicating Decreased Glomerular Size Selectivity, Are Not Affected by Dalteparin Therapy in Patients with Type 2 Diabetes.
Fifty-four type 2 diabetic patients with neuroischemic foot ulcers were randomised to treatment with 5000 IU of dalteparin, (n = 28), or physiological saline, (n = 26), once daily until ulcer healing or for a maximum of 6 months. Thirty-three patients had normo-, 15 micro-, and 6 macroalbuminuria. The urinary levels of IgM and IgG(2) were elevated in 47 and 50 patients, respectively. Elevated urinary levels of IgM and IgG(2) indicate decreased glomerular size selectivity. Urine IgM levels were associated with IGF-1/IGFBP-1 and IGFBP-1 levels. Dalteparin treatment increased urinary levels of glycosaminoglycans (P < 0.001) and serum IGFBP-1 (P < 0.05) while no significant effects were seen in any of the other studied parameters. In conclusion, dalteparin therapy in patients with type 2 diabetes had no effects on urinary levels of albumin, IgM, or IgG(2) despite significantly increased glycosaminoglycans in urine. Elevated urinary levels of IgM and IgG(2) might be more sensitive markers of renal disease than albuminuria in patients with type 2 diabetes and antihypertensive therapy.


The effect of maxillary sinus antrostomy size on the sinus microbiome.
The optimal maxillary antrostomy size to surgically treat sinusitis is not well known. In this study, we examined clinical metrics of disease severity and symptom scores, measured secreted inflammatory markers, and characterized the sinus microbiome to determine if there were significant differences in outcome between different maxillary ostial sizes. Prospective randomized, single-blinded clinical trial enrolling 12 individuals diagnosed with recurrent acute or chronic rhinosinusitis. Each patient was blinded and randomized to receive minimal maxillary ostial dilation via balloon sinuplasty on 1 side vs a mega-antrostomy on the contralateral side. Data collected included symptom scores (20-item Sino-Nasal Outcome Test [SNOT-20]), endoscopy, and radiologic Lund-Mackay scores. During surgery and at their postoperative visit swabs were obtained from each maxillary sinus, and 16S DNA and inflammatory cytokine levels analyzed. The use of each patient as their own control allowed us to minimize confounding variables. There was statistically significant improvement in SNOT-20 symptom scores postoperatively in all patients. There were no significant differences between maxillary ostial size in postoperative endoscopy scores, cytokine profile, or bacterial burden. There were statistically significant differences in relative postoperative abundance of Staphylococcus, Lactococcus, and Cyanobacteria between the mega-antrostomy and mini-antrostomy. The method used in surgical maxillary antrostomies had no effect on endoscopy scores or cytokine profiles. Microbiome analysis determined significant differences between the different antrostomy sizes in postoperative Staphylococcus, Lactococcus, and Cyanobacteria abundance. The clinical significance of these changes in the sinus microbiome are not known but may be a result of increased access to postoperative sinonasal irrigations.


Do children profit from looking beyond looks? From similarity-based to cue abstraction processes in multiple-cue judgment.
The authors investigated the ability of 9- to 11-year-olds and of adults to use similarity-based and rule-based processes as a function of task characteristics in a task that can be considered either a categorization task or a multiple-cue judgment task, depending on the nature of the criterion (binary vs. continuous). Both children and adults relied on similarity-based processes in the categorization task. However, adults relied on cue abstraction in the multiple-cue judgment task, whereas the majority of children continued to rely on similarity-based processes. Reliance on cue abstraction resulted in better judgments for adults but not for children in the multiple-cue judgment task. This suggests that 9- to 11-year-olds may have defaulted to similarity-based processes because they were not able to employ a cue abstraction process efficiently.


Gene structures, classification and expression models of the AP2/EREBP transcription factor family in rice.
We identified 163 AP2/EREBP (APETALA2/ethylene-responsive element-binding protein) genes in rice. We analyzed gene structures, phylogenies, domain duplication, genome localizations and expression profiles. Conserved amino acid residues and phylogeny construction using the AP2/ERF conserved domain sequence suggest that in rice the OsAP2/EREBP gene family can be classified broadly into four subfamilies [AP2, RAV (related to ABI3/VP1), DREB (dehydration-responsive element-binding protein) and ERF (ethylene-responsive factor)]. The chromosomal localizations of the OsAP2/EREBP genes indicated 20 segmental duplication events involving 40 genes; 58 redundant OsAP2/EREBP genes were involved in tandem duplication events. There were fewer introns after segmental duplication. We investigated expression profiles of this gene family under biotic stresses [infection with rice viruses such as rice stripe virus (RSV), rice tungro spherical virus (RTSV) and rice dwarf virus (RDV, three virus strains S, O and D84)], and various abiotic stresses. Symptoms of virus infection were more severe in RSV infection than in RTSV and RDV infection. Responses to biotic stresses are novel findings and these stresses enhance the ability to identify the best candidate genes for further functional analysis. The genes of subgroup B-5 were not induced under abiotic treatments whereas they were activated by the three RDV strains. None of the genes of subgroups A-3 were differentially expressed by any of the biotic stresses. Our 44K and 22K microarray results suggest that 53 and 52 non-redundant genes in this family were up-regulated in response to biotic and abiotic stresses, respectively. We further examined the stress responsiveness of most genes by reverse transcription-PCR. The study results should be useful in selecting candidate genes from specific subgroups for functional analysis.


WNT11, a new gene associated with early onset osteoporosis, is required for osteoblastogenesis.
Monogenic early onset osteoporosis (EOOP) is a rare disease defined by low bone mineral density (BMD) that results in increased risk of fracture in children and young adults. Although several causative genes have been identified, some of the EOOP causation remains unresolved. Whole-exome sequencing revealed a de novo heterozygous loss-of-function mutation in Wnt family member 11 (WNT11) (NM_004626.2:c.677_678dup p.Leu227Glyfs*22) in a 4-year-old boy with low BMD and fractures. We identified two heterozygous WNT11 missense variants (NM_004626.2:c.217G > A p.Ala73Thr) and (NM_004626.2:c.865G > A p.Val289Met) in a 51-year-old woman and in a 61-year-old woman, respectively, both with bone fragility. U2OS cells with heterozygous WNT11 mutation (NM_004626.2:c.690_721delfs*40) generated by CRISPR-Cas9 showed reduced cell proliferation (30%) and osteoblast differentiation (80%) as compared with wild-type U2OS cells. The expression of genes in the Wnt canonical and non-canonical pathways was inhibited in these mutant cells, but recombinant WNT11 treatment rescued the expression of Wnt pathway target genes. Furthermore, the expression of RSPO2, a WNT11 target involved in bone cell differentiation, and its receptor leucine-rich repeat containing G protein-coupled receptor 5 (LGR5), was decreased in WNT11 mutant cells. Treatment with WNT5A and WNT11 recombinant proteins reversed LGR5 expression, but Wnt family member 3A (WNT3A) recombinant protein treatment had no effect on LGR5 expression in mutant cells. Moreover, treatment with recombinant RSPO2 but not WNT11 or WNT3A activated the canonical pathway in mutant cells. In conclusion, we have identified WNT11 as a new gene responsible for EOOP, with loss-of-function variant inhibiting bone formation via Wnt canonical and non-canonical pathways. WNT11 may activate Wnt signaling by inducing the RSPO2-LGR5 complex via the non-canonical Wnt pathway.


[A model of experimental acute pancreatitis in the conscious rat].
We describe an experimental model of acute pancreatitis in conscious rats, based on biliary reflux; sodium deoxycholate was injected while maintaining the common duct (biliary and pancreatic) temporally occluded. A progressive increase of serum amylase, cardiac rate and hematocrit value, as well as a decrease of plasma proteins are characteristic of this experimental model. Blood pressure was maintained until shortly before death, which occurred after 85 minutes. This is a simple model, reliable and reproducible, which requires minimal surgical manipulation.


[Negative pressure system in the treatment of enterocutaneous fistulas in the pediatric population].
The management of children with enterocutaneous fistulas (EF) along with large abdominal wall defects secondary to multiple surgical interventions can be difficult and sometimes lead to intestinal failure (IF). The aim of this study is to present the results of negative pressure systems and their properties (edema reduction angiogenesis promotion and granulation tissue formation) in children with enterocutaneous fistulas (EF) and their prognosis. A retrospective analysis of children with refractory, high output EF treated with NPS between 2008-2014. Outcome variables were duration and effectiveness of treatment as well as complications associated with NPS. Eight patients met inclusion criteria and were treated with NPS during a median of 25 days (range 5-50). The aetiologies were volvulus (2), necrotizing enterocolitis (2), gastroschisis (2), Blue Rubber Bleb Nevus (1) and duodenopancreatic anastomosis fistula following hepatopancreatic transplantation (1). Most patients (n=7) had large abdominal wall defects that closed during treatment, though two patients required further laparotomies due to evisceration. Two patients developed a second EF that was also successfully treated with NPS. No complications were identified arising from the use of NPS. After a 5-yr follow up 3 patients had a multivisceral transplantation and survive, and 4 died due to encephalopathy (1), hemolytic anemia (1), catheter-related sepsis (1) and one while waiting for a multivisceral graft for transplantation. Despite of a limited series of patients we recommend NPS as a useful tool in the management of EF and/ or abdominal wall defects.


Trans-anethole prevents hypertension induced by chronic exposure to both restraint stress and nicotine in rats.
Chronic stress and smoking are major risk factors for hypertension, with stress also being a factor predisposing to smoking. Methods are needed to prevent and/or reduce hypertension induced by chronic exposure to both stress and nicotine. This study investigated whether trans-anethole would prevent hypertension induced by chronic exposure to both restraint stress and nicotine in rats. Rats received nicotine intraperitoneally for 21 days following restraint stress (2 h/day) and trans-anethole (62, 125, and 250 mg/kg) on days 4, 8, 12, 16 and 20. To confirm the preventive effects of trans-anethole, blood pressure and vascular tone were measured on the last day of the experiment, and compared with the results of nifedipine and aerobic exercise. The ability of trans-anethole, at doses of 125 mg/kg and 250 mg/kg, to prevent hypertension was comparable to that of aerobic exercise and nifedipine. Furthermore, nifedipine combined with aerobic exercise and trans-anethole reduced both blood pressure and vascular tone. These findings are the first to show that trans-anethole can prevent hypertension, suggesting that trans-anethole may be useful as a prophylactic antihypertensive agent.


Reduced surface expression of epithelial E-cadherin evoked by interferon-gamma is Fyn kinase-dependent.
Interferon gamma (IFNγ) is an important regulatory cytokine that can exert a pro-inflammatory effect in the gut, where it has been shown to increase epithelial permeability via disruption of the tight junctions. Here we investigated the potential for IFNγ to regulate the adherens junction protein E-cadherin, an important mediator of normal epithelial tissue function, using the model T84 human colonic epithelial cell line. IFNγ (10 ng/ml) stimulated increased internalization of E-cadherin as assessed by immunofluorescence microscopy; internalization was reversed when cells were treated with PP1 (125 nM), a Src kinase-selective inhibitor. Immunoprecipitation studies demonstrated loss of E-cadherin from membrane fractions following IFNγ treatment and a corresponding increase in cytosolic E-cadherin and its binding partners, p120-catenin and beta-catenin: effects that were Src-kinase dependent. E-cadherin and p120-catenin phosphorylation was increased by IFNγ treatment and siRNA studies showed this was dependent upon the Src-kinase isoform Fyn. E-cadherin ubiquitinylation and subsequent proteasomal degradation stimulated by IFNγ was found to be dependent upon Fyn and the E-cadherin-selective ubiquitin ligase, Hakai. Use of Fyn and Hakai siRNA inhibited the internalization of E-cadherin as shown by immunoblotting and confocal fluorescence microscopy. Finally, IFNγ treatment resulted in a more fragile T84 cell monolayer with increased cell detachment in response to physical stress, which was prevented by PP1 and siRNA targeting Fyn or Hakai. Collectively, these results demonstrate a Fyn kinase-dependent mechanism through which IFNγ regulates E-cadherin stability and suggest a novel mechanism of disruption of epithelial cell contact, which could contribute to perturbed epithelial barrier function.


The motion of model cells at capillary bifurcations.
The motion of a rigid particle suspended in a two-dimensional flow through a bifurcating channel is calculated numerically. The calculated particle trajectories depend on the partitioning of total volume at the bifurcation, the size of the particle relative to channel dimensions, and the upstream lateral position of the particle. Results are used to deduce the flux of particles in a dilute suspension entering each branch of a bifurcation as a function of the total volumetric flow entering each branch. The calculated results exhibit several qualitative features observed in experimental data for red cell flux at capillary bifurcations.


p15 promoter methylation - a novel prognostic marker in glioblastoma patients.
Glioblastomas are the most frequent and malignant brain tumors in adults. Surgical cure is virtually impossible and despite radiation and chemotherapy the clinical course is very poor. Epigenetic silencing of MGMT has been associated with a better response to temozolomide-chemotherapy. We previously showed that temozolomide increases the median survival time of patients with tumors harbouring deletions on 9p within the region for p15(INK4b), p16(INK4a), and 10q (MGMT). The aim of this study was to investigate the methylation status of p15, p16, p14ARF and MGMT in glioblastomas (n=27) and to correlate the results with the clinical data. Only patients with KPS >70, radical tumor resection, radiation and temozolomide-chemotherapy after recurrence were included. We observed promoter methylation of MGMT in 56% and of p15 in 37% of the tumors, whereas methylation of p16 and p14ARF were rare. Interestingly, methylation of p15 emerged as a significant predictor of shorter overall survival (16.9 vs. 23.8 months, p=0.025), whereas MGMT promoter methylation had no significant effect on median overall survival under this treatment regimen (22.5 vs. 22.1 months, p=0.49). In the presence of other clinically relevant factors, p15 methylation remains the only significant predictor (p=0.021). Although these results need to be confirmed in larger series as well as under different treatment conditions, our retrospective study shows clear evidence that p15 methylation is an important prognostic factor for survival and underlines that this tumor suppressor, involved in cell cycle control, is an attractive candidate for therapeutic approaches in glioblastomas.


Assessing oxidative stress in Steller sea lions (Eumetopias jubatus): Associations with mercury and selenium concentrations.
Selenium (Se) bioavailability is required for synthesis and function of essential Se-dependent antioxidants, including the enzyme glutathione peroxidase (GPx). Strong interactions between monomethyl mercury and Se impair the critical antioxidant role of Se. Approximately 20% of Steller sea lion (Eumetopias jubatus, SSL) pups sampled in the western Aleutian Islands, Alaska, had total Hg concentrations ([THg]) measured in hair and whole blood above thresholds of concern for adverse physiologic effects in pinnipeds. Importantly, low molar ratios of TSe:THg, in some cases < 1 in several tissues (hair, liver, pelt, muscle, spleen, intestine, heart, lungs, brain) were documented for one SSL pup with [THg] above threshold of concern, which may lead to antioxidant deficiency. Our aim with this study was to evaluate the relationship between circulating [THg], [MeHg+], [TSe] and TSe:THg molar ratio status relative to oxidative stress and antioxidants measured during general anesthesia in free-ranging SSL. We captured, anesthetized and sampled newborn SSL pups at rookeries located in the Aleutian Islands or Gulf of Alaska. Biomarkers analyzed for oxidative stress included 4-hydroxynenonal and thiobarbituric acid reactive substances (4-HNE and TBARS, respectively, lipid peroxidation), protein carbonyl content (PCC, protein oxidation), and GPx activity as a key indicator for Se-dependent antioxidant defense levels. We found a negative association between TBARS and [TSe], and SSL with low [TSe] had higher concentrations of 4-HNE than those with intermediate [TSe]. These results suggest that SSL with lower [TSe] experience increased lipid peroxidation potentially associated with [THg] status.


In vitro investigating of anticancer activity of new 7-MEOTA-tacrine heterodimers.
A combination of biochemical, biophysical and biological techniques was used to study calf thymus DNA interaction with newly synthesized 7-MEOTA-tacrine thiourea 12-17 and urea heterodimers 18-22, and to measure interference with type I and II topoisomerases. Their biological profile was also inspected in vitro on the HL-60 cell line using different flow cytometric techniques (cell cycle distribution, detection of mitochondrial membrane potential dissipation, and analysis of metabolic activity/viability). The compounds exhibited a profound inhibitory effect on topoisomerase activity (e.g. compound 22 inhibited type I topoisomerase at 1 µM concentration). The treatment of HL-60 cells with the studied compounds showed inhibition of cell growth especially with hybrids containing thiourea (14-17) and urea moieties (21 and 22). Moreover, treatment of human dermal fibroblasts with the studied compounds did not indicate significant cytotoxicity. The observed results suggest beneficial selectivity of the heterodimers as potential drugs to target cancer cells.


Effect of age, Helicobacter pylori infection, and gastritis with atrophy on serum gastrin and gastric acid secretion in healthy men.
Gastric acid secretion has been considered to decline with increasing age but this view is being re-evaluated as the importance of Helicobacter pylori infection emerges. This study aimed to determine the effect of age, H pylori, and gastritis with atrophy on the serum gastrin concentration, gastric secretory volumes, and acid output in healthy, asymptomatic men. Young men (mean (SD) age 22.9 (0.6) years; n = 22) were compared with old men (72.9 (1.2) years; n = 28) in respect of basal serum gastrin and basal, sham fed, pentagastrin stimulated maximal and peak acid secretion. Antral, corpus, and fundal biopsy specimens were taken for histology and H pylori status (histology, culture, and rapid urease test). H pylori associated gastritis was present in three of 22 young (13.6%) and 16 of 28 old (57.1%) men. Gastritis with atrophy was present in 11 old subjects, 10 of whom were H pylori positive. These subjects had higher mean (SD) serum gastrin concentrations than old subjects without atrophy and young subjects (61.8 (9.2); 40.0 (2.9); 36.8 (2.3) pmol/l respectively; p < 0.001). H pylori infected subjects had higher gastrin values than uninfected subjects, overall (55.3 (5.9); 36.0 (1.8) pmol/l; p < 0.001) and in subjects without atrophy (45.3 (4.2); 36.0 (1.8) pmol/l; p < 0.03). In subjects without H pylori infection, gastrin values did not differ with age (old 37.1 (1.7); young 35.4 (2.1) pmol/l). The maximal gastric secretory volume was lower in old subjects with atrophy. Acid output (mmol/h) in subjects with atrophy was lower than in subjects with no atrophy (basal: 3.0(1.1); 5.1(0.7); p=NS; sham led: 5.4 (1.4); 9.3 (0.8); p<0.02; maximal: 18.9 (4.0); 31.4(1.8); p<0.002; peak: 25.1(5.3); 43.4(2.7); p<0.003). However, acid secretion in old subjects without atrophy was not different to that in young subjects, irrespective of H pylori status. These results did not differ when acid output was expressed as mmol/h/kg lean body mass or mmol/h/kg fat free body weight. Using multiple linear regression analysis, gastritis with atrophy was the only factor that had an independent negative effect on acid secretion. In healthy men without atrophy, gastric acid secretion is preserved with ageing and is independent of H pylori status. Atrophy, which is closely related to H pylori infection, is associated with a decline in acid secretion. Increased basal serum gastrin is related to both atrophy and H pylori infection but not to ageing per se.


Distinct and different effects of the oncogenes v-myc and v-src on avian sympathetic neurons: retroviral transfer of v-myc stimulates neuronal proliferation whereas v-src transfer enhances neuronal differentiation.
Immature avian sympathetic neurons are able to proliferate in culture for a limited number of divisions albeit expressing several neuron-specific properties. The effect of avian retroviral transfer of oncogenes on proliferation and differentiation of sympathetic neurons was investigated. Primary cultures of 6-d-old quail sympathetic ganglia, consisting of 90% neuronal cells, were infected by Myelocytomatosis virus (MC29), which contains the oncogene v-myc, and by the v-src-containing Rous sarcoma virus (RSV). RSV infection, in contrast to findings in other cellular systems, resulted in a reduction of neuronal proliferation as determined by 3H-thymidine incorporation (50% of control 4 d after infection) and in increased morphological differentiation. This is reflected by increased neurite production, cell size, and expression of neurofilament protein. In addition, RSV-infected neurons, unlike uninfected cells, are able to survive in culture for time periods up to 14 d in the absence of added neurotrophic factors. In contrast, retroviral transfer of v-myc stimulated the proliferation of immature sympathetic neurons preserving many properties of uninfected cells. The neuron-specific cell surface antigen Q211 and the adrenergic marker enzyme tyrosine hydroxylase were maintained in MC29-infected cells and in the presence of chick embryo extract the cells could be propagated over several weeks and five passages. Within 7 d after infection, the number of Q211-positive neurons increased approximately 100-fold. These data demonstrate distinct and different effects of v-src and v-myc-containing retroviruses on proliferation and differentiation of sympathetic neurons: v-src transfer results in increased differentiation, whereas v-myc transfer maintains an immature status reflected by proliferation, immature morphology, and complex growth requirements. The possibility of expanding immature neuronal populations by transfer of v-myc will be of considerable importance for the molecular analysis of neuronal proliferation and differentiation.