A conserved virulence plasmidic region contributes to the virulence of the multiresistant Escherichia coli meningitis strain S286 belonging to phylogenetic group C.
Recent isolation of the non-K1 Escherichia coli neonatal meningitis strain S286, belonging to phylogroup C, which is closely related to major group B1, and producing an extended-spectrum beta-lactamase, encouraged us to seek the genetic determinants responsible for its virulence. We show that S286 belongs to the sequence O type ST23O78 and harbors 4 large plasmids. The largest one, pS286colV (~120 kb), not related to resistance, contains genes characteristic of a Conserved Virulence Plasmidic (CVP) region initially identified in B2 extra-intestinal avian pathogenic E. coli (APEC) strains and in the B2 neonatal meningitis E. coli strain S88. The sequence of this CVP region has a strong homology (98%) with that of the recently sequenced plasmid pChi7122-1 of the O78 APEC strain Chi7122. A CVP plasmid-cured variant of S286 was less virulent than the wild type strain in a neonatal rat sepsis model with a significant lower level of bacteremia at 24 h (4.1 ± 1.41 versus 2.60 ± 0.16 log CFU/ml, p = 0.001) and mortality. However, the mortality in the model of adult mice was comparable between wild type and variant indicating that pS286colV is not sufficient by itself to fully explain the virulence of S286. Gene expression analysis of pS286colV in iron depleted environment was very close to that of pS88, suggesting that genes of CVP region may be expressed similarly in two very different genetic backgrounds (group C versus group B2). Screening a collection of 178 human A/B1 extraintestinal pathogenic E. coli (ExPEC) strains revealed that the CVP region is highly prevalent (23%) and MLST analysis indicated that these CVP positive strains belong to several clusters and mostly to phylogroup C. The virulence of S286 is explained in part by the presence of CVP region and this region has spread in different clusters of human A/B1 ExPEC, especially in group C.


[Parameters and paradigms for meta-evaluation: an exploratory and reflective review].
The evaluation of an evaluation is not a mere play on words or simply an investigation into the person who evaluates the evaluation. It is far more than that. It is about how to evaluate the diverse components of the evaluation process and the evaluators per se. In this paper, we discuss some promising possibilities for meta-evaluation in the field of evaluation of programs and services. These include fostering interaction between theoretical and practical production in fieldwork and supporting the definition of methods and strategies in a sector imbued with political interests and a profusion of methodological possibilities, and promoting ethical and scientific rigor in the evaluation practices. We conclude this exploratory and reflective review by raising some historical and political questions related to evaluation and meta-evaluation of programs and services and by criticizing the universalist and egalitarian pretensions contained in some evaluation approaches.


Architecture of yolk-shell structured mesoporous silica nanospheres for catalytic applications.
We report the design and realization of yolk-shell structured nanospheres with periodic mesoporous organosilica (PMO) nanospheres or noble metal nanoparticles encapsulated in mesoporous silica shells via a selective etching method. These architectures have well controlled structure, size and morphology. The yolk-shell structured PMO@SiO2 nanoparticles can be precisely functionalized with different catalytic functionalities, even incompatible acidic and basic groups: the PMO core with amino (-NH2) groups and the mesoporous silica shell with sulfonic acid (-SO3H) groups. As a nanoreactor, the as-synthesized Au@SiO2 nanospheres show faster reduction of 4-nitrophenol than that of nitrobenzene. Furthermore, the prepared PMO-NH2@SiO2-SO3H nanoparticles can be used as bifunctional catalysts with highly efficient catalytic performance for catalyzing the deacetalization-Henry cascade reaction.


Need for Affect, Interpersonal Psychological Theory of Suicide, and Suicide Proneness.
The present study expands upon the Interpersonal Psychological Theory of Suicide (IPTS), examining its relationship with the need for affect (NFA), a construct explaining attitudes toward seeking both positive and negative emotions. A sample of 576 emerging adults completed measures of NFA, IPTS, suicide proneness, and demographics online. Findings include NFA accounting for a small significant amount of suicide proneness, a positive association between approaching emotions and one indicator of suicide proneness, negative association between approaching emotions and thwarted belongingness, and positive associations between avoidance of emotions with both perceived burdensomeness and thwarted belongingness. Findings are discussed concerning NFA as a theoretically and practically relevant factor for understanding suicide.


Bartholin Duct Cyst and Nodular Hyperplasia: A Clinicopathological Study of 32 Patients.
Background: Inflammatory lesions and cysts are the most common Bartholin glands pathologic diagnoses. There have been very few reports of benign solid lesions of Bartholin glands such as nodular hyperplasia. Materials and methods: Thirty-two lesions diagnosed as Bartholin duct cyst in the gynecologic specialized institute were reviewed by two expert gynecological pathologists. Results: Using specific criteria, 7 qualified as nodular hyperplasia, 25 as Bartholin duct cyst. The average age of the patients with nodular hyperplasia was 31 years (range, 24 to 43). These lesions were solid, solid and cystic or purely cystic, had a mean maximal dimension of 3.3 cm, and were frequently thought to be Bartholin cysts on clinical examination. Four lesions were diagnosed as Bartholin duct cysts, while in the review they were considered as nodular hyperplasia. Conclusion: Nodular hyperplasia of Bartholin gland may be more common than is reported in the literature. This study showed that the diagnosis of nodular hyperplasia has been overlooked due to limited literature sources.


Influence of nitrogen and potassium fertilization on leaf lifespan and allocation of above-ground growth in Eucalyptus plantations.
Eucalyptus grandis (W. Hill ex Maiden) leaf traits and tree growth were studied over 3 years after the establishment of two adjacent complete randomized block designs in southern Brazil. In a nitrogen (N) input experiment, a treatment with the application of 120 kg N ha(-1) was compared to a control treatment without N addition, and in a potassium (K) input experiment a control treatment without K addition was compared to a treatment with the application of 116 kg K ha(-1). Young leaves were tagged 9 months after planting to estimate the effect of N and K fertilizations on leaf lifespan. Leaf mass, specific leaf area and nutrient concentrations were measured on a composite sample per plot every 28 days until the last tagged leaf fell. Successive inventories, destructive sampling of trees and leaf litter fall collection made it possible to assess the effect of N and K fertilization on the dynamics of biomass accumulation in above-ground tree components. Whilst the effects of N fertilization on tree growth only occurred in the first 24 months after planting, K fertilization increased the above-ground net primary production from 4478 to 8737 g m(-2) over the first 36 months after planting. The average lifespan of tagged leaves was not modified by N addition but it increased from 111 to 149 days with K fertilization. The peak of leaf production occurred in the second year after planting (about 800 g m(-2) year(-1)) and was not significantly modified (P < 0.05) by N and K fertilizations. By contrast, K addition significantly increased the maximum leaf standing biomass from 292 to 528 g m(-2), mainly as a consequence of the increase in leaf lifespan. Potassium fertilization increased the stand biomass mainly through the enhancement in leaf area index (LAI) since growth efficiency (defined as the ratio between woody biomass production and LAI) was not significantly modified. A better understanding of the physiological processes governing the leaf lifespan is necessary to improve process-based models currently used in Eucalyptus plantations.


Refined Quantification of Infection Bottlenecks and Pathogen Dissemination with STAMPR.
Pathogen population dynamics during infection are critical determinants of infection susceptibility and define patterns of dissemination. However, deciphering these dynamics, particularly founding population sizes in host organs and patterns of dissemination between organs, is difficult because measuring bacterial burden alone is insufficient to observe these patterns. Introduction of allelic diversity into otherwise identical bacteria using DNA barcodes enables sequencing-based measurements of these parameters, in a method known as STAMP (Sequence Tag-based Analysis of Microbial Populations). However, bacteria often undergo unequal expansion within host organs, resulting in marked differences in the frequencies of barcodes in input and output libraries. Here, we show that these differences confound STAMP-based analyses of founding population sizes and dissemination patterns. We present STAMPR, a successor to STAMP, which accounts for such population expansions. Using data from systemic infection of barcoded extraintestinal pathogenic E. coli, we show that this new framework, along with the metrics it yields, enhances the fidelity of measurements of bottlenecks and dissemination patterns. STAMPR was also validated on an independent barcoded Pseudomonas aeruginosa data set, uncovering new patterns of dissemination within the data. This framework (available at https://github.com/hullahalli/stampr_rtisan), when coupled with barcoded data sets, enables a more complete assessment of within-host bacterial population dynamics. IMPORTANCE Barcoded bacteria are often employed to monitor pathogen population dynamics during infection. The accuracy of these measurements is diminished by unequal bacterial expansion rates. Here, we develop computational tools to circumvent this limitation and establish additional metrics that collectively enhance the fidelity of measuring within-host pathogen founding population sizes and dissemination patterns. These new tools will benefit future studies of the dynamics of pathogens and symbionts within their respective hosts and may have additional barcode-based applications beyond host-microbe interactions.


Do cardiologists have higher thresholds for recommending coronary arteriography than family physicians?
The purpose of this study was to use a new model of decision making to understand variability in physicians' utilization of diagnostic tests. We studied physicians' recommendations for coronary arteriography in hypothetical patients with chest pain by analyzing responses of 235 cardiologists and family physicians. Thresholds for testing were derived by obtaining estimates of the probability of disease and recommendations for coronary arteriography before and after an exercise test. We found that cardiologists compared with family practitioners had a significantly higher decision threshold and recommended coronary arteriography in fewer patients. These findings suggest that analyzing physicians' decision-making thresholds may be used to characterize differences in the practice behavior of groups of physicians.


Regional survival differences across Europe in HIV-positive people: the EuroSIDA study.
To analyse the survival differences between macro-regions of Europe (northern, central and southern Europe) between 1994 and early 1999, and their possible association with antiretroviral treatment differences. From September 1994 the EuroSIDA study has prospectively followed non-selected HIV-infected people from 50 clinical sites in 18 European countries (n = 7331). Cox proportional hazards models were used to compare death rates between regions and to investigate the relationship between treatment usage and regional mortality rates. Kaplan-Meier curves were used to compare survival from the first CD4 lymphocyte count of < 200 x 10(6)/l or < 50 x 10(6)/l. At the time of analysis, the median follow-up was 21 months and there was a total of 1544 deaths. In people with a CD4+ cell count that fell below 200 or 50 x 10(6)/l those from central Europe had a better prognosis compared with those from the two other regions (P < 0.05). Patients from central Europe were more frequently exposed to reverse transcriptase inhibitors and protease inhibitors compared with patients from other regions (P < 0.001). There was a significant difference in risk of death between regions after adjustment for baseline differences in demography, presence of AIDS and level of immunodeficiency (risk of death in central Europe was 37% lower than that in southern Europe (P < 0.0001) and 33% lower than in northern Europe (P < 0.0001)). After adjustment for use of individual antiretroviral agents, intensity of treatment regimen, CD4 lymphocyte count, weight, haemoglobin and development of AIDS as time-dependent covariates, the differences became much smaller (risk in central Europe 13% lower than that in southern Europe (P = 0.071) and 15% lower than in northern Europe (P = 0.054). Antiretroviral therapy has been used more aggressively in Europe in recent years, resulting in improved prognosis. In this study we observed that the HIV mortality rate in central Europe was significantly lower than those in northern and southern Europe in the period 1994 to early 1999. This finding appears to be due to the effect on survival of different treatment policies and drug availability in the three regions of Europe during this time period, with central European countries, on average, having introduced more aggressive treatment strategies earlier.


Ardnamurchan 3D cone-sheet architecture explained by a single elongate magma chamber.
The Palaeogene Ardnamurchan central igneous complex, NW Scotland, was a defining place for the development of the classic concepts of cone-sheet and ring-dyke emplacement and has thus fundamentally influenced our thinking on subvolcanic structures. We have used the available structural information on Ardnamurchan to project the underlying three-dimensional (3D) cone-sheet structure. Here we show that a single elongate magma chamber likely acted as the source of the cone-sheet swarm(s) instead of the traditionally accepted model of three successive centres. This proposal is supported by the ridge-like morphology of the Ardnamurchan volcano and is consistent with the depth and elongation of the gravity anomaly underlying the peninsula. Our model challenges the traditional model of cone-sheet emplacement at Ardnamurchan that involves successive but independent centres in favour of a more dynamical one that involves a single, but elongate and progressively evolving magma chamber system.


Structure and function of the chromogranin A gene. Clues to evolution and tissue-specific expression.
Chromogranin A is the index member of a family of acidic proteins stored and released throughout the neuroendocrine system with peptide hormones and neurotransmitters. To better understand its functional domains, its evolutionary lineage, and the basis of its tissue-specific pattern of expression, we obtained a mouse chromogranin A cDNA and used it to isolate the chromogrannin A gene from the mouse genome. We then characterized the gene's exon/intron structure, and the structure and function of its 5'-regulatory region (promoter/enhancer). The chromogranin A gene was complex, its eight exons and seven introns spanning about 11 kilobase pairs. The eight exons displayed some correspondence to putative functional domains suspected within the cDNA-deduced primary structure of the protein. Three exons also displayed both length and sequence homology to exons in another member of the chromogranin/secretogranin family, chromogranin B, suggesting an evolutionary relationship. A 1.2-kilobase pair genomic fragment just 5' of the coding region was able to program cell type-specific gene expression in a transfection/reporter system. Both pituitary corticotropes and adrenal chromaffin cells recognized this promoter. The promoter possessed some known consensus transcriptional control elements (TATA box, cyclic AMP-response element, and Sp1 site), but otherwise novel transcriptional control elements seem to be operative. The results suggest that this complex gene is encoded by exon modules with evolutionary links to homologous modules in other chromogranin/secretogranin protein family members, and that the 5'-flanking region of the gene is sufficient to confer neuroendocrine tissue-specific expression of the chromogranin A gene.


Breath [13CO2] recovery from an oral glucose load during exercise: comparison between [U-13C] and [1,2-13C]glucose.
The purpose of the present experiment was to compare 13CO2 recovery at the mouth, and the corresponding exogenous glucose oxidation computed, during a 100-min exercise at 63 +/- 3% maximal O2 uptake with ingestion of glucose (1.75 g/kg) in six active male subjects, by use of [U-13C] and [1,2-13C]glucose. We hypothesized that 13C recovery and exogenous glucose oxidation could be lower with [1,2-13C] than [U-13C]glucose because both tracers provide [13C]acetate, with possible loss of 13C in the tricarboxylic acid (TCA) cycle, but decarboxylation of pyruvate from [U-13C]glucose also provides 13CO2, which is entirely recovered at the mouth during exercise. The recovery of 13C (25.8 +/- 2.3 and 27.4 +/- 1.2% over the exercise period) and the amounts of exogenous glucose oxidized computed were not significantly different with [1,2-13C] and [U-13C]glucose (28.9 +/- 2.6 and 30.7 +/- 1.3 g, between minutes 40 and 100), suggesting that no significant loss of 13C occurred in the TCA cycle. This stems from the fact that, during exercise, the rate of exogenous glucose oxidation is probably much larger than the flux of the metabolic pathways fueled from TCA cycle intermediates. It is thus unlikely that a significant portion of the 13C entering the TCA cycle could be diverted to these pathways. From a methodological standpoint, this result indicates that when a large amount of [13C]glucose is ingested and oxidized during exercise, 13CO2 production at the mouth accurately reflects the rate of glucose entry in the TCA cycle and that no correction factor is needed to compute the oxidative flux of exogenous glucose.


Diaporine, a novel endophyte-derived regulator of macrophage differentiation.
Diaporine (1), an unprecedented symmetric polyketide, was characterized from the endophytic fungus. The structure was determined by extensive spectroscopic analyses. Diaporine can inhibit significantly the differentiation of macrophages and has potential to induce conversion from the M2 to the M1 phenotype, in addition to regulation of the TLR4-MAPK signal pathway and PPARγ activity.


Increased apoptosis and decreased density of medial smooth muscle cells in human abdominal aortic aneurysms.
To determine the increase of apoptosis and the decrease of smooth muscle cells (SMCs) density in human abdominal aortic aneurysms (AAA). In situ terminal transferase-mediated dUTP nick end labeling (TUNEL) was employed to detect apoptosis of SMCs in patients with AAA (n = 25) and normal abdominal aortae (n = 10). Positive cells were identified by specific cell marker in combination with immunohistochemistry. Meanwhile SMC counting was performed by anti-alpha-actin immunohistostaining to compare the SMC density. TUNEL staining revealed that there was significantly increased apoptosis in AAAs (average 8.6%) compared with normal abdominal aortae (average 0.95%, P < 0.01). Double staining showed that most of these cells were SMCs. Counting of alpha-actin positive SMCs revealed that medial SMC density of AAAs (37.5 +/- 7.6 SMCs/HPF) was reduced by 79.1% in comparison with that of normal abdominal aortae (179.2 +/- 16.1 SMCs/HPF, P < 0.01). Significantly increased SMCs of AAA bear apoptotic markers initiating cell death. Elevated apoptosis may result in a decreased density of SMCs in AAA, which may profoundly influence the development of AAA.


Cell membrane-associated MT1-MMP dependent activation of MMP-2 in SiHa (human cervical cancer) cells.
Among the soluble MMPs, MMP-2 (gelatinase A) is particularly important in the invasive property of tumor cells. Cell membrane-associated MMP-2 activation is one of the challenging areas in tumor biology. In the present communication, we studied the membrane dependent activation of MMP-2 in SiHa cells. Activation of pro-MMP-2 by membrane fraction, membrane extract, and live SiHa cells was studied by gelatin zymography. The role of MT1-MMP in MMP-2 activation was studied by incubating SiHa cells and cell membrane fractions with anti-MT1-MMP antibody. Activation of purified pro-MMP-2 by membrane fraction isolated from SiHa cells, by SiHa cell membrane extract and by SiHa cells, pro-MMP-2 from Con A treated HT-1080 conditioned medium by SiHa cells, and pro-MMP-2 from serum free culture medium of SiHa cells and cervical tissue homogenate by SiHa cell membrane fraction was shown by gelatin zymography. SiHa membrane fraction activated only pro-MMP-2 from purified MMP-9/MMP-2 mixture, indicating that the activation is specific for MMP-2. Inhibition of MMP-2 activation in the presence of anti-MT1-MMP antibody strongly indicated that the cell membrane mediated MMP-2 activation is MT1-MMP dependent. Immunocytochemistry of SiHa cells demonstrated expression of MT1-MMP at focal points. Invasion assay showed that invasiveness of anti-MT1-MMP antibody treated SiHa cells through Matrigel was drastically reduced compared to control SiHa cells. Our findings furnish an example of the cell membrane-associated MT1-MMP mediated MMP-2 activation in SiHa cells and suggest that this MT1-MMP mediated MMP-2 activation is of importance in tumor invasion and metastasis. This MT1-MMP mediated MMP-2 activation on tumor cell surface could be a realistic target for managing metastatic diseases.


Study on intracranial meningioma using PET ligand investigation during follow-up over years (SIMPLIFY).
Radiologic follow-up of patients with a meningioma at the skull base or near the venous sinuses with magnetic resonance imaging (MRI) after stereotactic radiotherapy (SRT) and neurosurgical resection(s) can be difficult to interpret. This study evaluates the addition of 11C-methionine positron emission tomography (MET-PET) to the regular MRI follow-up. This prospective pilot study included patients with predominantly WHO grade I meningiomas at the skull base or near large vascular structures. Previous SRT was part of their oncological treatment. A MET-PET in adjunct to their regular MRI follow-up was performed. The standardized uptake value (SUV) was determined for the tumor and the healthy brain, on the pre-SRT target delineation MET-PET and the follow-up MET-PET. Tumor-to-normal ratios were calculated, and 11C-methionine uptake over time was analyzed. Agreement between the combined MRI/MET-PET report and the MRI-only report was determined using Cohen's κ. Twenty patients with stable disease underwent an additional MET-PET, with a median follow-up of 84 months after SRT. Post-SRT SUV T/N ratios ranged between 2.16 and 3.17. When comparing the pre-SRT and the post-SRT MET-PET, five categories of SUV T/N ratios did not change significantly. Only the SUVpeak T/Ncortex decreased significantly from 2.57 (SD 1.02) to 2.20 (SD 0.87) [p = 0.004]. A κ of 0.77 was found, when comparing the MRI/MET-PET report to the MRI-only report, indicating no major change in interpretation of follow-up data. In this pilot study, 11C-methionine uptake remained remarkably high in meningiomas with long-term follow-up after SRT. Adding MET-PET to the regular MRI follow-up had no impact on the interpretation of follow-up imaging.


Peripheral GABA(A) receptors: evidence for peripheral primary afferent depolarization.
We propose that the primary afferent depolarization that follows GABA(A) receptor activation in the spinal cord also occurs in the periphery. As evidence, the present study localizes beta2/beta3 and alpha1 subunits of the GABA(A) receptor on 10-14% of the unmyelinated primary afferents axons in the glabrous skin of the cat paw. Behavioral studies demonstrate that local peripheral injection of the GABA(A) agonist muscimol at a low concentration (2.0 microM) attenuates, and at a high concentration (1 mM) enhances, formalin-induced nociceptive behaviors. Intraplantar injection of muscimol alone at a high dose evokes thermal hyperalgesia. Bicuculline, a GABA(A) antagonist, prevents these muscimol-induced changes in behavior. The muscimol-induced effects are due to local rather than systemic or central activation of GABA(A) receptors, as such effects are not observed in the contralateral paw. We interpret these findings to indicate that activation of GABA(A) receptors by low concentrations of muscimol depolarizes peripheral primary afferent terminals, a phenomenon we call peripheral primary afferent depolarization, in turn reducing the size of the peripheral action potentials and concomitantly reducing the amount of algogenic substances released from the peripheral terminals of these fibers. This sequence of events presumably results in a reduction in nociceptor activation. Higher concentrations of muscimol further depolarize GABA(A) receptor-containing terminals, which then initiates action potentials in nociceptors analogous to the appearance of dorsal root reflexes that arise following activation of GABA(A) receptors on central primary afferent terminals. These latter events reverse the analgesic effects of GABA(A) ligands and lead to potentiation of nociceptive input. Thus, the present study provides anatomical and behavioral evidence supporting a bimodal role for GABA(A) receptors in the modulation of peripheral nociceptive transmission.


Berberine: complex with DNA.
A complex of calf-thymus DNA with berberine sediments in the analytical ultracentrifuge. The DNA produced systematic changes in the absorption spectrum of berberine which suggest that single alkaloid molecules bind to DNA. Flow dichroism of purines and pyrimidines and of berberine in the complex with DNA had the same signs and magnitudes. Berberine shifted the thermal strand separation profile of DNA to higher temperatures. Therefore, the alkaloid forms a complex with DNA, probably by intercalation.


Mediating Effects of Social Support on Quality of Life for Parents of Adults with Autism.
The aim of this study was to examine the mediating effect of formal and informal social support on the relationship of caregiver burden and quality of life (QOL), using a sample of 320 parents (aged 50 or older) of adult children with autism spectrum disorder (ASD). Multiple linear regression and mediation analyses indicated that caregiver burden had a negative impact on QOL and that informal social support partially mediated the relationship between caregiver burden and parents' QOL. Formal social support did not mediate the relationship between caregiver burden and QOL. The findings underscored the need to support aging parents of adult children with ASD through enhancing their informal social support networks.


[Combination chemotherapy and chemoembolization in the treatment of primary inoperable hepatoblastoma].
A 33 months old girl and a 5 months old boy presented with unresectable hepatoblastoma (group III) and were treated with combination chemotherapy using Adriamycin, Cyclophosphamide, Cisplatinum (regimen I) and Adriamycin, Etoposide, Cisplatinum (regimen II). In both patients tumor size and alpha-fetoprotein levels decreased remarkably. Tumor regression was considerably enhanced by selective chemoembolization (Ethibloc, Adriamycin) of the right hepatic artery in one patient. After preoperative chemotherapy hepatoblastomas could be removed surgically. Local recurrence in the older patient was treated again successfully by surgery and chemotherapy. The patients are surviving disease-free 17 and 28 months from diagnosis. The efficacy of the treatment warrants prospective clinical trials in patients with unresectable hepatoblastoma.


Use of Antiplatelet Therapy/DAPT for Post-PCI Patients Undergoing Noncardiac Surgery.
Dual antiplatelet therapy (DAPT) is prescribed to millions of patients worldwide following coronary stenting. DAPT is indicated to lower the risk of ischemic events, such as myocardial infarction, including stent thrombosis, ischemic stroke, or death from cardiovascular causes. A significant number of these patients undergo noncardiac surgery and may require DAPT interruption. This poses a significant clinical dilemma because DAPT interruption exposes patients to the potential risk of stent thrombosis, perioperative myocardial infarction, or both. Conversely, continuing DAPT may be associated with excess bleeding complications. Observational data in this area are conflicting, and there are no randomized clinical trials to guide practitioner decision making. On the basis of predominantly consensus recommendations, various strategies for managing DAPT during the perioperative period have been proposed. This review presents 3 commonly encountered clinical scenarios that lead into an evidence-based discussion of practical strategies for managing perioperative antiplatelet therapy in patients following percutaneous coronary intervention.


Multi-parametric phospho-flow cytometry: a crucial tool for T lymphocyte signaling studies.
Tools such as protein immunoblotting have proven benefits for investigating T lymphocyte signaling but have several drawbacks such as the number of cells required and the difficulty of distinguishing subset-specific differences without expensive and invasive cell sorting. Recent advances in immunology and the identification of T lymphocyte sub-populations making up only a very small fraction of the total population highlight the importance of studying signaling in those small subsets in a feasible, cost-effective, high-throughput manner. To this end, we have developed a simplified protocol to study both intracellular phosphorylation patterns of important signal transduction molecules concomitantly with T cell surface marker expression. A multi-parametric analysis may allow the quantification of the phosphorylation of up to five signaling molecules in CD4 and CD8 T lymphocytes and their naïve, central memory, effector memory, and TEMRA subsets. This enables precise identification of subset-specific signaling and alterations of signaling pathways in physiological and pathological situations. The importance of such detailed analysis is discussed.


Werner Forssmann - A Nobel Prize Winner and His Political Attitude before and after 1945.
In 1956, the Nobel Prize in Physiology or Medicine was awarded jointly to Werner Forssmann, André Frédéric Cournand and Dickinson W. Richards for their development of cardiac catheterization. Forssmann performed a self-experiment in 1929 by inserting a urethral catheter into his right ventricular cavity via his antecubital vein. Despite his popularity as one of the first German Nobel Laureates after 1945, little is known about Forssmann personally. This contribution aims to close this gap regarding the Nazi period and early post-war Germany. Primary historical sources from Forssmann's private archive were examined, evaluated and interpreted for the first time. Additionally, a comparative analysis based on further archival and secondary sources was performed. Werner Forssmann joined the Nazi Party, the Sturmabteilung (Stormtroopers, SA) and the Nationalsozialistischer Deutscher Ärztebund (Nazi Doctors' Association) in 1932, a year before Hitler's assumption of power. In his autobiography, Forssmann referred to the political situation in passing. However, he expressed his personal thoughts in private letters which he wrote as a medical officer during the war. After World War II, Forssmann underwent denazification and was banned from practicing medicine for 3 years. He did not seem to be averse to Hitler's politics and in some ways expressed his approval. However, correspondence from the 1960s with 2 Jewish colleagues reveals that Forssmann may have changed his attitude toward National Socialism later. Werner Forssmann's political attitudes during the Third Reich and in the post-war era can be characterized as early agreement that gradually changed to a more critical distance to Nazi ideology. In this respect, Forssmann appears to be quite a typical example of a larger proportion of German medical doctors during these eras.


Unit activity in medial thalamus: comparative effects of caffeine an amphetamine.
The effects of caffeine and d-amphetamine sulfate were evaluated upon single unit activity of medial thalamus in chloral hydrate anesthetized rats. Single unit activity recorded extracellularly with platinum-iridium microelectrodes. It was found that caffeine, 0.1 to 0.5 mg/kg i.v., markedly suppressed and that amphetamine, 0.1 to 1 mg/kg i.v., markedly augmented the spontaneous firing rates of medial thalamic neurons. It was also demonstrated that amphetamine was capable of activating the medial thalamic neuronal activity while it was suppressed by caffeine. On the other hand, while the firing rate was increased by a previous amphetamine injection, administration of caffeine suppressed the amphetamine-induced augmentation of neuronal activity. The data imply that these two central nervous system (CNS) stimulants, caffeine and amphetamine, have different mechanisms of action at least at the medial thalamic site. Moreover, since medial thalamus participates in the recruitment phenomenon and exerts a stabilizing action on the cerebral cortex, our findings that caffeine suppressed this brain area suggest that the medial thalamus may be an important site of action for the arousal induced by caffeine. In addition, the demonstration of inhibitory effects of caffeine on the medial thalamus in this study and excitatory effects on brain stem reticular formation from our previous studies indicate that caffeine has differential actions on different brain areas. This observation broadens the generally held belief that caffeine excites at all levels of the CNS to specifically include inhibition of inhibitory influences in at least one functional system of the brain.


Modulation of antigen-induced B and T cell responses by antigen-specific IgE antibodies.
Ag-specific IgE Abs not only mediate immediate hypersensitivity through mast cell activation, but also enhance in vitro Ag presentation and in vivo specific Ab responses in mice. To delineate the role of IgE Ab in the modulation of Ag-specific responses, spleen cells from OVA-sensitized BALB/C mice were cultured together with OVA-specific IgE (or IgG isotypes). OVA-dependent proliferative responses and anti-OVA IgE production were enhanced in the presence of anti-OVA IgE. A significant decrease in IFN-gamma secretion in OVA-stimulated cultures was observed in the presence of anti-OVA IgE, but no changes in IL-4 production were detected. Anti-OVA IgG isotypes or anti-TNP IgE showed no significant effect on any of these Ag-dependent responses. Addition of anti-CD23 Ab abolished these effects of anti-OVA IgE. Further, OVA-sensitized spleen cells from CD23-deficient mice responded to in vitro stimulation with OVA, but demonstrated no modulation by anti-OVA IgE. These results demonstrate that Ag-specific IgE not only augments Ag presentation and T cell proliferation, but also alters the pattern of cytokine production and increases specific IgE synthesis. These modulatory effects of Ag-specific IgE appear to be mediated by binding to Fc epsilon RII/CD23.


Selective growth-inhibiting effects of compounds identified in Tabebuia impetiginosa inner bark on human intestinal bacteria.
The growth-inhibiting activity of anthraquinone-2-carboxylic acid and lapachol identified in the inner bark of taheebo, Tabebuia impetiginosa, toward 10 human intestinal bacteria was evaluated by using a paper disk diffusion bioassay and compared to those of seven lapachol congeners (1,4-naphthoquinone, naphthazarin, menadione, lawsone, plumbagin, juglone, and dichlone) as well as two commercially available antibiotics, chloramphenicol and tetracycline. Anthraquinone-2-carboxylic acid exhibited very strong growth inhibition of Clostridium paraputrificum at 1 microg/disk while 100 microg/disk of lapachol was needed for moderate growth inhibition of the same organism. These two isolates exhibited weak inhibition of Clostridium perfringens and Escherichia coli at 100 microg/disk while no adverse effects were observed on the growth of Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium infantis, Lactobacillus acidophilus, and Lactobacillus casei at 1000 microg/disk. Structure-activity relationships indicate that a methyl group in the C-2 position of 1,4-naphthoquinone derivatives might play an important role in antibacterial activity.


Biodegradation of acrylic based resins: A review.
The development of different types of materials with application in dentistry is an area of intense growth and research, due to its importance in oral health. Among the different materials there are the acrylic based resins that have been extensively used either in restorations or in dentures. The objective of this manuscript was to review the acrylic based resins biodegradation phenomena. Specific attention was given to the causes and consequences of materials degradation under the oral environment. Information from scientific full papers, reviews or abstracts published from 1963 to date were included in the review. Published material was searched in dental literature using general and specialist databases, like the PubMED database. Published studies regarding the description of biodegradation mechanisms, in vitro and in vivo release experiments and cell based studies conducted on acrylic based resins or their components were evaluated. Studies related to the effect of biodegradation on the physical and mechanical properties of the materials were also analyzed. Different factors such as saliva characteristics, chewing or thermal and chemical dietary changes may be responsible for the biodegradation of acrylic based resins. Release of potential toxic compounds from the material and change on their physical and mechanical properties are the major consequences of biodegradation. Increasing concern arises from potential toxic effects of biodegradation products under clinical application thus justifying an intensive research in this area.


[Contemporary dietotherapy of the irritable bowel syndrome].
Irritable bowel syndrome (IBS) is the most prevalent functional disease of the gastrointestinal tract. This highly prevalent condition is best diagnosed by assessing the constellation of symptoms with which patients present to their physicians. Because some critics have previously questioned whether irritable bowel syndrome and other functional gastrointestinal disorders truly exist because they do not have defining structural features, the Rome Foundation fostered the use of symptom-based criteria for universal use. In most cases treatment is reduced to symptomatic therapy because a lot of unknown in pathogenesis by irritable bowel syndrome. Irritable bowel syndrome leads to decrease of quality of life of the patients and could be one of the reasons of patients' disability. Food is believed by patients promotes symptoms and the diet or avoiding specific food can reduce symptoms. Possible role of different food and microbiota in the pathophysiology of irritable bowel syndrome, as well as the data from randomized, controlled clinical trials dedicated to the effects of diet in irritable bowel syndrome are summarized and discussed in this review. The efficacy of the diet, enriched by fiber, prebiotics, probiotics, peppermint oil, curcumin and vitamin B6 in irritable bowel syndrome patients was shown in numerous studies. In some studies restriction in consumption of fermented carbohydrates, coffee and alcohol, as well as diet with elimination IgG-sensed food was also shown to be effective in irritable bowel syndrome. Food intolerances, defined as non-toxic non-immune adverse reactions to food, include reactions to bioactive chemicals in foods and metabolic reactions to poorly absorbed dietary carbohydrates. New dietary approaches like polyunsaturated fatty acids intake correction and the low tryptophan intake are discussed.


Tryptophan and Kynurenine Metabolites: Are They Related to Depression?
Some previous studies found decreased concentrations of L-tryptophan (TRY) and increased L-kynurenine (KYN), or its metabolites, in the body fluids of subjects with major depressive disorder (MDD), sometimes in association with suicidal behavior. Such changes might indicate a shift of TRY away from serotonin production, possibly via the effects of inflammatory peptides which activate indoleamine-2,3-dioxygenase. However, these findings have been inconsistent and require replication. We used sensitive liquid-chromatography mass spectrometry methods to assay plasma concentrations of TRY, 5-hydroxyindoleacetic acid (5-HIAA), and KYN and its metabolites (anthranilic acid and xanthurenic acid). We compared 49 hospitalized, depressed subjects diagnosed with MDD (n = 37) or bipolar disorder (BD, n = 12), with (n = 22) or without (n = 27) previous suicide attempts, to 78 healthy, ambulatory controls of similar age and sex (total n = 127). Contrary to expectation, TRY plasma concentrations were higher, KYN plasma concentrations were lower, and their ratio much higher in depressed subjects, with no relationship to suicidal history. Concentrations of 5-HIAA and the kynurenine metabolites did not differ between depressed and healthy subjects. These findings are opposite to expectations and not consistent with a hypothesized increased conversion from TRY to KYN in depressed subjects. In addition, we found no evidence of altered production of serotonin as 5-HIAA concentration was unchanged. None of the observed changes was associated with a history of suicide attempt.


A case of acquired smooth muscle hamartoma on the sole.
A smooth muscle hamartoma is a benign proliferation of smooth muscle bundles within the dermis. It arises from smooth muscle cells that are located in arrector pili muscles, dartos muscles, vascular smooth muscles, muscularis mammillae and the areolae. Acquired smooth muscle hamartoma (ASMH) is rare, with only 10 such cases having been reported in the English medical literature to date. Most of these cases of ASMH were shown to have originated from arrector pili and dartos muscles. Only one case was reported to have originated from vascular smooth muscle cells. A 21 year-old woman presented with a tender pigmented nodule, with numbness, on the sole of her foot, and this lesion had developed over the previous 18 months. The lesion showed no hyperpigmentation or hypertrichosis, and the biopsies demonstrated increased smooth muscle bundles in the dermis, and especially around the blood vessels. Moreover, the specimens stained positive with Masson trichrome stain and alpha-smooth-muscle actin antibodies, thus supporting our diagnosis of ASMH of the foot sole. Herein, we report on a rare case of ASMH on the foot sole, and this lesion originated from vascular smooth muscle cells. This type of case has not been previously described in the English medical literature.


Rapid resolution liquid chromatography (RRLC) analysis for quality control of Rhodiola rosea roots and commercial standardized products.
A simple, sensitive and reliable reversed phase Rapid Resolution Liquid Chromatography (RRLC) method was developed and validated for six biologically active compounds (salidroside, tyrosol, rosarin, rosavin, rosin and rosiridin) in Rhodiola rosea L. roots and powder extracts. The method uses a Phenomenex C18 (2)-HST column at 40 degrees C with a neutral gradient system mobile phase (H20 and acetonitrile), a flow rate of 1.0 mL/min, and UV detection wavelengths set at 205 and 254 nm, simultaneously. Baseline separation of the six active compounds was achieved within 8 minutes. The average percentages of rosavins (rosarin, rosavin, and rosin) in authentic R. rosea roots and root powder extracts were quantitatively determined and a characteristic R. rosea roots RRLC profile was established. The RRLC method is accurate and sensitive; in addition, it effectively increases the sample analysis throughput compared with conventional HPLC.


Sale-based estimation of pharmaceutical concentrations and associated environmental risk in the Japanese wastewater system.
Information on sales and emission of selected pharmaceuticals were used to predict their concentrations in Japanese wastewater influent through a >300 of pharmaceuticals data sink. A combined wastewater-based epidemiology and environmental risk analysis follow was established. By comparing predicted environmental concentrations (PECs) of pharmaceuticals in wastewater influent against measured environmental concentrations (MECs) reported in previous studies, it was found that the model gave accurate results for 17 pharmaceuticals (0.5 < PEC/MEC < 2), and acceptable results for 32 out of 40 pharmaceuticals (0.1 < PEC/MEC < 10). Although the majority of pharmaceuticals considered in the model were antibiotics and analgesics, pranlukast, a receptor antagonist, was predicted to have the highest concentration in wastewater influent. With regard to the composition of wastewater effluent, the Estimation Program Interface (EPI) suite was used to predict pharmaceutical removal through activated sludge treatment. Although the performance of the EPI suite was variable in terms of accurate prediction of the removal of different pharmaceuticals, it could be an efficient tool in practice for predicting removal under extreme scenarios. By using the EPI suite with input data on PEC in the wastewater influent, the PEC values of pharmaceuticals in wastewater effluent were predicted. The concentrations of 26 pharmaceuticals were relatively high (>1 μg/L), and the PECs of 6 pharmaceuticals were extremely high (>10 μg/L) in wastewater effluent, which could be attributed to their high usage rates by consumers and poor removal rates in wastewater treatment plants (WWTPs). Furthermore, environmental risk assessment (ERA) was carried out by calculating the ratio of predicted no effect concentration (PNEC) to PEC of different pharmaceuticals, and it was found that 9 pharmaceuticals were likely to have high toxicity, and 54 pharmaceuticals were likely to have potential toxicity. It is recommended that this is further investigated in detail. The priority screening and environmental risk assessment results on pharmaceuticals can provide reliable basis for policy-making and environmental management.


Evaluation of mycobacteria growth indicator tube for direct and indirect drug susceptibility testing of Mycobacterium tuberculosis from respiratory specimens in a Siberian prison hospital.
The manual Mycobacteria Growth Indicator Tube (MGIT) method was evaluated for performing direct and indirect drug susceptibility testing (DST) of Mycobacterium tuberculosis for isoniazid and rifampin on 101 strongly smear-positive sputum specimens in a Siberian prison hospital. Using the indirect method of proportion (MOP) as the "gold standard," the accuracies of isoniazid and rifampin susceptibility testing by the direct MGIT system were 97.0 and 94.1%, respectively. The accuracy of the indirect MGIT system was 98.0% for both drugs. The turnaround times from specimen processing to reporting of the DST results ranged between 4 and 23 (mean, 9.2) days by the direct MGIT method, 9 and 30 (mean, 15.3) days by the indirect MGIT method, and 26 and 101 (mean, 59.6) days by the indirect MOP. MGIT appears to be a reliable, rapid, and convenient method for performing direct and indirect DSTs in low-resource settings, but further studies are required to refine the direct DST protocol. Cost is the only factor prohibiting widespread implementation of MGIT.


Factors extending the chronological lifespan of yeast: Ecl1 family genes.
Ecl1 family genes are conserved among yeast, in which their overexpression extends chronological lifespan. Ecl1 family genes were first identified in the fission yeast Schizosaccharomyces pombe; at the time, they were considered noncoding RNA owing to their short coding sequence of fewer than 300 base pairs. Schizosaccharomyces pombe carries three Ecl1 family genes, ecl1+, ecl2+ and ecl3+, whereas Saccharomyces cerevisiae has one, ECL1. Their overexpression extends chronological lifespan, increases oxidative stress resistance and induces sexual development in fission yeast. A recent study indicated that Ecl1 family genes play a significant role in responding to environmental zinc or sulfur depletion. In this review, we focus on Ecl1 family genes in fission yeast and describe the relationship between nutritional depletion and cellular output, as the latter depends on Ecl1 family genes. Furthermore, we present the roles and functions of Ecl1 family genes characterized to date.


Association of Functional Status, Cognition, Social Support, and Geriatric Syndrome With Admission From the Emergency Department.
The role of patient-level factors that are unrelated to the specific clinical condition leading to an emergency department (ED) visit, such as functional status, cognitive status, social supports, and geriatric syndromes, in admission decisions is not well understood, partly because these data are not available in administrative databases. To determine the extent to which patient-level factors are associated with rates of hospital admission from the ED. This cohort study analyzed survey data collected from participants (or their proxies, such as family members) enrolled in the Health and Retirement Study (HRS) from January 1, 2000, to December 31, 2018. These HRS data were linked to Medicare fee-for-service claims data from January 1, 1999, to December 31, 2018. Information on functional status, cognitive status, social supports, and geriatric syndromes was obtained from the HRS data, whereas ED visits, subsequent hospital admission or ED discharge, and other claims-derived comorbidities and sociodemographic characteristics were obtained from Medicare data. Data were analyzed from September 2021 to April 2023. The primary outcome measure was hospital admission after an ED visit. A baseline logistic regression model was estimated, with a binary indicator of admission as the dependent variable of interest. For each primary variable of interest derived from the HRS data, the model was reestimated, including the HRS variable of interest as an independent variable. For each of these models, the odds ratio (OR) and average marginal effect (AME) of changing the value of the variable of interest were calculated. A total of 42 392 ED visits by 11 783 unique patients were included. At the time of the ED visit, patients had a mean (SD) age of 77.4 (9.6) years, and visits were predominantly for female (25 719 visits [60.7%]) and White (32 148 visits [75.8%]) individuals. The overall percentage of patients admitted was 42.5%. After controlling for ED diagnosis and demographic characteristics, functional status, cognition status, and social supports all were associated with the likelihood of admission. For instance, difficulty performing 5 activities of daily living was associated with an 8.5-percentage point (OR, 1.47; 95% CI, 1.29-1.66) AME increase in the likelihood of admission. Having dementia was associated with an AME increase in the likelihood of admission of 4.6 percentage points (OR, 1.23; 95% CI, 1.14-1.33). Living with a spouse was associated with an AME decrease in the likelihood of admission of 3.9 percentage points (OR, 0.84; 95% CI, 0.79-0.89), and having children living within 10 miles was associated with an AME decrease in the likelihood of admission of 5.0 percentage points (OR, 0.80; 95% CI, 0.71-0.89). Other common geriatric syndromes, including trouble falling asleep, waking early, trouble with vision, glaucoma or cataract, use of hearing aids or trouble with hearing, falls in past 2 years, incontinence, depression, and polypharmacy, were not meaningfully associated with the likelihood of admission. Results of this cohort study suggest that the key patient-level characteristics, including social supports, cognitive status, and functional status, were associated with the decision to admit older patients to the hospital from the ED. These factors are critical to consider when devising strategies to reduce low-value admissions among older adult patients from the ED.


Formulation and biopharmaceutical evaluation of bitter taste masking microparticles containing azithromycin loaded in dispersible tablets.
The objective of this study was to prepare and evaluate some physiochemical and biopharmaceutical properties of bitter taste masking microparticles containing azithromycin loaded in dispersible tablets. In the first stage of the study, the bitter taste masking microparticles were prepared by solvent evaporation and spray drying method. When compared to the bitter threshold (32.43µg/ml) of azithromycin (AZI), the microparticles using AZI:Eudragit L100=1:4 and having a size distribution of 45-212µm did significantly mask the bitter taste of AZI. Fourier transform infrared spectroscopy (FTIR), and proton nuclear magnetic resonance spectroscopy (1H NMR) proved that the taste masking of microparticles resulted from the intermolecular interaction of the amine group in AZI and the carbonyl group in Eudragit L100. Differential scanning calorimeter (DSC) analysis was used to display the amorphous state of AZI in microparticles. Images obtaining from optical microscopy and scanning electron microscopy (SEM) indicated the existence of microparticles in regular cube shape with many layers. In the second stage, dispersible tablets containing microparticles (DTs-MP) were prepared by direct compression technique. Stability study was conducted to screen pH modulators for DTs-MP, and a combination of alkali agents (CaCO3:NaH2PO4, 2:1) was added into DTs-MP to create microenvironment pH of 5.0-6.0 for the tablets. The disintegration time of optimum DTs-MP was 53±5.29s and strongly depended on the kinds of lubricant and diluent. The pharmacokinetic study in the rabbit model using liquid chromatography tandem mass spectrometry showed that the mean relative bioavailability (AUC) and mean maximum concentration (Cmax) of DTs-MP were improved by 2.19 and 2.02 times, respectively, compared to the reference product (Zithromax®, Pfizer).


Subgroups of non-atypical hyperplasia of breast defined by proliferation of oestrogen receptor-positive cells.
In normal breast, there is a negative association between expression of oestrogen receptor (ER) and the proliferation marker Ki67, indicating that ER-positive (ER+) cells do not divide, or that the receptor is down-regulated when they do so. However, dual staining has been found in carcinomas and precancerous lesions, indicating that abnormal regulation of ER could be important in breast tumourigenesis. ER expression in relationship to cell proliferation was studied in 241 foci of hyperplasia of usual type (HUT), a lesion associated with a 1.5 to 2-fold increase in risk of developing breast cancer. Dual label immunofluorescence was employed, using the antibodies 1D5 and Ki67. Two hundred and thirteen foci of HUT contained ER+ cells, which were distributed singly or contiguously and increased with age. Most foci resembled normal breast, but 51 contained dual labelled cells, which did not increase with age. Some of these foci exhibited few, scattered ER+ cells with greater proliferation rates than the ER-negative (ER-) cells, whereas others contained many, contiguous ER+ cells, whose rate of proliferation was less than that of the ER- cells. The latter picture is similar to that which has previously been reported in atypical ductal hyperplasia and ductal carcinoma in situ. The first type of HUT may evolve into the second. The proportion of Ki67+ cells that was ER+ was similar in both types, suggesting a homeostatic mechanism that slows the proliferation of ER+ cells as they become confluent. Overriding this inhibition may be crucial in further progression. Non-atypical hyperplasia is thus heterogeneous in ER expression and proliferation and a significant proportion exhibit abnormal regulation of ER. These findings could have implications for pathological diagnosis, risk assessment, and prophylactic hormonal therapy.


The Escherichia coli heat shock protease HtrA participates in defense against oxidative stress.
The serine protease HtrA (DegP), which is indispensable for cell survival at elevated temperatures, is a peripheral membrane protein, localized on the periplasmic side of the inner membrane in Escherichia coli, and the biochemical and genetic evidence indicates that the physiological role of HtrA is to degrade denatured proteins formed in the cellular envelope during heat shock. The aim of this study was to find out if the HtrA protease contributes to protection of the cell against oxidative stress. We compared the influence of various oxidizing agents on htrA mutant cells with their effects on wild-type bacteria, and found that the htrA mutation did not increase sensitivity to hydrogen peroxide or paraquat but made the cell extremely sensitive to ferrous [Fe(II)] ions, which are known to enhance oxidation of proteins. Treatment with ferrous ions caused a larger increase in the level of protein carbonyl groups in the membrane fraction of the cell than in the periplasm and cytoplasm. Iron-induced oxidation of membrane proteins was enhanced in the htrA mutant relative to wild-type cells. Inhibition of the growth of the htrA mutant by iron could be alleviated more efficiently by a nitroxide antioxidant that localizes in the membranes (A-TEMPO) than by a derivative (40H-TEMPO) that acts mainly in the soluble fraction of the cell. Inhibition of the growth of the htrA mutant was more pronounced following treatment with cumene hydroperoxide, which partitions into membranes, than with t-butyl hydroperoxide, which forms radical mainly in the cytosol. Both ferrous ions and cumene hydroperoxide, but not hydrogen peroxide, paraquat or t-butyl hydroperoxide, induced synthesis of HtrA. Our results show that HtrA plays a role in defense against oxidative shock and support the hypothesis that HtrA participates in the degradation of oxidatively damaged proteins localized in the cell envelope, especially those associated with the membranes.


Digenean trematode infections of native freshwater snails and invasive Potamopyrgus antipodarum in the Grand Teton National Park/John D. Rockefeller Memorial Parkway Area.
Outside its native range, the invasive New Zealand mud snail (NZMS), Potamopyrgus antipodarum, is rarely reported to harbor parasites. To test this observation, 7 sites along the Snake River and Polecat Creek in the Grand Teton National Park/John D Rockefeller Memorial Parkway area (Wyoming) were surveyed for native aquatic snails, NZMS, and associated digenean trematodes, in July 2005. At 6 sites, native snails harbored patent digenean infections; within 2 hr, < or =10% of lymnaeid snails shed furcocercariae or xiphidiocercariae, and < or =42% of physid snails released furcocercariae or echinostome cercariae. Partial 18S rDNA sequences were recovered from several furcocercariae. Potamopyrgus antipodarum was present at, and collected from, 5 sites. Polymerase chain reaction assays targeting digenean rDNA sequences in DNA extracted from pools of 150 NZMS snails did not detect parasites. The examination of 960 NZMS by overnight shedding yielded 1 occurrence of (surface-encysted) metacercariae of an unclassified notocotylid (based on 18S and 28S rDNA sequences). The dissection of 150 ethanol-fixed NZMS (30/site) revealed 2 types of digenean metacercariae encysted in tissues of 5 snails from Polecat Creek. Thus, invasive NZMS may serve as first and second intermediate host for digenean parasites.


Overexpression of the DYRK1A Gene (Dual-Specificity Tyrosine Phosphorylation-Regulated Kinase 1A) Induces Alterations of the Serotoninergic and Dopaminergic Processing in Murine Brain Tissues.
Trisomy 21 (T21) or Down syndrome (DS) is the most common genetic disorder associated with intellectual disability and affects around 5 million persons worldwide. Neuroanatomical phenotypes associated with T21 include slight reduction of brain size and weight, abnormalities in several brain areas including spines dysgenesis, dendritic morphogenesis, and early neuroanatomical characteristics of Alzheimer's disease. Monoamine neurotransmitters are involved in dendrites development, functioning of synapses, memory consolidation, and their levels measured in the cerebrospinal fluid, blood, or brain areas that are modified in individuals with T21. DYRK1A is one of the recognized key genes that could explain some of the deficits present in individuals with T21. We investigated by high-performance liquid chromatography with electrochemical detection the contents and processing of monoamines neurotransmitters in four brain areas of female and male transgenic mice for the Dyrk1a gene (mBactgDyrk1a). DYRK1A overexpression induced dramatic deficits in the serotonin contents of the four brain areas tested and major deficits in dopamine and adrenaline contents especially in the hypothalamus. These results suggest that DYRK1A overexpression might be associated with the modification of monoamines content found in individuals with T21 and reinforce the interest to target the level of DYRK1A expression as a therapeutic approach for persons with T21.


[Cytomegalovirus infection and pneumocystis pneumonia after coronary stenting and cardiac transplantation for acute myocardial infarction].
The paper presents the clinical and anatomic data of combination treatment for acute myocardial infarction, by stenting the infarct-related artery, followed by cardiac transplantation, which have provided the optimal result of surgery and caused no severe rejection reaction. The immediate causes of death have been infectious complications (cytomegalovirus infection and pneumocystis pneumonia) developing in the presence of immunodeficiency state. The following diagnosis formulation is pathogenetically warranted: the underlying disease is "Secondary immunodeficiency due to immunosuppressive therapy" and infectious complications may be assigned to the individual rubric "Secondary disease" (i.e. occurring in immunodeficiency).


Drought response in the spikes of barley: gene expression in the lemma, palea, awn, and seed.
The photosynthetic organs of the barley spike (lemma, palea, and awn) are considered resistant to drought. However, there is little information about gene expression in the spike organs under drought conditions. We compared response of the transcriptome of the lemma, palea, awn, and seed to drought stress using the Barley1 Genome Array. Barley plants were exposed to drought treatment for 4 days at the grain-filling stage by withholding water. At the end of the stress, relative water content of the lemma, palea, and awn dropped from 85% to 60%. Nevertheless, the water content of the seed only decreased from 89% to 81%. Transcript abundance followed the water status of the spike organs; the awn had more drought-regulated genes followed by lemma and palea, and the seed showed very little change in gene expression. Despite expressing more drought-associated genes, many genes for amino acid, amino acid derivative, and carbohydrate metabolism, as well as for photosynthesis, respiration, and stress response, were down-regulated in the awn compared with the lemma, palea, and seed. This suggests that the lemma and the palea are more resistant to drought stress compared with the awn.


The perceptions and aspirations illicit drug users hold toward health care staff and the care they receive.
Over the 30 years of conflict, Northern Ireland escaped the worst excess of illegal drug trafficking and usage. However, the recent 'peace dividend' has brought with it an unprecedented rise in the availability and use of illicit drugs. With this, new problems and pressures have been brought to bear on the health service. The literature would suggest that drug users are loathed and feared by health care staff. Staff will also admit to be lacking in the knowledge and skills necessary for the delivery of appropriate support and treatment for this client group. Further, the literature has little to offer on the experiences and aspirations of drug users in relation to their treatment and the staff who care for them. In order to understand the drug users' experiences of health care and health staff, focus group methodology was employed to obtain qualitative data. A total of 20 illicit drug users from across Northern Ireland took part. Supporting the literature, all had experienced 'care' that they felt was filled with judgement, hostility and loathing. They recognized clearly the challenge they pose to health care staff. These findings indicate that there is obvious dissonance between those tasked to care for drug users and drug users themselves, with little respect being shown on either side. Results suggest that action needs to be taken to address the deficits in the knowledge, skills and values of health care professionals in relation to illicit drug users. The findings will be of interest to service providers within and outside the United Kingdom.


Urban air pollution and emergency room admissions for respiratory symptoms: a case-crossover study in Palermo, Italy.
Air pollution from vehicular traffic has been associated with respiratory diseases. In Palermo, the largest metropolitan area in Sicily, urban air pollution is mainly addressed to traffic-related pollution because of lack of industrial settlements, and the presence of a temperate climate that contribute to the limited use of domestic heating plants. This study aimed to investigate the association between traffic-related air pollution and emergency room admissions for acute respiratory symptoms. From January 2004 through December 2007, air pollutant concentrations and emergency room visits were collected for a case-crossover study conducted in Palermo, Sicily. Risk estimates of short-term exposures to particulate matter and gaseous ambient pollutants including carbon monoxide, nitrogen dioxide, and sulfur dioxide were calculated by using a conditional logistic regression analysis. Emergency departments provided data on 48,519 visits for respiratory symptoms. Adjusted case-crossover analyses revealed stronger effects in the warm season for the most part of the pollutants considered, with a positive association for PM10 (odds ratio = 1.039, 95% confidence interval: 1.020 - 1.059), SO2 (OR = 1.068, 95% CI: 1.014 - 1.126), nitrogen dioxide (NO2: OR = 1.043, 95% CI: 1.021 - 1.065), and CO (OR = 1.128, 95% CI: 1.074 - 1.184), especially among females (according to an increase of 10 μg/m3 in PM10, NO2, SO2, and 1 mg/m3 in CO exposure). A positive association was observed either in warm or in cold season only for PM10. Our findings suggest that, in our setting, exposure to ambient levels of air pollution is an important determinant of emergency room (ER) visits for acute respiratory symptoms, particularly during the warm season. ER admittance may be considered a good proxy to evaluate the adverse effects of air pollution on respiratory health.


Pathogenicity and immunogenicity of a quadruple gene-deleted pseudorabies virus variant as a vaccine candidate.
Since late 2011, the PRV variants have emerged in China, characterized by the increased virulence. The traditional attenuated vaccines have proven insufficient in providing complete protection, resulting in substantial economic losses to swine industry. In this study, a vaccine candidate strain, ZJ01-ΔgI/gE/TK/UL21, carrying the quadruple gene deletion was derived from the previously generated three gene-deleted virus ZJ01-ΔgI/gE/TK. As anticipated, piglets inoculated with ZJ01-ΔgI/gE/TK/UL21 exhibited normal body temperatures and showed no viral shedding, consistent with the observations from piglets treated with ZJ01-ΔgI/gE/TK. Importantly, a significant higher level of interferon induction was observed among piglets in the ZJ01-ΔgI/gE/TK/UL21 group compared to those in the ZJ01-ΔgI/gE/TK group. Upon challenge with the PRV variant ZJ01, piglets immunized with ZJ01-ΔgI/gE/TK/UL21 exhibited reduced viral shedding compared to the ZJ01-ΔgI/gE/TK group. Furthermore, piglets vaccinated with ZJ01-ΔgI/gE/TK/UL21 exhibited minimal pathological lesions in brain tissues, similar to those in the ZJ01-ΔgI/gE/TK group. These results underscore the potential of ZJ01-ΔgI/gE/TK/UL21 as a promising vaccine for controlling PRV infection.


Gut Microbial Signatures Can Discriminate Unipolar from Bipolar Depression.
Discriminating depressive episodes of bipolar disorder (BD) from major depressive disorder (MDD) is a major clinical challenge. Recently, gut microbiome alterations are implicated in these two mood disorders; however, little is known about the shared and distinct microbial characteristics in MDD versus BD. Here, using 16S ribosomal RNA (rRNA) gene sequencing, the microbial compositions of 165 subjects with MDD are compared with 217 BD, and 217 healthy controls (HCs). It is found that the microbial compositions are different between the three groups. Compared to HCs, MDD is characterized by altered covarying operational taxonomic units (OTUs) assigned to the Bacteroidaceae family, and BD shows disturbed covarying OTUs belonging to Lachnospiraceae, Prevotellaceae, and Ruminococcaceae families. Furthermore, a signature of 26 OTUs is identified that can distinguish patients with MDD from those with BD or HCs, with area under the curve (AUC) values ranging from 0.961 to 0.986 in discovery sets, and 0.702 to 0.741 in validation sets. Moreover, 4 of 26 microbial markers correlate with disease severity in MDD or BD. Together, distinct gut microbial compositions are identified in MDD compared to BD and HCs, and a novel marker panel is provided for distinguishing MDD from BD based on gut microbiome signatures.


Ethnobotanical survey of medicinal plants in Bozyazı district of Mersin, Turkey.
This research contributes momentous ethnopharmaceutical information on medicinal plants used by local people who live in the Bozyazı district of Mersin in Turkey. To arrange a list of herbal treatments from the Bozyazı region with their habitual use, plant parts and application methods besides computing some statistical indices. Plant specimens that have been collected by herbalists or local people from Bozyazı for therapeutic purposes were investigated by visiting the villages during the study period, and then conveyed to the laboratory to diagnose the species. In addition, the relative frequency citation (RFC), use values (UV), informant consensus factor (ICF) and fidelity level (FL) were calculated. In this survey conducted between 2013 and 2015, 159 taxa associated with 55 families, excluding 9 edible mushrooms, were detected as they have been utilized for centuries in terms of the healing by visiting 14 villages and 4 herbalist shops in Bozyazı. Lamiaceae, Asteraceae Orchidaceae, Rosaceae, Fabaceae and Geraniaceae have been determined as the most used families for therapeutic purposes. The study revealed that most of the residents concurred on the use of Sideritis erythrantha herba to cure respiratory diseases that demonstrated the highest fidelity level (89). 63 taxa have not matched with any studies conducted in and around the region. Throughout the research carried out Bozyazı district, it has been observed that people living in rural areas have more knowledge and experience about the use and benefits of plants, compared to the people living in the district center. It was detected that the medicinal plants have been used mostly in the therapy of Hemorrhoids. The most information is obtained from the mountain villagers and Yuruk nomads, having average ages of 46.3, during the whole study. In terms of ethnobotanical perspective, the district has stayed valuable due to the winding roads over the mountains and along the coast is very difficult to drive making Bozyazı too far from large cities and mass tourism centers.


Comparison study of two types of expander-based breast reconstruction: acellular dermal matrix-assisted versus total submuscular placement.
Recent reported complications have called some authors to express concern regarding the increased popularity of acellular dermal matrix (ADM)-based breast reconstruction, and its role as an alternative to traditional total submuscular approaches. To address this issue, we compared tissue expansion properties, complication rates, and patient satisfaction for both operative techniques at the same institution. A retrospective review was completed on 75 patients and 100 tissue expander/implant-based breast reconstructions at a single academic institution from 2007 to 2010. Of these cases, 31 patients were reconstructed with ADM and 44 with a submuscular coverage technique. Total complications including seroma, hematoma, infection, skin necrosis, and explantation did not significantly differ between groups (n = 13 for ADM vs. 17 for submuscular, P = 0.814). Consistent with prior reports, ADM-based reconstructions were associated with significantly increased intraoperative fill volumes and lower total number of sessions to achieve final volume. Submuscular reconstructions required a significantly higher tissue expander fill volume. Eight patients in the submuscular group required surgical revision of the breast and inframammary fold, compared with 4 in the ADM group; however, this difference was not significant. Patient satisfaction was equivalent between the 2 groups; however, it was higher in patients with bilateral reconstruction and lower among those who had received adjuvant radiation therapy. Satisfaction with nipple reconstruction was inversely proportional to time elapsed from the procedure to survey conduction. This is the first study to perform a head-to-head comparison on the basis of patient satisfaction, the results of which may be useful in preoperative planning and counseling.


Running kinematics and shock absorption do not change after brief exhaustive running.
Because of the nature of running, the forces encountered require a proper coordination of joint action of the lower extremity to dissipate the ground reaction forces and accelerations through the kinetic chain. Running-related muscle fatigue may reduce the shock absorbing capacity of the lower extremity and alter running kinematics. The purpose of this study was to determine if a bout of exhaustive running at a physiologically determined high intensity, changes running kinematics, impact accelerations, and alters shock attenuating capabilities. It was hypothesized that as a result of fatigue induced by an exhaustive run, running kinematics, impact accelerations at the head and shank, acceleration reduction, and shock attenuation would change. A within-subject, repeated-measures design was used for this study. Twelve healthy, competitive male and female distance runners participated. Subjects performed 2 testing sessions consisting of a VO2max treadmill protocol to determine the heart rate at ventilatory threshold and a fatigue-inducing running bout at the identified ventilatory threshold heart rate. Kinematic data included knee flexion, pronation, time to maximum knee flexion, and time to maximum pronation. Acceleration data included shank acceleration, head acceleration, and shock attenuation. No significant differences resulted for the kinematic or acceleration variables. Although the results of this study do not support the original hypotheses, the influence of running fatigue on kinematics and accelerations remains inconclusive. Future research is necessary to examine fatigue-induced changes in running kinematics and accelerations and to determine the threshold at which point the changes may occur.


Two-stage opening of the Dover Strait and the origin of island Britain.
Late Quaternary separation of Britain from mainland Europe is considered to be a consequence of spillover of a large proglacial lake in the Southern North Sea basin. Lake spillover is inferred to have caused breaching of a rock ridge at the Dover Strait, although this hypothesis remains untested. Here we show that opening of the Strait involved at least two major episodes of erosion. Sub-bottom records reveal a remarkable set of sediment-infilled depressions that are deeply incised into bedrock that we interpret as giant plunge pools. These support a model of initial erosion of the Dover Strait by lake overspill, plunge pool erosion by waterfalls and subsequent dam breaching. Cross-cutting of these landforms by a prominent bedrock-eroded valley that is characterized by features associated with catastrophic flooding indicates final breaching of the Strait by high-magnitude flows. These events set-up conditions for island Britain during sea-level highstands and caused large-scale re-routing of NW European drainage.


Smurf1 polyubiquitinates on K285/K282 of the kinases Mst1/2 to attenuate their tumor-suppressor functions.
Sterile 20-like kinases Mst1 and Mst2 (Mst1/2) and large tumor suppressor 1/2 are core kinases to mediate Hippo signaling in maintaining tissue homeostasis. We have previously demonstrated that Smad ubiquitin (Ub) regulatory factor 1 (Smurf1), a HECT-type E3 ligase, ubiquitinates and in turn destabilizes large tumor suppressor 1/2 to induce the transcriptional output of Hippo signaling. Here, we unexpectedly find that Smurf1 interacts with and polyubiquitinates Mst1/2 by virtue of K27- and K29-linked Ub chains, resulting in the proteasomal degradation of Mst1/2 and attenuation of their tumor-suppressor functions. Among the potential Ub acceptor sites on Mst1/2, K285/K282 are conserved and essential for Smurf1-induced polyubiquitination and degradation of Mst1/2 as well as transcriptional output of Hippo signaling. As a result, K285R/K282R mutation of Mst1/2 not only negates the transcriptional output of Hippo signaling but enhances the tumor-suppressor functions of Mst1/2. Together, we demonstrate that Smurf1-mediated polyubiquitination on K285/K282 of Mst1/2 destabilizes Mst1/2 to attenuate their tumor-suppressor functions. Thus, the present study identifies Smurf1-mediated ubiquitination of Mst1/2 as a hitherto uncharacterized mechanism fine-tuning the Hippo signaling pathway and may provide additional targets for therapeutic intervention of diseases associated with this important pathway.


Effect of ricinoleic acid in acute and subchronic experimental models of inflammation.
Observational studies indicate that topical application of ricinoleic acid (RA), the main component of castor oil, exerts remarkable analgesic and anti-inflammatory effects. Pharmacological characterization has shown similarities between the effects of RA and those of capsaicin, suggesting a potential interaction of this drug on sensory neuropeptide-mediated neurogenic inflammation. The aim of this study was to assess RA anti-inflammatory activities in comparison with capsaicin in several models of acute and subchronic inflammation. The acute inflammation was induced by intradermal injection of carrageenan in the mouse or by histamine in the guinea-pig eyelid. In either experiment, the extent of the oedema thickness was measured. Subchronic oedema was induced by complete Freund's adjuvant injection in the ventral right paw of mice. Tissue substance P (SP) was measured in the carrageenan experiments by radioimmunoassay (RIA). It was found that the acute topical application of RA (0.9 mg/mouse) or capsaicin (0.09 mg/mouse) significantly increased the mouse paw oedema induced by carrageenan, while an 8-day repeated topical treatment with the same doses of both compounds resulted in a marked inhibition of carrageenan-induced paw oedema matched by a reduction in SP tissue levels. Similar effects were found against histamine-induced eyelid oedema in guinea-pigs after acute or repeated application of RA or capsaicin. RA and capsaicin given for 1-3 weeks reduced the established oedema induced by Freund's adjuvant, a subchronic model of inflammation, particularly if given by the intradermal route. Either in mouse paw or in guinea-pig eyelid, capsaicin but not RA by itself produced a slight hyperemia and activation of a behavioural response (e.g. scratching of the eyelids). On the basis of the present results, RA may be seen as a new capsaicin-like, non-pungent anti-inflammatory agent suitable for peripheral application.


Design and Synthesis of α-Anomeric Diacetylene-Containing Glycosides as Photopolymerizable Molecular Gelators.
Glycolipids with diacetylene functional groups are fascinating compounds with many practical uses. Among these, diacetylene-containing gelators are especially important because they can form photopolymerizable gels, which are useful stimuli-responsive materials. Inspired by the unique properties of diacetylene-containing gelators and to understand the structural influences especially the location of the diacetylene functional groups on the self-assembling properties, a series of 15 novel N-acetyl-d-glucosamine derivatives with the diacetylene functional group introduced at the anomeric position were designed and synthesized. The diacetylene function is attached to the sugar through α-glycosylation with the distance from the anomeric oxygen being varied from one, two, and three methylene groups, and the other side contains hydroxyl, carboxyl, phenyl, and alkyl substituents. Remarkably, all compounds can form self-assembled gels in one or more selected solvents. A majority of these synthesized diacetylene glycosides are effective gelators for ethanol/water (v/v 1:1), dimethyl sulfoxide/water (v/v 1:1), and toluene, and one compound also formed a hydrogel at 1.0 wt %. Typically, these glycosides form gels that are photopolymerizable to afford red-colored gels. Scanning electronic microscopy indicated that the gelators formed helices, fibers, and planar sheet-like morphologies. The chemical structures of the derivatives affected their gelation properties and responses to UV treatment. The carboxylic acid-functionalized derivative 17 was able to immobilize basic solutions and form transparent gels. We expect that these diacetylene glycosides especially the hydroxyl and carboxylic acid derivatives will be useful as stimuli-responsive glycolipids for biomedical research.


Sorting signals required for trafficking of the cysteine-rich acidic repetitive transmembrane protein in Trypanosoma brucei.
In trypanosomatids, endocytosis and exocytosis are restricted to the flagellar pocket (FP). The cysteine-rich acidic repetitive transmembrane (CRAM) protein is located at the FP of Trypanosoma brucei and potentially functions as a receptor or an essential component for lipoprotein uptake. We characterized sorting determinants involved in efficient trafficking of CRAM to and from the FP of T. brucei. Previous studies indicated the presence of signals in the CRAM C terminus, specific for its localization to the FP and for efficient endocytosis (H. Yang, D. G. Russell, B. Zeng, M. Eiki, and M.G.-S. Lee, Mol. Cell. Biol. 20:5149-5163, 2000.) To delineate functional domains of putative sorting signals, we performed a mutagenesis series of the CRAM C terminus. Subcellular localization of CRAM mutants demonstrated that the amino acid sequence between -5 and -14 (referred to as a transport signal) is essential for exporting CRAM from the endoplasmic reticulum to the FP, and mutations of amino acids at -12 (V), -10 (V), or -5 (D) led to retention of CRAM in the endoplasmic reticulum. Comparison of the endocytosis efficiency of CRAM mutants demonstrated that the sequence from amino acid -5 to -23 (referred to as a putative endocytosis signal) is required for efficient endocytosis and overlaps with the transport signal. Apparently the CRAM-derived sorting signal can efficiently interact with the T. brucei micro1 adaptin, and mutations at amino acids essential for the function of the transport signal abolished the interaction of the signal with T. brucei micro1, strengthening the hypothesis of the involvement of the clathrin- and adaptor-dependent pathway in trafficking of CRAM via the FP.


Expression of an antigenic adenovirus epitope in a group B coxsackievirus.
Group B coxsackieviruses (CVB) cause human myocarditis, while human adenovirus type 2 (Ad2) is implicated as an agent of this disease. The L1 loop of the Ad2 hexon protein has been demonstrated to be antigenic in rabbits. To evaluate the feasibility of a multivalent vaccine strain against the CVB and Ad2, we cloned the sequence encoding the Ad2 hexon L1 loop, flanked by dissimilar sequences encoding the protease 2A (2Apro) recognition sites, into the genome of an attenuated strain of CVB type 3 (CVB3/0) at the junction of 2Apro and the capsid protein 1D. Progeny virus (CVB3-PL2-Ad2L1) was obtained following transfection of the construct into HeLa cells. Replication of CVB3-PL2-Ad2L1 in diverse cell cultures demonstrated that the yield of the chimeric virus was between 0.5 to 2 log units less than the parental strain. Western blot analyses of the CVB3 capsid protein 1D in CVB3-PL2-Ad2L1-infected HeLa cells demonstrated production of the expected capsid protein. Viral proteins were detected earlier and in approximately fourfold greater amounts in CVB3-PL2-Ad2L1-infected HeLa cells than in CVB3/0-infected cells. Cleavage of the CVB3-PL2-Ad2L1 polyprotein by 2Apro was slowed, accompanied by an accumulation of the fusion 1D-L1 loop protein. Reverse transcription-PCR sequence analysis of CVB3-PL2-Ad2L1 RNA demonstrated that the Ad2 hexon polypeptide coding sequence was maintained in the chimeric viral genome through at least 10 passages in HeLa cells. Mice inoculated with CVB3-PL2-Ad2L1 demonstrated a brief viremia with no replication detectable in the heart but prolonged replication of virus in the pancreas in the absence of pathologic changes in either organ. CVB3-PL2-Ad2L1 induced binding and neutralizing anti-Ad2 antibodies, in addition to antibodies against CVB3 in mice. CVB3-PL2-Ad2L1 was used to challenge mice previously inoculated with CVB3/0 and with preexisting anti-CVB3 neutralizing-antibody titers; anti-Ad2 neutralizing and binding antibodies were induced in these mice at higher levels than in mice without anti-CVB3 immunity. The data demonstrate that a CVB vector can stably express an antigenic polypeptide of Ad2 from within the CVB open reading frame that results in the induction of protective immune responses against both viruses.


Antigenic relatedness between the spiralins of Spiroplasma citri and Spiroplasma melliferum.
Four spiralins were compared by rocket immunoelectrophoresis, quantitative immunoblotting techniques, and the spiroplasma deformation test with the use of antispiralin (polyclonal) monospecific antibodies. This investigation revealed that the spiralins of Spiroplasma citri and S. melliferum are antigenically related and that probably no more than two epitopes simultaneously saturable with antibodies are shared by the two proteins. One at least of these epitopes is accessible to antibodies on the spiroplasma cell surface.


Collagen metabolism during right ventricular hypertrophy following induced lung injury.
Collagen synthesis and degradation rates were estimated in ventricles of normal rabbits and in those with right ventricular hypertrophy resulting from bleomycin-induced lung injury. Synthesis rates were estimated from the radioactivity in tissue [14C]hydroxyproline following a single intravenous injection of [14C]proline with a "flooding" dose of unlabeled proline (Biochem. J. 206: 535-544, 1982). The rate in the left ventricle was 5.8 +/- 1.0 compared with 2.9 +/- 0.4%/day in the right (P less than 0.02). Degradation rates, based on [14C]hydroxyproline levels in the tissue-free pool, indicated that in both normal ventricles about one-third of newly produced collagen was degraded rapidly following its synthesis. Six days after bleomycin, right ventricular collagen content fell by 35%, associated with a marked increase in tissue-free hydroxyproline levels. After 14 days the right ventricular weight and collagen content had increased by 49.4 +/- 3.7% (P less than 0.001) and 31.7 +/- 4.4% (P less than 0.05), respectively, and collagen synthesis rates increased to 9.7 +/- 1.8%/day (P less than 0.001). It is concluded that collagen is synthesized and degraded quite rapidly in normal heart tissue but that fractional rates differ between ventricles. The evidence also suggests that increased collagen synthesis and breakdown of mature collagen occurs as the connective tissue matrix is remodeled during adaptive cardiac growth.


Transjugular intrahepatic portosystemic shunt for Budd-Chiari syndrome: A comprehensive review.
Budd-Chiari syndrome (BCS) is a relatively rare clinical condition with a wide range of symptomatology, caused by the obstruction of the hepatic venous outflow. If left untreated, it has got an high mortality rate. Its management is based on a step-wise approach, depending on the clinical presentation, and includes different treatment from anticoagulation therapy up to Interventional Radiology techniques, such as transjugular intrahepatic portosystemic shunt (TIPS). TIPS is today considered a safe and highly effective treatment and should be recommended for BCS patients, including those awaiting orthotopic liver transplantation. In this review the pathophysiology, diagnosis and treatment options of BCS are presented, with a special focus on published data regarding the techniques and outcomes of TIPS for the treatment of BCS. Moreover, unresolved issues and future research will be discussed.


Identification of a siderophore utilization locus in nontypeable Haemophilus influenzae.
Haemophilus influenzae has an absolute aerobic growth requirement for either heme, or iron in the presence of protoporphyrin IX. Both iron and heme in the mammalian host are strictly limited in their availability to invading microorganisms. Many bacterial species overcome iron limitation in their environment by the synthesis and secretion of small iron binding molecules termed siderophores, which bind iron and deliver it into the bacterial cell via specific siderophore receptor proteins on the bacterial cell surface. There are currently no reports of siderophore production or utilization by H. influenzae. Comparative genomics revealed a putative four gene operon in the recently sequenced nontypeable H. influenzae strain R2846 that encodes predicted proteins exhibiting significant identity at the amino acid level to proteins involved in the utilization of the siderophore ferrichrome in other bacterial species. No siderophore biosynthesis genes were identified in the R2846 genome. Both comparative genomics and a PCR based analysis identified several additional H. influenzae strains possessing this operon. In growth curve assays strains containing the genes were able to utilize ferrichrome as an iron source. H. influenzae strains lacking the operon were unable to obtain iron from ferrichrome. An insertional mutation in one gene of the operon abrogated the ability of strains to utilize ferrichrome. In addition transcription of genes in the identified operon were repressible by high iron/heme levels in the growth media. We have identified an iron/heme-repressible siderophore utilization locus present in several nontypeable H. influenzae strains. The same strains do not possess genes encoding proteins associated with siderophore synthesis. The siderophore utilization locus may enable the utilization of siderophores produced by other microorganisms in the polymicrobial environmental niche of the human nasopharynx colonized by H. influenzae. This is the first report of siderophore utilization by H. influenzae.


Reliability and responsiveness of elbow trajectory tracking in chronic poststroke hemiparesis.
This study established the reliability of a novel upper-limb trajectory-tracking task for assessment of perceptual motor control in hemiparetic adults. Eleven persons with chronic poststroke hemiparesis (mean 58.6 months) and eleven nondisabled control subjects performed an elbow flexion-extension task against a low-resistance isotonic load at three speeds: 25 degrees/s, 45 degrees/s, and 65 degrees/s. Both arms (paretic and nonparetic or dominant and nondominant) were tested during two identical sessions separated by 1 week. Relative reliability (intraclass correlation coefficient [ICC]) ranged from 0.5 to 0.8 and absolute reliability (standard error of measurement [SEM%]) ranged between 19% to 36% across both subject groups. No systematic errors between test sessions were revealed. Smallest real differences (SRDs) were determined to be +/- 2 degrees to 3 degrees in nondisabled, +/- 2 degrees to 5 degrees in nonparetic and +/- 9 degrees in paretic arms. Responsiveness ratios derived with the use of the SRDs ranged between 1.91 to 2.45, indicating that this instrument is sensitive to clinically important change and suitable for demonstrating effects on upper-limb motor performance following clinical intervention.


Progressive symmetric erythrokeratodermia. Histological and ultrastructural study of patient before and after treatment with etretinate.
Progressive symmetric erythrokeratodermia is a rare inherited cornification disorder characterized by symmetric erythematous hyperkeratotic plaques. Approximately 25 cases have been reported, but the ultrastructural features have not been well characterized. In this article, we describe the clinical, histologic, and ultrastructural findings noted in our patient. Biopsy specimens taken before treatment with etretinate showed hyperkeratosis with a well-preserved granular layer and acanthosis. Ultrastructural examination disclosed that granular cells contained markedly swollen mitochondria and that corneocytes contained many lipidlike vacuoles. After initiation of treatment, considerable clinical improvement was observed, but without significant histologic modification. The principal posttreatment ultrastructural changes observed were the reduction of mitochondrial swelling in granular cells and the reduction in the number of lipidlike vacuoles in corneocytes.


Adaptive changes in tumor cells in response to reductive stress.
Reductive stress is characterized by an excess of cellular electron donors and can be linked with various human pathologies including cancer. We developed melanoma cell lines resistant to reductive stress agents: rotenone (ROTR), n-acetyl-L-cysteine, (NACR), or dithiothreitol (DTTR). Resistant cells divided more rapidly and had intracellular homeostatic redox-couple ratios that were shifted towards the reduced state. Resistance caused alterations in general cell morphology, but only ROTR cells had significant changes in mitochondrial morphology with higher numbers that were more isolated, fragmented and swollen, with greater membrane depolarization and decreased numbers of networks. These changes were accompanied by lower basal oxygen consumption and maximal respiration rates. Whole cell flux analyses and mitochondrial function assays showed that NACR and DTTR preferentially utilized tricarboxylic acid (TCA) cycle intermediates, while ROTR used ketone body substrates such as D, L-β-hydroxybutyric acid. NACR and DTTR cells had constitutively decreased levels of reactive oxygen species (ROS), although this was accompanied by activation of nuclear factor erythroid 2-related factor 2 (Nrf2), with concomitant increased expression of the downstream gene products such as glutathione S-transferase P (GSTP). Further adaptations included enhanced expression of endoplasmic reticulum proteins controlling the unfolded protein response (UPR). Although expression patterns of these UPR proteins were distinct between the resistant cells, a trend implied that resistance to reductive stress is accompanied by a constitutively increased UPR phenotype in each line. Overall, tumor cells, although tolerant of oxidative stress, can adapt their energy and survival mechanisms in lethal reductive stress conditions.


Evidence for kinks in DNA folding in the nucleosome.
The nucleosome subunit of chromatin consists of DNA folded around a histone core as a 1.8-turn left-handed solenoid. The crystal structure of the nucleosome core particle revealed that it has a dyad symmetry axis and that the minor helix groove faces outwards from the protein core. Richmond et al. noticed that the path traversed by the helix has severe bends at sites approximately one and four helix turns from the dyad axis. We have developed two photochemical methods to study the structure of DNA, and in particular that wrapped around the nucleosome core. One method depends on the sensitization of singlet oxygen production by an eosin analogue. We have monitored the rate at which excited state oxygen diffuses into contact with DNA base planes, and find that it attacks the nucleosome with high specificity. We have also mapped the DNA binding of the intercalating dye methylene blue, and conclude that it binds to the same sites accessible to oxygen by diffusion. On the basis of these results we suggest that the DNA in the nucleosome is bent or kinked at two sites, 1.5 helix turns from the dyad axis.