Protective effects of broccoli (Brassica oleracea) against oxidative damage in vitro and in vivo.
The antioxidative effect and protective potential against diabetes of the broccoli flower were investigated both in vitro and in a diabetic rat model. Among fractions of MeOH, CH2Cl2, BuOH, and H2O, the BuOH fraction exerted the strongest inhibitory activities on 1,1-diphenyl-2-picrylhydrazyl radical, radical-induced protein oxidation, and nitric oxide generation by sodium nitroprusside. The in vitro results suggest that the BuOH fraction from the broccoli flower has a protective potential against oxidative stress. The rat model with diabetes induced by streptozotocin was employed to evaluate the protective effect of the BuOH fraction in vivo. Diabetic rats showed reduced body weight gain and heavier kidney and liver weights than normal rats, while oral administration of the BuOH fraction at an oral dose of 100 or 200 mg/kg body weight/d for 20 d attenuated the physiological changes induced by diabetes. In addition, oral administration of the BuOH fraction to diabetic rats led to significant decreases in serum glucose and glycosylated protein, while it resulted in the increase of serum albumin, implying that the BuOH fraction improves the abnormal metabolism of glucose and protein that leads to oxidative stress. Moreover, it significantly reduced thiobarbituric acid-reactive substance levels in serum, hepatic and renal mitochondria. This suggests that the BuOH fraction would alleviate the oxidative stress associated with diabetes through the inhibition of lipid peroxidation. The present study demonstrates that the BuOH fraction has an antioxidative effect in vitro and it protects against oxidative stress induced by diabetes in an in vivo model.


Markerless gene replacement in Escherichia coli stimulated by a double-strand break in the chromosome.
A simple and efficient gene replacement method, based on the recombination and repair activities of the cell, was developed. The method permits the targeted construction of markerless deletions, insertions and point mutations in the Escherichia coli chromosome. A suicide plasmid, carrying the mutant allele and the recognition site of meganuclease I- Sce I, is inserted into the genome by homologous recombination between the mutant and the wild-type (wt) alleles. Resolution of this cointegrate by intramolecular recombination of the allele pair results in either a mutant or a wt chromosome which can be distinguished by allele-specific PCR screening. The resolution process is stimulated by introducing a unique double-strand break (DSB) into the chromosome at the I- Sce I site. Cleavage by the nuclease not only enhances the frequency of resolution by two to three orders of magnitude, but also selects for the resolved products. The DSB-stimulated gene replacement method can be used in recombination-proficient E.coli cells, does not require specific growth conditions, and is potentially applicable in other microorganisms. Use of the method was demonstrated by constructing a 17-bp and a 62-kb deletion in the MG1655 chromosome. Cleavage of the chromosome induces the SOS response but does not lead to an increased mutation rate.


Dewetting-induced membrane formation by adhesion of amphiphile-laden interfaces.
We introduce an approach for forming bilayer polymer membranes by adhesion of amphiphile-laden interfaces. This adhesion is induced by a reduction of solvent quality for the amphiphilic diblock copolymers through selective evaporation of good solvent in the solvent mixture. By combining this membrane formation mechanism with a double-emulsion-templated approach for vesicle formation, we fabricate monodisperse polymersomes that exhibit excellent membrane uniformity, and structural stability, using a method that has high encapsulation efficiency. Moreover, we also show that the technique is versatile and can be applied to different block copolymers. The ability to direct the assembly of amphiphiles into a membrane creates new opportunities to engineer the structures of vesicles on the level of the individual bilayer leaflets.


[Diagnosis of concomitant ischemic heart disease and selection of surgical tactics in arteriosclerosis of major arteries of the lower extremities in relation to heart function in patients with extensive arteriosclerosis].
The results of examination of 74 patients treated in the clinic for ischemia of the lower extremities with concomitant ischemic heart disease (IHD) diagnosed by the data of clinico-instrumental investigations are presented. In 10.8% of the patients, IHD was diagnosed in clinical examination, in 89.2%--in performing the loading test. The authors consider that the volume of intervention on the aorto-iliac segment should correspond to a functional state of the myocardium, on the femoro-popliteal segment--to severity of a lesion at this region and is not restricted by functional state of the myocardium. To the patients with a functional reserve corresponding to a functional class IV, the performance of coronarography for definition of the expediency to perform one-stage intervention on the coronary bed and major arteries of the lower extremities is indicated.


fMRI activity in the medial temporal lobe during recognition memory as a function of study-test interval.
The phenomenon of temporally graded retrograde amnesia (loss of information acquired before the onset of amnesia) suggests that the hippocampus, and possibly other medial temporal lobe (MTL) structures, have a time-limited role in memory. In three experiments, we made a first attempt to use fMRI to assess activity in the hippocampal region (the CA fields of the hippocampus, the dentate gyrus, and the subiculum) and the adjacent parahippocampal gyrus (parahippocampal, entorhinal, and perirhinal cortices) during recognition memory testing as a function of study-test interval. Experiment 1 (n = 5) demonstrated activity in the hippocampal region and parahippocampal gyrus for targets relative to foils during recognition memory performance following a single study-test delay of about one-half hour. In Experiment 2, 15 participants studied line drawings at each of three different times prior to scanning: one-half hour, 1 day, and 1 week. fMRI data were then collected during recognition memory testing, using targets from all three delays and foils. While an overall effect of targets vs. foils was found in both the hippocampal region and the parahippocampal gyrus, there was no effect of study-test interval on target activity. In Experiment 3 (n = 13), behavioral performance (reaction time and accuracy) was equated across the three delays. Again, no effect of study-test interval was observed. It is possible that the time span sampled in our study (one-half hour to 1 week) was too short to observe changes in activity. Alternatively, activity in the MTL during memory testing may occur even when these structures are not necessary for retrieval.


A role for the rice homeobox gene Oshox1 in provascular cell fate commitment.
The vascular tissues of plants form a network of interconnected cell files throughout the plant body. The transition from a genetically totipotent meristematic precursor to different stages of a committed procambial cell, and its subsequent differentiation into a mature vascular element, involves developmental events whose molecular nature is still mostly unknown. The rice protein Oshox1 is a member of the homeodomain leucine zipper family of transcription factors. Here we show that the strikingly precise onset of Oshox1 gene expression marks critical, early stages of provascular ontogenesis in which the developmental fate of procambial cells is specified but not yet stably determined. This suggests that the Oshox1 gene may be involved in the establishment of the conditions required to restrict the developmental potential of procambial cells. In support of this hypothesis, ectopic expression of Oshox1 in provascular cells that normally do not yet express this gene results in anticipation of procambial cell fate commitment, eventually culminating in premature vascular differentiation. Oshox1 represents the first example of a transcription factor whose function can be linked to specification events mediating provascular cell fate commitment.


Out-of-the tropics or trans-tropical dispersal? The origins of the disjunct distribution of the gooseneck barnacle Pollicipes elegans.
Studying species with disjunct distributions allows biogeographers to evaluate factors controlling species ranges, limits on gene flow, and allopatric speciation. Here, we use phylogeographic and population genetic studies of the barnacle Pollicipes elegans to discriminate between two primary hypotheses about the origin of disjunct distributions of extra-tropical populations: trans-tropical stepping-stone colonization versus an out-of-the tropics origin. Nucleotide diversity peaked in the centre of the species' range in samples from El Salvador and was lower in samples from higher latitudes at Mexico and Peru. Haplotypes from El Salvador samples also had a deeper coalescent, or an older time to a most recent common ancestor. A deep phylogeographical break exists between Mexico and all samples taken to the south (El Salvador and Peru). Isolation-with-migration analyses showed no significant gene flow between any of the three regions indicating that the difference in genetic differentiation among all three regions is explained primarily by differences in population separation times. Approximate Bayesian Computation model testing found strong support for an out-of-the tropics origin of extra-tropical populations in P. elegans. We found little evidence consistent with a stepping-stone history of trans-tropical colonization, but instead found strong evidence for a tropical origin model for the largely disjunct distribution of P. elegans. Sea surface temperature and habitat suitability are likely mechanisms driving decline of populations in tropical regions, causing the disjunct distribution.


A perspective from experimental studies of burning mouth syndrome.
Burning mouth syndrome (BMS) is one of the most frequently seen idiopathic pain conditions in a dental setting. Peri- and postmenopausal women are most frequently affected, and patients who experience BMS complain of persistent burning pain mainly at the tip and the bilateral border of the tongue. Recent studies have assessed whether BMS is a neuropathic pain condition, based on morphologic changes in biopsied tongue specimens, and whether there are abnormal pain responses in patients with this disease. Somatosensory studies have reported some abnormal findings in sensory and pain detection thresholds with inconsistency; however, the most distinct finding was exaggerated responses to painful stimuli. Imaging and electrophysiologic studies have suggested the possibility of dysregulation of the pain-modulating system in the central nervous system, which may explain the enhanced pain responses despite the lack of typical responses toward quantitative sensory tests. Basic studies have suggested the possible involvement of neuroprotective steroids, although the underlying mechanisms of this condition have not been elucidated. Experimental studies are looking for preferable supportive therapies for BMS patients despite the obscure pathogenesis.


Rapid determination of advanced glycation end products of proteins using MALDI-TOF-MS and PERL script peptide searching algorithm.
Advanced glycation end products (AGEs), which are composed of various glucose or carbohydrate adducts, are thought to be responsible for several diabetic and age-related complications. However, to date, specific sites on proteins that are modified by AGEs remain largely unknown. We report here the use of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) to determine the type and localization of several AGEs formed in vitro on human beta-2-microglobulin (beta2M), and in vivo on type 2 ryanodine receptor calcium-release channel (RyR2), and sarco(endo)plasmic reticulum (SERCA2a). A PERL script algorithm, developed in-house, makes searching the relatively large amount of data generated by the MALDI-MS more manageable. The outstanding sensitivity of MALDI-TOF-MS coupled with the PERL script algorithm allows such an approach to be a very useful tool in detecting AGEs and other post-translational modifications. We believe that this method could be an important tool when searching for post-translational modifications on proteins.


Fruit and vegetable consumption and mortality from all causes, cardiovascular disease, and cancer: systematic review and dose-response meta-analysis of prospective cohort studies.
To examine and quantify the potential dose-response relation between fruit and vegetable consumption and risk of all cause, cardiovascular, and cancer mortality. Medline, Embase, and the Cochrane library searched up to 30 August 2013 without language restrictions. Reference lists of retrieved articles. Prospective cohort studies that reported risk estimates for all cause, cardiovascular, and cancer mortality by levels of fruit and vegetable consumption. Random effects models were used to calculate pooled hazard ratios and 95% confidence intervals and to incorporate variation between studies. The linear and non-linear dose-response relations were evaluated with data from categories of fruit and vegetable consumption in each study. Sixteen prospective cohort studies were eligible in this meta-analysis. During follow-up periods ranging from 4.6 to 26 years there were 56,423 deaths (11,512 from cardiovascular disease and 16,817 from cancer) among 833,234 participants. Higher consumption of fruit and vegetables was significantly associated with a lower risk of all cause mortality. Pooled hazard ratios of all cause mortality were 0.95 (95% confidence interval 0.92 to 0.98) for an increment of one serving a day of fruit and vegetables (P=0.001), 0.94 (0.90 to 0.98) for fruit (P=0.002), and 0.95 (0.92 to 0.99) for vegetables (P=0.006). There was a threshold around five servings of fruit and vegetables a day, after which the risk of all cause mortality did not reduce further. A significant inverse association was observed for cardiovascular mortality (hazard ratio for each additional serving a day of fruit and vegetables 0.96, 95% confidence interval 0.92 to 0.99), while higher consumption of fruit and vegetables was not appreciably associated with risk of cancer mortality. This meta-analysis provides further evidence that a higher consumption of fruit and vegetables is associated with a lower risk of all cause mortality, particularly cardiovascular mortality.


The effect of trimethoprim on macromolecular synthesis in Escherichia coli. Regulation of ribonucleic acid synthesis by 'Magic Spot' nucleotides.
During the inhibition of RC(str), but not RC(rel) mutants of Escherichia coli by trimethoprim the unusual nucleotides MSI (guanosine tetraphosphate, ppGpp) and MSII rapidly accumulated. The production of these nucleotides was not dependent on the addition of nucleotide base supplements to RC(str) cultures before trimethoprim, and the MSI nucleotide concentrations in non-supplemented or purine-supplemented cultures were comparable with the concentrations obtained when the cells were inhibited with l-valine (1g/l). Rifampicin rapidly decreased MSI and MSII nucleotide concentrations in trimethoprim-inhibited cultures to the basal values. Several situations were noted, in which MS nucleotide concentrations in trimethoprim-inhibited RC(str) cells could be drastically lowered without giving rise to an immediate resumption of stable RNA accumulation. If RC(str) mutants were first inhibited with trimethoprim and then given purines 15min later, MS nucleotide concentrations fell rapidly, because of a temporarily enhanced rate of accumulation of stable RNA. However, after a further 5min, RNA accumulation stopped, though MS nucleotide concentrations remained low. Also, if either glycine or methionine were added to trimethoprim-inhibited cultures supplemented with purines, RNA accumulation did not resume, though MS nucleotide concentrations rapidly declined. With both amino acids present, there was both a decline in MS nucleotide concentration and a resumption in stable RNA synthesis. These findings suggest that MSI nucleotide concentrations in trimethoprim-inhibited bacteria are not the sole factors in the control of stable RNA synthesis. It is possible that, during the period when the RC(str) cells contained high concentrations of MS nucleotides, some factor important in the MSI-mediated control of stable RNA synthesis was irreversibly inactivated. However, as antibiotics (e.g. chloramphenicol) both abolished high MS nucleotide concentrations and permitted a rapid resumption of stable RNA accumulation in the same conditions, it is more likely that an additional control mechanism has come into play.


Screening of SLC26A4 Gene Hotspots in 2673 Patients Associated with Sensorineural Hearing Loss in Northwestern China.
This study aimed to investigate the incidence of the hotspot mutations c.919-2A>G and c.2168A>G in SLC26A4 in the northwestern Chinese population. A total of 2673 unrelated patients were recruited from northwestern China, and clinical information was obtained from all patients. Peripheral blood samples were acquired to detect the genotype of each patient by direct sequencing. Statistical analysis was conducted with Statistical Package for the Social Sciences 19.0 software. Overall, 118 patients (4.4%) were identified with biallelic mutations, including 84 (3.14%) homozygotes and 34 (1.27%) compound heterozygotes. Moreover, significant differences between Han and Uighur were identified regarding the frequencies of c.919-2A>G homozygous and biallelic mutations. This model for the rapid screening of hotspot mutations can identify the molecular cause for 4.4% of patients with severe to profound sensorineural hearing loss in northwestern China, and there may be distinctive hotspot mutations in different ethnic populations.


Teaching Microbiology by Celebrating Traditional Foods and Cultures from Morocco and Perú.
According to a report published by the Center for Community College Student Engagement and others, global learning is considered one of the high-impact practices used to increase student engagement and motivation to learn. Engagement and motivation have also been linked to increased learning gains and improvement in the overall learning experience. Furthermore, global learning helps students explore other people's cultures and worldviews, which is an important skill for students to gain in order to compete and adapt to solve the problems of our global society. Here, I discuss two class activities that faculty can adopt to implement global learning in their courses, with the purpose of engaging and motivating students to learn microbiology while celebrating some traditions from Morocco and Perú. Students researched traditional fermented foods and drinks from Perú and Morocco. Then, they answered guided questions to help them link the food items to microbiological concepts learned in class. For example: normal flora and fermentation were learned as students researched the process of making a Peruvian drink called "chicha de jora," which is made from chewed corn that becomes fermented as it mixes with oral bacteria from saliva. While engaging in global learning, students learned some microbiology concepts; they passed the knowledge on to the campus community with poster presentations held during International Education Week. Based on students' feedback and participation, I can conclude that teaching microbiology using global learning was engaging, promoted student learning, and motivated students to learn.


Biochemical recurrence after radical prostatectomy: what does it mean?
Radical prostatectomy (RP) has been used as the main primary treatment for prostate cancer (PCa) for many years with excellent oncologic results. However, approximately 20-40% of those patients has failed to RP and presented biochemical recurrence (BCR). Prostatic specific antigen (PSA) has been the pivotal tool for recurrence diagnosis, but there is no consensus about the best PSA threshold to define BCR until this moment. The natural history of BCR after surgical procedure is highly variable, but it is important to distinguish biochemical and clinical recurrence and to find the correct timing to start multimodal treatment strategy. Also, it is important to understand the role of each clinical and pathological feature of prostate cancer in BCR, progression to metastatic disease and cancer specific mortality (CSM). Review design: A simple review was made in Medline for articles written in English language about biochemical recurrence after radical prostatectomy. To provide an updated assessment of BCR definition, its meaning, PCa natural history after BCR and the weight of each clinical/pathological feature and risk group classifications in BCR, metastatic disease and CSM.


InsP6-sensitive variants of the Gle1 mRNA export factor rescue growth and fertility defects of the ipk1 low-phytic-acid mutation in Arabidopsis.
Myo-inositol-1,2,3,4,5,6-hexakisphosphate (InsP(6)), also known as phytic acid, accumulates in large quantities in plant seeds, serving as a phosphorus reservoir, but is an animal antinutrient and an important source of water pollution. Here, we report that Gle1 (GLFG lethal 1) in conjunction with InsP(6) functions as an activator of the ATPase/RNA helicase LOS4 (low expression of osmotically responsive genes 4), which is involved in mRNA export in plants, supporting the Gle1-InsP(6)-Dbp5 (LOS4 homolog) paradigm proposed in yeast. Interestingly, plant Gle1 proteins have modifications in several key residues of the InsP(6) binding pocket, which reduce the basicity of the surface charge. Arabidopsis thaliana Gle1 variants containing mutations that increase the basic charge of the InsP(6) binding surface show increased sensitivity to InsP(6) concentrations for the stimulation of LOS4 ATPase activity in vitro. Expression of the Gle1 variants with enhanced InsP(6) sensitivity rescues the mRNA export defect of the ipk1 (inositol 1,3,4,5,6-pentakisphosphate 2-kinase) InsP(6)-deficient mutant and, furthermore, significantly improves vegetative growth, seed yield, and seed performance of the mutant. These results suggest that Gle1 is an important factor responsible for mediating InsP(6) functions in plant growth and reproduction and that Gle1 variants with increased InsP(6) sensitivity may be useful for engineering high-yielding low-phytate crops.


Effect of climate and geography on worldwide fine resolution economic activity.
Geography, including climatic factors, have long been considered potentially important elements in shaping socio-economic activities, alongside other determinants, such as institutions. Here we demonstrate that geography and climate variables satisfactorily explain the worldwide economic activity as measured by the per capita Gross Cell Product (GCP-PC) at a fine geographical resolution, typically much higher than country average. A 1° by 1° GCP-PC dataset has been key for establishing and testing a direct relationship between 'local' geography/climate and GCP-PC. Not only have we tested the geography and climate hypothesis using many possible explanatory variables, importantly we have also predicted and reconstructed GCP-PC worldwide by retaining the most significant predictors. While this study confirms that latitude is the most important predictor for GCP-PC when taken in isolation, the accuracy of the GCP-PC prediction is greatly improved when other factors mainly related to variations in climatic variables, rather than average climatic conditions as typically used, are considered. However, latitude diminishes in importance when only the wealthier parts of the globe are considered. This work points to specific features of the climate system which explain economic activity, such as the variability in air pressure. Implications of these findings range from an improved understanding of why socio-economically better-off societies are geographically placed where they are in the present, past and future to informing where new economic activities could be established in order to yield favourable economic outcomes based on geography and climate conditions.


AT1R blockade in adverse milieus: role of SMRT and corepressor complexes.
ANG II type 1 receptor blockade (AT1R-BLK) is used extensively to slow down the progression of proteinuric kidney diseases. We hypothesized that AT1R-BLK provides podocyte protection through regulation of silencing mediator of retinoic acid and thyroid hormone receptor (SMRT) and vitamin D receptor (VDR) expression under adverse milieus such as high glucose and human immunodeficiency virus infection. Both AT1R-BLK and VDR agonists (VDAs) stimulated VDR complex formation that differed not only in their composition but also in their functionality. AT1R-BLK-induced VDR complexes contained predominantly unliganded VDR, SMRT, and phosphorylated histone deacetylase 3, whereas VDA-VDR complexes were constituted by liganded VDR and CREB-binding protein/p300. AT1R-BLK-induced complexes attenuated podocyte acetyl-histone 3 levels as well as cytochrome P-450 family 24A1 expression, thus indicating their deacetylating and repressive properties. On the other hand, VDA-VDR complexes not only increased podocyte acetyl-histone 3 levels but also enhanced cytochrome P-450 family 24A1 expression, thus suggesting their acetylating and gene activation properties. AT1R-BLK- induced podocyte SMRT inhibited expression of the proapoptotic gene BAX through downregulation of Wip1 and phosphorylation of checkpoint kinase 2 in high-glucose milieu. Since SMRT-depleted podocytes lacked AT1R-BLK-mediated protection against DNA damage, it appears that SMRT is necessary for DNA repairs during AT1R-BLK. We conclude that AT1R-BLK provides podocyte protection in adverse milieus predominantly through SMRT expression and partly through unliganded VDR expression in 1,25(OH)2D-deficient states; on the other hand, AT1R-BLK contributes to liganded VDR expression in 1,25(OH)2D-sufficient states.


Measurement of region-specific nitrate levels of the posterior chamber of the rat eye using low-flow push-pull perfusion.
The determination of the presence of nitric oxide metabolites in the rat vitreous cavity in a regioselective manner is complicated by the size and shape of the eye as well as the diffusivity of the molecule and its metabolites. In this work, in vivo low-flow push-pull perfusion sampling was utilized with a rapid capillary electrophoretic assay to monitor levels of the major NO metabolite, nitrate, at the vitreoretinal interface (VRI) of normal and aged rat models. The sampling probe tips were placed in three different positions in the posterior chamber through a 29-gauge guide needle. Sampling was performed along the VRI over the optic nerve head and regions peripheral to the optic nerve head. Additionally, samples were collected from the middle vitreous region to compare to VRI sampling. A significant (P < 0.05) difference in the perfusate nitrate concentration was observed in each location, which may be due to the source of NO production or the clearance mechanism of the molecule from the vitreous cavity. Infusion of L-NAME with physiological saline led to a significant decrease (35%) in the observed nitrate level. LFPPP was then utilized to observe nitrate levels after an average of 4.5 months of aging. A 3-fold increase in the mean level of nitrate over the optic nerve head was observed in mature animals compared to younger control animals. Precise measurement of NO metabolites along the VRI may provide insights into the function of NO in maintaining homeostatic conditions and the molecular changes at the diseased retina.


2.5-month-old infants' reasoning about when objects should and should not be occluded.
The present research examined 2.5-month-old infants' reasoning about occlusion events. Three experiments investigated infants' ability to predict whether an object should remain continuously hidden or become temporarily visible when passing behind an occluder with an opening in its midsection. In Experiment 1, the infants were habituated to a short toy mouse that moved back and forth behind a screen. Next, the infants saw two test events that were identical to the habituation event except that a portion of the screen's midsection was removed to create a large window. In one event (high-window event), the window extended from the screen's upper edge; the mouse was shorter than the bottom of the window and thus did not become visible when passing behind the screen. In the other event (low-window event), the window extended from the screen's lower edge; although the mouse was shorter than the top of the window and hence should have become fully visible when passing behind the screen, it never appeared in the window. The infants tended to look equally at the high- and low-window events, suggesting that they were not surprised when the mouse failed to appear in the low window. However, positive results were obtained in Experiment 2 when the low-window event was modified: a portion of the screen above the window was removed so that the left and right sections of the screen were no longer connected (two-screens event). The infants looked reliably longer at the two-screens than at the high-window event. Together, the results of Experiments 1 and 2 suggested that, at 2.5 months of age, infants possess only very limited expectations about when objects should and should not be occluded. Specifically, infants expect objects (1) to become visible when passing between occluders and (2) to remain hidden when passing behind occluders, irrespective of whether these have openings extending from their upper or lower edges. Experiment 3 provided support for this interpretation. The implications of these findings for models of the origins and development of infants' knowledge about occlusion events are discussed.


Phase I study of dose-dense alternating doublets in advanced non-small-cell lung cancer.
We performed a study to determine the feasibility of a rapidly alternating administration of cisplatin/vinorelbine (CV) and docetaxel/gemcitabine (DG) in the treatment of advanced non-small-cell lung cancer (NSCLC). Thirty-four patients with NSCLC (6% stage IIIB, 94% stage IV) were enrolled. The initial schema was to give CV on days 1 and 8 followed by DG on days 15 and 22, every 28 days. Granulocyte colony-stimulating factor (G-CSF) was used on days 9-14 and 23-28. Despite dose reductions, this sequence was not feasible. Therefore, the sequence was switched to give DG before CV, cycle duration was extended to 35 days, and G-CSF was given on days 2-7 and 23-28. Neutropenia and thrombocytopenia were dose-limiting toxicities. The recommended doses for phase II studies are docetaxel 75 mg/m2 on day 1, gemcitabine 1000 mg/m2 on days 1 and 8, cisplatin 60 mg/m2 on day 15, and vinorelbine 25 mg/m2 on days 15 and 22, every 35 days with G-CSF 5 microg/kg on days 2-7 and 23-28. However, treatment delays were required in subsequent cycles due to cumulative myelosuppression. A less intensive schedule is recommended for subsequent cycles for further testing. Overall response rate was 29% (95% confidence interval [CI], 14%-45%), and median survival for all patients was 11.8 months. One- and 2-year survival rates were 47% (95% CI, 30%-63%) and 14% (95% CI, 1.4%-40%), respectively. Although initial administration of this regimen was feasible, dose intensity could not be maintained in subsequent cycles due to cumulative myelosuppression. A sequential rather than alternating use of doublet regimens might be more readily feasible and may permit greater maintenance of dose intensity.


Insights into the Design of p97-targeting Small Molecules from Structural Studies on p97 Functional Mechanism.
p97, also known as valosin-containing protein or CDC48, is a member of the AAA+ protein family that is highly conserved in eukaryotes. It binds to various cofactors in the body to perform its protein-unfolding function and participates in DNA repair, degradation of subcellular membrane proteins, and protein quality control pathways, among other processes. Its malfunction can lead to many diseases, such as inclusion body myopathy, associated with Paget's disease of bone and/or frontotemporal dementia, amyotrophic lateral sclerosis disease, and others. In recent years, many small-molecule inhibitors have been deployed against p97, including bis (diethyldithiocarbamate)- copper and CB-5083, which entered the first phase of clinical tests but failed. One bottleneck in the design of p97 drugs is that its molecular mechanism remains unclear. This paper summarizes recent studies on the molecular mechanisms of p97, which may lead to insight into how the next generation of small molecules targeting p97 can be designed.


A review of the ongoing discussion about definition, diagnosis and pathomechanism of subclinical endometritis in dairy cows.
In the last decade, several new aspects of the inflammation of the bovine endometrium have been investigated and described, including a new definition of subclinical endometritis. This review summarizes the recent discussion about the definition, diagnosis and pathomechanism of subclinical endometritis. Subclinical endometritis also referred to as cytological endometritis is defined by findings of endometrial cytology, which is usually performed with the cytobrush-technique or by low-volume flushing of the uterus. The sampling procedure is minimally invasive and has no negative impact on subsequent conception rate. The suggested threshold value for polymorphonuclear cells (PMN) as diagnostic for subclinical endometritis depends on the time postpartum and varies from 5 to 18%. It has also been shown that a general threshold of 5% PMN is eligible for all cows between 21 and 62 days postpartum. Accuracy and repeatability of counting PMN under the microscope have been evaluated and can be regarded as reliable. The impact of subclinical endometritis on reproductive performance is characterized by decreased conception rates, and prolonged days to first service and days open. In addition, it has been demonstrated that subclinical endometritis has an impact on survival and quality of the embryo. Some studies, however, did not confirm this negative effect of subclinical endometritis on fertility. More detailed analyses of the cytobrush samples revealed higher mRNA expression of several cytokines in cows with subclinical endometritis compared with healthy cows, and contributed to the understanding of detrimental effects of subclinical endometritis on fertility. In contrast to clinical endometritis, there are no predominant bacteria related to subclinical endometritis, but associations between the presence of α-hemolytic streptococci and Trueperella pyogenes and subclinical endometritis have been found. For the treatment of subclinical endometritis, intrauterine infusions with cephapirin as well as the administration of PGF2α have been recommended. Other studies, however, did not confirm the efficiency of these treatments.


Consequences of inhibiting amyloid precursor protein processing enzymes on synaptic function and plasticity.
Alzheimer's disease (AD) is a neurodegenerative disease, one of whose major pathological hallmarks is the accumulation of amyloid plaques comprised of aggregated β-amyloid (Aβ) peptides. It is now recognized that soluble Aβ oligomers may lead to synaptic dysfunctions early in AD pathology preceding plaque deposition. Aβ is produced by a sequential cleavage of amyloid precursor protein (APP) by the activity of β- and γ-secretases, which have been identified as major candidate therapeutic targets of AD. This paper focuses on how Aβ alters synaptic function and the functional consequences of inhibiting the activity of the two secretases responsible for Aβ generation. Abnormalities in synaptic function resulting from the absence or inhibition of the Aβ-producing enzymes suggest that Aβ itself may have normal physiological functions which are disrupted by abnormal accumulation of Aβ during AD pathology. This interpretation suggests that AD therapeutics targeting the β- and γ-secretases should be developed to restore normal levels of Aβ or combined with measures to circumvent the associated synaptic dysfunction(s) in order to have minimal impact on normal synaptic function.


Taking the lead in fundholding.
The prevalence of rather negative, or at best mixed, attitudes of doctors towards participation in management has been reported in several previous research studies. This paper adds to that growing empirical literature, but complements most of the previous studies by focusing on general medical practitioners in primary care rather than on doctors in secondary care settings. The focus for the study were doctors who had agreed to undertake the lead partner role in the now-defunct general practitioner fundholding initiative within the UK National Health Service. The findings indicate widespread reluctance but the presence of a minority of enthusiasts, which resonates with previous studies of secondary care settings. A finding of potential significance for future research and practice was that some doctors who were initially reluctant became more keen on undertaking a management role once they had experienced it.


High WHO/ISUP Grade and Unfavorable Architecture, Rather Than Typing of Papillary Renal Cell Carcinoma, May Be Associated With Worse Prognosis.
Conflicting data have been published on the prognostic significance of histologic parameters in papillary renal cell carcinoma (PRCC). We conducted a comprehensive evaluation of clinical and histologic parameters in PRCC in nephrectomies and their impact on prognosis, with an emphasis on World Health Organization (WHO)/International Society of Urological Pathology (ISUP) grade, tumor architecture (solid, micropapillary, and hobnail), and PRCC type. A total of 185 PRCC cases were evaluated, 117 (63.2%) type 1, 45 (24.3%) type 2, and 11 (5.9%) mixed type 1 and type 2. Using WHO/ISUP grading criteria, PRCCs were graded as follows: 6 (3.2%) grade 1; 116 (62.7%) grade 2; 61 (33.0%) grade 3; and 2 (1.1%) grade 4. The solid architecture was present in 3 cases (1.6%) and comprised 10%, 10%, and 30% of the tumor area. Micropapillary architecture was present in 10 cases (5.4%), ranging from 5% to 30% of the tumor (mean=11%; median=10%). Hobnail architecture was seen in 9 cases (4.9%), with mean percentage of 23% (median=15%; range: 5% to 50%) involvement of tumor area. Parameters associated with worse disease-free survival (DFS) and overall survival (OS) in the univariate analysis included WHO/ISUP grade, pathologic stage, tumor size, and solid, micropapillary, or hobnail architecture (P<0.05). The pathologic stage and WHO/ISUP grade were significantly associated with both DFS and OS in stepwise multivariate Cox regression analysis (P<0.05). In addition, micropapillary architecture and type 1 histology were linked with an adverse impact on OS (P<0.05). We found no difference in DFS (P=0.8237) and OS (P=0.8222) for type 1 versus type 2 PRCC in our patient cohort. In addition, we performed a meta-analysis with data from studies with reported hazard ratios (HRs) on PRCC type in relation to DFS and OS. We identified 5 studies that reported DFS and found no significant effect for type 2 PRCC (P=0.30; HR=1.43; 95% confidence interval: 0.73-2.80). We identified 7 studies that reported OS and found no significant association between type 2 PRCC and worse OS (P=0.41; HR: 1.21; 95% confidence interval: 0.77-1.91). Our findings suggest that high WHO/ISUP grade and unfavorable architecture (solid, micropapillary, or hobnail), rather than typing of PRCC, are associated with worse outcomes.


Biosynthesis and structure of the Burkholderia cenocepacia K56-2 lipopolysaccharide core oligosaccharide: truncation of the core oligosaccharide leads to increased binding and sensitivity to polymyxin B.
Burkholderia cenocepacia is an opportunistic pathogen that displays a remarkably high resistance to antimicrobial peptides. We hypothesize that high resistance to antimicrobial peptides in these bacteria is because of the barrier properties of the outer membrane. Here we report the identification of genes for the biosynthesis of the core oligosaccharide (OS) moiety of the B. cenocepacia lipopolysaccharide. We constructed a panel of isogenic mutants with truncated core OS that facilitated functional gene assignments and the elucidation of the core OS structure in the prototypic strain K56-2. The core OS structure consists of three heptoses in the inner core region, 3-deoxy-d-manno-octulosonic acid, d-glycero-d-talo-octulosonic acid, and 4-amino-4-deoxy-l-arabinose linked to d-glycero-d-talo-octulosonic acid. Also, glucose is linked to heptose I, whereas heptose II carries a second glucose and a terminal heptose, which is the site of attachment of the O antigen. We established that the level of core truncation in the mutants was proportional to their increased in vitro sensitivity to polymyxin B (PmB). Binding assays using fluorescent 5-dimethylaminonaphthalene-1-sulfonyl-labeled PmB demonstrated a correlation between sensitivity and increased binding of PmB to intact cells. Also, the mutant producing a heptoseless core OS did not survive in macrophages as compared with the parental K56-2 strain. Together, our results demonstrate that a complete core OS is required for full PmB resistance in B. cenocepacia and that resistance is due, at least in part, to the ability of B. cenocepacia to prevent binding of the peptide to the bacterial cell envelope.


Establishment and characterisation of six human biliary tract cancer cell lines.
Human cell lines established from biliary tract cancers are rare, and only five have been reported previously. We report the characterisation of six new six biliary tract cancer cell lines (designated SNU-245, SNU-308, SNU-478, SNU-869, SNU-1079 and SNU-1196) established from primary tumour samples of Korean patients. The cell lines were isolated from two extrahepatic bile duct cancers (one adenocarcinoma of common bile duct, one hilar bile duct cancer), two adenocarcinomas of ampulla of Vater, one intrahepatic bile duct cancer (cholangiocarcinoma), and one adenocarcinoma of the gall bladder. The cell phenotypes, including the histopathology of the primary tumours and in vitro growth characteristics, were determined. We also performed molecular characterisation, including DNA fingerprinting analysis and abnormalities of K-ras, p15, p16, p53, hMLH1, hMSH2, DPC4, beta-catenin, E-cadherin, hOGG1, STK11, and TGF-betaRII genes by PCR-SSCP and sequencing analysis. In addition, we compared the genetic alterations in tumour cell lines and their corresponding tumour tissues. All lines grew as adherent cells. Population doubling times varied from 48-72 h. The culture success rate was 20% (six out of 30 attempts). All cell lines showed (i) relatively high viability; (ii) absence of mycoplasma or bacteria contamination; and (iii) genetic heterogeneity by DNA fingerprinting analysis. Among the lines, three lines had p53 mutations; and homozygous deletions in both p16 and p15 genes were found three and three lines, respectively; one line had a heterozygous missense mutation in hMLH1; E-cadherin gene was hypermethylated in two lines. Since the establishment of biliary tract cancer cell lines has been rarely reported in the literature, these newly established and well characterised biliary tract cancer cell lines would be very useful for studying the biology of biliary tract cancers, particularly those related to hypermethylation of E-cadherin gene in biliary tract cancer.


The implications of the 2010 Patient Protection and Affordable Care Act and the Health Care and Education Reconciliation Act on cancer care delivery.
In March 2010, President Obama signed into law the Patient Protection and Affordable Care Act and the Health Care and Education Reconciliation Act. This legislation attempts to address cost control and improve the quality of healthcare in the United States. Cancer is a major health problem in the United States and the leading cause of death for Americans under the age of 80. Therefore, cancer care providers need to be fully engaged in ongoing discussions regarding quality measurement and care delivery. With the optimum level of collaboration and support, the proposals in the legislation can be properly structured to deliver improved access to care via better delivery systems, as well as more appropriate reimbursement to advance the prevention and treatment of cancer.


A Pediatric Case of Persistent Thrombocytopenia in a Patient Who Developed All.
Inherited thrombocytopenia is a topic that was expanded greatly over the last decade and many new genes are being identified. However, inheritance patterns are not always easy to identify because sporadic cases from de novo mutations may in fact be more common. Few studies have assessed the relationship between thrombocytopenia and malignancies, specifically acute lymphoblastic leukemia (ALL). Here we present a pediatric case of persistent thrombocytopenia associated with T-cell ALL. Our patient was initially diagnosed with immune thrombocytopenic purpura with no evidence of malignancy on bone marrow biopsy but presented shortly after with ALL.


RUPE-phospho: Rapid Ultrasound-Assisted Peptide-Identification-Enhanced Phosphoproteomics Workflow for Microscale Samples.
Global phosphoproteome profiling can provide insights into cellular signaling and disease pathogenesis. To achieve comprehensive phosphoproteomic analyses with minute quantities of material, we developed a rapid and sensitive phosphoproteomics sample preparation strategy based on ultrasound. We found that ultrasonication-assisted digestion can significantly improve peptide identification by 20% due to the generation of longer peptides that can be detected by mass spectrometry. By integrating this rapid ultrasound-assisted peptide-identification-enhanced proteomic method (RUPE) with streamlined phosphopeptide enrichment steps, we established RUPE-phospho, a fast and efficient strategy to characterize protein phosphorylation in mass-limited samples. This approach dramatically reduces the sample loss and processing time: 24 samples can be processed in 3 h; 5325 phosphosites, 4549 phosphopeptides, and 1888 phosphoproteins were quantified from 5 μg of human embryonic kidney (HEK) 293T cell lysate. In addition, 9219 phosphosites were quantified from 1-2 mg of OCT-embedded mouse brain with 120 min streamlined RUPE-phospho workflow. RUPE-phospho facilitates phosphoproteome profiling for microscale samples and will provide a powerful tool for proteomics-driven precision medicine research.


Enzymatic synthesis of biphenyl-DNA oligonucleotides.
The incorporation of nucleotides equipped with C-glycosidic aromatic nucleobases into DNA and RNA is an alluring strategy for a number of practical applications including fluorescent labelling of oligonucleotides, expansion of the genetic alphabet for the generation of aptamers and semi-synthetic organisms, or the modulation of excess electron transfer within DNA. However, the generation of C-nucleoside containing oligonucleotides relies mainly on solid-phase synthesis which is quite labor intensive and restricted to short sequences. Here, we explore the possibility of constructing biphenyl-modified DNA sequences using enzymatic synthesis. The presence of multiple biphenyl-units or biphenyl residues modified with electron donors and acceptors permits the incorporation of a single dBphMP nucleotide. Moreover, templates with multiple abasic sites enable the incorporation of up to two dBphMP nucleotides, while TdT-mediated tailing reactions produce single-stranded DNA oligonucleotides with four biphenyl residues appended at the 3'-end.


Modeling optical memory effects with phase screens.
During the last decade, optical memory effects have been explored extensively for various applications. In this letter we propose phase screen models to facilitate the analysis and the simulation of wave propagation through optical media that exhibits memory effects. We show that the classical optical memory effect, which implies tilt wave correlations of the input and the scattered fields, can be readily modeled by a single random phase screen. For the recently discovered generalized optical memory effect, which implies the existence of shift wave correlations in addition to the tilt correlation, we propose an appropriate generalized random phase screen model.


The management of bipolar mania: a national survey of baseline data from the EMBLEM study in Italy.
Although a number of studies have assessed the management of mania in routine clinical practice, no studies have so far evaluated the short- and long-term management and outcome of patients affected by bipolar mania in different European countries. The objective of the study is to present, in the context of a large multicenter survey (EMBLEM study), an overview of the baseline data on the acute management of a representative sample of manic bipolar patients treated in the Italian psychiatric hospital and community settings. EMBLEM is a 2-year observational longitudinal study that evaluates across 14 European countries the patterns of the drug prescribed in patients with bipolar mania, their socio-demographic and clinical features and the outcomes of the treatment. The study consists of a 12-week acute phase and a < or = 24-month maintenance phase. Bipolar patients were included into the study as in- or out-patients, if they initiated or changed, according to the decision of their psychiatrist, oral antipsychotics, anticonvulsants and/or lithium for the treatment of an episode of mania. Data concerning socio-demographic characteristics, psychiatric and medical history, severity of mania, prescribed medications, functional status and quality of life were collected at baseline and during the follow-up period. In Italy, 563 patients were recruited in 56 sites: 376 were outpatients and 187 inpatients. The mean age was 45.8 years. The mean CGI-BP was 4.4 (+/- 0.9) for overall score and mania, 1.9 (+/- 1.2) for depression and 2.6 (+/- 1.6) for hallucinations/delusions. The YMRS showed that 14.4% had a total score < 12, 25.1% > or = 12 and < 20, and 60.5% > or = 20. At entry, 75 patients (13.7%) were treatment-naïve, 186 (34.1%) were receiving a monotherapy (of which haloperidol [24.2%], valproate [16.7%] and lithium [14.5%] were the most frequently prescribed) while 285 (52.2%) a combined therapy (of which 8.0% were represented by haloperidol/lithium combinations). After a switch to an oral medication, 137 patients (24.8%) were prescribed a monotherapy while the rest (415, 75.2%) received a combination of drugs. Data collected at baseline in the Italian cohort of the EMBLEM study represent a relevant source of information to start addressing the short and long-term therapeutic strategies for improving the clinical as well as the socio-economic outcomes of patients affected by bipolar mania. Although it's not an epidemiological investigation and has some limitations, the results show several interesting findings as a relatively late age of onset of bipolar disorder, a low rate of past suicide attempts, a low lifetime rate of alcohol abuse and drug addiction.


Infrequency of Rickettsia rickettsii in Dermacentor variabilis removed from humans, with comments on the role of other human-biting ticks associated with spotted fever group Rickettsiae in the United States.
From 1997 to 2009, the Tick-Borne Disease Laboratory of the U.S. Army Public Health Command (USAPHC) (formerly the U.S. Army Center for Health Promotion and Preventive Medicine) screened 5286 Dermacentor variabilis ticks removed from Department of Defense (DOD) personnel, their dependents, and DOD civilian personnel for spotted fever group rickettsiae using polymerase chain reaction and restriction fragment length polymorphism analysis. Rickettsia montanensis (171/5286 = 3.2%) and Rickettsia amblyommii (7/5286 = 0.1%) were detected in a small number of samples, but no ticks were found positive for Rickettsia rickettsii, the agent of Rocky Mountain spotted fever (RMSF) until May 2009, when it was detected in one D. variabilis male removed from a child in Maryland. This result was confirmed by nucleotide sequence analysis of the rickettsial isolate and of the positive control used in the polymerase chain reaction, which was different from the isolate. Lethal effects of rickettsiostatic proteins of D. variabilis on R. rickettsii and lethal effects of R. rickettsii infection on tick hosts may account for this extremely low prevalence. Recent reports of R. rickettsii in species Rhipicephalus sanguineus and Amblyomma americanum ticks suggest their involvement in transmission of RMSF, and other pathogenic rickettsiae have been detected in Amblyomma maculatum. The areas of the U.S. endemic for RMSF are also those where D. variabilis exist in sympatry with populations of A. americanum and A. maculatum. Interactions among the sympatric species of ticks may be involved in the development of a focus of RMSF transmission. On the other hand, the overlap of foci of RMSF cases and areas of A. americanum and A. maculatum populations might indicate the misdiagnosis as RMSF of diseases actually caused by other rickettsiae vectored by these ticks. Further studies on tick vectors are needed to elucidate the etiology of RMSF.


Nanobody Based Immunoassay for Human Soluble Epoxide Hydrolase Detection Using Polymeric Horseradish Peroxidase (PolyHRP) for Signal Enhancement: The Rediscovery of PolyHRP?
Soluble epoxide hydrolase (sEH) is a potential pharmacological target for treating hypertension, vascular inflammation, cancer, pain, and multiple cardiovascular related diseases. A variable domain of the heavy chain antibody (termed single domain antibody (sdAb), nanobody, or VHH) possesses the advantages of small size, high stability, ease of genetic manipulation, and ability for continuous manufacture, making such nanobody a superior choice as an immunoreagent. In this work, we developed an ultrasensitive nanobody based immunoassay for human sEH detection using polymeric horseradish peroxidase (PolyHRP) for signal enhancement. Llama nanobodies against human sEH were used as the detection antibody in sandwich enzyme linked immunosorbent assays (ELISA) with polyclonal anti-sEH as the capture antibody. A conventional sandwich ELISA using a horseradish peroxidase (HRP) labeled anti-hemeagglutinin (HA) tag as the tracer showed a marginal sensitivity (0.0015 optical density (OD)·mL/ng) and limit of detection (LOD) of 3.02 ng/mL. However, the introduction of the PolyHRP as the tracer demonstrated a 141-fold increase in the sensitivity (0.21 OD·mL/ng) and 57-fold decrease in LOD (0.05 ng/mL). Systematic comparison of three different tracers in four ELISA formats demonstrated the overwhelming advantage of PolyHRP as a label for nanobody based immunoassay. This enhanced sEH immunoassay was further evaluated in terms of selectivity against other epoxide hydrolases and detection of the target protein in human tissue homogenate samples. Comparison with an enzyme activity based assay and a Western blot for sEH detection reveals good correlation with the immunoassay. This work demonstrates increased competiveness of nanobodies for practical sEH protein detection utilizing PolyHRP. It is worthwhile to rediscover the promising potential of PolyHRP in nanobody and other affinity based methods after its low-profile existence for decades.


Helper roles of NK and CD8+ T cells in the induction of tumor immunity. Polarized dendritic cells as cancer vaccines.
The work in our laboratory addresses two interrelated areas of dendritic cell (DC) biology: (1) the role of DCs as mediators of feedback interactions between NK cells, CD8+ and CD4+ T cells; and (2) the possibility to use such feedback and the paradigms derived from anti-viral responses, to promote the induction of therapeutic immunity against cancer. We observed that CD8+ T cells and NK cells, the classical "effector" cells, also play "helper" roles, regulating ability of DCs to induce type-1 immune immunity, critical for fighting tumors and intracellular pathogens. Our work aims to delineate which pathways of NK and CD8+ T cell activation result in their helper activity, and to identify the molecular mechanisms allowing them to induce type-1 polarized DCs (DC1s) with selectively enhanced ability to promote type-1 responses and anti-cancer immunity. The results of these studies allowed us and our colleagues to design phase I/II clinical trials incorporating the paradigms of DC polarization and helper activity of effector cells in cancer immunotherapy.


Today's Students, Tomorrow's Physicians: Opinions on Enacted and Prospective Health Care Policies.
Future physicians will be key stakeholders in the formation, implementation, and success of health care policies enacted during their careers, though little is known of their opinions of enacted and proposed policies since the 2016 U.S. presidential election. This study aimed to understand the opinions of medical students related to policies including, but not limited to, protections for people with pre-existing conditions, a public option on the private exchange, and single-payer health care. Online surveys were completed by 1,660 medical students at 7 U.S. medical schools between October 2017 and November 2017. The authors used multiple logistic regression to examine associations between student characteristics and support of policies. In total, 1,660 of 4,503 (36.9%) eligible medical students completed the survey. A majority of respondents identified 4 extant Affordable Care Act policies as important, including its protections for patients with pre-existing conditions (95.3%) and Medicaid expansion (77.8%). With respect to prospective reforms, 82.6% supported a public insurance option, and 70.5% supported a single-payer health care system. Only 2.2% supported reducing funding for Medicaid. Although views varied by sex, anticipated specialty, and political affiliation, medical students largely supported prospective policies that would expand insurance coverage and access to health care.


In vitro interaction of artemisinin derivatives or the fully synthetic peroxidic anti-malarial OZ277 with thapsigargin in Plasmodium falciparum strains.
Semi-synthetic artemisinin derivatives are powerful peroxidic drugs in artemisinin-based combination therapy (ACT) recommended as first-line treatment of Plasmodium falciparum malaria in disease-endemic countries. Studies by Eckstein-Ludwig and co-workers showed both thapsigargin and artemisinin specifically inhibit the sarcoplasmic reticulum Ca²⁺-ATPase of Plasmodium falciparum (PfATP6). In the present study the type of interaction between thapsigargin and artemisinin derivatives as well as the ozonide OZ277 (RBx11160 or arterolane) was evaluated in parasite cultures. The latter compound is an adamantane-based peroxide and the first fully synthetic clinical candidate recently registered in India by Ranbaxy Laboratories Ltd. for anti-malarial combination therapy. Drug interaction studies were performed using a previously described fixed ratio method and anti-malarial activity measured using the [3H] hypoxanthine incorporation assay. The sum 50% and 90% fractional inhibitory concentration (∑FIC₅₀, ₉₀) of the interaction of thapsigargin with OZ277, artemether or artesunate, against NF54 and K1 strains of P. falciparum ranged from 0.9 to 1.4. The interaction of thapsigargin with OZ277, artesunate or artemether was additive, data consistent with previous observations indicating that activity of anti-malarial peroxides does not derive from reversible interactions with parasite targets.


Arsenic in Drinking Water and Diabetes.
Arsenic is ubiquitous in soil and water environments and is consistently at the top of the Agency for Toxic Substances Disease Registry (ATSDR) substance priority list. It has been shown to induce toxicity even at low levels of exposure. One of the major routes of exposure to arsenic is through drinking water. This review presents current information related to the distribution of arsenic in the environment, the resultant impacts on human health, especially related to diabetes, which is one of the most prevalent chronic diseases, regulation of arsenic in drinking water, and approaches for treatment of arsenic in drinking water for both public utilities and private wells. Taken together, this information points out the existing challenges to understanding both the complex health impacts of arsenic and to implementing the treatment strategies needed to effectively reduce arsenic exposure at different scales.


The association between obstructive sleep apnea and hypertension by race/ethnicity in a nationally representative sample.
The association between obstructive sleep apnea (OSA) and hypertension by race/ethnicity has not been well characterized in a national sample. Adult participants in the 2007-2008 National Health and Nutrition Examination Survey were reviewed by self-report of sleep apnea diagnosis, snorting, gasping or stopping breathing during sleep, and snoring to derive whether OSA was probable (pOSA). Multivariable logistic regression determined whether pOSA predicted hypertension in the overall cohort, and by body mass index (BMI) group and ethno-racial strata. pOSA predicted hypertension in several groups: (1) Within BMI strata, there was a significant association among overweight individuals [odds ratio [OR], 1.82; 95% confidence interval [CI], 1.26-2.62); (2) In race/ethnicity subgroups, the association was significant among Hispanic/Latinos (OR, 1.69; 95% CI, 1.13-2.53) and whites (OR, 1.40; 95% CI, 1.07-1.84); (3) In models stratified by both race/ethnicity and BMI, pOSA predicted hypertension among overweight black/African Americans (OR, 4.74; 95% CI, 1.86-12.03), overweight whites (OR, 1.65; 95% CI, 1.06-2.57), and obese Hispanic/Latino participants (OR, 2.01; 95% CI, 1.16-3.49). A simple, self-report tool for OSA was strongly associated with hypertension, and may serve as a potential future opportunity for OSA diagnosis.


Predictors of Clinically Suspected Intra-articular Hip Symptoms and Prevalence of Hip Pathomorphologies Presenting to Sports Medicine and Hip Preservation Orthopaedic Surgeons.
The first purpose of the study was to evaluate the prevalence of various radiographic parameters and pathomorphologies for patients presenting with the diagnosis of hip pain. The second purpose of this study was to identify those pathomorphologies and radiographic parameters that were predictive of clinically suspected intra-articular and hip joint-related symptoms. A total of 998 hips (499 patients, 228 males, 271 females, mean age 38 years) presented to 2 orthopaedic surgeons with the diagnosis of hip pain. Patients were retrospectively identified as intra-articular and hip joint-related symptoms or extra-articular and non-hip joint-related symptoms based on history, examination, injection response, and diagnosis listed on clinical notes. A detailed morphologic evaluation of anteroposterior and 45° modified Dunn lateral radiographs of both hips was performed for all patients. The presence of at least 1 finding consistent with femoroacetabular impingement (FAI) was noted in 96.6% of patients (89.9% of hips) and was bilateral in 83%. The prevalence of dysplasia was 10.6% in patients (6.7% of hips) and was bilateral in 2.8%. Cam-type morphology was more common in males (P < .001). Profunda and protrusio were more common in females (P < .001). Acetabular retroversion was more common in males (P = .02). Fifty-seven percent of hips (564/998) were diagnosed clinically with intra-articular and hip joint related symptoms. Cam-type FAI, mixed-type FAI, increasing alpha angle, and increasing Tönnis grade were independent predictors of clinically suspected intra-articular and hip joint symptoms (P < .001), whereas isolated Pincer-type morphology was not. FAI is highly prevalent (96.6%) and frequently bilateral (83%) in patients presenting to an orthopaedic clinic with hip pain. Cam-type morphology and acetabular retroversion are more frequent in men, whereas profunda and protrusio are more frequent in women. Cam-type morphology, increasing alpha angle (larger cam morphology), and increasing Tönnis grade were highly predictive of clinically suspected intra-articular symptoms, whereas isolated pincer-type morphology was not. Level III, case-control study.


Nomogram for predicting prolonged postoperative ileus after laparoscopic low anterior resection for rectal cancer.
Prolonged postoperative ileus (PPOI) is a common complication after colorectal surgery that increases patient discomfort, hospital stay, and financial burden. However, predictive tools to assess the risk of PPOI in patients undergoing laparoscopic low anterior resection have not been developed. Thus, the purpose of this study was to develop a nomogram to predict PPOI after laparoscopic low anterior resection for rectal cancer. A total of 548 consecutive patients who underwent laparoscopic low anterior resection for mid-low rectal cancer at a single tertiary medical center were retrospectively enrolled between January 2019 and January 2023. Univariate and multivariate logistic regression analysis was performed to analyze potential predictors of PPOI. The nomogram was constructed using the filtered variables and internally verified by bootstrap resampling. Model performance was evaluated by receiver operating characteristic curve and calibration curve, and the clinical usefulness was evaluated by the decision curve. Among 548 consecutive patients, 72 patients (13.1%) presented with PPOI. Multivariate logistic analysis showed that advantage age, hypoalbuminemia, high surgical difficulty, and postoperative use of opioid analgesic were independent prognostic factors for PPOI. These variables were used to construct the nomogram model to predict PPOI. Internal validation, conducted through bootstrap resampling, confirmed the great discrimination of the nomogram with an area under the curve of 0.738 (95%CI 0.736-0.741). We created a novel nomogram for predicting PPOI after laparoscopic low anterior resection. This nomogram can assist surgeons in identifying patients at a heightened risk of PPOI.


Nosocomial Pneumocystis jirovecii infections.
Airborne transmission of Pneumocystis sp. from host to host has been demonstrated in rodent models and several observations suggest that interindividual transmission occurs in humans. Moreover, it is accepted that the Pneumocystis organisms infecting each mammalian species are host specific and that the hypothesis of an animal reservoir for Pneumocystis jirovecii (P. jirovecii), the human-specific Pneumocystis species, can be excluded. An exosaprophytic form of the fungus cannot be strictly ruled out. However, these data point toward the potential for the specific host to serve as its own reservoir and for Pneumocystis infection in humans as an anthroponosis with humans as a reservoir for P. jirovecii. This review highlights the main data on host-to-host transmission of Pneumocystis in rodent models and in humans by the airborne route and provides a rationale for considering the occurrence of nosocomial infections and measures for their prevention


Detection of anthrax simulants with microcalorimetric spectroscopy: Bacillus subtilis and Bacillus cereus spores.
Recent advances in the development of ultrasensitive micromechnical thermal detectors have led to the advent of novel subfemtojoule microcalorimetric spectoscopy (CalSpec). On the basis of principles of photothermal IR spectroscopy combined with efficient thermomechanical transduction, CalSpec provides acquisition of vibrational spectra of microscopic samples and absorbates. We use CalSpec as a method of identifying nanogram quantities of biological micro-organisms. Our studies focus on Bacillus subtilis and Bacillus cereus spores as simulants for Bacillus anthracis spores. Using CalSpec, we measured IR spectra of B. subtilis and B. cereus spores present on surfaces in nanogram quantities (approximately 100-1,000 spores). The spectra acquired in the wavelength range of 690-4000 cm(-1) (2.5-14.5 microm) contain information-rich vibrational signatures that reflect the different ratios of biochemical makeup of the micro-organisms. The distinctive features in the spectra obtained for the two types of microorganism can be used to distinguish between the spores of the Bacillus family. As compared with conventional IR and Fourier-transform IR microscopic spectroscopy techniques, the advantages of the present technique include significantly improved sensitivity (at least a full order of magnitude), absence of expensive IR detectors, and excellent potential for miniaturization.


Brain functional correlates of working memory: reduced load-modulated activation and deactivation in aging without hyperactivation or functional reorganization.
We aimed to identify brain functional correlates of working memory performance in aging, in hopes of facilitating understanding of mechanisms that promote better versus worse working memory in late-life. Among 64 healthy adults, aged 23 to 78, we examined the relationship between age, working memory performance, and brain functional response during task performance. We focused on the association between working memory load-modulated functional response and individual differences in performance and whether these function-performance relationships differed with age. As expected, older age was associated with poorer working memory performance. Older age was also associated with reduced load-modulated activation including in bilateral prefrontal and parietal regions and left caudate as well as reduced deactivation including in the medial prefrontal cortex. Contrary to findings of hyperactivation in aging, we found no evidence of increased activation with older age. Positive associations identified between brain response and performance did not differ with age. Our findings suggest that the neural mechanisms underlying better versus worse working memory performance are age-invariant across adulthood, and argue against a pattern of functional reorganization in aging. Results are discussed within the broader literature, in which significant heterogeneity in findings between studies has been common.


Consumer channeling by health insurers: natural experiments with preferred providers in the Dutch pharmacy market.
Consumer channeling is an important element in the insurer-provider bargaining process. Health insurers can influence provider choice by offering insurance contracts with restricted provider networks. Alternatively, they can offer contracts with unrestricted access and use incentives to motivate consumers to visit preferred providers. Little is known, however, about the effectiveness of this alternative strategy of consumer channeling. Using data from two natural experiments in the Dutch pharmacy market, we examine how consumers respond to incentives used by health insurers to influence their choice of provider. We find that consumers are sensitive to rather small incentives and that temporary incentives may sort a long-term effect on provider choice. In addition, we find that both consumer and provider characteristics determine whether consumers are willing to switch to preferred pharmacies.


Resistance physical exercise alleviates lipopolysaccharide-triggered neuroinflammation in cortex and hippocampus of rats via purinergic signaling.
Resistance physical exercise has neuroprotective and anti-inflammatory effects on many known diseases and, therefore, it has been increasingly explored. The way in which this type of exercise exerts these actions is still under investigation. In this study, we aimed to analyze the enzymes and components of the purinergic system involved in the inflammatory process triggered by the P2X7R. Rats were divided into four groups: control, exercise (EX), lipopolysaccharide (LPS), and EX + LPS. The animals in the exercise groups were subjected to a 12-week ladder-climbing resistance physical exercise and received LPS after the last session for sepsis induction. Enzymes activities (NTPDase, 5'-nucleotidase, and adenosine deaminase), purinoceptors' density (P2X7R, A1, and A2A), and the levels of inflammatory indicators (pyrin domain-containing protein 3 (NLRP3), Caspase-1, interleukin (IL)- 6, IL-1B, and tumor necrosis factor (TNF) -α) were measured in the cortex and hippocampus of the animals. The results show that exercise prevented (in the both structures) the increase of: 1) nucleoside-triphosphatase (NTPDase) and 5'-nucleotidase activities; 2) P2X7R density; 3) NLRP3 and Caspase-1; and 4) IL-6, IL-1β, and TNF-α It is suggested that the purinergic system and the inflammatory pathway of P2X7R are of fundamental importance and influence the effects of resistance physical exercise on LPS-induced inflammation. Thus, the modulation of the P2X7R by resistance physical exercise offers new avenues for the management of inflammatory-related illnesses.


Oral health-related quality of life among HIV-infected and at-risk women.
Objective measures of dental diseases reflect only their clinical end-point. There is a need to use multidimensional measures of diseases that consider their psychosocial aspects and functional impact. The aim of this study is to compare the oral health-related quality of life (OHRQOL) between a group of HIV-infected women and a similar group of at-risk HIV-uninfected women, and to investigate the role of potential confounding clinical oral health and behavioral factors. Our sample included HIV-infected women (87%) and women at risk for HIV infection (13%) followed up for 5.5 years. OHRQOL was measured using the short version of the Oral Health Impact Profile (OHIP-14), which is a validated and reliable instrument. HIV-infected women averaged 10% poorer OHRQOL than HIV-uninfected women; this difference was not apparent after adjusting for the number of study visits attended and significant behavioral and clinical oral health factors. The OHRQOL was inversely related to dental and periodontal diseases and to smoking and freebase cocaine use; these relationships were not confounded by HIV status. The study identified specific clinical and behavioral factors where dental professionals can intervene to possibly improve the OHRQOL of HIV-infected or at-risk HIV-uninfected women.


Evolution and emergence of multidrug-resistant Mycobacterium tuberculosis in Chisinau, Moldova.
The evolution and emergence of drug-resistant tuberculosis (TB) has been studied extensively in some contexts, but the ecological drivers of these two processes remain poorly understood. This study sought to describe the joint evolutionary and epidemiological histories of a novel multidrug-resistant Mycobacterium tuberculosis strain recently identified in the capital city of the Republic of Moldova (MDR Ural/4.2), where genomic surveillance of drug-resistant M. tuberculosis has been limited thus far. Using whole genome sequence data and Bayesian phylogenomic methods, we reconstruct the stepwise acquisition of drug resistance mutations in the MDR Ural/4.2 strain, estimate its historical bacterial population size over time, and infer the migration history of this strain between Eastern European countries. We infer that MDR Ural/4.2 likely evolved (via acquisition of rpoB S450L, which confers resistance to rifampin) in the early 1990s, during a period of social turmoil following Moldovan independence from the Soviet Union. This strain subsequently underwent substantial population size expansion in the early 2000s, at a time when national guidelines encouraged inpatient treatment of TB patients. We infer exportation of this strain and its isoniazid-resistant ancestral precursor from Moldova to neighbouring countries starting as early as 1985. Our findings suggest temporal and ecological associations between specific public health practices, including inpatient hospitalization of drug-resistant TB cases from the early 2000s until 2013, and the evolution of drug-resistant M. tuberculosis in Moldova. These findings underscore the need for regional coordination in TB control and expanded genomic surveillance efforts across Eastern Europe.


Acute Chest Pain and Broad Complex Tachycardia. A Non-typical Case of Pre-excited Atrial Fibrillation.
Wolff-Parkinson-White syndrome is a common condition in the emergency department. A case is presented of a 76-year-old patient with acute chest pain and broad complex tachycardia. Despite the fact that previous and post cardioversion ECG tracings in sinus rhythm showed no signs of pre-excitation, the characteristic pattern of pre-excited atrial fibrillation (AF) is recognized and after successful DC cardioversion the patient is referred for catheter ablation of the accessory pathway. This case illustrates a non-typical presentation of the WPW syndrome, with an older patient than usual with slight signs of pre-excitation. We highlight the need for high grades of suspicion for the early recognition of pre-excited AF when attending patients with tachycardia and the obligation to know the distinctive aspects of its management for this potentially life-threatening arrhythmia.


Phylogenomic analyses reveal a single deep-water colonisation in Patellogastropoda.
Patellogastropoda, the true limpets, is a major group of gastropods widely distributed in marine habitats from the intertidal to deep sea. Though important for understanding their evolutionary radiation, the phylogenetic relationships among the patellogastropod families have always been challenging to reconstruct, with contradictory results likely due to insufficient sampling. Here, we obtained mitogenomic and phylogenomic data (transcriptomic or genomic) from six species representing the three predominantly deep-water patellogastropod families: Lepetidae, Neolepetopsidae, and Pectinodontidae. By using various phylogenetic methods, we show that mitogenome phylogeny recovers monophyly of eight families in most of the trees, though the relationships among families remain contentious. Meanwhile, a more robust family-level topology consistent with morphology was achieved by phylogenomics. This also reveals that these mainly deep-water families are monophyletic, suggesting a single colonisation of the deep water around the Jurassic. We also found a lack of significant correlation between genome size and habitat depth, despite some deep-water species exhibiting larger genome sizes. Our phylogenomic tree provides a stable phylogenetic backbone for Patellogastropoda that includes seven of the nine recognized families and paves the way for future evolutionary analyses in this major group of molluscs.


Effects of clinical factors on psychosocial variables in renal transplant recipients.
This paper is a report of a study investigating the effects of clinical factors (side effects of immunosuppressive medications, transplant-related hospitalizations, donor type, duration of dialysis before transplantation and time post-transplant) on cognitive appraisal of health, perceived self-efficacy, perceived social support, coping and health-related quality of life after renal transplantation. Some clinical factors such as hospitalizations, side effects of medications, donor type and dialysis, which influence the health-related quality of life of renal transplant recipients, have been investigated. However, the effects of these clinical factors on psychosocial variables after renal transplantation have not been well documented. Method. Using a descriptive cross-sectional design, a convenience sample of 160 renal transplant recipients was recruited (N = 55 < 1 year post-transplant; N = 105 1-3 years post-transplant) from May, 2005 to January, 2006. Standardized instruments were used to measure the key constructs. Multivariate analysis of variance was used to examine the effects of clinical factors on the psychosocial outcome measures. Participants reporting more (>17) immunosuppressive medication-associated side effects appraised their health more negatively, used more disengagement coping, had lower degrees of perceived self-efficacy, and reported lower physical and mental health-related quality of life than those with fewer symptoms (<or=17). Participants less than 1 year post-transplant used more engagement coping, reported higher degrees of perceived self-efficacy and perceived social support than participants in the later period post-transplant. Interventions aimed at alleviating bothersome medication side effects are needed to help these patients cope with transplantation and improve their health-related quality of life.


Skin surface biomarkers are associated with future development of atopic dermatitis in children with family history of allergic disease.
Atopic dermatitis (AD) is a common childhood chronic inflammatory skin disorder that can significantly impact quality of life and has been linked to the subsequent development of food allergy, asthma, and allergic rhinitis, an association known as the "atopic march." The aim of this study was to identify biomarkers collected non-invasively from the skin surface in order to predict AD before diagnosis across a broad age range of children. Non-invasive skin surface measures and biomarkers were collected from 160 children (3-48 months of age) of three groups: (A) healthy with no family history of allergic disease, (B) healthy with family history of allergic disease, and (C) diagnosed AD. Eleven of 101 children in group B reported AD diagnosis in the subsequent 12 months following the measurements. The children who developed AD had increased skin immune markers before disease onset, compared to those who did not develop AD in the same group and to the control group. In those enrolled with AD, lesional skin was characterized by increased concentrations of certain immune markers and transepidermal water loss, and decreased skin surface hydration. Defining risk susceptibility before onset of AD through non-invasive methods may help identify children who may benefit from early preventative interventions.


In vitro evaluation of nebivolol effects on nonadrenergic noncholinergic responses in rabbit corpus cavernosum.
The purpose of this study was to compare the effects of nebivolol on nonadrenergic noncholinergic (NANC) relaxation functions that are mediated by electric field stimulation (EFS) in rabbit corpus cavernosum smooth muscle by comparison with other beta-adrenergic receptor blockers and show the level on which its effects through nitric oxide take place. After the effects of nebivolol on the isolated corpus cavernosum tissues that were contracted through the alpha-adrenergic pathway and application of L-NAME' (NG -nitro-L-arginine methyl ester) which is a competitive inhibitor of nitric oxide synthase (NOS), the changes that occurred were recorded. Following the effect on the tissue that was contracted with phenylephrine in the presence of atropine and guanethidine that was created by EFS, nebivolol and other beta-blockers were added and the changes were recorded. After receiving relaxation responses with EFS-mediated NANC, no difference was observed between the relaxation responses due to addition of nebivolol and other beta-adrenergic blockers (p > 0.05). The finding that nebivolol which has a NO-mediated relaxation effect did not have an effect on EFS-mediated NANC relaxation but created relaxation on the tissue that was contracted by phenylephrine and the effect was reversed by L-NAME, shows that its effects are on a postsynaptic level.


Comparative proteome cataloging of Lactobacillus rhamnosus strains GG and Lc705.
The present study reports an in-depth proteome analysis of two Lactobacillus rhamnosus strains, the well-known probiotic strain GG and the dairy strain Lc705. We used GeLC-MS/MS, in which proteins are separated using 1-DE and identified using nanoLC-MS/MS, to generate high-quality protein catalogs. To maximize the number of identifications, all data sets were searched against the target databases using two search engines, Mascot and Paragon. As a result, over 1600 high-confidence protein identifications, covering nearly 60% of the predicted proteomes, were obtained from each strain. This approach enabled identification of more than 40% of all predicted surfome proteins, including a high number of lipoproteins, integral membrane proteins, peptidoglycan associated proteins, and proteins predicted to be released into the extracellular environment. A comparison of both data sets revealed the expression of more than 90 proteins in GG and 150 in Lc705, which lack evolutionary counterparts in the other strain. Differences were noted in proteins with a likely role in biofilm formation, phage-related functions, reshaping the bacterial cell wall, and immunomodulation. The present study provides the most comprehensive catalog of the Lactobacillus proteins to date and holds great promise for the discovery of novel probiotic effector molecules.


The prognostic impact of human papillomavirus status following treatment failure in oropharyngeal cancer.
Despite the human papillomavirus conferring a better prognosis in the primary treatment setting, the prognostic impact of viral status after treatment failure in oropharyngeal squamous cell carcinoma patients is poorly understood. We retrospectively identified 33 oropharyngeal squamous cell carcinoma (OPC) patients with local and/or distant disease recurrence post-treatment, and looked at metastatic patterns, time to failure and survival patterns by HPV status. Median overall survival following local failure was not significantly different by HPV status (17 months for HPV+ vs. 14 months for HPV-, p = 0.23). However, following distant failures, HPV+ patients lived significantly longer than HPV- patients (median 42 months vs. 11 months, p = 0.004). HPV- patients were more likely to have locoregional failures as compared to HPV+ patients (p = 0.005), but the difference in distant failure between both groups was not significant (p = 0.09). HPV+ patients were more likely to develop metastases to sites other than the lung and bones. HPV positivity predicts a favorable prognosis with the potential for long-term survival following distant, not locoregional, failures. These results have important implications for the aggressiveness of treatment and type of surveillance imaging performed.


Acute intermittent hypoxia in rat in vivo elicits a robust increase in tonic sympathetic nerve activity that is independent of respiratory drive.
Acute intermittent hypoxia (AIH) elicits long-term increases in respiratory and sympathetic outflow (long-term facilitation, LTF). It is still unclear whether sympathetic LTF is totally dependent on changes in respiration, even though respiratory drive modulates sympathetic nerve activity (SNA). In urethane-anaesthetized, vagotomized mechanically ventilated Sprague-Dawley rats, we investigated the effect of ten 45 s episodes of 10% O2-90% N(2) on splanchnic sympathetic nerve activity (sSNA) and phrenic nerve activity (PNA). We then tested whether or not hypoxic sympathetic chemoreceptor and baroreceptor reflexes were changed 60 min after AIH. We found that 17 animals manifested a sustained increase of sSNA (+51.2+/-4.7%) 60 min after AIH, but only 10 of these rats also expressed phrenic LTF compared with the time controls (rats not exposed to hypoxia, n=5). Inspiratory triggered averages of integrated sSNA showed respiratory modulation of SNA regardless of whether or not phrenic LTF had developed. The hypoxic chemoreceptor reflex was enhanced by 60 min after the development of AIH (peak change from 76.9+/-13.9 to 159.5+/-24.9%). Finally, sympathetic baroreceptor reflex sensitivity increased after sympathetic LTF was established (Gainmax from 1.79+/-0.18 to 2.60+/-0.28% mmHg1). Our findings indicate that respiratory-sympathetic coupling does contribute to sympathetic LTF, but that an additional tonic increase of sympathetic tone is also present that is independent of the level of PNA. Sympathetic LTF is not linked to the change in baroreflex function, since the baroreflex appears to be enhanced rather than impaired, but does play an important role in the enhancement of the hypoxic chemoreflex.


Asthma management at discharge from the emergency department: a descriptive study.
The management of asthma remains suboptimal despite the publication of Canadian asthma guidelines in 1999. A descriptive study was conducted to estimate the proportion of patients admitted to the emergency department (ED) for an asthma exacerbation who received a management plan at discharge that was in accordance with seven criteria stated in the Canadian asthma guidelines. The present study took place in two tertiary care hospitals in Montreal, Quebec. A total of 37 patients were enrolled. Three (8%) patients received a management plan at discharge that was in accordance with all seven criteria. Inhaled corticosteroids and oral corticosteroids were prescribed at discharge for 29 (78%) and 35 (95%) patients, respectively. Minimal asthma education was provided for 29 (78%) patients and a medical follow-up was recommended to 22 (60%) patients. Airflow obstruction was evaluated at discharge for only 20 (54%) patients. Overall, asthma management at discharge from the ED was generally not in accordance with the 1999 Canadian asthma guidelines. A standardized management plan should be implemented in the ED to improve the care of patients with asthma exacerbations.


Effect of Bronchoscopist-Directed Sedation and Other Factors on Patient Comfort during Diagnostic Flexible Bronchoscopy.
The factors affecting patient comfort during flexible bronchoscopy are not sufficiently addressed in resource-limited countries, and a need to devise strategies to improve patient experience is felt. The present study was undertaken to assess the effect of sedation and other factors on patient comfort during flexible bronchoscopy. A total of 374 patients, aged ≥18 years undergoing flexible bronchoscopy, were enrolled in this prospective, single-center, cross-sectional study. The baseline clinical data of sedation and nonsedation groups were recorded. Anxiety level was assessed using Visual Analog Scale (VAS). Postprocedure VAS score (primary outcome measure) assessed the discomfort related to flexible bronchoscopy. Patient-reported willingness for a repeat procedure and occurrence of adverse events were used as secondary outcome measures. Based on the median of VAS score, the study population was divided into low discomfort and high discomfort groups, and the factors affecting comfort levels in these two groups were noted. Postprocedural VAS score (median (Q1, Q3)) for sedation and nonsedation groups was 20 (15, 30) and 25 (20, 40), respectively (<0.001). Willingness for a repeat procedure was significantly more in the sedation group (p=0.002). In the low and high discomfort groups, the preprocedural anxiety level (median (Q1, Q3)) was 20 (10, 25) and 30 (20, 40), respectively (p < 0.001). Bronchoscopist-directed sedation significantly reduces patient discomfort and increases their willingness for a repeat procedure. Midazolam-fentanyl combination and lower prebronchoscopy anxiety are associated with lower discomfort levels during flexible bronchoscopy. This study is registered with the Clinical Trial Registry of India (CTRI/2018/11/016328).


Distinctive expansion of gene families associated with plant cell wall degradation, secondary metabolism, and nutrient uptake in the genomes of grapevine trunk pathogens.
Trunk diseases threaten the longevity and productivity of grapevines in all viticulture production systems. They are caused by distantly-related fungi that form chronic wood infections. Variation in wood-decay abilities and production of phytotoxic compounds are thought to contribute to their unique disease symptoms. We recently released the draft sequences of Eutypa lata, Neofusicoccum parvum and Togninia minima, causal agents of Eutypa dieback, Botryosphaeria dieback and Esca, respectively. In this work, we first expanded genomic resources to three important trunk pathogens, Diaporthe ampelina, Diplodia seriata, and Phaeomoniella chlamydospora, causal agents of Phomopsis dieback, Botryosphaeria dieback, and Esca, respectively. Then we integrated all currently-available information into a genome-wide comparative study to identify gene families potentially associated with host colonization and disease development. The integration of RNA-seq, comparative and ab initio approaches improved the protein-coding gene prediction in T. minima, whereas shotgun sequencing yielded nearly complete genome drafts of Dia. ampelina, Dip. seriata, and P. chlamydospora. The predicted proteomes of all sequenced trunk pathogens were annotated with a focus on functions likely associated with pathogenesis and virulence, namely (i) wood degradation, (ii) nutrient uptake, and (iii) toxin production. Specific patterns of gene family expansion were described using Computational Analysis of gene Family Evolution, which revealed lineage-specific evolution of distinct mechanisms of virulence, such as specific cell wall oxidative functions and secondary metabolic pathways in N. parvum, Dia. ampelina, and E. lata. Phylogenetically-informed principal component analysis revealed more similar repertoires of expanded functions among species that cause similar symptoms, which in some cases did not reflect phylogenetic relationships, thereby suggesting patterns of convergent evolution. This study describes the repertoires of putative virulence functions in the genomes of ubiquitous grapevine trunk pathogens. Gene families with significantly faster rates of gene gain can now provide a basis for further studies of in planta gene expression, diversity by genome re-sequencing, and targeted reverse genetic approaches. The functional validation of potential virulence factors will lead to a more comprehensive understanding of the mechanisms of pathogenesis and virulence, which ultimately will enable the development of accurate diagnostic tools and effective disease management.


Bilateral obstructive uropathy secondary to giant periureteral diverticulum.
Bladder diverticula are herniations of the mucosa through the fibers of the bladder muscle connected by necks of variable amplitude. They are often asymptomatic, although they may lead to complications that require a surgical therapeutic approach. We report the case of a patient with bilateral obstructive uropathy secondary to a giant periureteral diverticulum that was treated by diverticulectomy and reimplantation of the left ureter in the bladder.


Interacting neural ensembles in orbitofrontal cortex for social and feeding behaviour.
Categorically distinct basic drives (for example, for social versus feeding behaviour1-3) can exert potent influences on each other; such interactions are likely to have important adaptive consequences (such as appropriate regulation of feeding in the context of social hierarchies) and can become maladaptive (such as in clinical settings involving anorexia). It is known that neural systems regulating natural and adaptive caloric intake, and those regulating social behaviours, involve related circuitry4-7, but the causal circuit mechanisms of these drive adjudications are not clear. Here we investigate the causal role in behaviour of cellular-resolution experience-specific neuronal populations in the orbitofrontal cortex, a major reward-processing hub that contains diverse activity-specific neuronal populations that respond differentially to various aspects of caloric intake8-13 and social stimuli14,15. We coupled genetically encoded activity imaging with the development and application of methods for optogenetic control of multiple individually defined cells, to both optically monitor and manipulate the activity of many orbitofrontal cortex neurons at the single-cell level in real time during rewarding experiences (caloric consumption and social interaction). We identified distinct populations within the orbitofrontal cortex that selectively responded to either caloric rewards or social stimuli, and found that activity of individually specified naturally feeding-responsive neurons was causally linked to increased feeding behaviour; this effect was selective as, by contrast, single-cell resolution activation of naturally social-responsive neurons inhibited feeding, and activation of neurons responsive to neither feeding nor social stimuli did not alter feeding behaviour. These results reveal the presence of potent cellular-level subnetworks within the orbitofrontal cortex that can be precisely engaged to bidirectionally control feeding behaviours subject to, for example, social influences.


Vitamin D status among adolescents in Europe: the Healthy Lifestyle in Europe by Nutrition in Adolescence study.
An adequate vitamin D status is essential during childhood and adolescence, for its important role in cell growth, skeletal structure and development. It also reduces the risk of conditions such as CVD, osteoporosis, diabetes mellitus, infections and autoimmune disease. As comparable data on the European level are lacking, assessment of vitamin D concentrations was included in the Healthy Lifestyle in Europe by Nutrition in Adolescence (HELENA) study. Fasting blood samples were obtained from a subsample of 1006 adolescents (470 males; 46·8 %) with an age range of 12·5-17·5 years, selected in the ten HELENA cities in the nine European countries participating in this cross-sectional study, and analysed for 25-hydroxycholecalciferol (25(OH)D) by ELISA using EDTA plasma. As specific reference values for adolescents are missing, percentile distribution were computed by age and sex. Median 25(OH)D levels for the whole population were 57·1 nmol/l (5th percentile 24·3 nmol/l, 95th percentile 99·05 nmol/l). Vitamin D status was classified into four groups according to international guidelines (sufficiency/optimal levels ≥ 75 nmol/l; insufficiency 50-75 nmol/l; deficiency 27·5-49·99 nmol/l and severe deficiency < 27·5 nmol/l). About 80 % of the sample had suboptimal levels (39 % had insufficient, 27 % deficient and 15 % severely deficient levels). Vitamin D concentrations increased with age (P < 0·01) and tended to decrease according to BMI. Geographical differences were also identified. Our study results indicate that vitamin D deficiency is a highly prevalent condition in European adolescents and should be a matter of concern for public health authorities.