One-dimensional tomography: a comparison of Abel, onion-peeling, and filtered backprojection methods.
It is shown that the Abel inversion, onion-peeling, and filtered backprojection methods can be intercompared without assumptions about the object being deconvolved. If the projection data are taken at equally spaced radial positions, the deconvolved field is given by weighted sums of the projections divided by the data spacing. The weighting factors are independent of the data spacing. All the methods are remarkably similar and have Abelian behavior: the field at a radial location is primarily determined by the weighted differences of a few projections around the radial position. Onion-peeling and an Abel inversion using two-point interpolation are similar. When the Shepp-Logan filtered backprojection method is reduced to one dimension, it is essentially identical to an Abel inversion using three-point interpolation. The weighting factors directly determine the relative noise performance: the three-point Abel inversion is the best, while onion peeling is the worst with approximately twice the noise. Based on ease of calculation, robustness, and noise, the three-point Abel inversion is recommended.


Genomic analysis of sequence-dependent DNA curvature in Leishmania.
Leishmania major is a flagellated protozoan parasite of medical importance. Like other members of the Trypanosomatidae family, it possesses unique mechanisms of gene expression such as constitutive polycistronic transcription of directional gene clusters, gene amplification, mRNA trans-splicing, and extensive editing of mitochondrial transcripts. The molecular signals underlying most of these processes remain under investigation. In order to investigate the role of DNA secondary structure signals in gene expression, we carried out a genome-wide in silico analysis of the intrinsic DNA curvature. The L. major genome revealed a lower frequency of high intrinsic curvature regions as well as inter- and intra- chromosomal distribution heterogeneity, when compared to prokaryotic and eukaryotic organisms. Using a novel method aimed at detecting region-integrated intrinsic curvature (RIIC), high DNA curvature was found to be associated with regions implicated in transcription initiation. Those include divergent strand-switch regions between directional gene clusters and regions linked to markers of active transcription initiation such as acetylated H3 histone, TRF4 and SNAP50. These findings suggest a role for DNA curvature in transcription initiation in Leishmania supporting the relevance of DNA secondary structures signals.


Lumped parameter and feedback control models of the auto-regulatory response in the Circle of Willis.
The Circle of Willis (CoW) is a ring-like structure of blood vessels found beneath the hypothalamus at the base of the brain. Its main function is to distribute oxygen-rich arterial blood to the cerebral mass. A 1-dimensional model of the CoW has been created to simulate a series of possible clinical scenarios such as occlusions in afferent arteries, absent or string-like circulus vessels, or arterial infarctions. The model captures cerebral haemodynamic auto-regulation by using a proportional-integral-derivative (PID) controller to modify efferent resistances and maintain optimal efferent flowrates for a given circle geometry and afferent blood pressure. Results match limited clinical data and results obtained in prior studies to within 6%. In addition, a set of boundary conditions and geometry is presented for which the auto-regulated system cannot provide the necessary efferent flowrates and perfusion, representing a condition with increased risk of stroke and highlighting the importance of modelling the haemodynamics of the CoW. The system model created is computationally simple so it can be used to identify at-risk cerebral arterial geometries and conditions prior to surgery or other clinical procedures.


Transformation-tolerant object recognition in rats revealed by visual priming.
Successful use of rodents as models for studying object vision crucially depends on the ability of their visual system to construct representations of visual objects that tolerate (i.e., remain relatively unchanged with respect to) the tremendous changes in object appearance produced, for instance, by size and viewpoint variation. Whether this is the case is still controversial, despite some recent demonstration of transformation-tolerant object recognition in rats. In fact, it remains unknown to what extent such a tolerant recognition has a spontaneous, perceptual basis, or, alternatively, mainly reflects learning of arbitrary associative relations among trained object appearances. In this study, we addressed this question by training rats to categorize a continuum of morph objects resulting from blending two object prototypes. The resulting psychometric curve (reporting the proportion of responses to one prototype along the morph line) served as a reference when, in a second phase of the experiment, either prototype was briefly presented as a prime, immediately before a test morph object. The resulting shift of the psychometric curve showed that recognition became biased toward the identity of the prime. Critically, this bias was observed also when the primes were transformed along a variety of dimensions (i.e., size, position, viewpoint, and their combination) that the animals had never experienced before. These results indicate that rats spontaneously perceive different views/appearances of an object as similar (i.e., as instances of the same object) and argue for the existence of neuronal substrates underlying formation of transformation-tolerant object representations in rats.


Influence of a province-wide trauma system on motor vehicle collision process of trauma care and mortality: a 10-year follow-up evaluation.
Mature trauma systems have evolved to respond to major injury-related morbidity and mortality. Studies of mature trauma systems have demonstrated improved survival, especially among seriously injured patients. From 1995 to 1998, a province-wide trauma system was implemented in the province of Nova Scotia. We measured the proportion of admissions to a tertiary level trauma centre and the proportion of in-hospital deaths among patients with major injuries as a result of a motor vehicle collisions (MVCs) before and 10 years after provincial trauma systems implementation. We identified major trauma patients aged 16 years and older using external cause of injury codes pertaining to MVCs from population-based hospital claims and vital statistics data. Individuals who were admitted to hospital or died because of an MVC in 1993-1994 (preimplementation), were compared with those who were admitted to hospital or died in 2003-2005 (postimplementation). Postimplementation, there was a 9% increase in the number of seriously injured individuals with primary admission to tertiary care. This increase was statistically significant even after we adjusted for age, head injury and municipality of residence (relative risk [RR] 1.09, 95% confidence interval [CI] 1.04-1.14). The probability of dying while in hospital in the postimplementation period decreased by 29% (adjusted RR 0.57, 95% CI 0.32-1.03), although this difference was not statistically significant. Individuals seriously injured in MVCs in Nova Scotia were more likely to be admitted to tertiary care after the implementation of a province-wide trauma system. There was a trend toward decreased mortality, but further research is warranted to confirm the survival benefit and delineate other contributing factors.


Human endothelial cell-derived exosomal microRNA-99a/b drives a sustained inflammatory response during sepsis by inhibiting mTOR expression.
The pathophysiology of sepsis and its accompanying hyper-inflammatory response are key events that lead to multi-organ failure and death. A growing body of literature now suggests that the vascular endothelium plays a critical role in driving early events of sepsis progression. In this study, we demonstrate how endothelial-derived exosomes contribute to a successive pro-inflammatory phenotype of monocytes. Exosomes isolated from S. aureus infected endothelial cells drive both CD11b and MHCII expression in monocytes and contribute dysregulated cytokine production. Conversely, healthy endothelial exosomes had no major effect. microRNA (miRNA) profiling of exosomes identified miR-99 upregulation which we hypothesised as driving this phenotypic change through mechanistic target of rapamycin (mTOR). Knockdown of mTOR with miR-99a and miR-99b mimetics in S. aureus infected monocytes increased IL-6 and decreased IL-10 production. Interestingly, inhibition of miRNAs with antagomirs has the opposing effect. Collectively, endothelial exosomes are driving a pro-inflammatory phenotype in monocytes through dysregulated expression of miR-99a and miR-99b.


Use of the AO plate for immediate mandibular reconstruction in cancer patients.
Free vascularized bone grafts have revolutionized mandibular reconstruction, yet their use in all mandibulectomy patients is not always necessary. A recently developed alternative to bony reconstruction has been the use of the AO reconstruction plate. We compared the use of the AO reconstruction plate with immediate free bone graft mandibular reconstruction in 31 patients. Reconstruction plates were used in 20 and immediate free bone grafts were used in 11 patients. The overall success rate for use of the plate was 15 of 20 (75 percent). There were 6 anterior reconstructions, of which only 2 (33 percent) were successful. This is opposed to 13 of 14 (93 percent) lateral reconstructions that were successful in lateral plate placements. There were 11 immediate composite free flaps: 4 iliac crest, 4 scapula, 2 fibula, and 1 composite radial forearm flaps. Six repairs were for anterior defects, and there were 5 full-thickness defects, 3 of which were in the anterior position. All 11 flaps were successful. In conclusion, we believe the reconstruction plates are a useful adjunct for mandibular replacement in the head and neck cancer patient but should be reserved for lateral defects. For anterior reconstructions, even in patients with locally advanced disease, free-tissue transfer of composite osteocutaneous flaps is the reconstructive method of choice.


Fecal Microbiota Transplants for Inflammatory Bowel Disease Treatment: Synthetic- and Engineered Communities-Based Microbiota Transplants Are the Future.
The human intestine harbors a huge number of diverse microorganisms where a variety of complex interactions take place between the microbes as well as the host and gut microbiota. Significant long-term variations in the gut microbiota (dysbiosis) have been associated with a variety of health conditions including inflammatory bowel disease (IBD). Conventional fecal microbiota transplantations (FMTs) have been utilized to treat IBD and have been proved promising. However, various limitations such as transient results, pathogen transfer, storage, and reproducibility render conventional FMT less safe and less sustainable. Defined synthetic microbial communities (SynCom) have been used to dissect the host-microbiota-associated functions using gnotobiotic animals or in vitro cell models. This review focuses on the potential use of SynCom in IBD and its advantages and relative safety over conventional FMT. Additionally, this review reinforces how various technological advances could be combined with SynCom to have a better understanding of the complex microbial interactions in various gut inflammatory diseases including IBD. Some technological advances including the availability of a gut-on-a-chip system, intestinal organoids, ex vivo intestinal cultures, AI-based refining of the microbiome structural and functional data, and multiomic approaches may help in making more practical in vitro models of the human host. Additionally, an increase in the cultured diversity from gut microbiota and the availability of their genomic information would further make the design and utilization of SynCom more feasible. Taken together, the combined use of the available knowledge of the gut microbiota in health and disease and recent technological advances and the development of defined SynCom seem to be a promising, safe, and sustainable alternative to conventional FMT in treating IBD.


Cytoplasmic pH Regulation in Acer pseudoplatanus Cells: I. A P NMR Description of Acid-Load Effects.
Modifications of cytoplasmic pH in Acer pseudoplatanus L. cells cultivated in suspension have been induced by acid-loads and studied by using (31)P nuclear magnetic resonance spectroscopy. The initial drop of cytoplasmic pH, observed in the first minutes of exposure to weak lipophilic acids, was followed by a slow recovery to reach a plateau phase with a pH value lower than the initial one. Conversely, removal of the acid led to a sharp increase of cytoplasmic pH with in most cases an overshoot toward more alkaline values than the initial one and a subsequent decrease to more acidic values. This shows that A. pseudoplatanus cells powerfully regulate their cytoplasmic pH both on the acid side of their normal pH, during the acid-load, and on the alkaline side, after removal of acid. Similar results were obtained with different types of acid-loads, i.e. treatments with propionic or benzoic acid or bubbling with CO(2)-enriched air. This indicates that the occurrence of pH regulation does not depend upon the method used to acid-load the cells. The time courses of cytoplasmic pH observed for A. pseudoplatanus and also Catharanthus roseus cells are similar to those recorded for animal cells but different from those described for other plant materials for which no recovery phase was observed. This can be explained by different balances between the initial rate of proton influx brought in by the acids, and the capacity of proton consumption by the regulatory mechanisms. The existence of the recovery phase offers a unique possibility to study the regulation of the cytoplasmic pH of plant cells, as it has been done in animal systems.


Optical identification of single- and few-layer MoS₂ sheets.
A simple approach is developed to identify the layer number of 2D MoS₂ sheets. By using an optical imaging method combined with image analysis software, a high-contrast image of the MoS₂ sheets can be extracted from the red (R) channel of the color optical microscopy image. The value of the intensity difference in the grayscale image of the R channel between MoS₂ sheets (1-3 layers) and the SiO₂ substrate can be used to identify the layer number of the sheet.


Evaluation of occupational hazards in foundries.
The working environment of foundries is hazardous and characterized by multiple simultaneous chemical, physical and mechanical hazards exposure, which would lead to injuries of foundry workers. The aim of the present work is to evaluate occupational hazards in four foundries, two in Alexandria: El Nasr and Ramsis, and two in Behira: Misr Spinning and Weaving and Misr Rayon companies. Levels of total and respirable dust, free silica % in total dust and lead concentration in total and respirable dust; NO2, SO2 and CO concentrations; noise and heat stress levels have been determined in the present work. Occupational injuries data were analyzed in a three years period from 1998 to 2000. The results of the present work revealed; 1. The levels of total dust and respirable dust exceeded the threshold limit values at knockout and cleaning operations at El Nasr Company. 2. Free silica percentage exceeded permissible levels in all operations except pouring in El Nasr Company. 3. CO levels in Misr Spinning and Weaving Company were higher than threshold levels. 4. Noise levels in knockout and cleaning operations at the four companies were exceeding the threshold limit values. 5. Heat stress levels in melting and pouring operations in El Nasr and in pouring operation in Ramsis Company were higher than the maximum permissible levels. 6. The age group 31-40 years has recorded the highest average incidence rate of injuries of age groups (P<0.01). 7. Lower extremities and higher extremities have recorded the highest average incidence rate in the four companies (P<0.001-P<0.01 respectively). 8. Transportation and lifting was the main cause of injury in the four companies (P<0.05). 9. Faulty action and striking against was the main mean of injury in the four companies (P<0.01). 10. Ramsis Company has the highest average incidence rate in almost all injuries parameters and indices (frequency rate and severity rate) (P<0.05-P<0.001). The present work is a massive survey, which highlights the occupational hazards in Egyptian foundries.


Evaluation of saliva as a specimen for monitoring zidovudine therapy in HIV-infected patients.
To facilitate studies of the pharmacokinetic properties of zidovudine, the relationship between plasma and salivary concentrations of the drug was studied, after oral dosage, in 10 HIV-infected patients. Zidovudine concentrations were determined in plasma, unstimulated mixed saliva and citric-acid-stimulated mixed saliva over a period of 3 1/2 hours by high-performance liquid chromatography. Correlation coefficients were r = 0.97 (P less than 0.0001) for stimulated saliva compared with plasma and r = 0.89 (P less than 0.0001) for unstimulated saliva, with average values in unstimulated saliva being 113.8 +/- 44.6% in plasma and 67.8 +/- 25.4% in stimulated saliva. Stimulated saliva values found to be 70% of the total reflected the concentration of the unbound drug in plasma. Except for a shorter half-life time (t1/2) in saliva, pharmacokinetic parameters showed a good correlation in the three types of specimen. These findings and the convenience of sample collection suggest that citric-acid-stimulated saliva might be an appropriate specimen for monitoring zidovudine therapy.


Differential ligand binding affinities of human estrogen receptor-α isoforms.
Rapid non-genomic effects of 17β-estradiol are elicited by the activation of different estrogen receptor-α isoforms. Presence of surface binding sites for estrogen have been identified in cells transfected with full-length estrogen receptor-α66 (ER66) and the truncated isoforms, estrogen receptor-α46 (ER46) and estrogen receptor-α36 (ER36). However, the binding affinities of the membrane estrogen receptors (mERs) remain unknown due to the difficulty of developing of stable mER-transfected cell lines with sufficient mER density, which has largely hampered biochemical binding studies. The present study utilized cell-free expression systems to determine the binding affinities of 17β-estradiol to mERs, and the relationship among palmitoylation, membrane insertion and binding affinities. Saturation binding assays of human mERs revealed that [³H]-17β-estradiol bound ER66 and ER46 with Kd values of 68.81 and 60.72 pM, respectively, whereas ER36 displayed no specific binding within the tested concentration range. Inhibition of palmitoylation or removal of the nanolipoprotein particles, used as membrane substitute, reduced the binding affinities of ER66 and ER46 to 17β-estradiol. Moreover, ER66 and ER46 bound differentially with some estrogen receptor agonists and antagonists, and phytoestrogens. In particular, the classical estrogen receptor antagonist, ICI 182,780, had a higher affinity for ER66 than ER46. In summary, the present study defines the binding affinities for human estrogen receptor-α isoforms, and demonstrates that ER66 and ER46 show characteristics of mERs. The present data also indicates that palmitoylation and membrane insertion of mERs are important for proper receptor conformation allowing 17β-estradiol binding. The differential binding of ER66 and ER46 with certain compounds substantiates the prospect of developing mER-selective drugs.


Preparation and Investigation of Intelligent Polymeric Nanocapsule for Enhanced Oil Recovery.
Micro-/nanomotors colloidal particles have attracted increasing interest as composite surfactants, owing to the combined advantages of both Janus solid surfactants and micro-/nanomotors. Here we put micro-/nanomotors colloidal particles into hollow polymeric micro-encapsulates. An intelligent polymeric nanocapsule was prepared for enhanced oil recovery by the self-assembly method. The particle size range of the polymeric capsule can be controlled between 20 to 1000 nm by adjusting the cross-linking thickness of the capsule's outer membrane. The average particle size of polymeric capsules prepared in the study was 300 nm. The structure and properties of the Intelligent polymeric nanocapsule was characterized by a wide range of technics such as Fourier transform infrared spectroscopy, scanning electron microscopy by laser diffraction, fluorescence microscopy, pendant drop tensiometer, laser particle size instrument, and interface tension analyzer. It was found that the intelligent polymeric nanocapsule exhibited significant interfacial activity at the oil-water interface. When the Janus particles' concentration reached saturation concentration, the adsorption of the amphiphilic nanoparticles at the interface was saturated, and the equilibrium surface tension dropped to around 31 mN/m. When the particles' concentration reached a critical concentration of aggregation, the Gibbs stability criterion was fulfilled. The intelligent polymeric nanocapsule system has a better plugging and enhanced oil recovery capacity. The results obtained provide fundamental insights into the understanding of the assembly behavior and emulsifying properties of the intelligent polymeric nanocapsule, and further demonstrate the future potential of the intelligent polymeric nanocapsule used as colloid surfactants for enhanced oil recovery applications.


Structure activity relationships of monocyte chemoattractant proteins in complex with a blocking antibody.
Monocyte chemoattractant proteins (MCPs) are cytokines that direct immune cells bearing appropriate receptors to sites of inflammation or injury and are therefore attractive therapeutic targets for inhibitory molecules. 11K2 is a blocking mouse monoclonal antibody active against several human and murine MCPs. A 2.5 A structure of the Fab fragment of this antibody in complex with human MCP-1 has been solved. The Fab blocks CCR2 receptor binding to MCP-1 through an adjacent but distinct binding site. The orientation of the Fab indicates that a single MCP-1 dimer will bind two 11K2 antibodies. Several key residues on the antibody and on human MCPs were predicted to be involved in antibody selectivity. Mutational analysis of these residues confirms their involvement in the antibody-chemokine interaction. In addition to mutations that decreased or disrupted binding, one antibody mutation resulted in a 70-fold increase in affinity for human MCP-2. A key residue missing in human MCP-3, a chemokine not recognized by the antibody, was identified and engineering the preferred residue into the chemokine conferred binding to the antibody.


Myeloid progenitors in remission bone marrow in patients with malignant blood diseases: clues for slow hematopoietic regeneration after ABMT?
Slow hematopoietic recovery is a well-known feature in patients undergoing autologous bone marrow transplantation (ABMT), and here we demonstrate that compared with patients with other malignant blood diseases, the thrombocytopenia in acute myeloid leukemia (AML) patients stands out, with the median time to recovery of > 50 x 10(9) platelets being 101 days for AML patients and 22-37 days for other patient groups. We have consequently evaluated the content and fate of myeloid progenitor cells (BFU-E, CFU-GM, and CFU-GEMM) in bone marrow preparations from cancer patients in an attempt to uncover factors of importance for their slow hematopoietic recovery after ABMT using three experimental setups. First, we analyzed progenitors in marrow samples from 36 patients [18 AML, 5 acute lymphoblastic leukemia (ALL), and 13 non-Hodgkin's lymphoma (NHL)] exposed to multiple doses of severely myelotoxic cytoreductive regimens and observed that AML patients had suppressed numbers of CFU-GEMM and BFU-E (80% that of normal volunteers) but increased levels of CFU-GM. In contrast, the decrease in ALL patients was more pronounced (50% that of normal volunteers) and demonstrable for all progenitors. NHL patients exhibited normal progenitor frequencies of CFU-GM and CFU-GEMM, whereas their BFU-E counts were found to be increased nearly two-fold. Second, we followed the fate of progenitors through the laboratory manipulations of marrow grafts and found no evidence for selective losses in terms of disease (7 AML, 5 NHL, 1 ALL, and 2 testicular germ cell tumor patients) and progenitor type. Third, we retrospectively evaluated the cell yields of grafts from 121 cases [39 AML, 20 ALL, 22 NHL, 9 Burkitt's lymphoma (BL), 18 Hodgkin's disease (HD), and 13 testicular germ cell tumors] and found those of heavily treated AML, HD, and germinal tumors most affected, with a clear negative correlation in AML patients between age on the one hand and mononuclear cell yield and CFU-GM content on the other. We conclude that the amount of previous chemotherapy and age of patients appear to be among the determining factors on the number of myeloid progenitor cells in cancer patients and that thresholds for acceptability of grafts should take these variables into account.


Contribution of equilibrative nucleoside transporter(s) to intestinal basolateral and apical transports of anticancer trifluridine.
Trifluridine (FTD) exhibits anticancer activities after its oral administration despite its hydrophilic nature. It was previously reported that concentrative nucleoside transporter (CNT) 1 mediates the apical uptake of FTD in human small intestinal epithelial cells (HIECs). In the present study, FTD was also identified as a substrate for equilibrative nucleoside transporter (ENT) 1 and ENT2 in transporter gene-transfected cells. An immunocytochemical analysis revealed that ENT1 was expressed at the basolateral and apical membranes of HIECs. Cellular accumulation increased in the presence of S-(4-nitrobenzyl)-6-thioinosine (NBMPR), an ENT selective inhibitor. Cytotoxicity in HIEC monolayers at low FTD concentrations was increased by NBMPR, and this may have been due to inhibition of the ENT-mediated basolateral transport of FTD by NBMPR. These results suggest that ENTs reduce the intestinal cytotoxicity of FTD by facilitating its basolateral efflux. On the other hand, the intracellular accumulation and cytotoxicity of FTD in HIECs were decreased at higher concentrations of FTD by NBMPR, and this may have been due to the NBMPR inhibition of the apical uptake of FTD, which has been suggested to be mediated by CNTs and ENTs. In conclusion, ENTs were responsible for intestinal transepithelial permeation by mediating the basolateral efflux of FTD after its uptake by CNT1 from the apical side, resulting in decreases in its intracellular accumulation and intestinal toxicity in humans. Equilibrative nucleoside transporters may also partially contribute to the low-affinity uptake of FTD across the apical membrane along with high-affinity CNT1.


The significance of the C(α) substituent in the selective inhibition of matrix metalloproteinases 1 and 9.
Matrix metalloproteinases (MMPs) cleave and degrade most components of the extracellular matrix, and unregulated MMP activity has been correlated to cancer and metastasis. Hence there is a burgeoning need to develop inhibitors that bind selectively to structurally similar MMPs. The inhibition profiles of peptidomimetics containing C(α) substituents at the α,β unsaturated carbon were evaluated against the recombinant forms of ADAM17, MMP1, and MMP9. The dicarboxylic acid D2 and hydroxamate C2 inhibited MMP9 but not MMP1. The unsaturated compound E2 displayed selective inhibition for MMP1, compared with the saturated precursor C2, with an IC(50) value of 3.91 μm. The molecular basis for this selectivity was further investigated by the molecular docking of E2 and D2 into the active sites of MMP1 and MMP9. These data demonstrate hydrogen-bonding interactions between the carbonyl group of the C(α) substituent of E2 and the side chain of Asn180 present in the active site of MMP1. Conversely, the docked MMP9-D2 structure shows hydrophobic and hydrogen bonding between the ligand's morpholine substituent and second carboxylic acid group with Leu187 and an amide, respectively. This study suggests that substituents other than P(1)' and P(2)' may confer selectivity among MMPs and may aid in the search for novel lead compounds.


Mapping functional brain activation using [14C]-iodoantipyrine in male serotonin transporter knockout mice.
Serotonin transporter knockout mice have been a powerful tool in understanding the role played by the serotonin transporter in modulating physiological function and behavior. However, little work has examined brain function in this mouse model. We tested the hypothesis that male knockout mice show exaggerated limbic activation during exposure to an emotional stressor, similar to human subjects with genetically reduced transcription of the serotonin transporter. Functional brain mapping using [(14)C]-iodoantipyrine was performed during recall of a fear conditioned tone. Regional cerebral blood flow was analyzed by statistical parametric mapping from autoradiographs of the three-dimensionally reconstructed brains. During recall, knockout mice compared to wild-type mice showed increased freezing, increased regional cerebral blood flow of the amygdala, insula, and barrel field somatosensory cortex, decreased regional cerebral blood flow of the ventral hippocampus, and conditioning-dependent alterations in regional cerebral blood flow in the medial prefrontal cortex (prelimbic, infralimbic, and cingulate). Anxiety tests relying on sensorimotor exploration showed a small (open field) or paradoxical effect (marble burying) of loss of the serotonin transporter on anxiety behavior, which may reflect known abnormalities in the knockout animal's sensory system. Experiments evaluating whisker function showed that knockout mice displayed impaired whisker sensation in the spontaneous gap crossing task and appetitive gap cross training. This study is the first to demonstrate altered functional activation in the serotonin transporter knockout mice of critical nodes of the fear conditioning circuit. Alterations in whisker sensation and functional activation of barrel field somatosensory cortex extend earlier reports of barrel field abnormalities, which may confound behavioral measures relying on sensorimotor exploration.


Electronically Modified Cobalt Aminopyridine Complexes Reveal an Orthogonal Axis for Catalytic Optimization for CO2 Reduction.
The design of effective electrocatalysts for carbon dioxide reduction requires understanding the mechanistic underpinnings governing the binding, reduction, and protonation of CO2. A critical aspect to understanding and tuning these factors for optimal catalysis revolves around controlling the electronic environments of the primary and secondary coordination sphere. Herein we report a series of para-substituted cobalt aminopyridine macrocyclic catalysts 2-4 capable of carrying out the electrochemical reduction of CO2 to CO. Under catalytic conditions, complexes 2-4, as well as the unsubstituted cobalt aminopyridine complex 1, exhibit icat/ip values ranging from 144 to 781. Complexes 2 and 4 exhibit a pronounced precatalytic wave suggestive of an ECEC mechanism. A Hammett analysis reveals that ligand modifications with electron-donating groups enhance catalysis (ρ < 0), indicative of positive charge buildup in the transition state. This trend also extends to the CoI/0 potential, where complexes possessing more negative E(CoI/0) reductions exhibit greater icat/ip values. The reported modifications offer a synthetic lever to tune catalytic activity, orthogonal to our previous study of the role of pendant hydrogen bond donors.


HIV Diagnosis, Linkage to Care, and Retention among Men Who Have Sex with Men and Transgender Women in Guatemala City.
Men who have sex with men (MSM) and transgender women are disproportionately affected by HIV in Guatemala, yet little is known about their experiences with diagnosis, linkage to care, and retention. We conducted qualitative in-depth interviews with 26 MSM and transgender women living with HIV in Guatemala City. HIV diagnosis experiences changed over time with increasing asymptomatic testing at non-governmental organizations. Fear of the physical and social impacts of HIV delayed testing, acceptance of diagnosis, and linkage to HIV care. These fears were driven by layered stigma and discrimination due to non-normative gender expressions and / or sexual orientation. Retention-specific determinants included HIV clinic dynamics and limited employment opportunities. There is an urgent need to improve support systems for early testing and linkage to care and to expand employment opportunities. Stigma and discrimination must be addressed at the family, clinic and contextual levels to reduce fear of diagnosis and improve access to care.


Exposure to valproic acid is associated with less pulmonary infiltrates and improvements in diverse clinical outcomes and laboratory parameters in patients hospitalized with COVID-19.
Valproic acid (VPA) has shown beneficial effects in vitro against SARS-CoV-2 infection, but no study has analyzed its efficacy in the clinical setting. This multicenter, retrospective study included 165 adult patients receiving VPA at the time of admission to hospital, and 330 controls matched for sex, age and date of admission. A number of clinical, outcome and laboratory parameters were recorded to evaluate differences between the two groups. Four major clinical endpoints were considered: development of lung infiltrates, in-hospital respiratory worsening, ICU admissions and death. VPA-treated patients had higher lymphocyte (P<0.0001) and monocyte (P = 0.0002) counts, and lower levels of diverse inflammatory parameters, including a composite biochemical severity score (P = 0.016). VPA patients had shorter duration of symptoms (P<0.0001), were more commonly asymptomatic (P = 0.016), and developed less commonly lung infiltrates (65.8%/88.2%, P<0.0001), respiratory worsening (20.6%/30.6%, P = 0.019) and ICU admissions (6.1%/13.0%, P = 0.018). There was no difference in survival (84.8%/88.8%, P = 0.2), although death was more commonly related to non-COVID-19 causes in the VPA group (36.0%/10.8%, P = 0.017). The cumulative hazard for developing adverse clinical endpoints was higher in controls than in the VPA group for infiltrates (P<0.0001), respiratory worsening (P<0.0001), and ICU admissions (P = 0.001), but not for death (0.6). Multivariate analysis revealed that VPA treatment was independently protective for the development of the first three clinical endpoints (P = 0.0002, P = 0.03, and P = 0.025, respectively), but not for death (P = 0.2). VPA-treated patients seem to develop less serious COVID-19 than control patients, according to diverse clinical endpoints and laboratory markers.


Excess diacylglycerol at the endoplasmic reticulum disrupts endomembrane homeostasis and autophagy.
When stressed, eukaryotic cells produce triacylglycerol (TAG) to store nutrients and mobilize autophagy to combat internal damage. We and others previously reported that in yeast, elimination of TAG synthesizing enzymes inhibits autophagy under nitrogen starvation, yet the underlying mechanism has remained elusive. Here, we show that disruption of TAG synthesis led to diacylglycerol (DAG) accumulation and its relocation from the vacuolar membrane to the endoplasmic reticulum (ER). We further show that, beyond autophagy, ER-accumulated DAG caused severe defects in the endomembrane system, including disturbing the balance of ER-Golgi protein trafficking, manifesting in bulging of ER and loss of the Golgi apparatus. Genetic or chemical manipulations that increase consumption or decrease supply of DAG reversed these defects. In contrast, increased amounts of precursors of glycerolipid synthesis, including phosphatidic acid and free fatty acids, did not replicate the effects of excess DAG. We also provide evidence that the observed endomembrane defects do not rely on Golgi-produced DAG, Pkc1 signaling, or the unfolded protein response. This work identifies DAG as the critical lipid molecule responsible for autophagy inhibition under condition of defective TAG synthesis and demonstrates the disruption of ER and Golgi function by excess DAG as the potential cause of the autophagy defect.


Nuclear Magnetic Resonance Spectroscopy to Identify Metabolite Biomarkers of Nonresponsiveness to Targeted Therapy in Glioblastoma Tumor Stem Cells.
Glioblastoma is the most common and malignant brain tumor, and current therapies confer only modest survival benefits. A major obstacle is our ability to monitor treatment effect on tumors. Current imaging modalities are ambiguous, and repeated biopsies are not encouraged. To scout for markers of treatment response, we used NMR spectroscopy to study the effects of a survivin inhibitor on the metabolome of primary glioblastoma cancer stem cells. Applying high resolution NMR spectroscopy (1H resonance frequency: 800.03 MHz) to just 3 million cells per sample, we achieved sensitive and high resolving determinations of, e.g., amino acids, nucleosides, and constituents of the citric acid cycle. For control samples that were cultured, prepared, and measured at varying dates, peak area relative standard deviations were 15-20%. Analyses of unfractionated lysates were performed for straightforward compound identification with COLMAR and HMDB databases. Principal component analysis revealed that citrate levels were clearly upregulated in nonresponsive cells, while lactate levels substantially decreased following treatment for both responsive and nonresponsive cells. Hence, lactate and citrate may be potential markers of successful drug uptake and poor response to survivin inhibitors, respectively. Our metabolomics approach provided alternative biomarker candidates compared to spectrometry-based proteomics, underlining benefits of complementary methodologies. These initial findings make a foundation for exploring in vivo MR spectroscopy (MRS) of brain tumors, as citrate and lactate are MRS-visible. In sum, NMR metabolomics is a tool for addressing glioblastoma.


[Research on DNA fluorescence capillary biosensor marked by Goldview].
Goldview marked DNA fluorescence capillary biosensor was studied in the present paper. Based on fluorescence capillary analysis (FCA), the DNA biosensor uses capillary as immobilization carrier and detection carrier of DNA probe. Probes (20-mer-ssDNA) were immobilized on the inner wall of capillary by poly-l-lysine, and DNA fluorescence capillary biosensor (DNA-FCB) was made. After being hybridized with complementary target DNA and dyed by Goldview, the target DNA was qualified or quantified by detecting the fluorescence density of the Goldview using F-4500 spectrofluorometer. The sample volume was 12 microL. The concentration of the target DNA showed good linearity with the fluorescence intensity in the range of 0. 4-4 micromol x L(-1) (2.4-24 mg x L(-1)) (y = 65.911x + 3.9944, r = 0.9989). The RSD was lower than 3.5%. The concentration detection limit of the target DNA was 0.39 micromol x L(-1) (2.2 mg x L(-1)). The DNA-FCB can be used to qualify or quantify the target DNA. It's advantages are simplicity of manipulation, thimbleful of sample and reagent volumes, repeated use of capillary, and the lowest test cost. By using DNA-FCB to qualify the target DNA, we can consumedly decrease the pollution of the environment.


[Clinical laboratory characteristics of acute purulent lung abscesses].
The comparison of the clinical, laboratory and immunological indices in the patients with acute purulent lung abscess was carried out. The general and biochemical analysis of the blood was proved to reveal the signs of the acute inflammatory process presence independently on pathogenesis of the disease. The investigations of the immunological indices demonstrate the prevailing of humoral immune response to antigen. The suggested description of concomitant bronchitis was proved by the results of the cytological and the immunological study.


Bovine Leukaemia Virus: Current Epidemiological Circumstance and Future Prospective.
Bovine leukaemia virus (BLV) is a deltaretrovirus that is closely related to human T-cell leukaemia virus types 1 and 2 (HTLV-1 and -2). It causes enzootic bovine leukosis (EBL), which is the most important neoplastic disease in cattle. Most BLV-infected cattle are asymptomatic, which potentiates extremely high shedding rates of the virus in many cattle populations. Approximately 30% of them show persistent lymphocytosis that has various clinical outcomes; only a small proportion of animals (less than 5%) exhibit signs of EBL. BLV causes major economic losses in the cattle industry, especially in dairy farms. Direct costs are due to a decrease in animal productivity and in cow longevity; indirect costs are caused by restrictions that are placed on the import of animals and animal products from infected areas. Most European regions have implemented an efficient eradication programme, yet BLV prevalence remains high worldwide. Control of the disease is not feasible because there is no effective vaccine against it. Therefore, detection and early diagnosis of the disease are essential in order to diminish its spreading and the economic losses it causes. This review comprises an overview of bovine leukosis, which highlights the epidemiology of the disease, diagnostic tests that are used and effective control strategies.


Nickel: a micronutrient essential for higher plants.
Nickel was established as an essential micronutrient for the growth of temperate cereal crops. Grain from barley (Hordeum vulgare L. cv ;Onda'; containing 40 to 80 nanograms of Ni per gram dry weight) grown in solution culture with negligible Ni concentrations (< 30 nanograms of Ni per liter) exhibited greatly reduced germination rates (i.e. 50% less than grain from Ni-adequate plants) and seedling vigor of the viable grain was greatly depressed. Grain containing less than 30 nanograms per gram dry weight was inviable. Under Ni-deficient conditions, barley plants fail to produce viable grain because of a disruption of the maternal plant's normal grain-filling and maturation processes that occur following formation of the grain embryo. The observations that (a) barley plants fail to complete their life cycle in the absence of Ni and (b) addition of Ni to the growth medium completely alleviates deficiency symptoms in the maternal plants satisfies the essentiality criteria; thus, Ni should be considered a micronutrient for cereals. Because Ni is required by legumes, and is now established as essential for cereals, we conclude that Ni should be added to the list of micronutrients essential for all higher plant growth.


Balanced Leader Distribution Algorithm in Kubernetes Clusters.
Container-based virtualization is becoming a de facto way to build and deploy applications because of its simplicity and convenience. Kubernetes is a well-known open-source project that provides an orchestration platform for containerized applications. An application in Kubernetes can contain multiple replicas to achieve high scalability and availability. Stateless applications have no requirement for persistent storage; however, stateful applications require persistent storage for each replica. Therefore, stateful applications usually require a strong consistency of data among replicas. To achieve this, the application often relies on a leader, which is responsible for maintaining consistency and coordinating tasks among replicas. This leads to a problem that the leader often has heavy loads due to its inherent design. In a Kubernetes cluster, having the leaders of multiple applications concentrated in a specific node may become a bottleneck within the system. In this paper, we propose a leader election algorithm that overcomes the bottleneck problem by evenly distributing the leaders throughout nodes in the cluster. We also conduct experiments to prove the correctness and effectiveness of our leader election algorithm compared with a default algorithm in Kubernetes.


In Situ Determination of Speciation and Local Structure of NaCl-SrCl2 and LiF-ZrF4 Molten Salts.
Understanding the local environment of the metal atoms in salt melts is important for modeling the properties of melts and predicting their behavior and thus helping enable the development of technologies such as molten salt reactors and solar-thermal power systems and new approaches to recycling rare-earth metals. Toward that end, we have developed an in situ approach for measuring the coordination of metals in molten salt coupling X-ray absorption spectroscopy (XAS) and Raman spectroscopy. Our approach was demonstrated for two salt mixtures (1.9 and 5 mol % SrCl2 in NaCl, 0.8 and 5 mol % ZrF4 in LiF) at up to 1100 °C. Near-edge (X-ray absorption near-edge structure, XANES) and extended X-ray absorption fine structure (EXAFS) spectra were measured. The EXAFS response was modeled using ab initio FEFF calculations. Strontium's first shell is observed to be coordinated with chlorine (Sr2+-Cl-) and zirconium's first shell is coordinated by fluorine (Zr4+-F-), both having coordination numbers that decrease with increasing temperature. Multiple zirconium complexes are believed to be present in the melt, which may interfere and distort the EXAFS spectra and result in an anomalously low zirconium first shell coordination number. The use of boron nitride (BN) powder as a salt diluent for XAFS measurements was found to not interfere with measurements and thus can be used for investigations of such systems.


[A case of retroperitoneal fibrosis forming a pelvic tumor--a type of multifocal fibrosclerosis].
We experienced a case of retroperitoneal fibrosis with formation of a pelvic tumor. The patient was a 55-year-old female who showed right-side hydronephrosis on examination and was admitted to this department for consultation. Retrograde pyelography clearly showed that the hydronephrosis was due to lower ureteral obstruction resulting from a pelvic tumor. Ultrasound, CT and MRI showed a tumor adhering to the right posterior surface of the bladder, and it was thought that the tumor was probably malignant. Transdermal/transvesical needle biopsy and transvaginal needle biopsy showed a minimal invasion and allowed the diagnosis of retroperitoneal fibrosis to be made. Ureterolysis was not carried out, and the patient was treated with steroid monotherapy. She was begun on 30 mg of prednisolone, and administration was continued for approximately 3 months while gradually decreasing the dosage. On completion of administration, the intrapelvic tumor had completely disappeared, showing that treatment was markedly effective. The patient had a history of an orbital pseudotumor, and it was concluded that this was a rare case coming under the heading of multifocal fibrosclerosis.


Methylation of the promoter for an immediate-early frog virus 3 gene does not inhibit transcription.
Methylation of critical sites within the promoter region of eucaryotic genes has been shown to inhibit transcription by RNA polymerase II. However, although the large DNA virus frog virus 3 (FV3) has a highly methylated genome, it uses host RNA polymerase II for at least the immediate-early stage of transcription. We have previously shown that an FV3-induced trans-acting protein allows transcription from adenovirus promoters inactivated by methylation. Since FV3 immediate-early genes are transcribed in the absence of de novo protein synthesis, it appears that the virus-induced trans-acting protein that allows transcription from methylated templates is not required for transcription of the immediate-early FV3 genes, possibly because they are not methylated in critical regulatory sequences. In this study, we used site-directed mutagenesis to alter the three CpG dinucleotide sequences in the promoter region of an immediate-early FV3 gene and thereby created sites recognized by bacterial methylases. Transient-expression assays demonstrated that neither the mutations nor methylation of the mutated sites inhibited transcription from the FV3 promoter in FV3-infected cells. These findings support the hypothesis that the immediate-early genes of FV3 do not contain methylatable sites in regions critical for transcription. The function of the virus-induced trans-acting protein that can override the inhibitory effect of methylation may therefore be to facilitate the transcription of methylated delayed-early or late FV3 genes.


Poor specific antibody response immunodeficiency (dysgammaglobulinemia) predates systemic lupus erythematosus.
Poor specific antibody response is a well-known primary immunodeficiency that is related to hypogammaglobulinemia or common variable immunodeficiency (CVID). The co-existence of CVID or hypogammaglobulinemia and systemic lupus erythematosus (SLE) has been rarely described. In all reported cases, the diagnosis of SLE antedates CVID. We report a 15-year-old Saudi girl who was diagnosed with poor specific antibody response at age 6 years in the form of poor or no antibody response and dysgammaglobulinemia. She developed SLE with musculoskeletal and hematological manifestations, positive antinuclear antibody and high anti-dsDNA nine years later. She was treated with rituximab with good response.


[University nursing education and its consequences on the professor's indentity].
The modification of the statute and expansion of nursing training will also lead to a modification of the instructor's workload. Despite previous university training, the new research and service provider requirements (issued by the Law regarding Advanced and Specialized Schools), will confront the instructor with new activities that he/she may have little or no experience in performing. The instructor's work, and his/her conflict with the assigned work, would allow an observation of his/her teaching practices as well as any instability brought on by change of status in nursing training. The instructor would now have a new identity: that of researcher and promoter of professional knowledge. However, this transformation involves the creation of a site regrouping the ensemble of participants where the experimental and the practical can coexist and develop.


Osteoid osteoma of distal phalanx of toe: a rare cause of foot pain.
Osteoid osteoma is an uncommon benign tumor and causes severe pain, being worse at night, that responds dramatically to nonsteroidal anti-inflammatory medications. An osteoid osteoma of the toe is very rare and arising in a pedal phalanx may be difficult to diagnose. A 34-year-old male has local swelling and tenderness but there were no hyperemia, temperature increase, or clubbing. There was a 2-month history of antibiotic treatment with suspicion of soft tissue infection in another clinic. The osteoid osteoma was completely excised by curettage and nidus removal with open surgical technique. The patient was followed up for 63 months with annual clinical and radiographic evaluations. There was no relapse of the pain and no residual recurrent tumour. Osteoid osteoma may be difficult to distinguish from chronic infection or myxedema. The patients may be taken for unnecessary treatment. The aim of the treatment for osteoid osteoma is to remove entire nidus by open surgical excision or by percutaneous procedures such as percutaneous radiofrequency and laser ablation. Osteoid osteomas having radiologic and clinical features other than classical presentation of osteoid osteoma are called atypical osteoid osteomas. Atypical localized osteoid osteomas can be easily misdiagnosed and treatment is often complicated.


Ultrastructural deficiency in autonomic innervation in cremasteric muscle of boys with undescended testis.
The cremaster muscles (CM) associated with undescended testis reveal neurogenic alterations that mainly affect type 2 fibers. The ultrastructure of CM has been evaluated to define if further evidence to explain the alterations could be identified. CM of 8 boys with inguinal hernia and 8 boys with undescended testis at similar ages were biopsied. Samples were processed for electron microscopic evaluations. Semithin and thin sections were examined under an electron microscope. The CM associated with inguinal hernia showed normal ultrastructure. However, some alterations were encountered in CM associated with undescended testis. Unmyelinated fibers were diminished in number, and myelinated fibers were outnumbering the unmyelinated fibers. Marked disorientation of myofibers, redundant sarcolemma, empty sleeves of basal lamina, disarray of myofibrils, densely packed myofilaments, Z disk streaming, dilated sarcoplasmic reticulum, and dense-irregularly shaped mitochondria were repeatedly encountered. Satellite cells appeared inactive. Most of the fibers were contracted. The decrease in number of unmyelinated fibers appears to represent a decrease in autonomic nerve fibers. The alterations within muscle fibers may reflect a deficiency in autonomic innervation. Autonomic nervous system is highly responsive to circulating androgens. Factors decreasing the vulnerability of autonomic nervous system against androgenic effects may result in a CM with neurogenic alterations, thus inhibiting testicular descent. J Pediatr Surg 36:573-578.


Ventilation-perfusion inhomogeneity increases gas uptake in anesthesia: computer modeling of gas exchange.
Ventilation-perfusion (VA/Q) inhomogeneity was modeled to measure its effect on overall gas exchange during maintenance-phase N(2)O anesthesia with an inspired O(2) concentration of 30%. A multialveolar compartment computer model was used based on physiological log normal distributions of VA/Q inhomogeneity. Increasing the log standard deviation of the distribution of perfusion from 0 to 1.75 paradoxically increased O(2) uptake (VO(2)) where a low mixed venous partial pressure of N(2)O [high N(2)O uptake (VN(2)O)] was specified. With rising mixed venous partial pressure of N(2)O, a threshold was observed where VO(2) began to fall, whereas VN(2)O began to rise with increasing VA/Q inhomogeneity. This phenomenon is a magnification of the concentrating effects that VO(2) and VN(2)O have on each other in low VA/Q compartments. During "steady-state" N(2)O anesthesia, VN(2)O is predicted to paradoxically increase in the presence of worsening VA/Q inhomogeneity.


Effect of nuts on components of metabolic syndrome in healthy adults with overweight/obesity: A systematic review and meta-analysis.
Randomized controlled trials evaluating the effects of nut consumption on the metabolic profile of healthy adults with overweight/obesity have yielded conflicting results. This systematic review and meta-analysis aimed to summarize the effects of incorporating nuts into the diet on serum lipid profile, glycemic markers, and blood pressure in healthy adults with overweight/obesity. PubMed, Embase, Scopus, Web of Science, and Cochrane Library were searched up to April 2021. The random-effects model was used to determine the pooled effect sizes expressed as weighted mean difference (WMD) with % 95 confidence intervals (CIs). Ten eligible RCTs (with 12 arms) were included in the meta-analysis. The meta-analysis revealed that nut intake significantly decreased serum triglycerides (TG) (WMD: -13.19 mg/dL, 95% CI: - 25.90, - 0.48). Furthermore, subgroup analysis showed a significant reduction in serum LDL-cholesterol (LDL-C) following adherence to normocaloric, nut-enriched diets (WMD: - 4.56 mg/dL, 95% CI: - 8.24, - 0.88). However, nuts did not affect serum total cholesterol, high-density lipoprotein cholesterol, glycemic markers, and blood pressure. Overall, incorporating nuts into the diet of healthy adults with overweight/obesity have favorable effects on serum TG and LDL-C. Thus, nuts might exert protective effects against dyslipidemia in this population. PROPSPERO CRD42021250662.


Defining Meditation: Foundations for an Activity-Based Phenomenological Classification System.
Classifying different meditation techniques is essential for the progress of meditation research, as this will enable discerning which effects are associated with which techniques, in addition to supporting the development of increasingly effective and efficient meditation-based training programs and clinical interventions. However, both the task of defining meditation itself, as well as defining specific techniques, faces many fundamental challenges. Here we describe problems involved in this endeavor and suggest an integrated model for defining meditation. For classifying different meditation techniques, we draw on classical, contemporary, and holistic systems of classification. We analyze different techniques and propose that all meditation techniques are based on a specific set of activities, that is: focusing, releasing, imagining, and moving in relation to an object of meditation, including fields of experience. Meditative activities can be combined and unified in the activities of observing, producing, and being aware. All meditative activities are unified in awareness of awareness. Defining specific meditation techniques may be done by specifying which activities and objects are involved. The advantage of our approach is that it can potentially account for the inner workings of all current systems of classification and hence it lays the foundation for formulating an overarching system of meditation that can guide future research and practice.


Effect of variations in sucrose consumption on salivary Lactobacillus count and sucrase activity in man.
Dental students (n = 31) with either high salivary lactobacillus count (greater than 10(4) CFU/ml) or high salivary sucrase activity (greater than or equal to 10 mumol/min X mg protein X 10(-3), or both, were selected to participate in this dietary experiment. For 2 weeks the students avoided sucrose in their diet. Stimulated saliva samples were collected before and after the diet. An additional follow-up sample was collected after 2 weeks of normal diet. The lactobacillus count of undiluted saliva was determined by the Dentocult dip-slide technique. The sucrase activity was determined spectrophotometrically by measuring the sucrose-cleaving activity of centrifuged saliva supernatant. Both the reduction in dietary sucrose and the return to normal diet caused a significant change in these values.


A Chimeric UDP-glucose pyrophosphorylase produced by protein engineering exhibits sensitivity to allosteric regulators.
In bacteria, glycogen or oligosaccharide accumulation involves glucose-1-phosphate partitioning into either ADP-glucose (ADP-Glc) or UDP-Glc. Their respective synthesis is catalyzed by allosterically regulated ADP-Glc pyrophosphorylase (EC 22.214.171.124, ADP-Glc PPase) or unregulated UDP-Glc PPase (EC 126.96.36.199). In this work, we characterized the UDP-Glc PPase from Streptococcus mutans. In addition, we constructed a chimeric protein by cutting the C-terminal domain of the ADP-Glc PPase from Escherichia coli and pasting it to the entire S. mutans UDP-Glc PPase. Both proteins were fully active as UDP-Glc PPases and their kinetic parameters were measured. The chimeric enzyme had a slightly higher affinity for substrates than the native S. mutans UDP-Glc PPase, but the maximal activity was four times lower. Interestingly, the chimeric protein was sensitive to regulation by pyruvate, 3-phosphoglyceric acid and fructose-1,6-bis-phosphate, which are known to be effectors of ADP-Glc PPases from different sources. The three compounds activated the chimeric enzyme up to three-fold, and increased the affinity for substrates. This chimeric protein is the first reported UDP-Glc PPase with allosteric regulatory properties. In addition, this is a pioneer work dealing with a chimeric enzyme constructed as a hybrid of two pyrophosphorylases with different specificity toward nucleoside-diphospho-glucose and our results turn to be relevant for a deeper understanding of the evolution of allosterism in this family of enzymes.


In vitro models in biocompatibility assessment for biomedical-grade chitosan derivatives in wound management.
One of the ultimate goals of wound healing research is to find effective healing techniques that utilize the regeneration of similar tissues. This involves the modification of various wound dressing biomaterials for proper wound management. The biopolymer chitosan (beta-1,4-D-glucosamine) has natural biocompatibility and biodegradability that render it suitable for wound management. By definition, a biocompatible biomaterial does not have toxic or injurious effects on biological systems. Chemical and physical modifications of chitosan influence its biocompatibility and biodegradability to an uncertain degree. Hence, the modified biomedical-grade of chitosan derivatives should be pre-examined in vitro in order to produce high-quality, biocompatible dressings. In vitro toxicity examinations are more favorable than those performed in vivo, as the results are more reproducible and predictive. In this paper, basic in vitro tools were used to evaluate cellular and molecular responses with regard to the biocompatibility of biomedical-grade chitosan. Three paramount experimental parameters of biocompatibility in vitro namely cytocompatibility, genotoxicity and skin pro-inflammatory cytokine expression, were generally reviewed for biomedical-grade chitosan as wound dressing.


Case Report: Thymidine Kinase 2 (TK2) Deficiency: A Novel Mutation Associated With Childhood-Onset Mitochondrial Myopathy and Atypical Progression.
The nuclear gene TK2 encodes the mitochondrial thymidine kinase, an enzyme involved in the phosphorylation of deoxycytidine and deoxythymidine nucleosides. Biallelic TK2 mutations are associated with a spectrum of clinical presentations mainly affecting skeletal muscle and featuring muscle mitochondrial DNA (mtDNA) instability. Current classification includes infantile- ( ≤ 1 year), childhood- (1-12 years), and late-onset (≥12 years) forms. In addition to age at onset, these forms differ for progression, life expectancy, and signs of mtDNA instability (mtDNA depletion vs. accumulation of multiple mtDNA deletions). Childhood-onset TK2 deficiency typically causes a rapidly progressive proximal myopathy, which leads to wheelchair-bound status within 10 years of disease onset, and severe respiratory impairment. Muscle biopsy usually reveals a combination of mitochondrial myopathy and dystrophic features with reduced mtDNA content. Here we report the case of an Italian patient presenting childhood-onset, slowly progressive mitochondrial myopathy, ptosis, hypoacusis, dysphonia, and dysphagia, harboring the TK2 variants c.278A>G and c.543del, the latter unreported so far. Compared to other childhood-onset TK2-patients, our case displays atypical features, including slowly progressive muscle weakness and absence of respiratory failure, which are usually observed in late-onset forms. This report extends the genetic background of TK2-related myopathy, highlighting the clinical overlap among different forms.


Riboflavin synthesis genes ribE, ribB, ribH, ribA reside in the lux operon of Photobacterium leiognathi.
Nucleotide sequence of the riboflavin synthesis genes ribE, ribB, ribH, ribA (GenBank Accession No. AF364106) resided in the lux operon of Photobacterium leiognathi PL741 has been determined, and the amino acid sequences of riboflavin synthetase (RibE), DHBP synthetase (RibB), lumazine synthetase (RibH), GTP cyclohydrolase II (RibA) encoded by the riboflavin synthesis genes are deduced. Nucleotide sequence reveals that the ribE gene encodes the riboflavin synthetase responsible for converting lumazine to riboflavin, the ribB gene encodes the DHBP synthetase responsible for 3,4-dihydroxyl-2-butanone 4-phosphate synthesis, the ribH gene encodes the lumazine synthetase responsible for lumazine synthesis, and the ribA gene encodes the GTP cyclohydrolase II responsible for lumazine synthesis. Functional analysis illustrates that the specific segments lay behind the ribH and ribA genes might form potential loops Omega(oT) and Omega(TI)--Omega(TII); Omega(oT) is functioned as mRNA stability loop or/and for subregulation by alternative modulation, and Omega(TI)--Omega(TII) could be the transcriptional terminator of the lux operon. The gene order of the ribE, ribB, ribH, ribA genes resided in the lux operon and linked to the lum operon is <--ter*-lumQ-lumP-R&R-luxC-luxD-luxA-luxB-luxN-luxE-luxG-ribE-ribB-ribH-ribA-ter--> (R&R: regulatory region; ter: transcriptional terminator), whereas the R&R is the regulatory region for the lum and the lux operons, and ter and ter* are the transcriptional terminators for the lux and lum operons.


Effect of Multiple Sclerosis and Its Treatments on Male Fertility: Cues for Future Research.
Multiple sclerosis is a chronic disease that may lead to different types of symptoms and disabilities. with the better quality of life and decreased disability due to early diagnosis and the availability of disease-modifying therapies (DMTs), the treating physician is increasingly asked to counsel patients on its effects on fertility and reproduction. In particular, reproductive issues are still scarcely studied and discussed in men. Among the still open questions are the following: (a) Does multiple sclerosis cause infertility per sè? (b) Is multiple sclerosis correlated with conditions that increase the risk of infertility? (c) Do DMTs or other therapies for multiple sclerosis impact gonadal function in men? The aim of this review is to provide an overview on the available literature data about the reproductive issues unique to men with multiple sclerosis, underlining the numerous areas where evidence is lacking and, therefore, the priorities for future research.


Safety and efficacy of different anticoagulation regimens after left atrial appendage occlusion.
This study examined the safety and efficacy of different anticoagulation regimens in patients with atrial fibrillation (AF) after left atrial appendage occlusion (LAAC) using the Watchman closure device. AF patients who underwent LAAC using the Watchman closure device in the Department of Cardiology, Jiangsu Taizhou People's Hospital between March 2018 and November 2019 were retrospectively enrolled in this study. Patients were divided into two groups according to different postoperative anticoagulant regimens. One group was treated with anticoagulant therapy alone [oral anticoagulant (OAC)/new OAC (NOAC) group] and the other was given NOACs combined with single antiplatelet therapy (SPAT) (NOAC + SAPT group). The incidences of ischemic stroke/transient ischemic attack (TIA), major bleeding events, all-cause mortality, and device-related thrombosis (DRT) were recorded. Furthermore, multivariate regression was used to analyze the factors associated with the occurrence of early DRT. A total of 160 patients, including 42 (51%) females, with a mean age of 69.13±6.14 years were enrolled in this study. The mean CHA2DS2-VASc score was 3.66±1.11 and the mean HAS-BLED score was 4.16±0.82. During the postoperative follow-up period, there were no significant differences in the incidence of ischemic stroke/TIA [hazard ratio (HR) =0.616; 95% confidence interval (CI): 0.056 to 6.818; P=0.693], major bleeding events (HR =0.520; 95% CI: 0.047 to 5.764; P=0.594) nor all-cause mortality (HR =0.579; 95% CI: 0.052 to 6.394; P=0.656) between the two groups. However, the incidence of early DRT was higher in the OAC/NOAC group compared to the NOAC + SAPT group [odds ratio (OR) =0.120; 95% CI: 0.015 to 0.984; P=0.048]. Multivariate regression confirmed that atrial blood stasis (OR =11.367; 95% CI: 1.394 to 92.687; P=0.023) and peri-device leak (OR =9.337; 95% CI: 1.623 to 53.727; P=0.012) were independent predictors of early DRT. Short-term NOAC + SAPT after LAAC did not significantly increase the incidence of major bleeding events, ischemic stroke/TIA, nor all-cause mortality compared to OAC/NOAC therapy. Not only was NOAC + SAPT therapy found to be safe and effective in short-term follow-up, but it also lowered the risk of early DRT.


Solving the Martian meteorite age conundrum using micro-baddeleyite and launch-generated zircon.
Invaluable records of planetary dynamics and evolution can be recovered from the geochemical systematics of single meteorites. However, the interpreted ages of the ejected igneous crust of Mars differ by up to four billion years, a conundrum due in part to the difficulty of using geochemistry alone to distinguish between the ages of formation and the ages of the impact events that launched debris towards Earth. Here we solve the conundrum by combining in situ electron-beam nanostructural analyses and U-Pb (uranium-lead) isotopic measurements of the resistant micromineral baddeleyite (ZrO2) and host igneous minerals in the highly shock-metamorphosed shergottite Northwest Africa 5298 (ref. 8), which is a basaltic Martian meteorite. We establish that the micro-baddeleyite grains pre-date the launch event because they are shocked, cogenetic with host igneous minerals, and preserve primary igneous growth zoning. The grains least affected by shock disturbance, and which are rich in radiogenic Pb, date the basalt crystallization near the Martian surface to 187 ± 33 million years before present. Primitive, non-radiogenic Pb isotope compositions of the host minerals, common to most shergottites, do not help us to date the meteorite, instead indicating a magma source region that was fractionated more than four billion years ago to form a persistent reservoir so far unique to Mars. Local impact melting during ejection from Mars less than 22 ± 2 million years ago caused the growth of unshocked, launch-generated zircon and the partial disturbance of baddeleyite dates. We can thus confirm the presence of ancient, non-convecting mantle beneath young volcanic Mars, place an upper bound on the interplanetary travel time of the ejected Martian crust, and validate a new approach to the geochronology of the inner Solar System.


A technology-assisted approach for discontinuing contact isolation.
While contact isolation can be an effective tool in reducing the transmission of nosocomial methicillin-resistant Staphylococcus aureus (MRSA), it can increase costs and may decrease the quality of patient care. Therefore, it is important to insure that only patients who need contact precautions are isolated. We describe a simple automated report to help infection control practitioners pro-actively identify patients that may no longer need isolation.


Ontogenetic torsion in two basal gastropods occurs without shell attachments for larval retractor muscles.
Results of this study on two species of vetigastropods contradict the long-standing hypothesis, originally proposed by Garstang (1929), that the larval retractor muscles power the morphogenetic movement of ontogenetic torsion in all basal gastropods. In the trochid Calliostoma ligatum and the keyhole limpet Diodora aspera, the main and accessory larval retractor muscles failed to establish attachments onto the protoconch (larval shell) when the antibiotics streptomycin sulfate and penicillin G were added to cultures soon after fertilization. Defects in protoconch mineralization were also observed. Despite these abnormalities, developing larvae of these species accomplished complete or almost complete ontogenetic torsion, a process in which the head and foot rotate by 180 degrees relative to the protoconch and visceral mass. Analysis by using phalloidin-fluorophore conjugate and transmission electron microscopy showed that myofilaments differentiated within myocytes of the larval retractor muscles and adherens-like junctions formed between muscle and mantle epithelial cells in both normal and abnormal larvae. However, in abnormal larvae, apical microvilli of mantle cells that were connected to the base of the larval retractor muscles failed to associate with an extracellular matrix that normally anchors the microvilli to the mineralized protoconch. If morphogenesis among extant, basal gastropods preserves the original developmental alteration that created gastropod torsion, as proposed by Garstang (1929), then the alteration involved something other than the larval retractor muscles. Alternatively, the developmental process of torsion has evolved subsequent to its origin in at least some basal gastropod clades so that the original alteration is no longer preserved in these clades.


Ban the handshake in winter?
Gastrointestinal and respiratory viruses are particularly common during winter months, and reducing risk of transmission is challenging. Transfer of bacteria from the hands of one person to another is dramatically reduced with a fist bump compared to a handshake. If the same is true for viruses, should we recommend this change in greeting in the winter?


Investigations of antisense oligonucleotides targeted against bcl-2 RNAs.
Expression of the bcl-2 gene becomes deregulated in many non-Hodgkin lymphomas as the result of t(14;18) chromosomal translocations. Because bcl-2 regulates the survival of cells, and because its over-expression is associated with cellular resistance to killing by chemotherapeutic drugs and gamma-irradiation, this gene and its mRNA and protein products represent ideal targets for designing novel therapeutic strategies for the treatment of cancer. Here we describe the effects of an 18-mer phosphodiester oligonucleotide that is complementary to the first 6 codons of the bcl-2 mRNA's open reading frame. When tested for inhibition of in vitro protein synthesis using RNAse-H-supplemented reticulocyte lysates and RNA prepared by in vitro transcription of a human bcl-2 cDNA, the bcl-2 antisense (AS) oligomer completely abolished Bcl-2 protein production at 10 microM, but had no effect on the in vitro translation of a chicken bcl-2 RNA that contained three mismatches relative to the oligomer binding site on the human bcl-2 RNA. A control 18-mer having the same base composition as the AS oligomer but with scrambled order (SC) was not inhibitory. Addition of AS and SC oligomers to cultures of a NIH-3T3 fibroblast cell line that had been stably infected with a recombinant retrovirus containing the same human bcl-2 cDNA used for in vitro transcription/translation experiments revealed concentration-dependent reductions in the relative levels of the 26-kD human Bcl-2 protein (as determined by immunoblotting) by the AS but not by the SC oligomer. Similar results were obtained when AS and SC oligomers were applied to a t(14;18)-containing lymphoma cell line SU-DHL-4 that was cultured in low-serum media. When used at 200 microM, the bcl-2 AS oligomer produced 84-95% reductions in Bcl-2 protein levels in SU-DHL-4 cells but had relatively little effect on the levels of other mitochondrial control proteins, suggesting that the inhibitory effects were specific. Treatment of SU-DHL-4 cells with AS oligomer lead to essentially complete loss of bcl-2 mRNA from cells within 1 day of addition to cultures, but presumably because of the long half-life of the Bcl-2 protein (approximately 14 h), commensurate reductions in Bcl-2 protein levels did not occur until 3 days.(ABSTRACT TRUNCATED AT 400 WORDS)


Associations between amino acids in the evolution of HIV type 1 protease sequences under indinavir therapy.
Significant diversity exists in amino acid sequences encoding HIV-1 protease in individuals naive for protease inhibitors, which could influence the rate of evolution of resistance. High-level resistance to indinavir requires multiple substitutions among at least 11 amino acid sites, and no single substitution was observed in all of 29 resistant isolates obtained from patients on long-term indinavir monotherapy. We have analyzed the evolution of PR in these sequences. The divergence from the baseline amino acid sequence by week 24 was 4%, increasing more than 7% by week 60. The mean difference between sequences from different patients at baseline was 6% (3-9%), rising to 10% after 40 weeks (3-16%), although at all time points nonsynonymous substitutions were less frequent than synonymous nucleotide changes. Analysis of associations between variants at different amino acid sites using a mutual information statistic revealed four pairs of sites to be significantly associated. In three cases these associations included residue 82. Clusters of baseline and week 24 amino acid sequences identified by maximum parsimony did not correlate significantly with the IC95 to indinavir, although a weak correlation of baseline clusters with phenotype at the week 24 time point was suggested.


Cross-ethnic group comparisons of HLA class II alleles and insulin dependent diabetes mellitus.
HLA class II associations with IDDM in populations of non-Caucasoid origin can provide important insights into the nature of the HLA and disease association. Firstly, HLA class II alleles that are rare in Caucasoids but common in other populations can be assessed for their contributory role in IDDM. Secondly, the different HLA class II gene linkage arrangements in different populations can help map the IDDM susceptibility determinants. This chapter reviews studies of HLA class II associations with IDDM in Asian Indians, Chinese, Japanese, Africans and black Americans. Most of these studies have been based on HLA-DR serology. However, DNA analyses, based on restriction fragment length polymorphism, sequence specific oligonucleotide hybridizations of polymerase chain reaction products and DNA sequencing, have made clear the identity of genes contributing to susceptibility or resistance to IDDM in populations of non-Caucasoid origin. DNA sequence analysis of the variable regions of the HLA-DQA, DQB and DRB genes has revealed at least eight alleles at HLA-DQA, 13 at HLA-DQB and 34 at HLA-DRB1. This chapter correlates HLA-DR and DQ allelic diversity with inherited predisposition to IDDM on a global basis. IDDM is strongly associated with the serological specificities of HLA-DQ, rather than with particular amino acid substitutions in class II alleles. DQw8 has a high risk for IDDM, DQw4, DQw5 and DQw9 have a lesser risk, while DQw6 and DQw7 are protective in IDDM. DQw2 is permissive for IDDM, depending on the presence of other HLA class II alleles. Increased heterozygosity at HLA is observed in Oriental patients, as it is in Caucasoid IDDM patients. The nature of this synergism is examined in terms of possible interactive effects between DQA and DQB alleles or DRB and DQB alleles; both effects could be operating. The conclusion from this genetic analysis is that molecular mimicry at HLA-DQ, with either foreign or autoantigens, may be an important mechanism in IDDM. Additionally, the anomalous role of DQw2 in IDDM suggests that a further mechanism, such as T cell activation, may control the ability to mount an immune response against autoantigens. Further studies, possibly with transfectant cell lines, are necessary to clarify the functional role of HLA class II genes in IDDM.


Response to Electrostimulation Is Impaired in Muscle Cells from Patients with Chronic Obstructive Pulmonary Disease.
Among the comorbidities associated with chronic obstructive pulmonary disease (COPD), skeletal muscle weakness and atrophy are known to affect patient survival rate. In addition to muscle deconditioning, various systemic and intrinsic factors have been implicated in COPD muscle dysfunction but an impaired COPD muscle adaptation to contraction has never been extensively studied. We submitted cultured myotubes from nine healthy subjects and nine patients with COPD to an endurance-type protocol of electrical pulse stimulation (EPS). EPS induced a decrease in the diameter, covered surface and expression of MHC1 in COPD myotubes. Although the expression of protein degradation markers was not affected, expression of the protein synthesis marker mTOR was not induced in COPD compared to healthy myotubes after EPS. The expression of the differentiation markers p16INK4a and p21 was impaired, while expression of Myf5 and MyoD tended to be affected in COPD muscle cells in response to EPS. The expression of mitochondrial biogenesis markers PGC1α and MFN2 was affected and expression of TFAM and COX1 tended to be reduced in COPD compared to healthy myotubes upon EPS. Lipid peroxidation was increased and the expression of the antioxidant enzymes SOD2 and GPx4 was affected in COPD compared to healthy myotubes in response to EPS. Thus, we provide evidence of an impaired response of COPD muscle cells to contraction, which might be involved in the muscle weakness observed in patients with COPD.


Evaluation of Pharmacokinetic/Pharmacodynamic Model-Based Optimized Combination Regimens against Multidrug-Resistant Pseudomonas aeruginosa in a Murine Thigh Infection Model by Using Humanized Dosing Schemes.
We previously optimized imipenem and tobramycin combination regimens against a double-resistant clinical Pseudomonas aeruginosa isolate by using in vitro infection models, mechanism-based pharmacokinetic/pharmacodynamic modeling (MBM), and Monte Carlo simulations. The current study aimed to evaluate these regimens in a neutropenic murine thigh infection model and to characterize the time course of bacterial killing and regrowth via MBM. We studied monotherapies and combinations of imipenem with tobramycin in vivo against the double-resistant clinical P. aeruginosa isolate by using humanized dosing schemes. Viable count profiles of total and resistant populations were quantified over 24 h. Tobramycin monotherapy (7 mg/kg every 24 h [q24h] as a 0.5-h infusion) was ineffective. Imipenem monotherapies (continuous infusion of 4 or 5 g/day with a 1-g loading dose) yielded 2.47 or 2.57 log10 CFU/thigh killing at 6 h. At 24 h, imipenem at 4 g/day led to regrowth up to the initial inoculum (4.79 ± 0.26 log10 CFU/thigh), whereas imipenem at 5 g/day displayed 1.75 log10 killing versus the initial inoculum. The combinations (i.e., imipenem at 4 or 5 g/day plus tobramycin) provided a clear benefit, with bacterial killing of ≥2.51 or ≥1.50 log10 CFU/thigh compared to the respective most active monotherapy at 24 h. No colonies were detected on 3×MIC agar plates for combinations, whereas increased resistance (at 3×MIC) emerged for monotherapies (except imipenem at 5 g/day). MBM suggested that tobramycin considerably enhanced the imipenem target site concentration up to 2.6-fold. The combination regimens, rationally optimized via a translational modeling approach, demonstrated substantially enhanced bacterial killing and suppression of regrowth in vivo against a double-resistant isolate and are therefore promising for future clinical evaluation.


Focal segmental glomerulosclerosis in malignant hypertension.
Focal segmental glomerulosclerosis (FSG) may occur in primary malignant hypertension (MHT) either as a result of glomerular hyperfiltration or fibrinoid necrosis (FN), and may contribute to renal dysfunction. To determine the frequency of occurrence and distribution of FSG in primary MHT we studied renal biopsy specimens from 38 black Africans--30 postmortem and 8 needle-biopsy specimens. There were 31 male subjects and 7 female, with a mean age of 46 (+/- 7) years. Mean blood pressure (BP) was 206 +/- 15/137 +/- 9 mmHg, median 24-hour proteinuria (interquartile (IQ) range) was 5.1 g (3.3-6.5 g), median serum albumin 3.4 g (3.2-3.8 g) and median serum creatinine 540 mumol/l (425-752 mumol/l). Mucoid intimal proliferation was present in all the sections but FN was seen in 29 (76%). Glomerulosclerosis was present in all the sections, and was axially distributed in 7 (18%), segmentally in 22 (58%), and globally in 9 (24%). Median 24-hour proteinuria was 2.8 g (0.8-3.5 g IQ range), 5.6 g (1.7-8.1 g) and 3.4 g (2.6-4.0 g) respectively, and corresponding values of serum creatinine were 770 mumol/l (106-1,274 mumol/l IQ range), 522 mumol/l (248-991 mumol/l) and 1,230 mumol/l (920-1,558 mumol/l) respectively. The distribution of glomerulosclerosis did not appear to relate to proteinuria or serum creatinine, although cases with segmentally distributed glomerulosclerosis appeared to have the highest proteinuria, and those with global glomerulosclerosis appeared to have the highest serum creatinine levels. FSG therefore occurs prominently in primary MHT and may contribute to renal dysfunction.


SIRT3 alleviates neuropathic pain by deacetylating FoxO3a in the spinal dorsal horn of diabetic model rats.
The underlying mechanisms of neuropathic pain remain unclear. This work aimed to investigate the role of Sirtuin3 (SIRT3), an nicotinamide adenosine dinucleotide+-dependent histone deacetylase, in the development of neuropathic pain induced by type 2 diabetes mellitus (T2DM) and to explore the associated mechanisms. Diabetic neuropathic pain (DNP) in rats was induced by high-fat diet/low-dose streptozotocin. The pain behaviors were examined using the von Frey and Hargreaves tests. The levels of SIRT3, manganese superoxide dismutase (MnSOD) and catalase (CAT) were determined using Western blot and RT-qPCR. The acetylation, phosphorylation and ubiquitination of forkhead box class O3a (FoxO3a) were analyzed by immunoprecipitation and Western blot. SIRT3 expression and activity were significantly reduced in the spinal dorsal horn of DNP model rats. Overexpression of spinal SIRT3 reversed the pain hypersensitivity in the DNP model rats, but knockdown of spinal SIRT3 mimicked the pain effect, eliciting pain hypersensitivity in normal rats. Moreover, overexpression of spinal SIRT3 in DNP model rats increased the FoxO3a level and upregulated the antioxidant genes MnSOD and CAT by deacetylating FoxO3a and inhibiting FoxO3a phosphorylation and ubiquitination. Knockdown of spinal SIRT3 in normal rats decreased the FoxO3a level and downregulated MnSOD and CAT by inhibiting the deacetylation of FoxO3a and further increasing FoxO3a phosphorylation and ubiquitination. These results suggest that, by deacetylating FoxO3a and further reducing its phosphorylation, ubiquitination and degradation in the spinal dorsal horn, SIRT3 stabilizes FoxO3a protein and inhibits oxidative stress, resulting in pain alleviation in T2DM model rats.


The ripening disorder berry shrivel affects anthocyanin biosynthesis and sugar metabolism in Zweigelt grape berries.
Timescale analyses suggest the berry shrivel (BS) disorder is induced before veraison with strong effects on anthocyanin biosynthesis, and minor effects on sugar transport and metabolism. Berry shrivel (BS)-affected grapes have low sugar contents, high acidity, less anthocyanins and flaccid berries. To date no pathogenic causes are known, and studies to elucidate the molecular basis leading to symptom induction and development are limited. Here we present a study on pre-symptomatic as well as symptomatic BS berries to characterize early metabolic changes, with focus on anthocyanin biosynthesis and sugars metabolism. Healthy and BS berries from six sampling time points were used (BBCH79-BBCH89). Our objectives are (1) to search for the beginning of BS-related physiological processes; (2) to search for key enzymes and sugar transporters involved in BS induction and development and (3) to understand the consequences on polyphenol biosynthesis. We employed high performance anion exchange chromatography coupled with pulsed amperometric detection (HPAEC-PAD) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) technology for sugar and polyphenol analyses, respectively. Additionally we conducted expression analyses (qPCR) of key genes and enzymatic activity assays. Our results show that BS-related processes start before veraison, as determined by slightly reduced hexose contents and reduced expression levels of a vacuolar invertase (VviGIN1), two monosaccharide transporters (VviTMT2, VviTMT3) and the anthocyanin biosynthesis (VviUFGT, VviMYBA1/2) genes. Lower amounts of delphinidin and cyanidin glycosidic forms were determined, while caftaric acid, quercetin-3-O-glucuronide and (+)-catechin were increased in BS berries. Although not all results were conclusive, especially for the sugar metabolism, our data provide important knowledge to improve the understanding of the highly complex berry shrivel ripening disorder.


Regulation of MHC molecules on MBP positive oligodendrocytes in mice by IFN-gamma and TNF-alpha.
The expression of class I and class II histocompatibility antigens by myelin basic protein (MBP)-positive oligodendrocytes, in response to exogenous cytokines, has been investigated in vitro. It has been found that interferon-gamma (IFN-gamma), although capable of class I induction, does not induce class II on oligodendrocytes. Furthermore, tumour necrosis factor-alpha (TNF-alpha), which was shown to induce class I MHC on other neural cells, failed to induce class I on oligodendrocytes. A combination of IFN-gamma and TNF-alpha also failed to facilitate the expression of class II antigens on oligodendrocytes, nor did it amplify the expression of class I seen with IFN-gamma alone. Thus it appears that MBP+ murine oligodendrocytes are refractory to class II induction, and express class I in response to IFN-gamma but not TNF-alpha. The differential regulation and class of MHC expression may have implications in terms of the initiation and targeting of immune responses directed toward the oligodendrocyte.


Comparison of low dose versus ultra-low dose hormone therapy in menopausal symptoms and quality of life in perimenopause women.
The study was to compare the efficacy, safety, and tolerability of low dose versus ultra-low dose hormone therapy (HT) in the management of perimenopause symptoms and quality of life. Retrospective analysis of perimenopause patients prescribed for 25 weeks HT in the outpatient clinic of menopause. A total of 132 perimenopause women were included in two treatment regimens: one with low dose HT (LD-HT) and one with ultra-low dose HT (ULD-HT). Changes in serum levels of follicle-stimulating hormone, estradiol as well as transvaginal ultrasound (TVUS), the 36-item Short Form Health Survey (SF-36), the Kupperman Index (KI), and adverse effects were assessed at baseline, 4, 13, and 25 weeks. By the end of 25 weeks of treatment, each score of SF-36 domains for both LD-HT and ULD-HT groups were increased, the KI decreased, and the endometrial thickness increased in both groups and there was no statistical difference between two groups. Both groups have negligible differences in incidence of adverse effects. Low dose and ultra-low dose HT both can serve in improving symptoms of perimenopause, thereby offering a better quality of life with decreased incidence of side effects. Ultra-low dose treatment may have a better advantage on safety and tolerance.


Quercetin Alleviates Oxidative Damage by Activating Nuclear Factor Erythroid 2-Related Factor 2 Signaling in Porcine Enterocytes.
Oxidative stress has been implicated in the etiology of multiple gastrointestinal disorders, such as irritable bowel syndrome and inflammatory bowel disease. This study was conducted to evaluate effects of natural product quercetin on diquat-induced oxidative stress in porcine enterocytes and underlying mechanisms. Intestinal porcine epithelial cell line 1 (IPEC-1) cells pretreated with or without quercetin (5 μM, 24 h) were incubated with vehicle or diquat (100 μM) for 6 h. The results showed that diquat treatment induced apoptosis in a caspase-3-dependent manner, as accompanied by elevated reactive oxygen species (ROS) production, increased mitochondrial depolarization, and reduced the abundance of tight junction proteins. These adverse effects of diquat were remarkably abrogated by quercetin administration. Further study indicated that the protective effect of quercetin was associated with elevated protein abundance of nuclear factor erythroid 2-related factor 2 (Nrf2) and increased intracellular glutathione (GSH) content. Interestingly, the beneficial effects of quercetin on diquat-induced oxidative damage were abolished by all-trans-retinoic acid (Atra), a specific inhibitor of Nrf2, indicating a Nrf2-dependent regulation manner. The results show that quercetin attenuates diquat-induced cell injury by promoting protein abundance of Nrf2 and regulating GSH-related redox homeostasis in enterocytes. These findings provide new insights into a function role of quercetin in maintaining intestinal homeostasis.


Positive feedback interactions between microtubule and actin dynamics during cell motility.
The migration of tissue cells requires interplay between the microtubule and actin cytoskeletal systems. Recent reports suggest that interactions of microtubules with actin dynamics creates a polarization of microtubule assembly behavior in cells, such that microtubule growth occurs at the leading edge and microtubule shortening occurs at the cell body and rear. Microtubule growth and shortening may activate Rac1 and RhoA signaling, respectively, to control actin dynamics. Thus, an actin-dependent gradient in microtubule dynamic-instability parameters in cells may feed back through the activation of specific signalling pathways to perpetuate the polarized actin-assembly dynamics required for cell motility.


Prognostic value of monosomal karyotype in comparison to complex aberrant karyotype in acute myeloid leukemia: a study on 824 cases with aberrant karyotype.
In acute myeloid leukemia (AML) the subset with complex karyotype (CK) is traditionally regarded as the worst prognostic group. However, ≥ 3, ≥ 4, or ≥ 5 abnormalities have been variably used for its definition. Recently, monosomal karyotype (MSK) was suggested to indicate an even inferior outcome. We tested which definition fits best to identify the most unfavorable subgroup. After excluding patients with t(15;17)/PML-RARA, t(8;21)/RUNX1-RUNX1T1, inv (16)/t(16;16)/CBFB-MYH11, and normal karyotype, 824 patients with AML with cytogenetic abnormalities were analyzed. Patients with MSK or CK defined as ≥ 3, ≥ 4, or ≥ 5 abnormalities showed an inferior overall survival compared with the respective remaining patients not fulfilling these criteria (for all, P < .001). Hazard ratios were 1.93, 1.68, 1.94, and 1.92. CK ≥ 4 as a single parameter identified the largest proportion of patients with very poor risk. However, combining CK ≥ 4 and MSK detected an even larger number of patients with very unfavorable outcome (261 of 824; 31.7%).


[Indirect evaluation of respiratory muscle strength with the help of markers of maximal inspiratory and expiratory pressure in the mouth of healthy individuals].
The aim of the study was the evaluation of respiratory muscle strength by measurement of maximal inspiratory (PImax) and expiratory (PEmax) pressures values. Results for 166 clinically normal subjects (79 female and 87 male) were obtained. The results were as follow: in woman PImax ranged 38-104 cm H2O, average 60 cm H2O, PEmax ranged 46-140 cm H2O average 87.5 cm H2O, in men PImax was 40-120 cm H2O, average 73.2 cm H2O, PEmax ranged 46-140 cm H2O, average 115.9 cm H2O. PImax was negatively correlated with age in both groups. There was no correlation between age and PImax or PEmax in both groups (p > 0.05). There was no correlation between PImax and PEmax and height in women group and men group treated apart. The correlation was found between PImax as well as PEmax and height for whole group (p = 0.00019). There were observed positive correlation between PImax, PEmax and weight in both (male and female) groups. The comparison of results of present study with those obtained in former studies reveals important differences of norms for different populations. The normal values of maximal inspiratory and expiratory pressures in the mouth (PImax, PEmax) should be qualified individually for studied population. The normal values recommended by producers of medical equipment might be inadequate for studied population.


Blood access in daily hemodialysis.
Frequent dialyses are sometimes perceived as increasing the risk of blood access malfunction and decreased longevity. This review of the literature, however, indicates that the failure rates and overall fistula survival appear to be better with more frequent dialyses than with routine dialysis frequency, although the reasons for this phenomenon are not clear. One of the possible explanations is that frequent dialyses are associated with fewer intradialytic hypotensive episodes, which are very detrimental to the blood access. Another possible explanation is the generally lower blood flow used with more frequent hemodialyses, particularly long nocturnal hemodialysis. Finally, a decreased clotting tendency and decreased rates of hematoma formation at the puncture sites are additional possible explanations. Complication rates with bridge grafts are not higher with more frequent compared to routine thrice-weekly hemodialysis sessions. No such comparative data are available, however, for central-vein catheters. This lack of comparisons seems to stem from the intuitive assumption by nephrologists that hemodialyses that are more frequent should not adversely impact catheter complication rates and survival. No data at all are available on the use of the Dialock(R) hemodialysis system (Biolink Corp., Norwell, MA, USA) and LifeSite hemodialysis access system (Vasca, Inc., Tewksbury, MA, USA), two newer forms of hybrid access in patients undergoing frequent hemodialyses. Current evidence shows that the perceived risk of blood access malfunction and decreased longevity when patients undergo more frequent hemodialysis is not supported by the current literature.


Post-renal acute renal failure during pelvis irradiation for prostate cancer.
A case of post renal acute renal failure secondary to radiation therapy for prostate cancer is described. Severe bladder inflammatory reaction, leading to bilateral ureteric obstruction, occurred after a moderate dose of radiotherapy (6400 cGy in 35 fractions). At a follow-up time of 8 years, the patient has fully recovered and remains disease-free. To our knowledge, this complication has never been described previously. Given the thrust in prostate cancer treatment toward dose escalation this complication could become more frequent.