Combined lateral closing and medial opening-wedge high tibial osteotomy.
Long-term studies have indicated that the clinical success of high tibial osteotomy deteriorates with time. The purpose of this study was to evaluate the long-term results of a combined lateral closing and medial opening-wedge technique for high tibial osteotomy with a minimum follow-up of fifteen years. From January 1981 to June 1990, ninety-two patients (ninety-four knees) had a high tibial valgus osteotomy. The average preoperative varus deformity was 13.5 degrees . The surgical technique consisted of a proximal lateral closing-wedge osteotomy and use of the lateral wedge as a graft on the medial side of the osteotomy. No internal fixation was used. A knee brace was used to maintain the 8 degrees to 10 degrees of valgus overcorrection. Seventy-two knees in seventy patients with at least fifteen years of follow-up were evaluated. Clinical evaluation was done with The Hospital for Special Surgery knee-rating scale. The femorotibial alignment, posterior tibial slope, and the Insall-Salvati ratio were measured on radiographs. The mean initial postoperative correction (and standard deviation) for all knees was to 8.3 degrees +/- 2.7 degrees of valgus alignment. Survivorship analysis showed that the probability of survival (and 95% confidence interval), with conversion to total knee arthroplasty as the end point, was 100% at one year, 92% +/- 5.8% at ten years, 80% +/-7.7% at fifteen years, and 58% +/- 4.3% at twenty years. The survivorship, with a Hospital for Special Surgery knee score of <70 points as the end point, was 80% +/- 4.5% at ten years, 72% +/- 5.6% at fifteen years, and 42% +/- 4.2% at twenty years. Twenty-six knees underwent an arthroplasty at an average of 15.6 years after the index procedure. For the forty-six knees that had not undergone an arthroplasty, the knee score improved from an average of 67 points preoperatively to 82 points at the time of the most recent follow-up. There were two superficial wound infections and one delayed union. We believe that our technique of a combined lateral closing and medial opening-wedge high tibial osteotomy can provide good long-term outcomes because of the off-loading of the diseased medial compartment with minimal complications.


Impact of dry eye syndrome on vision-related quality of life in a non-clinic-based general population.
Dry eye syndrome (DES) is a common ocular disorder occurring in general population. The purpose of this study is to evaluate the impact of DES on vision-related quality of life (QoL) in a non-clinic-based general population. This population-based cross-sectional study enrolled subjects older than 40 years, who took part in an epidemiological study on dry eye in Sanle Community, Shanghai. Apart from the collection of sociodemographics, dry eye symptoms, and other clinical data, a Chinese version of the 25-item National Eye Institute Visual Functioning Questionnaire (NEI VFQ-25) was administered to all subjects. Comparisons of the NEI VFQ-25 subscale item scores and composite score were made among subgroups divided according to the presence of dry eye symptoms or signs. Multivariate regression analysis was performed to investigate the relationship between the clinical variables and the VFQ-25 composite score. A total of 229 participants were enrolled in the study, with an average age of (60.7±10.1) years old. Majority of these participants were female (59.8 %, 137/229). The total DES symptom scores (TDSS) in subjects either with definite DES or only with dry eye symptoms were significantly higher (F=60.331, P<0.001). The values of tear break-up time (TBUT) and Schirmer test were significantly lower in participants with DES and those with dry eye signs only (F=55.158 and 40.778, P<0.001). The composite score of the NEI VFQ-25 was significantly lower in subjects with DES (F=4.901, P=0.003). Moreover, the subscale scores of ocular pain and mental health were significantly lower in those with either DES or dry eye symptoms only (F=10.962 and 7.362 respectively, both P<0.001). The multiple regression analysis showed that the TDSS had a significant negative correlation with the VFQ-25 composite score as well as with the subscale score for ocular pain and mental health, even after the adjustment of all other factors (all P<0.01). The symptoms of dry eye are associated with an adverse impact on vision-related QoL in non-clinic-based general population, which is mainly represented as more ocular pain and discomfort, and impaired mental health as well. Apart from clinical examination, it is also important to refer to subjective symptoms and QoL scores when assessing the severity of DES.


Anti-B-cell Maturation Antigen Chimeric Antigen Receptor T cell Function against Multiple Myeloma Is Enhanced in the Presence of Lenalidomide.
Anti-B-cell maturation antigen (BCMA) chimeric antigen receptor (CAR) T cells have shown promising clinical responses in patients with relapsed/refractory multiple myeloma. Lenalidomide, an immunomodulatory drug, potentiates T cell functionality, drives antimyeloma activity, and alters the suppressive microenvironment; these properties may effectively combine with anti-BCMA CAR T cells to enhance function. Using an anti-BCMA CAR T, we demonstrated that lenalidomide enhances CAR T cell function in a concentration-dependent manner. Lenalidomide increased CAR T effector cytokine production, particularly under low CAR stimulation or in the presence of inhibitory ligand programmed cell death 1 ligand 1. Notably, lenalidomide also enhanced CAR T cytokine production, cytolytic activity, and activation profile relative to untreated CAR T cells in chronic stimulation assays. This unique potentiation of both short-term CAR T activity and long-term functionality during chronic stimulation prompted investigation of the molecular profile of lenalidomide-treated CAR T cells. Signatures from RNA sequencing and assay for transposase-accessible chromatin using sequencing indicated that pathways associated with T-helper 1 response, cytokine production, T cell activation, cell-cycle control, and cytoskeletal remodeling were altered with lenalidomide. Finally, study of lenalidomide and anti-BCMA CAR T cells in a murine, disseminated, multiple myeloma model indicated that lenalidomide increased CAR T cell counts in blood and significantly prolonged animal survival. In summary, preclinical studies demonstrated that lenalidomide potentiated CAR T activity in vivo in low-antigen or suppressive environments and delayed onset of functional exhaustion. These results support further investigation of lenalidomide and anti-BCMA CAR T cells in the clinic.


[Immunologic methods of study for evaluating the risk of anaphylactic reactions in intradisk chymopapain injections].
According to results available to date, skin tests are of limited value in determining the risk of anaphylactic shock. By identifying specific IgE antibodies, e.g. via the Chymo-FAST test or the corresponding RAST, the risk of anaphylactic shock of 0.2%-1% in the total group of patients can be reduced to 0.05% in patients in whom the test was negative, whereas in test-positive patients the risk is above 60%. Previous intolerance reactions among the remaining 0.05% are probably due to a non-testable pseudoallergic reaction, just like the 6% mild late reactions after the injection. For the remaining risk of 0.05% in IgE-negative patients drug prophylaxis with antihistamines or if necessary even steroids will continue to be mandatory. Examination of histamine release from leucocytes, which can be conducted in very few centres only, may possibly be able to clarify in future to what extent reinjection is permissible if a disc on a different level is affected, besides other scientific problems.


[Reference values of clinical donor blood analysis].
Basic hemogram parameters were analyzed in 835 blood donors (421 men and 414 women) aged 18 to 67 years) on an ADVIA 120 hematology analyzer. Reference ranges were calculated for each parameter in relation to their distribution (as X mean +/- 2alpha for normally distributed data and 2.5-97.5 per thousand for others). The findings suggest that the normal range should be extended for platelets, red blood cells, hemoglobin, and packed cell volume as compared with the earlier unified normal values, which must be taken into account when deciding whether a subject may be allowed to donate blood.


DNMT1 inhibition by pUG-fold quadruplex RNA.
Aberrant DNA methylation is one of the earliest hallmarks of cancer. DNMT1 is responsible for methylating newly replicated DNA, but the precise regulation of DNMT1 to ensure faithful DNA methylation remains poorly understood. A link between RNA and chromatin-associated proteins has recently emerged, and several studies have shown that DNMT1 can be regulated by a variety of RNAs. In this study, we have confirmed that human DNMT1 indeed interacts with multiple RNAs, including its own nuclear mRNA. Unexpectedly, we found that DNMT1 exhibits a strong and specific affinity for GU-rich RNAs that form a pUG-fold, a noncanonical G-quadruplex. We find that pUG-fold-capable RNAs inhibit DNMT1 activity by inhibiting binding of hemimethylated DNA, and we additionally provide evidence for multiple RNA binding modes with DNMT1. Together, our data indicate that a human chromatin-associated protein binds to and is regulated by pUG-fold RNA.


Ecology of Listeria spp. in a fish farm and molecular typing of Listeria monocytogenes from fish farming and processing companies.
This study focused on the ecology of Listeria monocytogenes in a fish farm by following the changes in its occurrence in different types of samples for a three year period. In addition, L. monocytogenes isolates from different seafood industry areas were compared with pulsed field gel electrophoresis (PFGE) typing to discover possible associations between primary production, further processing and final products. Weather conditions were found to have a strong influence on the probability of finding Listeria spp. in a fish farm environment. The number of samples contaminated with Listeria spp. was typically bigger after rainy periods. Brook and river waters as well as other runoff waters seemed to be the main contamination source at the farm studied. The farmed fish originally found to carry L. monocytogenes become gradually Listeria free. The time needed for the purification of the fish was several months. The sea bottom soil samples were the ones that preserved the L. monocytogenes contamination the longest time. It can be stated that the fish and fish farm equipment studied did not spread listeria contamination. On the contrary, they were found to suffer from listeria contamination coming from outside sources like the brook water. There was a wide range of different L. monocytogenes PFGE-pulsotypes (30) found at 15 Finnish fish farms and fish processing factories. L. monocytogenes isolates from the final products often belonged to the same pulsotypes as did the isolates from the processing environment as well as from the raw fish. This suggests that, in addition to the fish processing factory environment, the fish raw materials are important sources of L. monocytogenes contamination in final products.


Experiences of time loss among videogame players: an empirical study.
Playing videogames is now a major leisure pursuit, yet research in the area is comparatively sparse. Previous correlational evidence suggests that subjective time loss occurs during playing videogames. This study examined experiences of time loss among a relatively large group of gamers (n = 280). Quantitative and qualitative data were collected through an online survey. Results showed that time loss occurred irrespective of gender, age, or frequency of play, but was associated with particular structural characteristics of games such as their complexity, the presence of multi-levels, missions and/or high scores, multiplayer interactions, and plot. Results also demonstrated that time loss could have both positive and negative outcomes for players. Positive aspects of time loss included helping players to relax and temporarily escape from reality. Negative aspects included the sacrificing of other things in their lives, guilty feelings about wasted time, and social conflict. It is concluded that for many gamers, losing track of time is a positive experience and is one of the main reasons for playing videogames.


Enhancing diffraction-limited images using properties of the point spread function.
We propose an algorithm to enhance diffraction-limited images based on pixel-to-pixel correlations introduced by the finite width of the Point Spread Function (PSF). We simulate diffraction-limited images of point sources by convolving the PSF of a diffraction-limited lens with simulated images, and enhance the blurred images with our algorithm. Our algorithm reduces the PSF width, increases the contrast, and reveals structure on a length scale half of that resolvable in the unenhanced image. Our enhanced images compare favorably with images enhanced by conventional Tikhonov regularization.


A common partitivirus infection in United States and Czech Republic isolates of bat white-nose syndrome fungal pathogen Pseudogymnoascus destructans.
The psychrophilic (cold-loving) fungus Pseudogymnoascus destructans was discovered more than a decade ago to be the pathogen responsible for white-nose syndrome, an emerging disease of North American bats causing unprecedented population declines. The same species of fungus is found in Europe but without associated mortality in bats. We found P. destructans was infected with a mycovirus [named Pseudogymnoascus destructans partitivirus 1 (PdPV-1)]. The virus is bipartite, containing two double-stranded RNA (dsRNA) segments designated as dsRNA1 and dsRNA2. The cDNA sequences revealed that dsRNA1 dsRNA is 1,683 bp in length with an open reading frame (ORF) that encodes 539 amino acids (molecular mass of 62.7 kDa); dsRNA2 dsRNA is 1,524 bp in length with an ORF that encodes 434 amino acids (molecular mass of 46.9 kDa). The dsRNA1 ORF contains motifs representative of RNA-dependent RNA polymerase (RdRp), whereas the dsRNA2 ORF sequence showed homology with the putative capsid proteins (CPs) of mycoviruses. Phylogenetic analyses with PdPV-1 RdRp and CP sequences indicated that both segments constitute the genome of a novel virus in the family Partitiviridae. The purified virions were isometric with an estimated diameter of 33 nm. Reverse transcription PCR (RT-PCR) and sequencing revealed that all US isolates and a subset of Czech Republic isolates of P. destructans were infected with PdPV-1. However, PdPV-1 appears to be not widely dispersed in the fungal genus Pseudogymnoascus, as non-pathogenic fungi P. appendiculatus (1 isolate) and P. roseus (6 isolates) tested negative. P. destructans PdPV-1 could be a valuable tool to investigate fungal biogeography and the host-pathogen interactions in bat WNS.


Cytokine-mediated regulation of activating and inhibitory Fc gamma receptors in human monocytes.
Fc gamma receptors (Fc gammaR) trigger inflammatory reactions in response to immunoglobulin-opsonized pathogens and antigen-antibody complexes. The coordinate expression of activating and inhibitory Fc gammaR ensures the homeostasis of immune complex-driven inflammatory responses. In this study, we used antibodies with preferential binding for activating Fc gammaRIIa and inhibitory Fc gammaRIIb receptors to investigate the expression and regulation of Fc gammaRII isoforms in human monocytes. Cross-linking of Fc gammaRIIa triggered phagocytosis and cytokine production. Cross-linking of Fc gammaRIIb was associated with phosphorylation of the immunoreceptor tyrosine-based inhibitory motif and with a marked reduction in monocyte effector functions. Our study revealed that tumor necrosis factor alpha (TNF-alpha), interleukin (IL)-10, and IL-13 altered the transcriptional activity of the Fc gammaRIIB promoter in transfected cell lines and skewed the balance of activating versus inhibitory Fc gammaR in human monocytes. TNF-alpha decreased the expression of inhibitory Fc gammaRIIb. IL-10 up-regulated all classes of Fc gammaR and induced alternative activation in monocytes, an effect that was synergistic with that of TNF-alpha. In contrast, IL-4 and IL-13, in combination with TNF-alpha, decreased the expression of activating Fc gammaR and markedly down-regulated Fc gammaR-mediated function. Our findings suggest that the cytokine milieu can induce changes in the relative expression of Fc gammaR with opposing function and thus, may regulate the amplitude of Fc gammaR-mediated uptake and inflammation.


Application of Metabolomics to Epidemiologic Studies of Breast Cancer: New Perspectives for Etiology and Prevention.
To provide an overview on how the application of metabolomics (high-throughput characterization of metabolites from cells, organs, tissues, or biofluids) to population-based studies may inform our understanding of breast cancer etiology. We evaluated studies that applied metabolomic analyses to prediagnostic blood samples from prospective epidemiologic studies to identify circulating metabolites associated with breast cancer risk, overall and by breast cancer subtype and menopausal status. We provide some important considerations for the application and interpretation of metabolomics approaches in this context. Overall, specific lipids and amino acids were indicated as the most common metabolite classes associated with breast cancer development. However, comparison of results across studies is challenging because of heterogeneity in laboratory techniques, analytical methods, sample size, and applied statistical methods. Metabolomics is being increasingly applied to population-based studies for the identification of new etiologic hypotheses and/or mechanisms related to breast cancer development. Despite its success in applications to epidemiology, studies of larger sample size with detailed information on menopausal status, breast cancer subtypes, and repeated biologic samples collected over time are needed to improve comparison of results between studies and enhance validation of results, allowing potential clinical translation of findings.


Nucleolin functions in the first step of ribosomal RNA processing.
The first processing step of precursor ribosomal RNA (pre-rRNA) involves a cleavage within the 5' external transcribed spacer. This processing requires sequences downstream of the cleavage site which are perfectly conserved among human, mouse and Xenopus and also several small nucleolar RNAs (snoRNAs): U3, U14, U17 and E3. In this study, we show that nucleolin, one of the major RNA-binding proteins of the nucleolus, is involved in the early cleavage of pre-rRNA. Nucleolin interacts with the pre-rRNA substrate, and we demonstrate that this interaction is required for the processing reaction in vitro. Furthermore, we show that nucleolin interacts with the U3 snoRNP. Increased levels of nucleolin, in the presence of the U3 snoRNA, activate the processing activity of a S100 cell extract. Our results suggest that the interaction of nucleolin with the pre-rRNA substrate might be a limiting step in the primary processing reaction. Nucleolin is the first identified metazoan proteinaceous factor that interacts directly with the rRNA substrate and that is required for the processing reaction. Potential roles for nucleolin in the primary processing reaction and in ribosome biogenesis are discussed.


H9 Inhibits Tumor Growth and Induces Apoptosis via Intrinsic and Extrinsic Signaling Pathway in Human Non-Small Cell Lung Cancer Xenografts.
H9, a novel herbal extract, demonstrated cytotoxicity in A549 non-small cell lung cancer (NSCLC) cell lines. In this study, we investigated whether H9, and/or co-treatment with an anticancer drug, pemetrexed (PEM), inhibited tumor growth in BALB/c nude mice models bearing A549 NSCLC cells. The mice were separated into groups and administered H9 and PEM for 2 weeks. Protein and mRNA levels were detected using western blotting and reverse transcription polymerase chain reaction, respectively; immunohistochemistry (IHC) was also performed on the tumor tissues. H9 and co-treatment with PEM induced the cleavage of proapoptotic factors, such as caspase-3, caspase-8, caspase-9, and poly(ADP)-ribose polymerase (PARP). Expression levels of cell-death receptors involving Fas/FasL, TNF-related apoptosisinducing ligands (TRAIL), and TRAIL receptors were increased by H9 and co-treatment with PEM. Furthermore, analysis of levels of cell-cycle modulating proteins indicated that tumor cells were arrested in the G1/S phase. In addition, the phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K)/Akt survival signaling pathways were inhibited by H9 and co-treatment with PEM. In conclusion, H9 and co-treatment with PEM inhibited tumor growth in BALB/c nude mice models bearing A549 NSCLC cells. These results indicate that H9 and co-treatment with PEM can be used as an anticancer therapy in NSCLC.


EUS in cirrhotic patients with and without prior variceal hemorrhage in comparison with noncirrhotic control subjects.
Endoscopic ultrasound (EUS) was used to evaluate cirrhotic patients with and without prior variceal hemorrhage. The findings were compared with those of EUS in noncirrhotic control subjects to determine EUS features indicative of cirrhosis and of a risk for variceal hemorrhage. Patients with cirrhosis undergoing indicated endoscopic screening for varices or surveillance after endoscopic therapy for variceal hemorrhage were studied and compared with healthy noncirrhotic control patients undergoing EUS for benign conditions. Sixty-six cirrhotic patients (31 with prior hemorrhage) and 32 control patients were studied. Nonhemorrhage cirrhotic patients had more severe liver disease by Child's class (p = 0.02) and less beta-adrenergic blocker usage (p < 0.0001). Paraesophageal varices were detected in 97% of cirrhotic patients and 3% of control patients (p < 0.001) and were a more sensitive predictor of cirrhosis than varices at endoscopy (74%, p < 0.0001). Azygos vein and thoracic duct diameters, and gastric mucosa and submucosa thickness were greater for cirrhotic than control patients (p < 0.001). More hemorrhage patients had large (5 mm or greater) paraesophageal varices (odds ratio 3.1: 95% CI [1.1, 8.3]; p < 0.05) and paragastric varices (odds ratio 3.7: 95% CI [1.3, 10.2]; p = 0.01). Paraesophageal varix size correlated with ascites (p = 0.03) and, for nonhemorrhage patients, with Child's class (p < 0.01). Paraesophageal and paragastric varices correlate with the presence and severity of liver disease and portal hypertension. These data support the hypothesis that large paraesophageal and paragastric varices (5 mm or greater) may be risk factors for variceal hemorrhage, an observation that merits further prospective study. Cirrhosis causes dilation of the azygos vein and thoracic duct and thickening of gastric mucosa and submucosa.


Exercise and the platelet activator calcium chloride both influence the growth factor content of platelet-rich plasma (PRP): overlooked biochemical factors that could influence PRP treatment.
There is strong evidence that exercise affects platelet haemostasis factors, but this potential effect on growth factor concentrations in platelet-rich plasma (PRP) has never been studied. In addition, there is a paucity of studies focusing on the effects of activating agents used in conjunction with PRP. The first aim of this study was to evaluate the effect of exercise on platelet and platelet-derived growth factors (PDGF)-AB, hepatocyte growth factor (HGF), insulin-like growth factor-1 (IGF-1) and vascular endothelial growth factor (VEGF) concentrations in PRP. The second aim was to study the effect of the activating agent calcium chloride (CaCl2) on growth factor concentration in relation to different exercise states. Controlled laboratory study. Ten healthy participants performed 1 h of submaximal exercise with blood being withdrawn immediately pre, post and 18 h following. PRP was prepared in each condition in both an activated CaCl2 and non-activated form. Concentrations of PDGF-AB, HGF, IGF-1 and VEGF were evaluated using standard ELISA systems. Exercise had no significant effect on platelet concentration, but significantly suppressed both VEGF and PDGF-AB concentrations. Exercise state had no significant effect on IGF-1 or HGF concentration. Activation with CaCl2 resulted in a significant increase in PDGF-AB and IGF-1 concentrations, unchanged VEGF and significantly reduced HGF concentrations. Exercise significantly impacts on PDGFs in PRP with significantly reduced concentrations of VEGF and PDFG-AB. Furthermore, the activation of PRP with CaCl2 results in a differentiated GF release from platelets. These relevant factors can potentially influence outcome in daily clinical practice and are recommended to be accounted for in future study design.


[Purification and characterization of cAMP-dependent protein kinases of yeasts in a Saccharomyces cerevisiae wild strain and selected mutants of cAMP metabolism].
Protein kinases represent a diverse family of enzymes that play a critical role in regulation. Among nearly 100 known protein kinases, the cAMP-dependent enzyme is best understood biochemically. Unlike other protein kinases, cAMP-dependent protein kinase consists of two different types of subunits that dissociate, a regulatory subunit (R), which is the receptor for cAMP, and a catalytic subunit (C). In the absence of cAMP, the enzyme exists as an inactive tetramer, R2C2. The binding of intracellular cAMP to the R subunit decreases the affinity of the R subunit for the C subunit by approximately four orders of magnitude and, under physiological conditions, leads to dissociation of the holoenzyme into R2(cAMP)4 dimer and two free C subunits that are catalytically active. Mutants of the cAMP metabolism, adenylate cyclase and cell cycle mutants, provided further information about protein synthesis and cellular growth in Saccharomyces cerevisiae. The purified protein kinases were divided into different types according to their elution profiles from the DEAE-cellulose matrix. Two types of cAMP-dependent and two types of cAMP-independent protein kinases were isolated from the wild strain. Differences in the activities of the kinases in the mutants showed a close relationship to the locus of the respective mutations in the cell-cycle. Some properties of the protein kinases are discussed with respect to individual mutations.


GalaxyDock2: protein-ligand docking using beta-complex and global optimization.
In this article, an enhanced version of GalaxyDock protein-ligand docking program is introduced. GalaxyDock performs conformational space annealing (CSA) global optimization to find the optimal binding pose of a ligand both in the rigid-receptor mode and the flexible-receptor mode. Binding pose prediction has been improved compared to the earlier version by the efficient generation of high-quality initial conformations for CSA using a predocking method based on a beta-complex derived from the Voronoi diagram of receptor atoms. Binding affinity prediction has also been enhanced by using the optimal combination of energy components, while taking into consideration the energy of the unbound ligand state. The new version has been tested in terms of binding mode prediction, binding affinity prediction, and virtual screening on several benchmark sets, showing improved performance over the previous version and AutoDock, on which the GalaxyDock energy function is based. GalaxyDock2 also performs better than or comparable to other state-of-the-art docking programs. GalaxyDock2 is freely available at http://galaxy.seoklab.org/softwares/galaxydock.html.


Do patients consult the doctor less often than they used to?
Analysis of nationally representative surveys of adults and older teenagers has identified a small but steady decrease in consultation rates following the transient increase which occurred after the introduction of the National Health Service. Visiting rates decreased substantially after 1950 but no trend has been identified in attendance rates. However, between 1950 and 1978 visits have represented a decreasing proportion, and attendances an increasing proportion, of all consultations.


Recurrent CDC25C mutations drive malignant transformation in FPD/AML.
Familial platelet disorder (FPD) with predisposition to acute myelogenous leukaemia (AML) is characterized by platelet defects with a propensity for the development of haematological malignancies. Its molecular pathogenesis is poorly understood, except for the role of germline RUNX1 mutations. Here we show that CDC25C mutations are frequently found in FPD/AML patients (53%). Mutated CDC25C disrupts the G2/M checkpoint and promotes cell cycle progression even in the presence of DNA damage, suggesting a critical role for CDC25C in malignant transformation in FPD/AML. The predicted hierarchical architecture shows that CDC25C mutations define a founding pre-leukaemic clone, followed by stepwise acquisition of subclonal mutations that contribute to leukaemia progression. In three of seven individuals with CDC25C mutations, GATA2 is the target of subsequent mutation. Thus, CDC25C is a novel gene target identified in haematological malignancies. CDC25C is also useful as a clinical biomarker that predicts progression of FPD/AML in the early stage.


[Effect of monosomy of the autosomes on the preimplantation stages of embryogenesis in laboratory mice].
The effects of monosomy for the autosomes 1, 2, 3, 5, 6, 16 and 19 were studied in mice with single or double Robertsonian translocations. The monosomy for different autosomes affects the preimplantation development of the mouse embryos in different ways. The monosomy for the autosomes 1, 3, 6, 16 or 19 does not affect cleavage, compactization or blastulation and is, in some cases, even compatible with the implantation. The most these embryos are eliminated at the blastocyst stage (monosomy for the autosomes 3, 6 or 19) or, sometimes, at the postimplantation stages (1 or 16). The monosomy for the autosomes 2 or 5 is realized during cleavage causing the developmental delay, pathological changes in the nuclei of blastomeres and elimination at the morula stage. The results obtained suggest differential activity of chromosomes at the preimplantation stages of embryogenesis. Possible reasons for the early death of the embryos with particular types of monosomy are discussed. A hypothesis of mutual activation of the homologous autosomes at the early developmental stages is put forward. According to this hypothesis, the loci of a single unpaired autosome, especially of paternal origin, remain inactive during the early embryogenesis.


Cognitive flexibility-related prefrontal activation in preschoolers: A biological approach to temperamental effortful control.
Individual differences in temperament have been theorized to be supported by differential recruitment of key neural regions, resulting in the distinct patterns of behavior observed throughout life. Although a compelling model, its rigorous and systematic testing is lacking, particularly within the heightened neuroplasticity of early childhood. The current study tested a model of the link between temperament, the brain, and behavior for cognitive flexibility in a sample of 4-5-year-old children (N = 123) using functional near-infrared spectroscopy (fNIRS) to assess prefrontal cortex (PFC) activation. Structural Equation Modeling (SEM) was used to explore the link between survey reports of temperamental effortful control, and both performance-based and neuroimaging measures of cognitive flexibility. Results indicated that greater parent-reported temperamental effortful control was associated with better performance on a cognitive flexibility task, and less activation of the DLPFC in preschoolers. These findings support the theorized model of the interrelatedness between temperamental tendencies, behavior, and brain activation and suggest that better temperamentally regulated children use the DLPFC more efficiently for cognitive flexibility.


Comparison of carotid baroreflex control of plasma AVP concentration in conscious and anesthetized dogs.
We compared carotid sinus baroreflex control of endogenous plasma arginine vasopressin (AVP) in chronically prepared conscious and acutely prepared anesthetized dogs. The carotid sinuses of both conscious and pentobarbital-anesthetized dogs were isolated bilaterally and perfused at constant pressures. Carotid sinus pressure (CSP) was changed between 200 and 50 mmHg in 25-mmHg steps in intact conscious and anesthetized dogs. Similar runs were repeated after vagotomy. Mean arterial pressure (MAP) and heart rate (HR) were monitored. At each interval of CSP, blood was withdrawn for AVP analysis by radioimmunoassay. MAP responses to changes in CSP were not different in the four experimental groups. Both anesthesia and vagotomy increased the HR responses to changes in CSP. With vagi intact, AVP increased at high CSP in conscious but not in anesthetized dogs. After vagotomy, low CSP led to an increase in plasma AVP that did not differ between conscious and anesthetized dogs. The results suggest that the release of AVP is modulated by the action of the carotid baroreflex as a normal component of an integrated efferent response. The response is similar in conscious and pentobarbital-anesthetized dogs and is normally buffered by reflexes with vagal afferents.


Assessment of epicardial adipose tissue thickness and total calcium score in sarcoidosis patients.
Increased thickness of epicardial adipose tissue (EAT) and the total coronary artery calcium score (TCACS) are independent predictors of atherosclerosis. The aim of this study was to investigate whether EAT thickness, measured using thoracic computed tomography, and TCACS were greater in patients with sarcoidosis. This was a retrospective study. The details of participants who presented at the cardiology and pulmonology outpatient clinics between January 2011 and December 2018 with dyspnea, chest pain, or palpitations from the hospital data system were reviewed. Patients with transthoracic echocardiography and thorax computed tomography (CT) (CT) records were identified, and those who were diagnosed with sarcoidosis, had no other health problems, and did not take any medication were included in the study. A total of 45 controls and 78 sarcoidosis patients were enrolled. The mean age of the controls was 46.15±13.1 years, while it was 46.26±12.37 years in the sarcoidosis group, which represented no significant difference between the groups (p>0.05). When the groups were compared in terms of a fasting blood test, erythrocyte sedimentation rate (ESR), TCACS, EAT thickness, levels of C-reactive protein (CRP), total cholesterol, low-density lipoprotein (LDL), and triglycerides, it was observed that CRP and EAT thickness were higher in the sarcoidosis group. The results of this study indicated that the thickness of EAT calculated using thorax CT was greater in sarcoidosis patients; however, the TCACS was similar in both groups. In addition, there was a positive correlation between EAT thickness and the level of total cholesterol, LDL, triglycerides, CRP, and the sedimentation rate. These findings suggest that atherosclerosis may start earlier in those with sarcoidosis than in the healthy population.


Subcellular localization of CD80 receptors is dependent on an intact cytoplasmic tail and is required for CD28-dependent T cell costimulation.
CD28 provides a major costimulatory signal to T cells when it is cross-linked with mAb, immobilized recombinant ligand (CD80Ig or CD86Ig), or ligand-bearing cells but not when it is bound by specific Fab fragments or monomeric ligand. We wanted to determine how monomeric CD80 could cross-link CD28 since CD80 is expressed as a monomer on the surface of APC. We found that CD80 may interact with the actin-based cytoskeleton. To test whether the interaction of CD80 with the cytochalasin B-sensitive cytoskeleton was necessary for T cell costimulation through CD28, we constructed a tailless form of CD80 and generated stable transfectants of Chinese hamster ovary epithelial cells and Reh B cells expressing either the tailless or wild-type CD80 molecules. Unlike control cells expressing wild-type CD80, the tailless CD80 transfectants expressing equivalent levels of surface CD80 were not able to provide a costimulatory signal for anti-CD3-induced T cell proliferation, up-regulation of CD25 (IL-2Ralpha) expression, or the induction of IL-2 secretion. Thus, the cytoplasmic tail of CD80 apparently is required to signal T cells. Confocal microscopic studies revealed that wild-type CD80 and tailless CD80 have different patterns of subcellular distribution in both epithelial and lymphoid cells. Furthermore, T cell contact induces more patching and capping of CD80 in wild-type CD80-expressing cells than in tailless CD80-expressing cells. This suggests that the cytoplasmic region of CD80 functions to localize CD80 in complexes required for effective T cell costimulation.


Self-awareness of traumatically brain injured patients in the acute inpatient rehabilitation setting.
The self-awareness of brain injured patients in an acute inpatient rehabilitation programme was measured using the Functional Self-Appraisal Scale (FSAS), which compares patient and staff ratings of patient performance on tasks relevant for acute rehabilitation and in a format that is easy to administer. There was a significant difference between patients' and staffs ratings near admission, consistent with previous findings in post-acute settings that brain injured patients tend to overestimate their abilities relative to other raters. Patient and staff ratings tended to converge at time of discharge, likely reflecting patients' improvements on rehabilitation tasks rather than increased self-awareness. The average difference between patients' and staffs ratings did not correlate with neuropsychological functioning on admission, supporting evidence that self-awareness early in the recovery process is a unique construct. Future research is outlined, including investigating the relationship of early levels of self-awareness following TBI to functional outcome.


Perinatal testosterone exposure and cerebral lateralisation in adult males: evidence for the callosal hypothesis.
Two competing theories address the influence of foetal testosterone on cerebral laterality: one proposing exposure to high foetal testosterone concentrations is related to atypical lateralisation (Geschwind-Galaburda hypothesis), the other that high foetal testosterone concentrations exaggerate typical lateralisation (callosal hypothesis). The current study examined the relationship between cord testosterone concentrations and cerebral laterality for language and spatial memory in adulthood. Male participants with high (>0.15nmol) and low (<0.10nmol) cord testosterone levels were invited to take part in the study (n=18 in each group). Cerebral laterality was measured using functional Transcranial Doppler ultrasonography, while participants completed word generation and visual short-term memory tasks. Typical left lateralisation of language was more common in the high-testosterone group than in the low-testosterone group, χ(2)=4.50, df=1, p=034. Spatial memory laterality was unrelated to cord testosterone level. Our findings indicate that foetal testosterone exposure is related to language laterality in a direction that supports the callosal hypothesis.


Enantioselective conjugate addition to nitroolefins catalysed by helical peptides with a single remote stereogenic centre.
Two short pentapeptides rich in α-aminoisobutyric acid (Aib) residues have been shown to act as enantioselective organocatalysts for the conjugate addition of nucleophiles to nitroolefins. An L-alanine terminated peptide, (Aib)4(L-Ala)NHtBu, which has neither functionalised sidechains nor a highly designed reactive site, used an exposed N-terminal primary amine and the amide bonds of the backbone to mediate catalysis. Folding of this peptide into a 310 helical structure was observed by crystallography. Folding into a helix relays the conformational preference of the chiral alanine residue at the C-terminus to the primary amine at the N-terminus, 0.9 nm distant. The chiral environment and defined shape produced by the 310 helix brings the amine site into proximity to two exposed amide NHs. Reaction scope studies implied that the amine acts as a Brønsted base and the solvent-exposed NH groups of the helix, shown to weakly bind β-nitrostyrene, are needed to obtain an enantiomeric excess. Replacement of L-alanine with D-phenylalanine gave (Aib)4(D-Phe)NHtBu, a peptide that now catalysed the benchmark reaction with the opposite enantioselectivity. These studies show how achiral residues can play a key role in enantioselective catalysis by peptides through the promotion of folding.


Economic review of point-of-care EEG.
Aims: Point-of-care electroencephalogram (POC-EEG) is an acute care bedside screening tool for the identification of nonconvulsive seizures (NCS) and nonconvulsive status epilepticus (NCSE). The objective of this narrative review is to describe the economic themes related to POC-EEG in the United States (US).Materials and methods: We examined peer-reviewed, published manuscripts on the economic findings of POC-EEG for bedside use in US hospitals, which included those found through targeted searches on PubMed and Google Scholar. Conference abstracts, gray literature offerings, frank advertisements, white papers, and studies conducted outside the US were excluded.Results: Twelve manuscripts were identified and reviewed; results were then grouped into four categories of economic evidence. First, POC-EEG usage was associated with clinical management amendments and antiseizure medication reductions. Second, POC-EEG was correlated with fewer unnecessary transfers to other facilities for monitoring and reduced hospital length of stay (LOS). Third, when identifying NCS or NCSE onsite, POC-EEG was associated with greater reimbursement in Medical Severity-Diagnosis Related Group coding. Fourth, POC-EEG may lower labor costs via decreasing after-hours requests to EEG technologists for conventional EEG (convEEG).Limitations: We conducted a narrative review, not a systematic review. The studies were observational and utilized one rapid circumferential headband system, which limited generalizability of the findings and indicated publication bias. Some sample sizes were small and hospital characteristics may not represent all US hospitals. POC-EEG studies in pediatric populations were also lacking. Ultimately, further research is justified.Conclusions: POC-EEG is a rapid screening tool for NCS and NCSE in critical care and emergency medicine with potential financial benefits through refining clinical management, reducing unnecessary patient transfers and hospital LOS, improving reimbursement, and mitigating burdens on healthcare staff and hospitals. Since POC-EEG has limitations (i.e. no video component and reduced montage), the studies asserted that it did not replace convEEG.


The Role of Sodium Bicarbonate in the Management of Some Toxic Ingestions.
Adverse reactions to commonly prescribed medications and to substances of abuse may result in severe toxicity associated with increased morbidity and mortality. According to the Center for Disease Control, in 2013, at least 2113 human fatalities attributed to poisonings occurred in the United States of America. In this article, we review the data regarding the impact of systemic sodium bicarbonate administration in the management of certain poisonings including sodium channel blocker toxicities, salicylate overdose, and ingestion of some toxic alcohols and in various pharmacological toxicities. Based on the available literature and empiric experience, the administration of sodium bicarbonate appears to be beneficial in the management of a patient with the above-mentioned toxidromes. However, most of the available evidence originates from case reports, case series, and expert consensus recommendations. The potential mechanisms of sodium bicarbonate include high sodium load and the development of metabolic alkalosis with resultant decreased tissue penetration of the toxic substance with subsequent increased urinary excretion. While receiving sodium bicarbonate, patients must be monitored for the development of associated side effects including electrolyte abnormalities, the progression of metabolic alkalosis, volume overload, worsening respiratory status, and/or worsening metabolic acidosis. Patients with oliguric/anuric renal failure and advanced decompensated heart failure should not receive sodium bicarbonate.


Neuroaesthetics: a coming of age story.
Neuroaesthetics is gaining momentum. At this early juncture, it is worth taking stock of where the field is and what lies ahead. Here, I review writings that fall under the rubric of neuroaesthetics. These writings include discussions of the parallel organizational principles of the brain and the intent and practices of artists, the description of informative anecdotes, and the emergence of experimental neuroaesthetics. I then suggest a few areas within neuroaesthetics that might be pursued profitably. Finally, I raise some challenges for the field. These challenges are not unique to neuroaesthetics. As neuroaesthetics comes of age, it might take advantage of the lessons learned from more mature domains of inquiry within cognitive neuroscience.


The emerging role of perivascular cells (pericytes) in viral pathogenesis.
Viruses may exploit the cardiovascular system to facilitate transmission or within-host dissemination, and the symptoms of many viral diseases stem at least in part from a loss of vascular integrity. The microvascular architecture is comprised of an endothelial cell barrier ensheathed by perivascular cells (pericytes). Pericytes are antigen-presenting cells (APCs) and play crucial roles in angiogenesis and the maintenance of microvascular integrity through complex reciprocal contact-mediated and paracrine crosstalk with endothelial cells. We here review the emerging ways that viruses interact with pericytes and pay consideration to how these interactions influence microvascular function and viral pathogenesis. Major outcomes of virus-pericyte interactions include vascular leakage or haemorrhage, organ tropism facilitated by barrier disruption, including viral penetration of the blood-brain barrier and placenta, as well as inflammatory, neurological, cognitive and developmental sequelae. The underlying pathogenic mechanisms may include direct infection of pericytes, pericyte modulation by secreted viral gene products and/or the dysregulation of paracrine signalling from or to pericytes. Viruses we cover include the herpesvirus human cytomegalovirus (HCMV, Human betaherpesvirus 5), the retrovirus human immunodeficiency virus (HIV; causative agent of acquired immunodeficiency syndrome, AIDS, and HIV-associated neurocognitive disorder, HAND), the flaviviruses dengue virus (DENV), Japanese encephalitis virus (JEV) and Zika virus (ZIKV), and the coronavirus severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2; causative agent of coronavirus disease 2019, COVID-19). We touch on promising pericyte-focussed therapies for treating the diseases caused by these important human pathogens, many of which are emerging viruses or are causing new or long-standing global pandemics.


The anti-inflammatory agents used in rheumatic disorders.
The objectives of nonspecific anti-inflammatory therapy are, for the most part, to relieve symptoms. In selected patients such therapy may also slow, prevent, or reverse the destructive effects of the inflammatory process. It is well to remember that none of the wide variety of anti-inflammatory agents is without some inherent toxicity, although some agents are obviously much more toxic than others. The physician prescribing these agents must keep in mind both the potential benefits and the potential risks attendant to their use. In this way he can balance potential benefit against potential risk and arrive at the most rational drug therapy for any given rheumatic disorder.


Sperm entry is sufficient to trigger division of the central cell but the paternal genome is required for endosperm development in Arabidopsis.
Fertilization in flowering plants involves two sperm cells and two female gametes, the egg cell and the central cell, progenitors of the embryo and the endosperm, respectively. The mechanisms triggering zygotic development are unknown and whether both parental genomes are required for zygotic development is unclear. In Arabidopsis, previous studies reported that loss-of-function mutations in CYCLIN DEPENDENT KINASE A1 (CDKA;1) impedes cell cycle progression in the pollen leading to the production of a single sperm cell. Here, we report that a significant proportion of single cdka;1 pollen delivers two sperm cells, leading to a new assessment of the cdka;1 phenotype. We performed fertilization of wild-type ovules with cdka;1 mutant sperm cells and monitored in vivo the fusion of the male and female nuclei using fluorescent markers. When a single cdka;1 sperm was delivered, either female gamete could be fertilized leading to similar proportions of seeds containing either a single endosperm or a single embryo. When two cdka;1 sperm cells were released, they fused to each female gamete. Embryogenesis was initiated but the fusion between the nuclei of the sperm cell and the central cell failed. The failure of karyogamy in the central cell prevented incorporation of the paternal genome, impaired endosperm development and caused seed abortion. Our results thus support that the paternal genome plays an essential role during early seed development. However, sperm entry was sufficient to trigger central cell mitotic division, suggesting the existence of signaling events associated with sperm cell fusion with female gametes.


Effects of membrane-bound glucose dehydrogenase overproduction on the respiratory chain of Gluconobacter oxydans.
The acetic acid bacterium Gluconobacter oxydans incompletely oxidizes carbon sources as a natural part of its metabolism, and this feature has been exploited for many biotechnological applications. The most important enzymes used to harness the biocatalytic oxidative capacity of G. oxydans are the pyrroloquinoline quinone (PQQ)-dependent dehydrogenases. The membrane-bound PQQ-dependent glucose dehydrogenase (mGDH), encoded by gox0265, was used as model protein for homologous membrane protein production using the previously described Gluconobacter expression vector pBBR1p452. The mgdh gene had ninefold higher expression in the overproduction strain compared to the parental strain. Furthermore, membranes from the overexpression strain had a five- and threefold increase of mGDH activity and oxygen consumption rates, respectively. Oxygen consumption rate of the membrane fraction could not be increased by the addition of a substrate combination of glucose and ethanol in the overproduction strain, indicating that the terminal quinol oxidases of the respiratory chain were rate limiting. In contrast, addition of glucose and ethanol to membranes of the control strain increased oxygen consumption rates approaching the observed rates with G. oxydans overproducing mGDH. The higher glucose oxidation rates of the mGDH overproduction strain corresponded to a 70 % increase of the gluconate production rate compared to the control strain. The high rate of glucose oxidation may be useful in the industrial production of gluconates and ketogluconates, or as whole-cell biosensors. Furthermore, mGDH was purified to homogeneity by one-step strep-tactin affinity chromatography and characterized. To our knowledge, this is the first report of a membrane integral quinoprotein being purified by affinity chromatography and serves as a proof-of-principle for using G. oxydans as a host for membrane protein expression and purification.


Wrist fusion in posttraumatic brachial plexus palsy.
Devastating brachial plexus injury with multiple root avulsions results in wrist instability, imbalance, and inability of patients to control the placement of their hand in space. In this study, results of wrist arthrodesis were analyzed in relation to factors such as age, severity score, and additional hand reanimation procedures. Indices of patient satisfaction were also measured using the Terzis and Disabilities of the Arm, Shoulder and Hand questionnaires. Between January of 1978 and January of 2006, 97 patients with posttraumatic plexopathies underwent secondary procedures for hand reanimation. Sixty-one of these patients had wrist fusion. Arthrodesis was performed between the radius and third metacarpal using mainly a rigid plate; iliac bone graft was used in the majority of patients. Thirty-one patients underwent muscle transfers (45 free muscles) and 11 had tendon transfers in the wrist fusion group to enhance finger function. Patient self-assessment was carried out using the Terzis and Disabilities of the Arm, Shoulder and Hand questionnaires. All patients' wrists fused. Patients with free-muscle transfer for finger flexion and extension achieved superior muscle grading compared with patients without wrist fusion, but this was not significant. A total of 35 patients (57.37 percent) answered the questionnaires; 97.14 percent were satisfied with wrist stability and 88.57 percent reported that the procedure enhanced the overall upper limb function. The Disabilities of the Arm, Shoulder and Hand score was 59.14 +/- 12.9, which means moderate ability in daily activities. Wrist fusion in patients with brachial plexus palsy is recommended as a complementary procedure, offering a stable, painless carpus, with improvement of overall upper limb function and appearance.


Vertebral level of Tuffier's line measured by ultrasonography in parturients in the lateral decubitus position.
It is known that Tuffier's line intersects the spine at the L4 spinous process or at the L4-L5 intervertebral space. Full term parturient women undergo various physical changes. Therefore, determining the vertebral level with Tuffier's line based on palpation inevitably is not very accurate. The aim of this study was to use ultrasound to verify the difference between vertebral levels for the palpated Tuffier's line in parturient and non-parturient women in the lateral decubitus position. We consecutively enrolled 40 parturient women at 37-41 weeks of gestation and 40 non-parturient women scheduled for regional anesthesia. In the left lateral position, the location of the vertebra was identified using ultrasonography. We marked every intervertebral space from L5 to L2 vertebra, divided each spinous process into two equal parts, and numbered the spaces sequentially from 1 to 9. We drew a Tuffier's line by palpating, recorded the vertebral level that this line intersected. The mean value of an arbitrary number of vertebral level of Tuffier's line was 6.4 ± 0.9 in the non-pregnant group and this represents L4-lower vertebral level. In the pregnant group, the mean value was 3.0 ± 1.0 which represents L3-lower vertebral level. There was a significant difference between the two groups (P < 0.05). We compared using an available ultrasound technique the vertebral levels intersected by the palpated Tuffier's line between parturient and non-parturient women and found that the vertebral levels were more cephalad in the parturient women compared to the non-parturient women.


Challenges in the use of atomistic simulations to predict solubilities of drug-like molecules.
Background: Solubility is a physical property of high importance to the pharmaceutical industry, the prediction of which for potential drugs has so far been a hard task. We attempted to predict the solubility of acetylsalicylic acid (ASA) by estimating the absolute chemical potentials of its most stable polymorph and of solutions with different concentrations of the drug molecule. Methods: Chemical potentials were estimated from all-atom molecular dynamics simulations. We used the Einstein molecule method (EMM) to predict the absolute chemical potential of the solid and solvation free energy calculations to predict the excess chemical potentials of the liquid-phase systems. Results: Reliable estimations of the chemical potentials for the solid and for a single ASA molecule using the EMM required an extremely large number of intermediate states for the free energy calculations, meaning that the calculations were extremely demanding computationally. Despite the computational cost, however, the computed value did not agree well with the experimental value, potentially due to limitations with the underlying energy model. Perhaps better values could be obtained with a better energy model; however, it seems likely computational cost may remain a limiting factor for use of this particular approach to solubility estimation. Conclusions: Solubility prediction of drug-like solids remains computationally challenging, and it appears that both the underlying energy model and the computational approach applied may need improvement before the approach is suitable for routine use.


MMP-9 Concentration in Peritoneal Fluid Is a Valuable Biomarker Associated with Endotoxemia in Equine Colic.
The purpose of the study was to compare the results of sepsis scoring (clinical examination and clinical pathology) to the concentrations of matrix-metalloproteinases (MMPs) -2, -8, and -9; tissue-inhibitor of metalloproteinases (TIMPs) -1 and -2; and inflammatory chemokines interleukin (IL) 1β and tumor-necrosis-factor-alpha (TNF-α) in plasma and peritoneal fluid of equine colic patients. A modified sepsis scoring including general condition, heart and respiratory rate, rectal temperature, mucous membranes, white blood cell count (WBC), and ionized calcium was applied in 47 horses presented with clinical signs of colic. Using this scoring system, horses were classified as negative (n = 32, ≤6/19 points), questionable (n = 9, 7-9/19 points), or positive (n = 6, ≥10/19 points) for sepsis. MMPs, TIMPs, IL-1β, and TNF-α concentrations were evaluated in plasma and peritoneal fluid using species-specific sandwich ELISA kits. In a linear discriminant analysis, all parameters of sepsis scoring apart from calcium separated well between sepsis severity groups (P < 0.05). MMP-9 was the only biomarker of high diagnostic value, while all others remained insignificant. A significant influence of overall sepsis scoring on MMP-9 was found for peritoneal fluid (P = 0.005) with a regression coefficient of 0.092, while no association was found for plasma (P = 0.085). Using a MMP-9 concentration of >113 ng/ml in the peritoneal fluid was found to be the ideal cutoff to identify positive sepsis scoring (≥10/19 points; sensitivity of 83.3% and specificity of 82.9%). In conclusion, MMP-9 was found to be a biomarker of high diagnostic value for sepsis and endotoxemia in equine colic. The evaluation of peritoneal fluid seems preferable in comparison to plasma. As abdominocentesis is commonly performed in the diagnostic work-up of equine colic, a pen-side assay would be useful and easy-to-perform diagnostic support in the decision for therapeutic intervention.


Toxoplasma development - turn the switch on or off?
Toxoplasma gondii exhibits a complex, multi-stage life cycle in which the need for parasite expansion is balanced with the production of transmissible forms. For human disease the key developmental switch is from the tachyzoite to the mature bradyzoite, which is not well understood at the molecular level. This review highlights the role of the tachyzoite in regulating the initiation of bradyzoite differentiation through newly discovered transcription factors of the ApiAP2 family that must be turned off for development to unfold. Exit from the tachyzoite cell cycle is also tightly co-ordinated with the induction of bradyzoite gene expression, which is strongly influenced by the host cell environment. New evidence suggests a parasite casein kinase II and host anti-growth factor CDA1 can influence specific pathways that are responsible for sensing the host cell environment and informing the parasites decision to continue replication or to develop into bradyzoites. These results indicate tachyzoite gene expression mechanisms and signal transduction pathways likely hold the keys to tissue cyst formation in Toxoplasma.


Mucilaginibacter lutimaris sp. nov., isolated from a tidal flat sediment.
A Gram-staining-negative, non-spore-forming, facultatively aerobic, non-motile, rod-shaped bacterial strain, BR-3(T), was isolated from a tidal flat on the western coast of Korea, and subjected to a polyphasic study. Strain BR-3(T) grew optimally at 25 °C, at pH 6.5-7.0 and in the absence of NaCl. Phylogenetic analyses based on 16S rRNA gene sequences revealed that strain BR-3(T) fell within the clade comprising species of the genus Mucilaginibacter, joining the type strain of Mucilaginibacter rigui, with which it exhibited highest 16S rRNA gene sequence similarity (98.2%). 16S rRNA gene sequence similarity values between strain BR-3(T) and the type strains of the other species of the genus Mucilaginibacter were in the range 93.8-95.9%. A mean DNA-DNA relatedness value between strain BR-3(T) and M. rigui KCTC 12534(T) was 21%. Strain BR-3(T) contained MK-7 as the predominant menaquinone and C(16:1)ω7c and/or iso-C(15:0) 2-OH and iso-C(15:0) as the major fatty acids. The major polar lipids were phosphatidylethanolamine and an unidentified aminophospholipid. The DNA G+C content was 49.8 mol%. Differential phenotypic properties and phylogenetic and genetic distinctiveness of strain BR-3(T) demonstrated that this strain is separate from M. rigui as well as the other species of the genus Mucilaginibacter. On the basis of the data presented, strain BR-3(T) is considered to represent a novel species of the genus Mucilaginibacter, for which the name Mucilaginibacter lutimaris sp. nov. is proposed. The type strain is BR-3(T) (=KCTC 23461(T) =CCUG 60742(T)).


High cyclophilin D content of synaptic mitochondria results in increased vulnerability to permeability transition.
Mitochondria isolated from synaptosomes are more sensitive to Ca2+ overload and the resultant opening of the mitochondrial permeability transition pore (mPTP) than nonsynaptic mitochondria. To identify the mechanisms underlying these differences in Ca2+ dynamics, we examined relative levels of mPTP components in synaptic versus nonsynaptic mitochondria. Synaptic mitochondria had higher levels of cyclophilin D when compared with nonsynaptic mitochondria, whereas levels of the voltage-dependent anion channel and the adenine nucleotide translocase were similar in the two mitochondrial fractions. These differences in Ca2+ handling between synaptic and nonsynaptic mitochondria were greatly reduced in cyclophilin D null [Ppif-/- (peptidylprolyl isomerase F)] mice. Higher concentrations of cyclosporine A, which interacts with cyclophilin D to delay mPTP opening, were necessary to increase the Ca2+ uptake capacity of synaptic versus nonsynaptic mitochondria. To determine whether the differences in Ca2+ handling might reflect the relative abundance of neuronal and glial mitochondria in the two mitochondrial fractions, we compared cyclophilin D levels in primary cortical neurons and astrocytes. Primary rat cortical neurons possess higher cyclophilin D levels than do primary astrocytes. In the adult rat brain, cyclophilin D immunoreactivity was abundant in neurons but sparse in astrocytes. Together, these results demonstrate that the Ca2+ handling differences observed in synaptic versus nonsynaptic mitochondria are primarily the result of the high levels of cyclophilin D in synaptic mitochondria, reflecting the greater proportion of neuronal mitochondria in this fraction. The high levels of cyclophilin D in neuronal mitochondria result in their greater vulnerability to mPT and in higher levels of cyclosporine A being required to inhibit mPTP opening.


Insulin Sensitivity and Renal Hemodynamic Function in Metformin-Treated Adults With Type 2 Diabetes and Preserved Renal Function.
Impaired insulin sensitivity is associated with hyperfiltration (i.e., elevated glomerular filtration rate [GFR]) in adolescents with type 2 diabetes (T2D) and adults with prediabetes. Yet, these relationships are based on studies that relied on estimated GFR (eGFR), estimates of insulin sensitivity, or both. We aimed to verify the relationship between insulin sensitivity and renal hemodynamic function by gold standard methods in adults with T2D. Insulin sensitivity was assessed by hyperinsulinemic-euglycemic clamp (M value) (glucose infusion rate in mg/kglean/min) and renal hemodynamic function by urinary inulin (GFR) and para-aminohippuric acid (effective renal plasma flow [ERPF]) clearances in participants with T2D without overt kidney disease. Filtration fraction (FF) (GFR/ERPF) was calculated. Relationships between insulin sensitivity and renal hemodynamic parameters were examined by multivariable linear regression. Renal hemodynamic parameters were examined across tertiles of M values. We tested 44 adults with T2D, of whom 77% were male, with mean ± SD age 63 ± 7 years, BMI 31.2 ± 4.0 kg/m2, and HbA1c 7.4 ± 0.6%. Average GFR was 110 ± 26 mL/min, with an FF of 22.1 ± 2.8% and median 24-h urinary albumin excretion of 11.3 mg (interquartile range 5.8-17.0). Average M value was 5.6 ± 2.9 mg/kglean/min. Insulin sensitivity inversely correlated with GFR (r = -0.44, P < 0.01) and FF (r = -0.40, P < 0.01), and these associations remained significant after multivariable adjustments for age, sex, renin-angiotensin system inhibitor use, and HbA1c. In addition, GFR, FF, and urinary albumin excretion were highest in the participants in the lowest M value tertile. For the first time, we demonstrate that impaired insulin sensitivity is associated with intrarenal hemodynamic dysfunction by gold standard techniques in adults with T2D treated with metformin monotherapy.


The association of neuropsychiatric symptoms with regional brain volumes from patients in a tertiary multi-disciplinary memory clinic.
To examine the interaction between structural brain volume measures derived from a clinical magnetic resonance imaging (MRI) and occurrence of neuropsychiatric symptoms (NPS) in outpatient memory clinic patients. Clinical and neuroimaging data were collected from the medical records of outpatient memory clinic patients who were seen by neurologists, geriatric neuropsychiatrists, and geriatricians. MRI scan acquisition was carried out on a 3 T Siemens Verio scanner at Johns Hopkins Bayview Medical Center. Image analyses used an automated multi-label atlas fusion method with a geriatric atlas inventory to generate 193 anatomical regions from which volumes were measured. Regions of interest were generated a priori based on previous literature review of NPS in dementia. Regional volumes for agitation, apathy, and delusions were carried forward in a linear regression analysis. Seventy-two patients had clinical and usable neuroimaging data that were analyzed and grouped by Mini-Mental State Exam (MMSE). Neuropsychiatric Inventory Questionnaire (NPI-Q) agitation was inversely associated with rostral anterior cingulate cortex (ACC) bilaterally and left subcallosal ACC volumes in the moderate severity group. Delusions were positively associated with left ACC volumes in both severe and mild groups but inversely associated with the right dorsolateral prefrontal cortex (DLPFC) in the moderate subgroup. Agitation, apathy, and delusions are associated with volumes of a priori selected brain regions using clinical data and clinically acquired MRI scans. The ACC is an anatomic region common to these symptoms, particularly agitation and delusions, which closely mirror the findings of research-quality studies and suggest its importance as a behavioral hub.


Characterization of the heat shock response in Mycobacterium bovis BCG.
We have for the first time characterized the heat shock response in mycobacteria both at the level of transcription, by RNA extraction, Northern (RNA) blotting, and hybridization with gene-specific probes for the Mycobacterium tuberculosis 65- and 71-kDa heat shock proteins (HSPs), and at the level of translation, by [35S]methionine labelling, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and autoradiography. We observed increased synthesis of 40-, 65-, 71-, and 90-kDa proteins, which appear to be major HSPs in mycobacteria. The 40-, 71-, and 90-kDa HSPs are coordinately regulated in terms of temperature requirements and kinetics of induction but differ in the levels of expression. The 65- and 71-kDa HSPs are differentially regulated in response to temperature, with different kinetics and levels of induction. mRNA transcript sizes for the 71-, 65-, 40-, and 30-kDa proteins were found to be broadly consistent with DNA sequence open reading frames. A maximum increase of about 69-fold in the levels of mRNA for the 71-kDa HSP after 45 min of heat shock at 45 degrees C was observed, whereas the 65-kDa HSP mRNA increased only 5-fold. It was also found that in M. bovis BCG, as in Escherichia coli, a major control mechanism of the heat shock response is operative at the level of transcription. An ability to characterize the heat shock response in mycobacteria provides an experimental model with which to study environmentally regulated gene expression and an opportunity to identify virulence genes, which may coregulate as part of the heat shock regulon.


Save the young--the elderly have lived their lives: ageism in marriage and family therapy.
The paucity of literature addressing mental health issues concerning geriatric populations represents the perpetuation of ageist practices and beliefs in the field of marriage and family therapy. The purpose of this study was to assess whether client age and clinical training relate to the evaluation of couples who present for conjoint therapy. Written vignettes describing two couples, one older and one younger, who report issues involving the absence of sexual intimacy, increased frequency of arguments, and increased use of alcohol were evaluated by practicing marriage and family therapists, therapists-in-training, and individuals with no clinical background. It was hypothesized that respondents' views would vary in connection with the age of the couple and with the three levels of participant training. Results indicate that client age and participant training are associated with perceptions of individual and couple functioning. Our findings suggest that the relational and mental health concerns experienced by elder couples are not perceived as seriously as are identical concerns experienced by younger couples. Contrary to our expectations the observed differences between views of the two age conditions did not significantly differ between levels of participant training. Training and experience in marriage and family therapy may not significantly mitigate vulnerability to age-discrepant views.


Relationship between maternal dietary patterns and hypospadias.
Little is known about the role of maternal nutrition in the development of hypospadias, which is the most common urogenital congenital anomaly. This study investigated the relationship between maternal nutrition and the risk of hypospadias, particularly focusing on maternal food patterns. We compared 471 hypospadias cases with 490 controls in the United Kingdom. A questionnaire including information on life style, occupation, usual maternal diet and dietary supplements was administered using telephone interviews. Cases and controls were compared for individual food item intake and food patterns derived by cluster analysis. Multivariable logistic regression analysis adjusted for income, maternal age, low birthweight, smoking and folic acid supplement use was used to assess the relationship between maternal nutrition and hypospadias. Three food patterns were created with the labels 'health conscious', 'mixed' and 'non-health conscious'. 'Non-health conscious' subjects (low frequency of consumption of yoghurt, cheese, eggs, fruit and vegetables, fish, beans and pulses, olive oil and organic food) had a higher risk of hypospadias (odds ratio 1.54; 95% confidence interval 1.06, 2.26) compared with 'health conscious' subjects (high frequency of consumption of fresh fruit and vegetables, dried fruit, fresh or frozen fish, beans, pulses, soya products, olive oil and organic food), after adjustment for potential confounders. Intakes of individual foods were not strongly associated with hypospadias. We could not exclude the possibility of residual confounding, and this needs to be further investigated. We found an association between food pattern and hypospadias, with those with less health conscious food patterns having a higher risk. Further study is needed to confirm this association.


Diversity evaluation based on morphological, physiological and isozyme variation in genetic resources of garlic (Allium sativum L.) collected worldwide.
The aim of this study was to obtain primary information about the global diversity of garlic (Allium sativum L.) by evaluating morphological, physiological and isozyme variation. A total of 107 garlic accessions collected worldwide were grown in Yamaguchi, Japan. Five morphological traits (bulb weight, bulb diameter, number of cloves per bulb, number of bulbils and scape length) and one physiological trait (bolting period) of the collected garlic showed wide variation. Meanwhile, a total of 140 garlic accessions, including the 107 mentioned above, were characterized by leucine aminopeptidase (LAP) and phosphoglucoisomerase (PGI) isozyme analyses; they clearly showed polymorphisms in putative isozyme loci (Lap-1, Lap-2 and Pgi-1). Allelic frequencies were estimated in each group of accessions categorized by their geographical origin, and the observed (Ho) and expected (He) heterozygosities were calculated. The allelic frequencies differed between groups. A principal component analysis based on morpho-physiological data indicated a grouping of the garlic accessions into Central Asian and Northern Mediterranean groups as well as others. We discuss the roles of artificial and natural selection that may have caused differentiation in these traits, on the assumption that ancestral domesticated garlic populations have adapted in various regions using standing variation or mutations that accumulated during expansion, and have evolved along with human-preferred traits over a long history of cultivation.


Regulation of lens cyclic nucleotide metabolism by Ca2+ plus calmodulin.
Adenylate cyclase activity was identified in membranes isolated from bovine lens fiber cells. Basal activity, in the presence of microM Ca2+ was stimulated by either sodium fluoride, guanosine 5'-[alpha,beta-imido]triphosphate (Gpp(NH)p), or forskolin; ethylene glycolbis(2-aminoethylether) tetraacetic acid (EGTA) markedly inhibited both the basal activity and the extent of stimulation by these agents. Exogenous calmodulin enhanced the Ca2+-dependent stimulation of adenylate cyclase activity. In the presence of optimal concentrations of Ca2+ plus calmodulin, adenylate cyclase activity was approximately 15 times greater than that in the presence of EGTA. Adenylate cyclase activity was not stimulated by a number of potential agonists that included carbachol, serotonin, prostaglandin E1 (PGE1), prostaglandin E2 (PGE2), adenosine, isoproterenol epinephrine, dopamine, and phenylephrine. The presence of the Ns and Ni guanine nucleotide regulatory complexes was indicated by two observations: Cholera toxin catalyzed the adenosine diphosphate (ADP) ribosylation of a number of lens membrane proteins, including a 46,500-dalton component (likely the alpha-subunit of Ns), and Pertussis toxin catalyzed the ADP ribosylation of a single 41,000-dalton lens membrane component (likely the alpha-subunit of Ni). However, that Gpp(NH)p did not inhibit either the forskolin-activated or the calmodulin-activated adenylate cyclase activities does not indicate a role for Ni in regulating this enzyme. Both cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) phosphodiesterase activities were identified in a supernate fraction derived from bovine lens. The cAMP phosphodiesterase activity appeared to be predominantly the low Km form of the enzyme. The cGMP phosphodiesterase activity, which was Ca2+-dependent, was partly inhibited maximally by 7 microM R24571, indicating its probable calmodulin dependence.(ABSTRACT TRUNCATED AT 250 WORDS)


Topology of bonding in pi-electron systems.
Delocalization in conjugated pi-electron systems is shown to be closely related to the topological concept of connectivity. A topological bond order is defined as a quantitative measure of the extent to which the subspace of a pair of adjacent atoms approximates a disconnected component of the molecular topological space. The agreement between this measure and molecular-orbital bond orders in the ordering of bond strengths within molecules is near perfect. There is also excellent quantitative agreement between these two measures of bond strength. Bond orders appear to be primarily determined by molecular topology. A new type of intramolecular equivalence relation between bonds, called "coresiduality," emerges from the topological analysis. Two bonds are coresidual if the residues remaining when each pair of adjacent atoms is excised from the molecule are isomorphic. Coresidual bonds have equal bond orders. Coresiduality includes, but is more general than, equivalence due to symmetry.


Pseudomonas aeruginosa-induced apoptosis is defective in respiratory epithelial cells expressing mutant cystic fibrosis transmembrane conductance regulator.
Chronic lung infection with Pseudomonas aeruginosa constitutes the most severe manifestation of cystic fibrosis, a scenario that results from defects in early clearance of the microbe. Early clearance involves epithelial cell ingestion of bacteria, rapid activation of nuclear factor-kappa B and cellular desquamation within minutes of P. aeruginosa infection, processes that are deficient in cells with mutant alleles of Cftr. Analyzing the effect of Cftr genotype on the apoptotic response of airway epithelial cells to P. aeruginosa, we found that human bronchial epithelial cells expressing Delta F508 cystic fibrosis transmembrane conductance regulator (CFTR) underwent significantly delayed apoptosis compared with cells expressing wild-type (WT) CFTR. Mice with a WT Cftr allele had apoptotic cells in their lungs after P. aeruginosa infections, whereas mice homozygous for the Delta F508 or G551D Cftr alleles showed little apoptosis in response to acute infection. Pseudomonal infection induced expression of CD95 and CD95 ligand, a response that was also delayed in cells homozygous for mutant Cftr alleles. Thus, WT CFTR expression promotes a rapid expression of CD95/CD95 ligand and apoptotic response to P. aeruginosa infection. Prompt apoptosis of infected epithelial cells may be critical for clearance of P. aeruginosa, and CFTR-associated defects in apoptosis may contribute to the pathogenesis of the lung disease in cystic fibrosis.


Impaired spatial working memory after anterior thalamic lesions: recovery with cerebrolysin and enrichment.
Lesions to the anterior thalamic nuclei (ATN) in rats produce robust spatial memory deficits that reflect their influence as part of an extended hippocampal system. Recovery of spatial working memory after ATN lesions was examined using a 30-day administration of the neurotrophin cerebrolysin and/or an enriched housing environment. As expected, ATN lesions in standard-housed rats given saline produced severely impaired reinforced spatial alternation when compared to standard-housed rats with sham lesions. Both cerebrolysin and enrichment substantially improved this working memory deficit, including accuracy on trials that required attention to distal cues for successful performance. The combination of cerebrolysin and enrichment was more effective than either treatment alone when the delay between successive runs in a trial was increased to 40 s. Compared to the intact rats, ATN lesions in standard-housed groups produced substantial reduction in c-Fos expression in the retrosplenial cortex, which remained low after cerebrolysin and enrichment treatments. Evidence that multiple treatment strategies restore some memory functions in the current lesion model reinforces the prospect for treatments in human diencephalic amnesia.


Polymeric carriers for gene delivery: chitosan and poly(amidoamine) dendrimers.
Gene therapy is a potential medical solution that promises new treatments and may hold the cure for many different types of diseases and disorders of the human race. However, gene therapy is still a growing medical field and the technology is still in its infancy. The main challenge for gene therapy is to find safe and effective vectors that are able to deliver genes to the specific cells and get them to express inside the cells. Due to safety concerns, synthetic delivery systems, rather than viral vectors, are preferred for gene delivery and significant efforts have been focused on the development of this field. However, we are faced with problems like low gene transfer efficiency, cytotoxicity and lack of cell-targeting capability for these synthetic delivery systems. Over the years, we have seen a variety of new and effective polymers which have been designed and synthesized specifically for gene delivery. Moreover, various strategies that aimed at enhancing their physicochemical properties, improving transfection efficiency, reducing cytotoxicity as well as incorporating functional groups that offer better targetability and higher cellular uptake are established. Here, we look at two potential polymeric carriers, chitosan and poly(amidoamine) dendrimers, which have been widely reported for gene delivery. For chitosan, the interest arises from their availability, excellent non-cytotoxicity profile, biodegradability and ease of modification. For poly(amidoamine) dendrimers, the interest arises from their ease of synthesis with controlled structure and size, minimal cytotoxicity, biodegradability and high transfection efficiencies. The latest developments on these polymers for gene delivery will be the main focus of this article.


Imaging in gynecological disease (5): clinical and ultrasound characteristics in fibroma and fibrothecoma of the ovary.
To describe the clinical and ultrasound features of fibroma and fibrothecoma of the ovary. Sixty-eight women with a histological diagnosis of fibroma or fibrothecoma of the ovary who had undergone a preoperative ultrasound examination between 1999 and 2007 were identified from the databases of four ultrasound centers. The tumors were characterized on the basis of ultrasound images, ultrasound reports and research protocols (when applicable) using the terms and definitions of the International Ovarian Tumor Analysis (IOTA) group. In 51 patients, ultrasound information had been collected prospectively; in the remaining 17 cases it was retrieved retrospectively from ultrasound reports and images. In 44 cases, electronic ultrasound images of good quality were available. These were reviewed by two observers, who described them using pattern recognition. Of the 68 patients identified, 53 had fibroma and 15 had fibrothecoma. The mean patient age was 54 (range, 17-80) years. Sixty-three percent (41/65) were postmenopausal and 60% (39/65) had no symptoms. Most (75%; 51/68) fibromas/fibrothecomas were solid tumors and most (75%; 51/68) manifested minimal or moderate blood flow on color Doppler examination. Using pattern recognition, all solid fibromas/fibrothecomas were described as round, oval or slightly lobulated tumors. Most (66%; 29/44) were solid tumors, with regular or slightly irregular internal echogenicity with stripy shadows, and some contained cystic spaces. Others (23%, 10/44) were solid tumors with regular or slightly irregular internal echogenicity without stripy shadows and with or without cystic spaces. Two were solid tumors that were so dense it was difficult to assess their internal echogenicity, two were multilocular solid tumors with large cystic spaces and one was described as being mainly cystic. Half of the women with fibroma/fibrothecoma had fluid in the pouch of Douglas and 16% (11/68) had ascites; CA 125 titers > or =35 U/mL were found in 34% (17/50) of the cases in which CA 125 results were available. Most fibromas and fibrothecomas are round, oval or lobulated solid tumors that cast stripy shadows and are associated with fluid in the pouch of Douglas, and most manifest minimal to moderate vascularization. A fibroma/fibrothecoma with atypical ultrasound appearance may be mistaken for a malignancy, in particular if associated with fluid in the pouch of Douglas or ascites, high color content and raised CA 125 levels.


Computer simulations of EPSP-spike (E-S) potentiation in hippocampal CA1 pyramidal cells.
Long-term potentiation of hippocampal excitatory synapses is often accompanied by an increase in the probability of spiking to an EPSP of fixed strength (E-S potentiation). We used computer simulations of a CA1 pyramidal neuron to test the plausibility of the hypothesis that E-S potentiation is caused by changes in dendritic excitability. These changes were simulated by adding "hot spots" of noninactivating voltage-sensitive Ca2+ conductance to various dendritic compartments. This typically caused spiking in response to previously subthreshold synaptic inputs. The magnitude of the simulated E-S potentiation depended on the passive electrical properties of the cell, the excitability of the soma, and the relative locations on the dendrites of the synaptic inputs and hot spots. The specificity of the simulated E-S potentiation was quantified by colocalizing the hot spots with a subset (40 of 80) of the synaptic contacts, denoted "tetanized," and then comparing the effects of the hot spots on these and the remaining (untetanized) synaptic contacts. The simulated E-S potentiation tended to be specific to the tetanized input if the untetanized contacts were, on average, electrically closer to the soma than the tetanized contacts. Specificity was also high if the tetanized and untetanized contacts were segregated to different primary dendrites. The results also predict, however, that E-S potentiation by this mechanism will appear to be nonspecific (heterosynaptic) if the synapses of the untetanized input are sufficiently far from the soma relative to the tetanized synapses. Experimental confirmation of this prediction would support the hypothesis that changes in postsynaptic excitability can contribute to hippocampal E-S potentiation.


Substrate hydrophobicity and cell composition influence the extent of rate limitation and masking of isotope fractionation during microbial reductive dehalogenation of chlorinated ethenes.
This study investigated the effect of intracellular microscale mass transfer on microbial carbon isotope fractionation of tetrachloroethene (PCE) and trichloroethene (TCE). Significantly stronger isotope fractionation was observed for crude extracts vs intact cells of Sulfurospirillum multivorans, Geobacter lovleyi, Desulfuromonas michiganensis, Desulfitobacterium hafniense strain PCE-S, and Dehalobacter restrictus. Furthermore, carbon stable isotope fractionation was stronger for microorganisms with a Gram-positive cell envelope compared to those with a Gram-negative cell envelope. Significant differences were observed between model organisms in cellular sorption capacity for PCE (S. multivorans-K(d-PCE) = 0.42-0.51 L g(-1); D. hafniense-K(d-PCE) = 0.13 L g(-1)), as well as in envelope hydrophobicity (S. multivorans 33.0° to 72.2°; D. hafniense 59.1° to 60.8°) when previously cultivated with fumarate or PCE as electron acceptor, but not for TCE. Cell envelope properties and the tetrachloroethene reductive dehalogenase (PceA-RDase) localization did not result in significant effects on observed isotope fractionation of TCE. For PCE, however, systematic masking of isotope effects as a result of microscale mass transfer limitation at microbial membranes was observed, with carbon isotope enrichment factors of -2.2‰, -1.5 to -1.6‰, and -1.0‰ (CI95% < ± 0.2‰) for no membrane, hydrophilic outer membrane, and outer + cytoplasmic membrane, respectively. Conclusively, rate-limiting mass transfer barriers were (a) the outer membrane or cell wall and (b) the cytoplasmic membrane in case of a cytoplasmic location of the RDase enzyme. Overall, our results indicate that masking of isotope fractionation is determined by (1) hydrophobicity of the degraded compound, (2) properties of the cell envelope, and (3) the localization of the reacting enzyme.


Hip resurfacing arthroplasty: risk factors for failure over 25 years.
Many early metal-on-polyethylene hip resurfacing arthroplasty designs were abandoned after reports of high short-term and midterm failure rates. To investigate factors associated with failure, we retrospectively reviewed our experience with early-design hip resurfacing implants in 75 patients during a 25-year period (median followup, 7.9 years; range, 0.1-25.2 years). Implant failure was defined as revision for any reason. One of 75 patients was lost to followup. The estimated rate of implant survival was 73% at 5 years, 34% at 10 years, 27% at 15 years, 12% at 20 years, and 8% at 25 years. Of the many clinical and radiographic factors considered, only age, implant type, and gender were associated with implant survival independent of other variables considered. Hip resurfacing arthroplasty showed poor overall long-term survival in this series. Particular attention should be paid to the identified risk factors as long-term followup data become available for modern designs. Level IV, therapeutic study. See the Guidelines for Authors for a complete description of levels of evidence.


Altered neurotransmitter release machinery in mice deficient for the deubiquitinating enzyme Usp14.
Homozygous ataxic mice (ax(J)) express reduced levels of the deubiquitinating enzyme Usp14. They develop severe tremors by 2-3 wk of age, followed by hindlimb paralysis, and death by 6-8 wk. While changes in the ubiquitin proteasome system often result in the accumulation of ubiquitin protein aggregates and neuronal loss, these pathological markers are not observed in the ax(J) mice. Instead, defects in neurotransmission were observed in both the central and peripheral nervous systems of ax(J) mice. We have now identified several new alterations in peripheral neurotransmission in the ax(J) mice. Using the two-microelectrode voltage clamp technique on diaphragm muscles of ax(J) mice, we observed that under normal neurotransmitter release conditions ax(J) mice lacked paired-pulse facilitation and exhibited a frequency-dependent increase in rundown of the end plate current at high-frequency stimulation (HFS). Combined electrophysiology and styryl dye staining revealed a significant reduction in quantal content during the initial and plateau portions of the HFS train. In addition, uptake of styryl dyes (FM dye) during HFS demonstrated that the size of the readily releasable vesicle pool was significantly reduced. Destaining rates for styryl dyes suggested that ax(J) neuromuscular junctions are unable to mobilize a sufficient number of vesicles during times of intense activity. These results imply that ax(J) nerve terminals are unable to recruit a sufficient number of vesicles to keep pace with physiological rates of transmitter release. Therefore, ubiquitination of synaptic proteins appears to play an important role in the normal operation of the neurotransmitter release machinery and in regulating the size of pools of synaptic vesicles.


Blunt chest trauma: case report.
Care of patients with blunt cardiac trauma is challenging for bedside nurses because of the potentially elusive and subtle nature of clinical findings associated with such trauma. An understanding of the forces exerted during the trauma episode will assist nurses in the assessment and search for hidden injuries. A high index of suspicion and continued cautious assessment and attention to subtle changes in the patient's physical condition are essential. Keen attention to changes in the patient's vital signs, cardiac output, ECG findings, pulses, and fluid volume status alert nurses to potential injuries associated with blunt cardiac trauma ranging from myocardial contusion to cardiac tamponade, aortic tears, and cardiac rupture. Survival of patients with blunt cardiac trauma depends on early intervention.


Non-linear, adaptive array processing for acoustic interference suppression.
A method is introduced where blind source separation of acoustical sources is combined with spatial processing to remove non-Gaussian, broadband interferers from space-time displays such as bearing track recorder displays. This differs from most standard techniques such as generalized sidelobe cancellers in that the separation of signals is not done spatially. The algorithm performance is compared to adaptive beamforming techniques such as minimum variance distortionless response beamforming. Simulations and experiments using two acoustic sources were used to verify the performance of the algorithm. Simulations were also used to determine the effectiveness of the algorithm under various signal to interference, signal to noise, and array geometry conditions. A voice activity detection algorithm was used to benchmark the performance of the source isolation.


Dietary ω3-and ω6-Polyunsaturated fatty acids reconstitute fertility of Juvenile and adult Fads2-Deficient mice.
Polyunsaturated fatty acids (PUFAs), including essential fatty acids linoleic and α-linolenic acid and derived long chain and very long chain ω3-and ω6-polyunsaturated fatty acids, are vital structures in mammalian membrane systems and signaling molecules, pivotal in brain development, lipid, and energy metabolism and in female and male fertility during human evolution. Numerous nutritional studies suggest imbalance of PUFA metabolism as a critical factor in the pathogenesis of several human lifestyle diseases: dyslipoproteinemia, obesity, cardiovascular and neurodegenerative diseases, and infertility. The lack of unbiased animal models impedes molecular interpretation of the role of synthesized and dietary supplied PUFAs in these conditions. In this study, we used a Δ6 fatty acid desaturase (FADS2) deficient mouse mutant lacking key enzyme activity in the biosynthesis of ω3-and ω6-PUFAs from EFAs to address the molecular role of PUFAs in female and male fertility. Infertility is a hallmark of the pleiotropic but auxotrophic fads2-/- phenotype and is therefore helpful for stringent dietary studies on the role of individual PUFAs. Feeding regimens: Age- and gender-matched infertile fads2-/- mice were maintained on defined diets, normal diet containing essential fatty acids, and supplemented with ω6-arachidonic acid, ω3-docosahexaenoic acid, and arachidonic/docosahexaenoic acid, starting (a) after weaning and (b) initiated in 4-month-old female and male fads2-/- mice. Phospho- and sphingolipidomes of ovarian and testicular membrane lipid bilayers in each cohort were established and the impact on the expression and topology of membrane marker proteins, membrane morphology, germ cell development, and female and male fertility in the respective cohorts was elaborated. PUFA synthesis deficiency caused a halt to folliculogenesis, atresia of oocytes, and infertility of fads2-/- female mice. A PUFA-deficient membrane lipid bilayer core structure led to the disassembly of the gap junction network of the follicular granulosa cells. In fads2-/- testis, the blood-testis barrier was disrupted and spermatogenesis arrested, leading to infertility. Sustained supply of combined AA and DHA remodeled the PUFA-deficient ovarian and testicular membrane lipidomes, facilitating the reassembly of the functional gap junction network for regular ovarian cycles and the reconstitution of the blood-testis barrier in Sertoli cells, reconstituting fertility not only in developing newborns, but surprisingly also in adult infertile fads2-/- mice. These findings demonstrate the previously unrecognized membrane structure-based molecular link between nutrient ω3-and ω6-PUFAs, gonadal membrane structures, and female and male fertility and might foster studies of the pivotal role of dietary PUFAs in human fertility.


Neuroprotective effect of Cubebin: A dibenzylbutyrolactone lignan on scopolamine-induced amnesia in mice.
Acetylcholinesterase (AChE) inhibitors represent a major class of drugs which provide symptomatic relief and improvement in cognitive function in Alzheimer's disease (AD). In this study, cubebin, a dibenzylbutyrolactone lignan, was isolated from Piper cubeba and investigated for its AChE inhibitory activity in an attempt to explore its potential for memory-enhancing activities in mice. Molecular docking of cubebin was carried out followed by in vitro AChE activity. Mice were treated with cubebin (25 & 50 mg/kg; i.p.), for three days and memory impairment was induced by scopolamine (3 mg/kg; i.p.). Memory function was evaluated by Morris water maze (MWM) test. Biochemical parameters of oxidative stress and cholinergic function were estimated in brain. Molecular docking study revealed that cubebin was well bound within the binding site of the AChE enzyme showing interactions such as π-π stacking and hydrogen bonding with residues present therein. Cubebin inhibited AChE enzyme in an in vitro assay with IC50value of 992 μM. Scopolamine administration caused a significant impairment of learning and memory in mice, as indicated by a marked decrease in MWM performance. Scopolamine administration also produced a significant enhancement of brain AChE activity and oxidative stress in mice brain. Pre-treatment of cubebin (25 and 50 mg/kg; i.p.) significantly prevented scopolamine-induced learning and memory deficits along with attenuation of scopolamine-induced rise in brain AChE activity and oxidative stress level. Cubebin showed promising protective activity in scopolamine-induced spatial memory impairment in mice. This could be attributed to its brain AChE inhibition and antioxidant activity.