Clinical efficacy of cefpodoxime in respiratory tract infection.
Acute otitis media (AOM), sinusitis and tonsillopharyngitis are respiratory tract infections frequently encountered by primary-care physicians. Increasing bacterial resistance, particularly in Streptococcus pneumoniae, which is one of the most important respiratory tract bacteria implicated in community-acquired respiratory tract infections, has led to concern about the current options for empirical antibiotic treatment and has prompted a search for effective alternative treatments. Data from in vitro studies show that cefpodoxime has good activity against the main respiratory tract pathogens, S. pneumoniae, Haemophilus influenzae, Moraxella catarrhalis and Streptococcus pyogenes. Clinical studies confirm the efficacy of cefpodoxime in AOM, sinusitis and tonsillopharyngitis. As with all broad-spectrum antibiotics, there is the risk of promotion of bacterial resistance associated with overuse. However, if used with care, cefpodoxime can be considered as an alternative for empirical treatment of bacterial respiratory tract infections encountered in general practice, particularly where penicillins and macrolides have reduced efficacy against the main bacterial pathogens.


Abnormal neurofilament transport caused by targeted disruption of neuronal kinesin heavy chain KIF5A.
To test the hypothesis that fast anterograde molecular motor proteins power the slow axonal transport of neurofilaments (NFs), we used homologous recombination to generate mice lacking the neuronal-specific conventional kinesin heavy chain, KIF5A. Because null KIF5A mutants die immediately after birth, a synapsin-promoted Cre-recombinase transgene was used to direct inactivation of KIF5A in neurons postnatally. Three fourths of such mutant mice exhibited seizures and death at around 3 wk of age; the remaining animals survived to 3 mo or longer. In young mutant animals, fast axonal transport appeared to be intact, but NF-H, as well as NF-M and NF-L, accumulated in the cell bodies of peripheral sensory neurons accompanied by a reduction in sensory axon caliber. Older animals also developed age-dependent sensory neuron degeneration, an accumulation of NF subunits in cell bodies and a reduction in axons, loss of large caliber axons, and hind limb paralysis. These data support the hypothesis that a conventional kinesin plays a role in the microtubule-dependent slow axonal transport of at least one cargo, the NF proteins.


An independent relationship between plasma leptin and heart rate in untreated patients with essential hypertension.
To test the hypothesis that hemodynamic measurements in patients with essential hypertension are related independently to plasma leptin levels. We measured plasma leptin, insulin, office and ambulatory blood pressure and heart rate in 60 men with untreated mild hypertension. Plasma leptin correlated significantly with body mass index (r = 0.43, P = 0.001), 24 h heart rate (r = 0.35, P = 0.006) and 24 h diastolic blood pressure (r = 0.27, P = 0.04) but not with age (r = 0.03; P = 0.85) or 24 h systolic blood pressure (r = -0.08, P = 0.56). Plasma leptin levels adjusted for body mass index correlated significantly with 24 h heart rate (r = 0.36, P = 0.005) but not with 24 h diastolic blood pressure (r = 0.19, P = 0.15). We divided the patient population into tertiles of body mass index-adjusted plasma leptin levels. Age, plasma insulin, blood pressure, smoking status and physical activity habits were similar across the adjusted leptin tertiles. Patients from the third tertile of adjusted plasma leptin distribution (those with leptin levels higher than would be expected on the basis of body mass index) had significantly faster ambulatory heart rates than subjects from both the first and the second tertiles. The difference in heart rate across the three tertiles was most pronounced for the night-time values. In patients with essential hypertension, heart rate is faster in those patients with higher plasma leptin levels. This relationship is independent of age, body mass index, insulin levels, blood pressure level, smoking status and physical activity.


The effect of interferon on the formation of virus polyribosomes in L cells infected with vaccinia virus.
The effect of interferon treatment of mouse L cells on the fate of virus messenger RNA following infection with vaccinia virus has been studied. The polyribosomes of interferon-treated, infected cells are found to be disaggregated and it is proposed that htis results from inhibition of the initiation of virus polypeptide snythesis. Evidence is presented that inhibition of polypeptide chain elongation also occurs. The block in initiation appears to be due to the failure of the small ribosome subunit to attach to the virus messenger ribonucleoprotein complex. The translation of the different vaccinia messenger species is inhibited to a comparable extent.


Plasma aldosterone level within the normal range is less associated with cardiovascular and cerebrovascular risk in primary aldosteronism.
Previous studies showed higher risk of cardiovascular and cerebrovascular (CCV) events in primary aldosteronism compared with essential hypertension, but the patients of these studies were limited to primary aldosteronism patients with high plasma aldosterone concentration (PAC). The introduction of the aldosterone-renin ratio as the screening test for primary aldosteronism led to the recognition of primary aldosteronism patients with normal PAC (nPA). However, there is no information on the risk of primary aldosteronism including nPA. In this retrospectively and cross-sectional study, the clinical features and CCV event risk of primary aldosteronism at diagnosis including nPA were investigated and compared with essential hypertension. The study included 292 consecutive primary aldosteronism patients and 498 essential hypertension outpatients. All primary aldosteronism patients were diagnosed by autonomous aldosterone secretion using confirmatory tests, and then divided into nPA (n = 130) and primary aldosteronism patients with high PAC (hPA: n = 162) using a PAC cutoff level of less than 443 pmol/l (16 ng/dl), representing the normal upper limit of PAC. nPA patients were significantly older at diagnosis of primary aldosteronism and at onset of hypertension compared with hPA patients. They had milder hypokalemia and easier-to-control blood pressure. The results suggested that nPA could be considered a mild type of primary aldosteronism but not an early-stage hPA. Moreover, the risk of all CCV events in nPA was significantly lower than that in hPA (odds ratio 0.42, 95% confidence interval 0.18-0.90, P < 0.05) and not significantly higher than that in essential hypertension (odds ratio 0.95, 95% confidence interval 0.43-1.94, P = 0.899). This study suggests that aggressive diagnostic workout for nPA is less effective to prevent CCV events.


Artemisia annua L.: evidence of sesquiterpene lactones' fraction antinociceptive activity.
Artemisia annua L. has been used for many centuries in Chinese traditional medicine. Artemisinin, the active principle was first isolated and identified in the 1970s becoming the global back bone to the fight against malaria. Our research group previously developed an economic and ecological friendly process to obtain this compound. In the pursuit to also exploit the residue generated throughout the process we further evaluated the pharmacological potential of that extract. The alcoholic crude extract after artemisinin precipitation maintained an enriched sesquiterpene lactones content with residue artemisinin (1.72%) and deoxyartemisinin (0.31%), used as chemical markers for this sample. This study evaluated the pharmacological potential of the enriched sesquiterpene lactone fraction (Lac-FR) on different nociceptive and inflammatory experimental animal models. Previous findings on the biological properties of lactones obtained from natural products permitted us to explore the antinociceptive activities of these compounds based on in vivo chemical-induced behavioral assays. The enriched sesquiterpene lactone fraction (Lac-FR) was administrated by intraperitoneal injection producing a relevant reduction in the reaction time of the animals in both phases of the formalin test, significantly reduced the sensitivity to mechanical allodynia stimulus, reduced the paw edema caused by carrageenan injection and promoted high antinociceptive activity in tail flick model suggesting relationship with the opioid system. Further studies are being undertaken to elucidate the active components involved with the antinociceptive activity as well as evaluation of synergy effect that is seen by combination of substances that is greater than would be expected from consideration of individual contributions. For the first time, results presented herein provided consistent data to support the potential use of these lactones for pain relief as revealed by chemical-induced nociception assays in mice.


Mechanisms driving the lactate switch in Chinese hamster ovary cells.
The metabolism of Chinese Hamster Ovary (CHO) cells in a production environment has been extensively investigated. However, a key metabolic transition, the switch from lactate production to lactate consumption, remains enigmatic. Though commonly observed in CHO cultures, the mechanism(s) by which this metabolic shift is triggered is unknown. Despite this, efforts to control the switch have emerged due to the association of lactate consumption with improved cell growth and productivity. This review aims to consolidate current theories surrounding the lactate switch. The influence of pH, NAD+ /NADH, pyruvate availability and mitochondrial function on lactate consumption are explored. A hypothesis based on the cellular redox state is put forward to explain the onset of lactate consumption. Various techniques implemented to control the lactate switch, including manipulation of the culture environment, genetic engineering, and cell line selection are also discussed.


Effect of the systemic fungicide benodanil on the growth and development of six spring barley varieties in the presence or absence of brown rust infection in the glasshouse.
When applied at the juvenile growth stage, benodanil significantly reduced the occurrence of rust (Puccinia bordei) on barley plants. This was found in the susceptible varieties Midas, Zephyr, and Mazurka as well as in the slow-rusting varieties Tyra, Emir, and Vada. Benodanil increased the growth of non-infected plants, it reduced however tillering and the total leaf number of the plants. The effect on the growth, development, and yield of infected plants differed between the varieties, with the susceptible varieties reacting better to fungicide application.


[Parent-doctor relations in oncology: a qualitative approach].
We want to learn how parents of children with cancer perceive their relationship with hospital staff, especially with doctors. We used group-based qualitative methodology. The sample is composed of 14 mothers/fathers whose children contracted the disease more than two years previously. All parents want information that is both intelligible and detailed. The word cancer has a strong social stigma and is avoided when giving information to parents and to children. Communication between doctors and parents can lead to situations of tension during diagnosis and relapses. Parents trust the professionalism of doctors. Parents also want doctors to be competent and to have human qualities. The preparation of reports by physicians is the task most criticized by parents.


Impact of vascular endothelial growth factor single nucleotide polymorphism association on acute renal allograft rejection.
Vascular endothelial growth factor (VEGF) is a cytokine which plays an important role in the division, proliferation and migration of endothelial cells. In the kidney, VEGF expression is found in glomerular podocytes and in tubular epithelial cells, which may result in acute inflammatory reactions. The role of VEGF gene polymorphisms (-2578 C/A, -2549 18 bp Ins/Del, -1154 G/A and +936 C/T) was investigated in 272 patients who underwent renal transplantation. ARMS-PCR and PCR-RFLP were used. Patients were categorized into acute allograft rejection (n = 76) and nonrejection (n = 196). The VEGF -1154 GG genotype and the +936 T allele were found to be susceptible to acute rejection (AR). T-A-A-I, T-A-A-D, T-G-C-I and C-A-A-I haplotypes revealed a predisposition among AR cases. In silico analysis revealed +936 T as a significant allele involved in the transcription regulation. These results highlight the role of VEGF polymorphisms in acute allograft rejection.


360-Degree Feedback Model to Enhance Interprofessional Learning.
This article was migrated. The article was marked as recommended. Background: Providing meaningful feedback in interprofessional education (IPE) requires knowledge of discipline specific responsibilities and a method for measuring team dynamics while capturing individual performance. Methods: We implemented a 360-degree performance feedback model for a large-scale IPE simulation with standardized patients (SP) who transitioned from primary care to the emergency department. 293 medical, nursing, and pharmacy students were divided into 72 teams. We conducted a retrospective study evaluating feedback from 108 facilitators on individual and team based competencies, 12 SP patient satisfaction surveys and 293 student self-appraisals. We analyzed data using descriptive statistics and ANOVA for multiple group comparisons. Results: More than 94% of SP indicated they would return to the same student team to seek care. However, SP reported that the students did not summarize or clarify information, adapt to their level of understanding or encourage questions. Facilitators noted all disciplines were involved in formulating and implementing treatment plans. Student teams performed highest in mutual support and situational monitoring, and lowest in leadership and team structure. Students across all disciplines rated their teams as requiring light to no supervision. Conclusion: Applying the 360-degree performance model is feasible in IPE and provides multidimensional, qualitative feedback to enhance student learning.


On the origin of the unusual behavior in the stretching of single-stranded DNA.
Force-extension curves (FECs), which quantify the response of a variety of biomolecules subject to mechanical force (f), are often quantitatively fit using worm-like chain (WLC) or freely jointed chain (FJC) models. These models predict that the chain extension, x, normalized by the contour length increases linearly at small f and at high forces scale as x ~ (1 - f(-α)), where α = 0.5 for WLC and unity for FJC. In contrast, experiments on single-stranded DNA (ssDNA) show that over a range of f and ionic concentration, x scales as x ~ ln f, which cannot be explained using WLC or FJC models. Using theory and simulations we show that this unusual behavior in FEC in ssDNA is due to sequence-independent polyelectrolyte effects. We show that the x ~ ln f arises because in the absence of force the tangent correlation function, quantifying chain persistence, decays algebraically on length scales on the order of the Debye length. Our theory, which is most appropriate for monovalent salts, quantitatively fits the experimental data and further predicts that such a regime is not discernible in double-stranded DNA.


Blood Pressure in 6-Year-Old Children Born Extremely Preterm.
Advances in perinatal medicine have increased infant survival after very preterm birth. Although this progress is welcome, there is increasing concern that preterm birth is an emerging risk factor for hypertension at young age, with implications for the lifetime risk of cardiovascular disease. We measured casual blood pressures (BPs) in a population-based cohort of 6-year-old survivors of extremely preterm birth (<27 gestational weeks; n=171) and in age- and sex-matched controls born at term (n=172). Measured BP did not differ, but sex, age-, and height-adjusted median z scores were 0.14 SD higher (P=0.02) for systolic BP and 0.10 SD higher (P=0.01) for diastolic BP in children born extremely preterm than in controls. Among children born extremely preterm, shorter gestation, higher body mass index, and higher heart rate at follow-up were all independently associated with higher BP at 6 years of age, whereas preeclampsia, smoking in pregnancy, neonatal morbidity, and perinatal corticosteroid therapy were not. In multivariate regression analyses, systolic BP decreased by 0.10 SD (P=0.08) and diastolic BP by 0.09 SD (P=0.02) for each week-longer gestation. Six-year-old children born extremely preterm have normal but slightly higher BP than their peers born at term. Although this finding is reassuring for children born preterm and their families, follow-up at older age is warranted.


Choanal adenoid syndrome--a preliminary report.
A series of 19 patients with long-lasting nasal stenosis despite repeated treatment attempts, including, in many cases, repeated adenoidectomy, was found to have ectopic adenoid tissue inside the choanae. The choanal adenoid tissue could not be removed by curettage, only transnasally using forceps. The treatment was successful in all 19 patients. It is estimated that in at least 2% of the patients with long-lasting nasal stenosis, attention should be focused on the bottleneck of the nasopharyngeal airway represented by the choanae, primarily by fibre optic endoscopic examination.


Single molecule analysis of c-myb alternative splicing reveals novel classifiers for precursor B-ALL.
The c-Myb transcription factor, a key regulator of proliferation and differentiation in hematopoietic and other cell types, has an N-terminal DNA binding domain and a large C-terminal domain responsible for transcriptional activation, negative regulation and determining target gene specificity. Overexpression and rearrangement of the c-myb gene (MYB) has been reported in some patients with leukemias and other types of cancers, implicating activated alleles of c-myb in the development of human tumors. Alternative RNA splicing can produce variants of c-myb with qualitatively distinct transcriptional activities that may be involved in transformation and leukemogenesis. Here, by performing a detailed, single molecule assay we found that c-myb alternative RNA splicing was elevated and much more complex in leukemia samples than in cell lines or CD34+ hematopoietic progenitor cells from normal donors. The results revealed that leukemia samples express more than 60 different c-myb splice variants, most of which have multiple alternative splicing events and were not detectable by conventional microarray or PCR approaches. For example, the single molecule assay detected 21 and 22 splice variants containing the 9B and 9S exons, respectively, most of which encoded unexpected variant forms of c-Myb protein. Furthermore, the detailed analysis identified some splice variants whose expression correlated with poor survival in a small cohort of precursor B-ALL samples. Our findings indicate that single molecule assays can reveal complexities in c-myb alternative splicing that have potential as novel biomarkers and could help explain the role of c-Myb variants in the development of human leukemia.


Single-Cell Genomics Reveals a Diverse Metabolic Potential of Uncultivated Desulfatiglans-Related Deltaproteobacteria Widely Distributed in Marine Sediment.
Desulfatiglans-related organisms comprise one of the most abundant deltaproteobacterial lineages in marine sediments where they occur throughout the sediment column in a gradient of increasing sulfate and organic carbon limitation with depth. Characterized Desulfatiglans isolates are dissimilatory sulfate reducers able to grow by degrading aromatic hydrocarbons. The ecophysiology of environmental Desulfatiglans-populations is poorly understood, however, possibly utilization of aromatic compounds may explain their predominance in marine subsurface sediments. We sequenced and analyzed seven Desulfatiglans-related single-cell genomes (SAGs) from Aarhus Bay sediments to characterize their metabolic potential with regard to aromatic compound degradation and energy metabolism. The average genome assembly size was 1.3 Mbp and completeness estimates ranged between 20 and 50%. Five of the SAGs (group 1) originated from the sulfate-rich surface part of the sediment while two (group 2) originated from sulfate-depleted subsurface sediment. Based on 16S rRNA gene amplicon sequencing group 2 SAGs represent the more frequent types of Desulfatiglans-populations in Aarhus Bay sediments. Genes indicative of aromatic compound degradation could be identified in both groups, but the two groups were metabolically distinct with regard to energy conservation. Group 1 SAGs carry a full set of genes for dissimilatory sulfate reduction, whereas the group 2 SAGs lacked any genetic evidence for sulfate reduction. The latter may be due to incompleteness of the SAGs, but as alternative energy metabolisms group 2 SAGs carry the genetic potential for growth by acetogenesis and fermentation. Group 1 SAGs encoded reductive dehalogenase genes, allowing them to access organohalides and possibly conserve energy by their reduction. Both groups possess sulfatases unlike their cultured relatives allowing them to utilize sulfate esters as source of organic carbon and sulfate. In conclusion, the uncultivated marine Desulfatiglans populations are metabolically diverse, likely reflecting different strategies for coping with energy and sulfate limitation in the subsurface seabed.


The suppression of testis-brain RNA binding protein and kinesin heavy chain disrupts mRNA sorting in dendrites.
Ribonucleoprotein particles (RNPs) are thought to be key players in somato-dendritic sorting of mRNAs in CNS neurons and are implicated in activity-directed neuronal remodeling. Here, we use reporter constructs and gel mobility shift assays to show that the testis brain RNA-binding protein (TB-RBP) associates with mRNPs in a sequence (Y element) dependent manner. Using antisense oligonucleotides (anti-ODN), we demonstrate that blocking the TB-RBP Y element binding site disrupts and mis-localizes mRNPs containing (alpha)-calmodulin dependent kinase II (alpha)-CAMKII) and ligatin mRNAs. In addition, we show that suppression of kinesin heavy chain motor protein alters only the localization of (alpha)-CAMKII mRNA. Thus, differential sorting of mRNAs involves multiple mRNPs and selective motor proteins permitting localized mRNAs to utilize common mechanisms for shared steps.


Kinetics of M1 muscarinic receptor and G protein signaling to phospholipase C in living cells.
G protein-coupled receptors (GPCRs) mediate responses to external stimuli in various cell types. Early events, such as the binding of ligand and G proteins to the receptor, nucleotide exchange (NX), and GTPase activity at the Galpha subunit, are common for many different GPCRs. For G(q)-coupled M(1) muscarinic (acetylcholine) receptors (M(1)Rs), we recently measured time courses of intermediate steps in the signaling cascade using Förster resonance energy transfer (FRET). The expression of FRET probes changes the density of signaling molecules. To provide a full quantitative description of M(1)R signaling that includes a simulation of kinetics in native (tsA201) cells, we now determine the density of FRET probes and construct a kinetic model of M(1)R signaling through G(q) to activation of phospholipase C (PLC). Downstream effects on the trace membrane lipid phosphatidylinositol 4,5-bisphosphate (PIP(2)) and PIP(2)-dependent KCNQ2/3 current are considered in our companion paper in this issue (Falkenburger et al. 2010. J. Gen. Physiol. doi:10.1085/jgp.200910345). By calibrating their fluorescence intensity, we found that we selected transfected cells for our experiments with approximately 3,000 fluorescently labeled receptors, G proteins, or PLC molecules per microm(2) of plasma membrane. Endogenous levels are much lower, 1-40 per microm(2). Our kinetic model reproduces the time courses and concentration-response relationships measured by FRET and explains observed delays. It predicts affinities and rate constants that align well with literature values. In native tsA201 cells, much of the delay between ligand binding and PLC activation reflects slow binding of G proteins to receptors. With M(1)R and Gbeta FRET probes overexpressed, 10% of receptors have G proteins bound at rest, rising to 73% in the presence of agonist. In agreement with previous work, the model suggests that binding of PLC to Galpha(q) greatly speeds up NX and GTPase activity, and that PLC is maintained in the active state by cycles of rapid GTP hydrolysis and NX on Galpha(q) subunits bound to PLC.


In situ electrostimulation drives a regenerative shift in the zone of infarcted myocardium.
Electrostimulation represents a well-known trophic factor for different tissues. In vitro electrostimulation of non-stem and stem cells induces myogenic predifferentiation and may be a powerful tool to generate cells with the capacity to respond to local areas of injury. We evaluated the effects of in vivo electrostimulation on infarcted myocardium using a miniaturized multiparameter implantable stimulator in rats. Parameters of electrostimulation were organized to avoid a direct driving or pacing of native heart rhythm. Electrical stimuli were delivered for 14 days across the scar site. In situ electrostimulation used as a cell-free, cytokine-free stimulation system, improved myocardial function, and increased angiogenesis through endothelial progenitor cell migration and production of vascular endothelial growth factor (VEGF). In situ electrostimulation represents a novel means to stimulate repair of the heart and other organs, as well as to precondition tissues for treatment with cell-based therapies.


[Computer aided, examiner independent screening for inapparent eye misalignment among 590 infants and preschoolers--a pilot study with an orthoptic gold standard].
The purpose of the study was to investigate the computer aided screening method using Purkinje image I and IV reflection patterns for the detection of inapparent eye misalignment and to compare this to an orthoptic examination. 590 subjects up to 72 months of age with inapparent eye misalignment were recruited from the orthoptic outpatient department and externally. The computer aided screening consisted of taking a series of still video pictures with Purkinje reflection patterns. These were evaluated in an examiner independent way to reach a recommendation whether the child needed an ophthalmological referral or not. As gold standard, an orthoptic examination was performed. For analysis, the data were split by age groups. The orthoptic results were tested for certainty and repeatability. The computer aided examination had the highest sensitivity of 0.82 in the age group up to 2.5 years of age, and a specificity of 0.90. With an estimated prevalence for microtropia of 0.01, the extrapolated positive predictive value was 0.08, and the negative predictive value was 0.998. In the age group up to 2.5 years of age, the percentage of orthoptic examinations without clear result (neither non-referral, nor strabismic) was 22.4%, and 6.1% in the screening examination. Among the cases which were examined repeatedly, some were classified as "strabismic" in the beginning, and as "non-referral" in the end in the orthoptic examinations. The examiner independent, computer aided screening method is a cost effective option for the screening for inapparent eye misalignment, especially in the age group up to 2.5 years of age. If an orthoptic examination was carried out for screening, one should expect a higher rate of false positives, which entails more costly ophthalmological checks. Future studies should assess the validity of the single ortoptic examination as the gold standard in this age group.


RsaL-driven negative regulation promotes heterogeneity in Pseudomonas aeruginosa quorum sensing.
In its canonical interpretation, quorum sensing (QS) allows single cells in a bacterial population to synchronize gene expression and hence perform specific tasks collectively once the quorum cell density is reached. However, growing evidence in different bacterial species indicates that considerable cell-to-cell variation in the QS activation state occurs during growth, often resulting in coexisting subpopulations of cells in which QS is active (quorate cells) or inactive (non-quorate cells). Heterogeneity has been observed in the las QS system of the opportunistic pathogen Pseudomonas aeruginosa. However, the molecular mechanisms underlying this phenomenon have not yet been defined. The las QS system consists of an incoherent feedforward loop in which the LasR transcriptional regulator activates the expression of the lasI synthase gene and rsaL, coding for the lasI transcriptional repressor RsaL. Here, single-cell-level gene expression analyses performed in ad hoc engineered biosensor strains and deletion mutants revealed that direct binding of RsaL to the lasI promoter region increases heterogeneous activation of the las QS system. Experiments performed with a dual-fluorescence reporter system showed that the LasR-dependent expression of lasI and rsaL does not correlate in single cells, indicating that RsaL acts as a brake that stochastically limits the transition of non-quorate cells to the quorate state in a subpopulation of cells expressing high levels of this negative regulator. Interestingly, the rhl QS system that is not controlled by an analogous RsaL protein showed higher homogeneity with respect to the las system. IMPORTANCE Single-cell analyses can reveal that despite experiencing identical physico-chemical conditions, individual bacterial cells within a monoclonal population may exhibit variations in gene expression. Such phenotypic heterogeneity has been described for several aspects of bacterial physiology, including QS activation. This study demonstrates that the transition of non-quorate cells to the quorate state is a graded process that does not occur at a specific cell density and that subpopulations of non-quorate cells also persist at high cell density. Here, we provide a mechanistic explanation for this phenomenon, showing that a negative feedback regulatory loop integrated into the las system has a pivotal role in promoting cell-to-cell variation in the QS activation state and in limiting the transition of non-quorate cells to the quorate state in P. aeruginosa.


[A clinico-morphologico-functional study of the kidneys in glomerulonephritis].
The article deals with the results of a clinico-functional-morphological study of the kidneys in glomerulonephritis (50 observations) which was carried out with the use of the method of puncture biopsy of the kidneys. It was shown that clinical forms of glomerulonephritis, accompanied with hematuria, were characterized by the absence of fixation of immune complexes in the basal membrane of the glomeruli. According to the electron microscopy data, this corresponds to the dissappearance of deposits from the subendothelial parts of the basal membrane. The extramembranous glomerulonephritis, revealed with the help of electron microscopy technique, was characterized by the nephrotic syndrome. An increased tension of immunological processes in the glomeruli was accompanied by a more grave clinical course of glomerulonephritis, by impairment of not only glomerulous but tubular functions as well. A greater dicrease in the function of osmotic concentration of the urine in fibroplastic types of glomerulonephritis, as compared with non-firboplastic ones, was apparently connected not only with lesiones of tubules but rather with more expressed sclerotic changes in the interstitial tissue of the kidney.


BioVLAB-MMIA-NGS: microRNA-mRNA integrated analysis using high-throughput sequencing data.
It is now well established that microRNAs (miRNAs) play a critical role in regulating gene expression in a sequence-specific manner, and genome-wide efforts are underway to predict known and novel miRNA targets. However, the integrated miRNA-mRNA analysis remains a major computational challenge, requiring powerful informatics systems and bioinformatics expertise. The objective of this study was to modify our widely recognized Web server for the integrated mRNA-miRNA analysis (MMIA) and its subsequent deployment on the Amazon cloud (BioVLAB-MMIA) to be compatible with high-throughput platforms, including next-generation sequencing (NGS) data (e.g. RNA-seq). We developed a new version called the BioVLAB-MMIA-NGS, deployed on both Amazon cloud and on a high-performance publicly available server called MAHA. By using NGS data and integrating various bioinformatics tools and databases, BioVLAB-MMIA-NGS offers several advantages. First, sequencing data is more accurate than array-based methods for determining miRNA expression levels. Second, potential novel miRNAs can be detected by using various computational methods for characterizing miRNAs. Third, because miRNA-mediated gene regulation is due to hybridization of an miRNA to its target mRNA, sequencing data can be used to identify many-to-many relationship between miRNAs and target genes with high accuracy. http://epigenomics.snu.ac.kr/biovlab_mmia_ngs/.


Individual differences in preferences for and responses to four nicotine replacement products.
With several different forms of nicotine replacement therapy available for smokers, it is useful to know about factors that may influence choice of form. To examine factors associated with preference for different nicotine replacement products and effectiveness of these products. The simple comparison of success rates between the products has been reported previously. Five hundred and four smokers were invited to rank order four products (gum, patch, nasal spray and inhaler) in terms of preference and were then randomly allocated to receive one of the products for a smoking cessation attempt. They were followed up 1 week, 4 weeks, 12 weeks and 15 weeks later. Ratings of the products were repeated 1 week after the quit date. Mood and physical symptoms were assessed at baseline and at each follow-up visit. Urges to smoke were assessed at all post-quit date sessions. Abstinence was assessed by self-report with expired air carbon monoxide verification. The patch was the most popular product initially, followed by the spray and inhaler and then the gum. The difference was greater for women than men. However, all smokers quickly came to like whichever product they had been allocated. Smokers who initially preferred the spray or inhaler were heavier smokers than those who preferred the gum or patch. Prior experience of the gum was associated with lower initial preference for it, but did not affect outcome. Prior experience with the patch had no effect on preference or outcome. Being allocated to the preferred product did not increase success at stopping smoking. Women were more successful at stopping smoking on the inhaler than the gum. Among highly dependent smokers, those on the inhaler had a lower relapse rate than those on the other products. Regardless of initial preferences, whether patients obtain their preferred form of NRT or one selected for them did not seem to affect outcome. This may be because smokers came to like particular products as they got used to them. Other things being equal, women may be better advised to use inhaler rather than gum and men vice versa.


Wheat bran affects the site of fermentation of resistant starch and luminal indexes related to colon cancer risk: a study in pigs.
Recent studies suggest that resistant starch (effective in producing butyrate and lowering possibly toxic ammonia) is rapidly fermented in the proximal colon; the distal colon especially would, however, benefit from these properties of resistant starch. To determine whether wheat bran (a rich source of insoluble non-starch polysaccharides), known to hasten gastrointestinal transit, could carry resistant starch through to the distal colon and thus shift its site of fermentation. Twenty four pigs were fed four human type diets: a control diet, or control diet supplemented with resistant starch, wheat bran, or both. Intestinal contents and faeces were collected after two weeks. Without wheat bran, resistant starch was rapidly fermented in the caecum and proximal colon. Supplementation with wheat bran inhibited the caecal fermentation of resistant starch, resulting in an almost twofold increase (from 12.9 (2.5) to 20.5 (2.1) g/day, p<0.05) in resistant starch being fermented between the proximal colon and faeces. This resulted in higher butyrate (133%, p<0.05) and lower ammonia (81%, p<0.05) concentrations in the distal colonic regions. Wheat bran can shift the fermentation of resistant starch further distally, thereby improving the luminal conditions in the distal colonic regions where tumours most commonly occur. Therefore, the combined consumption of resistant starch and insoluble non-starch polysaccharides may contribute to the dietary modulation of colon cancer risk.


Methanotrophic contribution to biodegradation of phenoxy acids in cultures enriched from a groundwater-fed rapid sand filter.
Drinking water supply is in many parts of the world based on groundwater. Groundwater often contains methane, which can be oxidized by methanotrophs upon aeration. Sand from rapid sand filters fed with methane-rich groundwater can remove some pesticides (Hedegaard and Albrechtsen in Water Res 48:71-81, 2014). We enriched methanotrophs from filter sand and investigated whether they could drive the degradation of various pesticides. To enrich for methanotrophs, we designed and operated four laboratory-scale, continuously methane-fed column reactors, inoculated with filter sand and one control column fed with tap water. When enrichments were obtained, methane was continuously supplied to three reactors, while the fourth was starved for methane for 1 week, and the reactors were spiked with ten pesticides at groundwater-relevant concentrations (2.1-6.6 μg/L). Removal for most pesticides was not detected at the investigated contact time (1.37 min). However, the degradation of phenoxy acids was observed in the methanotrophic column reactor starved for methane, while it was not detected in the control column indicating the importance of methanotrophs. Phenoxy acid removal, using dichlorprop as a model compound, was further investigated in batch experiments with methanotrophic biomass collected from the enrichment reactors. Phenoxy acid removal (expressed per gram of matrix sand) was substantially improved in the methanotrophic enrichment compared to parent filter sand. The presence of methane did not clearly impact dichlorprop removal but did impact mineralization. We suggest that other heterotrophs are responsible for the first step in dichlorprop degradation, while the subsequent steps including ring-hydroxylation are driven by methanotrophs.


The Mycobacterium tuberculosis phagosome interacts with early endosomes and is accessible to exogenously administered transferrin.
Previous studies have demonstrated that the Mycobacterium tuberculosis phagosome in human monocyte-derived macrophages acquires markers of early and late endosomes, but direct evidence of interaction of the M. tuberculosis phagosome with the endosomal compartment has been lacking. Using the cryosection immunogold technique, we have found that the M. tuberculosis phagosome acquires exogenously added transferrin in a time-dependent fashion. Near-maximal acquisition of transferrin occurs within 15 min, kinetics of acquisition consistent with interaction of the M. tuberculosis phagosome with early endosomes. Transferrin is chased out of the M. tuberculosis phagosome by incubation of the infected macrophages in culture medium lacking human transferrin. Phagosomes containing latex beads or heat-killed M. tuberculosis, on the other hand, do not acquire staining for transferrin. These and other findings demonstrate that M. tuberculosis arrests the maturation of its phagosome at a stage at which the phagosome interacts with early and late endosomes, but not with lysosomes. The transferrin endocytic pathway potentially provides a novel route for targeting antimicrobials to the M. tuberculosis phagosome.


Reactivity and selectivity differences between catecholate and catechothiolate Ru complexes. Implications regarding design of stereoselective olefin metathesis catalysts.
The origins of the unexpected finding that Ru catechothiolate complexes, in contrast to catecholate derivatives, promote exceptional Z-selective olefin metathesis reactions are elucidated. We show that species containing a catechothiolate ligand, unlike catecholates, preserve their structural integrity under commonly used reaction conditions. DFT calculations indicate that, whereas alkene coordination is the stereochemistry-determining step with catecholate complexes, it is through the metallacyclobutane formation that the identity of the major isomer is determined with catechothiolate systems. The present findings suggest that previous models for Z selectivity, largely based on steric differences, should be altered to incorporate electronic factors as well.


Bis(1,10-phenanthroline-κ(2) N,N')(sulfato-κO)zinc(II) propane-1,2-diol monosolvate.
In the title compound, [Zn(SO4)(C12H8N2)2]·C3H8O2, the Zn(II) ion is in a distorted square-pyramidal coordination environment composed of four N atoms from two chelating 1,10-phenanthroline ligands and one O atom from a monodentate sulfate ligand. The Zn(II) ion lies on a twofold rotation axis. The sulfate ligand and propane-1,2-diol mol-ecules are disordered across the twofold rotation axis. The dihedral angle between the two chelating N2C2 groups is 83.26 (13)°. In the crystal, the complex mol-ecule and the propane-1,2-diol mol-ecule are connected through a pair of O-H⋯O hydrogen bonds.


Publication Bias Among Conference Abstracts Reporting on Pediatric Quality Improvement Projects.
This study evaluated progress to publication of pediatric quality improvement (QI) projects initially presented as national conference abstracts, according to project findings and other characteristics. QI abstracts were identified among presentations at the 2010-2015 American Academy of Pediatrics National Conference & Exhibition, and publications were tracked through June 2018. Positive findings (improvement on at least 1 quantitative project outcome), interventions, and analyses were correlated with journal publication. Of 142 abstracts, 128 (90%) reported positive findings. Forty-nine positive abstracts and 3 abstracts reporting negative results resulted in publication (38% vs 21%, respectively; P = .256). Median time to publication was 1.2 years for projects with positive findings, compared to >3 years for abstracts with negative findings (P = .029). Ninety percent of abstracts reported positive findings, and these abstracts progressed to publication more quickly. Overcoming publication bias for pediatric QI projects may enhance selection of promising interventions as new projects are designed.


Neonatal undernutrition alters responsiveness to morphine in mature rats: a possible source of epiphenomena in developmental drug studies.
We studied the effect of neonatal undernutrition and its attendant stresses on the behavior and thermoregulation of adult rats in the absence and presence of morphine. Undernutrition was accomplished by fostering half the pups in each litter to a nonpregnant, nonlactating female rat every other day for the first 6 days of life. As a control, the remaining pups were fostered to lactating rats. Significant alterations in body and brain weight and in brain DNA, RNA and protein were noted in undernourished rats at 10 and 21 days of age. At 6 months of age, no difference was observed in unconditioned exploratory behavior or in the acquisition and performance of a conditioned autoshaped lever touch response. Differential sensitivity to morphine was observed in nourished and undernourished rats performing the autoshaped response at asymptotic levels. At 1 year of age, undernourished rats displayed lower rectal temperatures than controls and showed an altered thermic response to morphine. These data demonstrate that poor nutritional status and other nondrug factors may be responsible for the altered thermoregulation and opiate sensitivity observed in rodents perinatally addicted to opioids, effects generally regarded as specific consequences of early opiate exposure.


GRAF1 integrates PINK1-Parkin signaling and actin dynamics to mediate cardiac mitochondrial homeostasis.
The serine/threonine kinase, PINK1, and the E3 ubiquitin ligase, Parkin, are known to facilitate LC3-dependent autophagosomal encasement and lysosomal clearance of dysfunctional mitochondria, and defects in this process contribute to a variety of cardiometabolic and neurological diseases. Although recent evidence indicates that dynamic actin remodeling plays an important role in PINK1/Parkin-mediated mitochondrial autophagy (mitophagy), the underlying signaling mechanisms remain unknown. Here, we identify the RhoGAP GRAF1 (Arhgap26) as a PINK1 substrate that regulates mitophagy. GRAF1 promotes the release of damaged mitochondria from F-actin anchors, regulates mitochondrial-associated Arp2/3-mediated actin remodeling and facilitates Parkin-LC3 interactions to enhance mitochondria capture by autophagosomes. Graf1 phosphorylation on PINK1-dependent sites is dysregulated in human heart failure, and cardiomyocyte-restricted Graf1 depletion in mice blunts mitochondrial clearance and attenuates compensatory metabolic adaptations to stress. Overall, we identify GRAF1 as an enzyme that coordinates cytoskeletal and metabolic remodeling to promote cardioprotection.


Smoking and colorectal cancer: different effects by type of cigarettes?
Although smoking is suggested to be a risk factor for colorectal cancer, the evidence to date is conflicting and may be confounded. Moreover, the effect of tobacco smoke may vary by time since initiation, type of tobacco product, anatomic subsites, and among ethnic groups. Data were derived from two consecutive population-based case-control studies conducted among Caucasians, Japanese, Native Hawaiians, Filipinos, and Chinese in Hawaii, including 1,959 ethnicity-, sex-, and age-matched case-control pairs. A lifetime history of smoking for different tobacco products and information on other risk factors were obtained by in-person interviews. Odds ratios (OR) and corresponding 95% confidence intervals (95% CI) were estimated using conditional logistic regression models with adjustment for potential confounders. Subjects who ever smoked were at an increased risk of colorectal cancer compared with never smokers (OR, 1.23; 95% CI, 0.99-1.52 for men and OR, 1.27; 95% CI, 1.01-1.59 for women). Increasing quartiles of pack-years over all tobacco products showed a clear dose-dependent association in men [for the highest quartile, Q4 (>40 pack-years) versus never smokers: OR, 1.48; 95% CI, 1.12-1.96; P(trend) = 0.002]. The dose-response trend was also present in women [for the highest quartile, Q4 (>30 pack-years) versus never smokers: OR, 1.38; 95% CI, 0.91-1.95; P(trend) = 0.04] and each ethnic group. There was a suggestion of a difference in risk with type of tobacco product. Non-filtered cigarettes increased risk of both colon and rectal cancer [for Q4 versus never smokers: OR, 1.59; 95% CI, 1.15-2.21; P(trend) = 0.001 and OR, 1.84; 95% CI, 1.18-2.86; P(trend) = 0.02, respectively], whereas filtered cigarettes seemed to increase risk of rectal but not colon cancer (OR, 1.37; 95% CI, 0.88-2.13; P(trend) = 0.06 and OR, 1.05; 95% CI, 0.79-1.39; P(trend) = 0.98, respectively). The effect of smoking was not limited to the distant past, and accumulated pack-years of smoking seemed to be more important than the time in which smoking occurred. The data from this large study corroborate previous reports of a positive association between smoking and colorectal cancer and suggest that the association may vary by type of cigarette.


Distinguishing the role of conscious and unconscious knowledge in evaluative conditioning.
Evaluative conditioning (EC) refers to a change in liking of a conditioned stimulus (CS) subsequent to its repeated pairing with a valent stimulus (US). Two studies that bring new light on the highly debated question of the role of awareness in EC were conducted. We developed an innovative method motivated by higher order and integration theories of consciousness to distinguish between the role of conscious and unconscious knowledge about the pairings. On each trial of the awareness test, participants had to indicate the valence of the US associated with a given CS and to make a 'structural knowledge attribution' by reporting the basis of their response. Valence identification accuracy was used to evaluate knowledge while the knowledge attribution was used to measure the conscious status of knowledge. Memory attribution indicated conscious knowledge about the pairings while feeling-based and random attributions indicated unconscious knowledge. A meta-analysis of the two studies revealed that valence identification accuracy was above chance level for memory and feeling-based attributions but not for the random attribution. EC was found in the three attributions. While EC effect size was medium for the memory attribution it was small for feeling-based and random attributions. Moreover, Experiment 2 included a delayed test. EC was still present 24 h after the conditioning took place. The results obtained for memory and feeling-based attributions suggest that both conscious and unconscious knowledge may underlie EC. The results obtained for random attribution suggest that EC may also occur without any knowledge of US valence.


Flexible transbronchial needle aspiration biopsy for histologic specimens.
Flexible transbronchial needle aspiration (TBNA) biopsy has been used to obtain cytologic specimens from mediastinal lymph nodes for the diagnosis and staging of bronchogenic carcinoma. The dependency on a sophisticated cytologic laboratory, the skill required to perform this procedure, and the inability to obtain enough tissue to diagnose noncarcinomatous processes in the mediastinum by smaller needles has limited its use. The application of TBNA would be greatly enhanced if a histologic specimen could be obtained from the mediastinal lymph node safely. For this reason, an 18-gauge needle was developed and used through a rigid bronchoscope. In ten patients, adequate tissue for histology was obtained in nine, and a specific diagnosis was made in eight patients without any complication. A similar technique was developed and is now reported using the flexible bronchoscope. We conclude that using a flexible transbronchial needle is an effective and safe way of obtaining histologic specimens from the mediastinum and can provide a diagnosis, both in malignant and benign mediastinal disease. Hopefully, this technique will further reduce the need for more invasive surgical procedures.


Short-term depression at the reciprocal synapses between a retinal bipolar cell terminal and amacrine cells.
Visual adaptation is thought to occur partly at retinal synapses that are subject to plastic changes. However, the locus and properties of this plasticity are not well known. Here, we studied short-term plasticity at the reciprocal synapse between bipolar cell terminals and amacrine cells in goldfish retinal slices. Depolarization of a single bipolar cell terminal for 100 ms triggers the release of glutamate onto amacrine cell processes, which in turn leads to GABAergic feedback from amacrine cells onto the same terminal. We find that this release of GABA undergoes paired-pulse depression (PPD) that recovers in <1 min (single exponential time constant, tau approximately = 12 s). This disynaptic PPD is independent of mGluR-mediated plasticity and depletion of glutamatergic synaptic vesicle pools, because exocytosis assayed via capacitance jumps (deltaC(m)) recovered completely after 10 s (tau approximately = 2 s). Fast application of GABA (10 mM) onto outside-out patches excised from bipolar cell terminals showed that the recovery of GABA(A) receptors from desensitization depends on the duration of the application [fast recovery (<2 s) for short applications; slow (tau approximately = 12 s) for prolonged applications]. We thus blocked GABA(A) receptors and retested the GABAergic response mediated by nondesensitizing GABA(C) receptors to two rapid glutamate puffs onto the bipolar cell terminal. These responses consistently displayed PPD. Furthermore, blocking AMPA-receptor desensitization with cyclothiazide, or evoking GABA release with NMDA receptors, did not reduce PPD. We thus suggest that depletion of synaptic vesicle pools in GABAergic amacrine cells is a major contributor to PPD.


Fluid dynamics, wall mechanics, and oxygen transfer in peripheral bypass anastomoses.
Intimal hyperplasia at vascular anastomoses seems to be promoted by altered flow conditions and stress distributions within the anastomotic region. In order to gain deeper insight into postoperative disease processes, and subsequently, to contribute to the development of improved vascular reconstructions, detailed studies, also on local flow dynamics and related mass transport and wall mechanical effects, are required. In context with in vivo studies, computer simulation based on casts of femoro-popliteal bypasses implanted into sheep were performed to analyze the flow dynamics, the oxygen transport, and the wall and suture mechanics in anatomically correct bypass configurations related to three established surgical techniques and resulting geometries (conventional type anastomosis, Taylor-patch and Miller-cuff anastomoses with venous interposition grafts of different modifications). The influence of geometry, compliance of the graft, the interponated vein patch and vein cuff, and of the artery was included. Time-dependent, three-dimensional Navier-Stokes equations describing the flow field, and a nonlinear shell structure for the vessel walls were coupled using finite element methods. The numerical results demonstrate nonphysiological flow patterns in the anastomotic region. Strongly skewed axial velocity profiles and secondary velocities occur in the junction region. In the Miller-cuff a vortex may induce a wash-out effect which protects the host artery. On the artery floor opposite the junction flow separation and zones of recirculation were found. The analysis of oxygen transport illustrates a correlation between zones of low wall shear stress and reduced oxygen flux into the wall. Wall mechanics show that increased compliance mismatch leads to increased and discontinuous intramural stresses. Comparison to histomorphological findings on intimal hyperplasia shows certain correlations, particularly increased compliance mismatch has a proliferate influence on suture line hyperplasia. The reduction of compliance mismatch using vein interposition results in decreased generation of intimal hyperplasia, and therefore, contributes to improvement of patency rates, while the geometrical modification and the resulting change of the flow pattern seems to be less important for the growth of anastomotic intimal hyperplasia.


Does changing to brighter road lighting improve road safety? Multilevel longitudinal analysis of road traffic collision frequency during the relighting of a UK city.
A step change in the night environment is taking place, with the large-scale installation of bright, broad-spectrum road lighting such as white light-emitting diodes (LEDs). One justification for this is a reduction in road traffic collisions (RTCs). This study aimed to estimate the effect of new lighting on personal injury RTCs within a large UK city. We analysed a 9-year time series of weekly RTC personal injury counts in 132 areas of the city using multilevel modelling. The RTC rate over a full 24-hour period was the primary outcome; darkness and daylight RTC rates were secondary. The background change in RTC rate was separated from the change associated with the number of newly installed bright lamps by including a polynomial underlying time trend for the logarithm of the mean number of collisions per week for each area. The study was based on a rigorous, predesigned and archived protocol. Within-area coefficients for the broad lighting effect were positive; as the number of bright lamps in an area increased, so did the RTC rate. The estimate for the increase in the within-area 24-hour RTC rate is 11% (95% CI 2% to 20%). The estimate of darkness-only RTCs is 16% (95% CI 2% to 32%). If the effect of lighting on darkness RTC rate is adjusted by that for daylight, one obtains 4% (95% CI -12% to +23%). No evidence was found for bright lamps leading to an improvement in road safety in any of the analyses. For this city, introducing brighter road lighting may have compromised safety rather than reducing harm.


Effects of Exercise and Sport in Solid Organ Transplant Recipients: A Review.
Solid organ transplantation is the criterion standard treatment for many with end-organ failure and can offer a new independence from the burden of disease. However solid organ transplant recipients (SOTRs) remain at high risk of cardiovascular (CV) disease, and poor quality of life and physical functioning. Increasing physical activity and exercise can improve the health of the general population; however, the effects on those with a transplant remain unclear. Intensive exercise and sporting activity has the potential to be beneficial, although there remain concerns particularly around the effects on immune function and the CV system. This review summarizes what is known about the effects of exercise on determinants of health in SOTRs and then collates the available literature investigating the consequences of intensive exercise and sport on the health of SOTR. There is a paucity of high-quality research, with most evidence being case studies or anecdotal; this is understandable given the relatively few numbers of SOTRs who are performing sport and exercise at a high level. However, if suitable evidence-based guidelines are to be formed and SOTRs are to be given reassurances that their activity levels are not detrimental to their transplanted organ and overall health, then more high-quality studies are required.


Inferring predominant pathways in cellular models of breast cancer using limited sample proteomic profiling.
Molecularly targeted drugs inhibit aberrant signaling within oncogenic pathways. Identifying the predominant pathways at work within a tumor is a key step towards tailoring therapies to the patient. Clinical samples pose significant challenges for proteomic profiling, an attractive approach for identifying predominant pathways. The objective of this study was to determine if information obtained from a limited sample (i.e., a single gel replicate) can provide insight into the predominant pathways in two well-characterized breast cancer models. A comparative proteomic analysis of total cell lysates was obtained from two cellular models of breast cancer, BT474 (HER2+/ER+) and SKBR3 (HER2+/ER-), using two-dimensional electrophoresis and MALDI-TOF mass spectrometry. Protein interaction networks and canonical pathways were extracted from the Ingenuity Pathway Knowledgebase (IPK) based on association with the observed pattern of differentially expressed proteins. Of the 304 spots that were picked, 167 protein spots were identified. A threshold of 1.5-fold was used to select 62 proteins used in the analysis. IPK analysis suggested that metabolic pathways were highly associated with protein expression in SKBR3 cells while cell motility pathways were highly associated with BT474 cells. Inferred protein networks were confirmed by observing an up-regulation of IGF-1R and profilin in BT474 and up-regulation of Ras and enolase in SKBR3 using western blot. When interpreted in the context of prior information, our results suggest that the overall patterns of differential protein expression obtained from limited samples can still aid in clinical decision making by providing an estimate of the predominant pathways that underpin cellular phenotype.


Acute effects of pea protein and hull fibre alone and combined on blood glucose, appetite, and food intake in healthy young men--a randomized crossover trial.
Whether pulse components can be used as value-added ingredients in foods formulated for blood glucose (BG) and food intake (FI) control requires investigation. The objective of this study was to examine of the effects of pea components on FI at an ad libitum meal, as well as appetite and BG responses before and after the meal. In a repeated-measures crossover trial, men (n = 15) randomly consumed (i) pea hull fibre (7 g), (ii) pea protein (10 g), (iii) pea protein (10 g) plus hull fibre (7 g), (iv) yellow peas (406 g), and (v) control. Pea hull fibre and protein were served with tomato sauce and noodles, while yellow peas were served with tomato sauce. Control was noodles and tomato sauce. FI was measured at a pizza meal (135 min). Appetite and BG were measured pre-pizza (0-135 min) and post-pizza (155-215 min). Protein plus fibre and yellow peas led to lower pre-pizza BG area under the curve compared with fibre and control. At 30 min, BG was lower after protein plus fibre and yellow peas compared with fibre and control, whereas at 45 and 75 min, protein plus fibre and yellow peas led to lower BG compared with fibre (p < 0.05). Following the pizza meal (155 min), yellow peas led to lower BG compared with fibre (p < 0.05). No differences were observed in FI or appetite. This trial supports the use of pea components as value-added ingredients in foods designed to improve glycemic control.


Protein kinase C activation causes neurite retraction via cyclinD1 and p60-katanin increase in rat hippocampal neurons.
Neurons are differentiated postmitotic cells residing in G0 phase of the cell cycle and are unable to proceed through G1 phase, in which cyclinD1 needs to be up-regulated for initiation. Yet, a growing body of evidence has shown that cell cycle re-activation via cyclinD1 up-regulation drives neurons into apoptosis. By contrast, there is also evidence demonstrating cell cycle proteins playing roles in neuronal differentiation. cyclinD1 has been shown to be differently regulated by protein kinase C alpha (PKC-α) in various mitotic cells. Based on these different effects, we investigated the role of PKC-α on cyclinD1 regulation in hippocampal neurons. Neurons were treated with PKC activator, PMA, and analysed for subcellular distributions of PKC-α and cyclinD1. Remarkably, PMA treatment increased nuclear PKC-α and cyclinD1, but not PKC-ε in hippocampal neurons. Increases in nuclear PKC-α and cyclinD1 were accompanied by microtubule re-organisation via increases in tau and retinoblastoma protein phosphorylation levels. Increased p60-katanin and p53 changed the neuronal morphology into neurons with shorter, but increased number of side branches. Since up-regulation of cell cycle is associated with apoptosis in neurons, we also analysed changes in Bax, Bcl-2 early and PARP (poly(ADP-ribose)polymerase), caspase3 late apoptotic markers. However, we did not observe any indication of apoptosis. These data suggest that in addition to their previously known roles in mitotic cells on cell cycle regulation, PKC-α and cyclinD1 seem to be important for differentiation, and nuclear PKC-α and cyclinD1 interfere with differentiation by promoting microtubule re-organisation through PKC signaling without triggering apoptosis.


Bio-inspired Carbon Hole Transporting Layer Derived from Aloe Vera Plant for Cost-Effective Fully Printable Mesoscopic Carbon Perovskite Solar Cells.
Herein, we introduce a new ecofriendly naturally extracted cross-linked carbon nanoparticles as a hole transporting layer (C-HTL) prepared by an ancient Indian method for carbon based printable mesoscopic perovskite solar cells (C-PSCs), which is low-cost so far used for fully printable PSCs. The fabricated PSCs having Glass/FTO/mp-TiO2/ZrO2/perovskite/AV-C configuration exhibited current density ( JSC) of 20.50 ± 0.5 mAcm-2, open circuit voltage ( VOC) of 0.965 ± 0.02 V and fill factor (FF) of 58 ± 2%, resulting in 12.3 ± 0.2% power conversion efficiency (PCE) for MAPbI3 perovskite absorber. The aloe-vera processed carbon C-HTL based PSCs yields up to 12.50% power conversion efficiency and 15.80% efficiency for conventional spiro-MeOTAD based HTM. The air and moisture stability >1000 h at >45% relative humidity (RH) for cross-linked AV-C nanoparticle-based PSCs. This stability is very high compared to conventional spiro-MeOTAD HTM-based PSCs. The prepared carbon nanoparticles facilitate an excellent penetration of perovskite absorber in triple-layer-based scaffold, which enables a high-quality perovskite crystal and results in high PCE. This novel bio-inspired AV-C cross-linked nanoparticle-based natural carbon C-HTL is low-cost until date. We believe this technique would be suitable for and helpful toward fully printable and air-moisture-stable PSCs.


A donor-acceptor (D-A) conjugated polymer for fast storage of anions.
Organic electrode materials have attracted a lot interest in batteries in recent years. However, most of them still suffer from low performance such as low electrode potential, slow reaction kinetics, and short cycle life. In this work, we report a strategy of fabricating donor-acceptor (D-A) conjugated polymers for facilitating the charge transfer and therefore accelerating the reaction kinetics by using the copolymer (p-TTPZ) of dihydrophenazine (PZ) and thianthrene (TT) as a proof-of-concept. The D-A conjugated polymer as p-type cathode could store anions and exhibited high discharge voltages (two plateaus at 3.82 V, 3.16 V respectively), a reversible capacity of 152 mAh g-1 at 0.1 A g-1, excellent rate performance with a high capacity of 124.2 mAh g-1 at 10 A g-1 (~50C) and remarkable cyclability. The performance, especially the rate capability was much higher than that of its counterpart homopolymers without D-A structure. As a result, the p-TTPZ//graphite full cells showed a high output voltage (3.26 V), a discharge specific capacity of 139.1 mAh g-1 at 0.05 A g-1 and excellent rate performance. This work provides a novel strategy for developing high performance organic electrode materials through molecular design and will pave a way towards high energy density organic batteries.


Divergent cortical generators of MEG and EEG during human sleep spindles suggested by distributed source modeling.
Sleep spindles are approximately 1-second bursts of 10-15 Hz activity, occurring during normal stage 2 sleep. In animals, sleep spindles can be synchronous across multiple cortical and thalamic locations, suggesting a distributed stable phase-locked generating system. The high synchrony of spindles across scalp EEG sites suggests that this may also be true in humans. However, prior MEG studies suggest multiple and varying generators. We recorded 306 channels of MEG simultaneously with 60 channels of EEG during naturally occurring spindles of stage 2 sleep in 7 healthy subjects. High-resolution structural MRI was obtained in each subject, to define the shells for a boundary element forward solution and to reconstruct the cortex providing the solution space for a noise-normalized minimum norm source estimation procedure. Integrated across the entire duration of all spindles, sources estimated from EEG and MEG are similar, diffuse and widespread, including all lobes from both hemispheres. However, the locations, phase and amplitude of sources simultaneously estimated from MEG versus EEG are highly distinct during the same spindles. Specifically, the sources estimated from EEG are highly synchronous across the cortex, whereas those from MEG rapidly shift in phase, hemisphere, and the location within the hemisphere. The heterogeneity of MEG sources implies that multiple generators are active during human sleep spindles. If the source modeling is correct, then EEG spindles are generated by a different, diffusely synchronous system. Animal studies have identified two thalamo-cortical systems, core and matrix, that produce focal or diffuse activation and thus could underlie MEG and EEG spindles, respectively. Alternatively, EEG spindles could reflect overlap at the sensors of the same sources as are seen from the MEG. Although our results generally match human intracranial recordings, additional improvements are possible and simultaneous intra- and extra-cranial measures are needed to test their accuracy.


Long peptide vaccination can lead to lethality through CD4+ T cell-mediated cytokine storm.
The optimization of anticancer therapeutic vaccines can lead to better efficacy but also to stronger adverse effects. In a mouse model of antitumor vaccination using a long peptide (LP), which included MHC class I- and II-restricted male (H-Y) epitopes, we observed unexpected mortality. Mice with an increased frequency of anti-H-Y CD4 T cells were primed with LP+CpG and boosted 10 d later. Within hours of boost, they displayed shock-like signs with high mortality. Serum cytokine levels were high. TNF-alpha secreted by the CD4 T cells was identified as the key effector molecule. Priming with a short peptide (SP), which included the MHC class II-restricted epitope, was a more efficient primer than LP, but did not lead to mortality when used as boost. The high mortality induced by LP compared with SP was probably related to its specific ability to be presented by B cells. Finally, targeting the LP sequence to dendritic cells allowed tumor protection without side effects. Our data: 1) confirm that the immune system can be very dangerous; 2) caution against the use of systemic activation of high-frequency Ag-specific T cells as induced by high doses of LP; and 3) underline the benefit of targeting Ag to dendritic cells.


Topical gabapentin solution for the management of burning mouth syndrome: A retrospective study.
The aim of this retrospective study was to evaluate the effectiveness and safety of topical gabapentin solution (250 mg/mL) for the management of burning mouth syndrome (BMS). A retrospective chart review was conducted of all patients diagnosed with BMS and managed with gabapentin 250 mg/mL solution (swish and spit) between January 2021 and October 2022. Patient-reported outcomes included changes in burning score ranked on a 10-point numeric rating scale (NRS) and reported adverse drug reactions (ADR). Wilcoxon signed-rank test was used to assess differences in the oral burning score ranked on a NRS (0-10) between the baseline visit and the second visit. A total of 19 patients (68.4% females) with BMS were included and evaluated for follow-up at a median of 86 days (range: 29-195). Overall, patients reported a median 2-point burning decrease on a 0-10 NRS between the baseline visit and the second visit (p < 0.01). ADRs were reported by 3 patients (15.8%). Although this was a small retrospective study, BMS management with topical gabapentin (250 mg/mL) appears to be effective and well-tolerated. Future randomized prospective studies are needed to verify these preliminary findings.


Antibodies against potassium channel interacting protein 2 induce necrosis in isolated rat cardiomyocytes.
Auto-antibodies against cardiac proteins have been described in patients with dilated cardiomyopathy. Antibodies against the C-terminal part of KChIP2 (anti-KChIP2 [C-12]) enhance cell death of rat cardiomyocytes. The underlying mechanisms are not fully understood. Therefore, we wanted to explore the mechanisms responsible for anti-KChIP2-mediated cell death. Rat cardiomyocytes were treated with anti-KChIP2 (C-12). KChIP2 RNA and protein expressions, nuclear NF-κB, mitochondrial membrane potential Δψm, caspase-3 and -9 activities, necrotic and apoptotic cells, total Ca(2+) and K(+) concentrations, and the effects on L-type Ca(2+) channels were quantified. Anti-KChIP2 (C-12) induced nuclear translocation of NF-κB. Anti-KChIP2 (C-12)-treatment for 2 h significantly reduced KChIP2 mRNA and protein expression. Anti-KChIP2 (C-12) induced nuclear translocation of NF-κB after 1 h. After 6 h, Δψm and caspase-3 and -9 activities were not significantly changed. After 24 h, anti-KChIP2 (C-12)-treated cells were 75 ± 3% necrotic, 2 ± 1% apoptotic, and 13 ± 2% viable. Eighty-six ± 1% of experimental buffer-treated cells were viable. Anti-KChIP2 (C-12) induced significant increases in total Ca(2+) (plus 11 ± 2%) and K(+) (plus 18 ± 2%) concentrations after 5 min. Anti-KChIP2 (C-12) resulted in an increased Ca(2+) influx through L-type Ca(2+) channels. In conclusion, our results suggest that anti-KChIP2 (C-12) enhances cell death of rat cardiomyocytes probably due to necrosis.


Hemodynamic correlates of the third heart sound and systolic time intervals.
Bedside diagnostic tools remain important in the care of patients with heart failure. Over the past two centuries, cardiac auscultation and phonocardiography have been essential in understanding cardiac pathophysiology and caring for patients with heart disease. Diastolic heart sounds (S3 and S4) and systolic time intervals have been particularly useful in this regard. Unfortunately, auscultation skills have declined considerably, and systolic time intervals have traditionally required carotid pulse tracings. Newer technology allows the automated detection of heart sounds and measurement of systolic time intervals in a simple, inexpensive, noninvasive system. Using the newer system, the authors present data on the hemodynamic correlates of the S3 and abnormal systolic time intervals. These data serve as the foundation for using the system to better understand the test characteristics and pathophysiology of the S3 and systolic time intervals, and help to define their use in improving the bedside diagnosis and management of patients with heart failure.


Teaching Initiated Question Asking to Children with Autism Spectrum Disorder Through a Short-Term Parent-Mediated Program.
This study investigated whether a brief parent-mediated intervention would increase the frequency of question asking in children with ASD. Mothers participated in a 3-week training consisting of 2-h sessions twice weekly. Data were collected in the context of concurrent multiple baseline design. Results demonstrate all three children increased frequency of question asking with two children maintaining gains. All three children demonstrated generalization of question asking to novel items, family members, and/or settings. Affect improved for two of the three children. Overall, mothers were able to reach Fidelity of Implementation during most sessions and rated the intervention as highly acceptable. Results are discussed in regard to the feasibility of providing a short-term parent-implemented intervention to increase social initiations through question asking.


[The people of the black waters: the Amazon caboclo of the Negro river].
The article constructs a historically contextualized description of the people who live along the Negro river, a Brazilian affluent in the Amazon basin. Drawing on information about the daily social experience of the participants from the dawn of the twentieth century through the mid-1990s, the processes by which the population and communities took shape are identified. On the Negro river, contact between Brazilian society and the autochthonous, catechized indigenous groups living there was determinant in shaping the territory's caboclo identity. Starting in the last quarter of the nineteenth century, this nomenclature took root and entered the popular lexicon. Extractivist activities played a major role in spreading the term, within a context where the predominant social relations derived from the 'cultura do barracão'.


Fatal scorpion sting in a child.
Fatal scorpion stings are rare in Nigeria. Hitherto, there has been no report from Nigeria of death following scorpion stings. This report is that of a 2-year-old boy who was stung by a scorpion while playing outside his home environment in Osogbo, South West Nigeria. He subsequently presented to the Children Emergency Unit of Ladoke Akintola University of Technology Teaching Hospital, Osogbo, in pain and with features of shock. He died within 2 h of admission despite all treatment given to relieve pain and manage shock. The case is reported in order to share the important lessons learned.


Paired box 6 gene delivery preserves beta cells and improves islet transplantation efficacy.
Loss of pancreatic beta cells is the central feature of all forms of diabetes. Current therapies fail to halt the declined beta cell mass. Thus, strategies to preserve beta cells are imperatively needed. In this study, we identified paired box 6 (PAX6) as a critical regulator of beta cell survival. Under diabetic conditions, the human beta cell line EndoC-βH1, db/db mouse and human islets displayed dampened insulin and incretin signalings and reduced beta cell survival, which were alleviated by PAX6 overexpression. Adeno-associated virus (AAV)-mediated PAX6 overexpression in beta cells of streptozotocin-induced diabetic mice and db/db mice led to a sustained maintenance of glucose homeostasis. AAV-PAX6 transduction in human islets reduced islet graft loss and improved glycemic control after transplantation into immunodeficient diabetic mice. Our study highlights a previously unappreciated role for PAX6 in beta cell survival and raises the possibility that ex vivo PAX6 gene transfer into islets prior to transplantation might enhance islet graft function and transplantation outcome.


Integrating Curriculum-Based Dynamic Assessment in Computerized Adaptive Testing: Development and Predictive Validity of the EDPL-BAI Battery on Reading Competence.
In recent decades there have been significant changes in the conceptualization of reading as well as in the perception of how this activity should be assessed. Interest in the analysis of reading processes has led to the emergence of new explanatory models based primarily on the contributions of cognitive psychology. In parallel, there have been notable advances in measurement procedures, especially in models based on Item Response Theory (IRT), as well as in the capacity and performance of specific software programs that allow data to be managed and analyzed. These changes have contributed significantly to the rise of testing procedures such as computerized adaptive tests (CATs), whose fundamental characteristic is that the sequence of items presented in the tests is adapted to the level of competence that the subject manifests. Likewise, the incorporation of elements of dynamic assessment (DA) as the prompts are gradually offered allows for obtaining information about the type and degree of support required to optimize the subject's performance. In this sense, the confluence of contributions from DA and CATs offers a new possibility for approaching the assessment of learning processes. In this article, we present a longitudinal research developed in two phases, through which a computerized dynamic adaptive assessment battery of reading processes (EDPL-BAI) was configured. The research frame involved 1,831 students (46% girls) from 13 public schools in three regions of Chile. The purpose of this study was to analyze the differential contribution on reading competence of dynamic scores obtained in a subsample composed of 324 (47% girls) students from third to sixth grade after the implementation of a set of adaptive dynamic tests of morpho-syntactic processes. The results achieved in the structural equation modeling indicate a good global fit. Individual relationships show a significant contribution of calibrated score that reflects estimated knowledge level on reading competence, as well as dynamic scores based on the assigned value of graduated prompts required by the students. These results showed significant predictive values on reading competence and incremental validity in relation to predictions made by static criterion tests.


[Molecular characterization and phylogenetic analysis of enteroviruses inducing the outbreaks and seasonal rises of morbidity in different regions of the Republic of Belarus].
As shown by the results of the analysis, viruses ECHO 30 circulating over the period of the last 8 years in Belarus, belonged to 3 different genetic subtypes which earlier or simultaneously circulated in other European states. The outbreaks of enterovirus infections (EVI) were facilitated by the appearance of a genetic viral subtype, relatively "new" for human population, and which had not earlier circulated on this territory. Thus, the development of outbreak morbidity in 2003 was caused by a change in the dominating subtype of virus ECHO 30, which caused the outbreak of 1997. The relatively "soft" rise of morbidity in 2004 was due to the continued circulation of the same subtype of virus ECHO 30, that in 2003. The largest outbreaks of EVI in the Republic of Belarus had a number of considerable differences: the outbreak of 1997 in Gomel was characterized by the genetic heterogeneity of infective agents, being simultaneously geographically localized within the limits of one city. However, during the outbreaks of 2003 the circulation of genetically closely related viruses of the one subtype among the population of geographically remote regions of the country was registered.


Autophagy and ER-stress contribute to photoreceptor degeneration in cultured adult porcine retina.
The aim of this study was to investigate rod and cone photoreceptor degeneration in organotypic cultures of adult porcine retina. Our hypothesis was that the photoreceptors accumulate opsins, which, together with exposure to cyclic dim light illumination, induce autophagy and endoplasmic reticulum stress (ER-stress) to overcome damaging protein overload. For this purpose, retinas were cultured for 48 h and 72 h during which they were illuminated with dim light for 8h/day; specimens were analyzed by means of immunohistochemistry, Western blot, real-time polymerase chain reaction (PCR) and transmission electron microscopy. ER-stress and photoreceptor degeneration was observed in conventionally cultured retinas. The additional stress in the form of dim light illumination for 8h/day resulted in increased levels of the ER-stress markers GRP78/BiP and CHOP, as well as increased level of active caspase-12. Increased autophagic processes in cone and rod photoreceptors were detected by LC3B-II increases and occurrence of autophagosomes at the ultrastructural level. Illumination also resulted in altered protein expression for autophagy inducers such as p62 and Beclin-1. Moreover, there was a decrease in phosphorylated mammalian target of rapamycin (mTOR), which further indicate an increase of autophagy. Rod and cone photoreceptors in retinas from a diurnal animal that were exposed to dim light illumination in vitro displayed autophagy and ER-stress processes. As no alteration of rhodopsin mRNA was observed, autophagy and ER-stress are suggested to decrease rhodopsin protein at the posttranscriptional level.


Early extubation versus prophylactic ventilation in the high risk patient: a comparison of postoperative management in the prevention of respiratory complications.
To evaluate whether prophylactic ventilation during the early postoperative period diminishes pulmonary complications, 35 high risk, elderly patients undergoing major, elective abdominal aortic reconstruction were prospectively randomized into either an early extubation group or a prophylactic ventilation group. The 17 patients assigned to the prophylactic ventilation group received mechanical ventilation by assist/control mode until 8 a.m. of the first postoperative day. The 17 patients assigned to the early extubation group were extubated after the operation as soon as they could maintain a pH of 7.35, with a spontaneous respiratory rate of less than 30. Preoperative measurements of functional residual capacity, intrapulmonary shunt, and oxygen delivery were compared to similar measurements during the initial two postoperative days. There were no significant differences between the groups with respect to age, length of operation, duration of anesthesia, operative blood loss, intraoperative fluid administration, or number of intraoperative transfusions. Patients in the prophylactic ventilation group were ventilated for an average of 18.3 +/- 0.5 hours. Patients in the early extubation group were ventilated for an average of 3.3 +/- 0.5 hours (p < 0.0005). No patient in either group required reintubation. Intrapulmonary shunt and oxygen delivery were not significantly different between the groups at any time during the study period. There was no mortality or significant morbidity in either group. These findings suggest that in high risk surgical patients, prophylactic ventilation, per se, may not diminish respiratory complications or improve gas exchange.


Flavor perception in human infants: development and functional significance.
Foods people consume impact on their health in many ways. In particular, excess intake of salty, sweet and fatty foods and inadequate intake of fruits and vegetables have been related to many diseases including diabetes, hypertension, cardiovascular disease and some cancers. The flavor of a food determines its acceptability and modulates intake. It is thus critical to understand the factors that influence flavor preferences in humans. To outline several of the important factors that shape flavor preferences in humans. We review a series of studies, mainly from our laboratories, on the important role of early experiences with flavors on subsequent flavor preference and food intake. Some taste preferences and aversions (e.g. liking for sweet, salty and umami; disliking for bitter) are innately organized, although early experiences can modify their expression. In utero events may impact on later taste and flavor preferences and modulate intake of nutrients. Both before and after birth, humans are exposed to a bewildering variety of flavors that influence subsequent liking and choice. Fetuses are exposed to flavors in amniotic fluid modulating preferences later in life and flavor learning continues after birth. Experience with flavors that are bitter, sour or have umami characteristics, as well as volatile flavors such as carrot and garlic, occurs through flavorings in breast milk, infant formula and early foods. These early experiences mold long-term food and flavor preferences which can impact upon later health.


N-methyl-D-aspartate antagonists prevent kainate neurotoxicity in rat retinal ganglion cells in vitro.
Under defined culture conditions, exogenous glutamate (Glu), NMDA, or an endogenous Glu-related toxin is lethal to rat retinal ganglion cells; these detrimental effects are NMDA receptor mediated because specific NMDA antagonists can prevent cellular injury. In the presence of an endogenous Glu-like toxin, 125 microM kainate (KA) increases the proportion of retinal ganglion cells that die, but the toxicity (due to both KA and the endogenous toxin) is totally prevented by 2-amino-5-phosphonovalerate (APV), a specific NMDA receptor antagonist. These findings indicate that the KA-induced portion of retinal ganglion cell death also appears to be mediated via NMDA receptors. There are at least 2 possible mechanisms for this lethal effect. In addition to KA receptors, KA could directly stimulate NMDA receptors. Alternatively, KA might activate its own specific receptor, which in turn leads to a net increase in the release of an endogenous Glu-related toxin; this endogenous substance would then activate NMDA receptors. Patch-clamp electrophysiology experiments have helped to distinguish between these possibilities. Concentrations of APV that completely block the current elicited by maximal nondesensitizing doses of NMDA exert no detectable inhibition of KA-evoked currents. Hence, at the concentrations used, it appears unlikely that KA directly activates NMDA receptors in this preparation. Furthermore, the fraction of toxicity attributed to the addition of KA can be blocked by the relatively specific non-NMDA antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX). This finding is consistent with the hypothesis that KA adds an increment of toxicity in this system by directly interacting with KA receptors.(ABSTRACT TRUNCATED AT 250 WORDS)


Optimization and simplification of antiretroviral therapy for adults and children.
The review reflects on opportunities and challenges for HIV treatment optimization for the next 5 years. Considering all currently available options, the fixed-dose combination of tenofovir + lamivudine (or emtricitabine) + efavirenz is considered as the best option for first-line treatment for the short to medium term. Second-line therapy will likely continue to be comprised of a boosted protease inhibitor in combination with two nucleoside/nucleotide reverse transcriptase inhibitors (NRTIs), with potential for combining with integrase inhibitors. For children, there is potential for simplification and harmonization with adult antiretroviral regimens. First-line therapy for children younger than 3 years of age may be best delivered using two nucleoside reverse transcriptase inhibitors (NRTIs) and a boosted protease inhibitor; above 3 years of age, the standard of care is two NRTIs and a non-nucleoside reverse transcriptase inhibitor (NNRTI) as recommended for adults. Important research questions include the dosing and safety of new antiretroviral agents and formulations, particularly once-daily fixed-dose combinations, the role of integrase inhibitors and the optimal second-line regimen for NNRTI-exposed children who fail protease inhibitor-containing first-line regimens. Treatment simplification is critical to further antiretroviral therapy scaling-up and support long-term retention in care. Future guidance should consider the broader benefits of earlier antiretroviral therapy initiation beyond potential AIDS mortality reduction, notably mitigation of short- and long-term HIV-associated comorbidities, reduction of HIV transmission, increased retention in care, and enhancing programme simplification.


BET Bromodomain Inhibitors with One-Step Synthesis Discovered from Virtual Screen.
Chemical inhibition of epigenetic regulatory proteins BrdT and Brd4 is emerging as a promising therapeutic strategy in contraception, cancer, and heart disease. We report an easily synthesized dihydropyridopyrimidine pan-BET inhibitor scaffold, which was uncovered via a virtual screen followed by testing in a fluorescence anisotropy assay. Dihydropyridopyimidine 3 was subjected to further characterization and is highly selective for the BET family of bromodomains. Structure-activity relationship data and ligand deconstruction highlight the importance of the substitution of the uracil moiety for potency and selectivity. Compound 3 was also cocrystallized with Brd4 for determining the ligand binding pose and rationalizing subsequent structure-activity data. An additional series of dihydropyridopyrimidines was synthesized to exploit the proximity of a channel near the ZA loop of Brd4, leading to compounds with submicromolar affinity and cellular target engagement. Given these findings, novel and easily synthesized inhibitors are being introduced to the growing field of bromodomain inhibitor development.


Is Computerized Working Memory Training Effective in Healthy Older Adults? Evidence from a Multi-Site, Randomized Controlled Trial.
Developing effective interventions to attenuate age-related cognitive decline and prevent or delay the onset of dementia are major public health goals. Computerized cognitive training (CCT) has been marketed increasingly to older adults, but its efficacy remains unclear. Working memory (WM), a key determinant of higher order cognitive abilities, is susceptible to age-related decline and a relevant target for CCT in elders. To evaluate the efficacy of CCT focused on WM compared to an active control condition in healthy older adults. Eighty-two cognitively normal adults from two sites (USA and Sweden) were randomly assigned to Cogmed Adaptive or Non-Adaptive (active control) CCT groups. Training was performed in participants' homes, five days per week over five weeks. Changes in the performance of the Cogmed trained tasks, and in five neuropsychological tests (Trail Making Test Part A and Part B, Digit Symbol, Controlled Oral Word Association Test and Semantic Fluency) were used as outcome measures. The groups were comparable at baseline. The Adaptive group showed robust gains in the trained tasks, and there was a time-by-group interaction for the Digit Symbol test, with significant improvement only after Adaptive training. In addition, the magnitude of the intervention effect was similar at both sites. Home-based CCT Adaptive WM training appears more effective than Non-Adaptive training in older adults from different cultural backgrounds. We present evidence of improvement in trained tasks and on a demanding untrained task dependent upon WM and processing speed. The benefits over the active control group suggest that the Adaptive CCT gains were linked to providing a continuously challenging level of WM difficulty.


Triple therapy with budesonide/glycopyrrolate/formoterol fumarate with co-suspension delivery technology versus dual therapies in chronic obstructive pulmonary disease (KRONOS): a double-blind, parallel-group, multicentre, phase 3 randomised controlled trial.
Inhaled corticosteroids have been used in patients with chronic obstructive pulmonary disease (COPD), but the potential benefits of their use in triple therapy are not well known. We aimed to compare the efficacy of a triple therapy with corresponding dual therapies in symptomatic patients with moderate to very severe COPD, without a requirement for a history of exacerbations. In this double-blind, parallel-group, multicentre phase 3 randomised controlled trial, we recruited patients from hospitals and care centres in Canada, China, Japan, and the USA. Eligible patients were 40-80 years of age, were current or former smokers (with a smoking history of ≥10 pack-years), had an established clinical history of COPD, and were symptomatic for COPD, despite receiving two or more inhaled maintenance therapies for at least 6 weeks before screening. We randomly assigned patients (2:2:1:1) using an interactive web response system to receive budesonide/glycopyrrolate/formoterol fumarate metered-dose inhaler 320/18/9·6 μg (BGF MDI), glycopyrrolate/ formoterol fumarate metered-dose inhaler 18/9·6 μg (GFF MDI), budesonide/formoterol fumarate metered-dose inhaler 320/9·6 μg (BFF MDI), or open-label budesonide/formoterol fumarate dry-powder inhaler 400/12 μg (BUD/ FORM DPI). Primary endpoints for the Europe/Canada statistical analysis approach were FEV1 area under the curve from 0-4 h (AUC0-4) for BGF MDI versus BFF MDI and BGF MDI versus BUD/FORM DPI over 24 weeks; and change from baseline in morning pre-dose trough FEV1 for BGF MDI versus GFF MDI and non-inferiority of BFF MDI versus BUD/FORM DPI (margin of -50 mL from lower bound of 95% CI) over 24 weeks. Comparisons with BUD/FORM DPI were made for the Europe/Canada statistical analysis approach only. This study is registered with ClinicalTrials.gov, number NCT02497001. Between Aug 20, 2015, and Jan 5, 2018, 3047 patients were screened from 215 sites, and 1902 were randomly assigned to receive BGF MDI (n=640), GFF MDI (n=627), BFF MDI (n=316), or BUD/FORM DPI (n=319). Over 24 weeks, BGF MDI significantly improved FEV1 AUC0-4 versus BFF MDI (least squares mean difference 104 mL, 95% CI 77 to 131; p<0·0001) and BUD/FORM DPI (91 mL, 64 to 117; p<0·0001). BGF MDI also significantly improved pre-dose trough FEV1 versus GFF MDI (22 mL, 4 to 39; p=0·0139) and was non-inferior to BUD/FORM DPI (-10 mL, -36 to 16; p=0·4390). At week 24, patients in the BGF MDI group had a significantly improved FEV1 AUC0-4 compared with patients receiving BFF MDI (116 mL, 95% CI 80 to 152; p<0·0001); there was a non-significant improvement in the change from baseline in morning pre-dose trough FEV1 at week 24 versus GFF MDI (13 mL, -9 to 36 mL; p=0·2375). The most common treatment-emergent adverse events were nasopharyngitis (n=49 [8%] in the BGF MDI group; n=41 [7%] in the GFF MDI group; n=26 [8%] in the BFF MDI group; and n=30 [9%] in the BUD/FORM DPI group) and upper respiratory tract infection (n=65 [10%]; n=38 [6%]; n=18 [6%]; and n=22 [7%]). Pneumonia incidence was low (<2%) and similar across treatments. There were two treatment-related deaths, both in the GFF MDI group. BGF MDI was efficacious, well tolerated, and could be a more appropriate treatment than the corresponding dual therapies for symptomatic patients with moderate to very severe COPD, irrespective of exacerbation history. Pearl-a member of the AstraZeneca Group.


Interactions of surfactants with a water treatment protein from Moringa oleifera seeds in solution studied by zeta-potential and light scattering measurements.
Protein extracted from Moringa oleifera (MO) seeds has been advocated as a cheap and environmental friendly alternative to ionic flocculants for water purification. However, the nature and mechanism of its interaction with particles in water, as well as with dissolved surface-active molecules, are not well understood. In this article, we report studies of the protein and its interaction with four surfactants using dynamic light scattering (DLS), zeta-potential and turbidity measurements. Zeta-potential measurements identified points of charge reversal and the turbidity and DLS measurements were used to characterize the microstructure and size of protein-surfactant complexes. From the points of charge reversal, it was estimated that 7 anions are required to neutralize the positive charges of each protein molecule at pH 7. For protein mixtures with sodium dodecyl sulfate and dodecyl di-acid sodium salt, the peak in turbidity corresponds to concentrations with a large change in zeta-potential. No turbidity was observed for protein mixtures with either the nonionic surfactant Triton X-100 or the zwitterionic surfactant N-dodecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate. Changes of pH in the range 4-10 have little effect on the zeta-potential, turbidity, and the hydrodynamic radius reflecting the high isoelectric point of the protein. Addition of small amounts of salt has little effect on the size of protein in solution. These results are discussed in the context of the use of the MO protein in water treatment.