Cadmium stress antioxidant responses and root-to-shoot communication in grafted tomato plants.
Many aspects related to ROS modulation of signaling networks and biological processes that control stress responses still remain unanswered. For this purpose, the grafting technique may be a powerful tool to investigate stress signaling and specific responses between plant organs during stress. In order to gain new insights on the modulation of antioxidant stress responses mechanisms, gas-exchange measurements, lipid peroxidation, H2O2 content, proline, superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), ascorbate peroxidase (APX) and guaiacol peroxidase (GPOX) were analyzed in Micro-Tom grafted plants submitted to cadmium (Cd). The results observed revealed that higher amounts of Cd accumulated mainly in the roots and rootstocks when compared to leaves and scions. Macronutrients uptake (Ca, S, P and Mg) decreased in non-grafted plants, but differed among plant parts in all grafted plants. The results showed that the accumulation of proline observed in scions of grafted plants could be associated to the lower MDA contents in the scions of grafted plants. In the presence of Cd, non-grafted plants displayed increased CAT, GR, GPOX and APX activities for both tissues, whilst grafted plants revealed distinct trends that clearly indicate signaling responses from the rootstocks, allowing sufficient time to activate defense mechanisms in shoot. The information available concerning plants subjected to grafting can provide a better understanding of the mechanisms of Cd detoxification involving root-to-shoot signaling, opening new possibilities on strategies which can be used to manipulate heavy metal tolerance, since antioxidant systems are directly involved in such mechanism.


Effect of graded heart rate reduction with ivabradine on myocardial oxygen consumption and diastolic time in exercising dogs.
Lowering heart rate reduces myocardial oxygen consumption (MVO2) and produces potent anti-ischemic effects. The development of selective heart rate-reducing agents represents an alternative approach to the use of beta-blockers. Therefore, our goal was to establish the dose-response curve of the effects of ivabradine (If channel inhibitor) on MVO2 and diastolic time. Seven conscious and chronically instrumented dogs were investigated during exercise at spontaneous and paced heart rate (250 beats/min) after administration of increasing doses of ivabradine (0.25, 0.5, and 1 mg/kg i.v.). During exercise, ivabradine dose dependently and significantly reduced the exercise-induced tachycardia (-17, -21, and -32% at 0.25, 0.5, and 1 mg/kg, respectively, versus saline) without altering myocardial contractility nor mean ejection wall stress. A linear relationship between heart rate (HR) and MVO2 was demonstrated (MVO2 = 0.044 x HR - 1.4; r = 0.987). These effects of ivabradine on MVO2 were abolished by atrial pacing. Similarly, ivabradine dose dependently increased diastolic time without altering the inverse and non linear relationship between diastolic time and heart rate observed with saline. In conclusion, selective heart rate reduction with ivabradine dose dependently increases diastolic time and reduces MVO2 with a linear relationship between heart rate and MVO2. The lack of "on-off" pharmacological profile will predict the possibility of using a wide range of dose regimen.


[High sense of coherence can have health enhancing effects].
Sense of coherence (SOC), e.g. meaningfulness of life, is according to the sociologist A. Antonovsky a health-promoting construct, which has been used in a number of descriptive studies in various populations. The aim of this population-based cross-sectional study was to investigate if middle-aged women with a self-reported low level of SOC also differed in biological variables, e.g. related to glucose and lipid metabolism, from women with a high level of SOC. These variables could potentially represent links in the pathophysiological chain of events causing somatic disease. We found significant differences according to dyslipidaemia (low HDL cholesterol and increased triglyceride levels) between groups of women with differing SOC scores, most negative findings were found in the group with lowest SOC. Women with medium-high or high SOC also reported less symptoms and need of regular medical controls. The results support a potential association between cognitive processes (SOC) and biological mechanisms causing adverse health effects.


Agonist-induced desensitization and endocytosis of heterodimeric GABAB receptors in CHO-K1 cells.
gamma-Aminobutyric acid B (GABA(B)) receptor is the first discovered G protein-coupled receptor that requires two subunits, GB1 and GB2, to form a functional receptor. Whereas the molecular and functional characteristics of GABA(B) receptors have been recently extensively studied, the mechanisms underlying receptor desensitization and endocytosis are still poorly understood. We have investigated the effect of continuous agonist exposure on the human GABA(B) receptor functional response and redistribution when expressed in Chinese hamster ovary (CHO-K1) cells. The wild-type GABA(B) receptor-mediated inhibition of the adenylate cyclase activity appeared desensitized after 2 h in the presence of GABA (100 microM). Fusion proteins were generated by attachment of cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP) to GB1 and GB2, respectively, and confocal microscopy experiments in intact living cells semi-stably expressing the constructs were performed. Incubation of co-expressing CFP-GB1 and YFP-GB2 cells in the presence of GABA (100 microM) for 2 h induced a profound receptor internalization, and CFP-GB1 and YFP-GB2 appeared co-localized in the endosome (labelled with Cy3-transferrin). The internalization was blocked by a selective GABA(B) receptor antagonist. These results represent the first clear visualization of agonist-induced internalization of the unique heterodimeric GABA(B) receptor.


Safety-catch linker strategies for the production of radiopharmaceuticals labeled with positron-emitting isotopes.
A novel synthetic stratetegy for compounds labeled with the positron-emitting isotope carbon-11 is described. The use of precursors attached to a solid support via safety-catch linkers allows selective release of radiolabeled material, leaving unreacted precursor attached to the support. Two different linkers demonstrate the application to the preparation of radiolabeled N-alkyl tertiary amines and N-alkylsulfonamides. This technique is expected to lead to more widespread use of positron emission tomography for the in vivo analysis of compound behavior.


Understanding the Sorghum-Colletotrichum sublineola Interactions for Enhanced Host Resistance.
Improving sorghum resistance is a sustainable method to reduce yield losses due to anthracnose, a devastating disease caused by Colletotrichum sublineola. Elucidating the molecular mechanisms of sorghum-C. sublineola interactions would help identify biomarkers for rapid and efficient identification of novel sources for host-plant resistance improvement, understanding the pathogen virulence, and facilitating resistance breeding. Despite concerted efforts to identify resistance sources, the knowledge about sorghum-anthracnose interactions remains scanty. Hence, in this review, we presented an overview of the current knowledge on the mechanisms of sorghum-C. sublineola molecular interactions, sources of resistance for sorghum breeding, quantitative trait loci (QTL), and major (R-) resistance gene sequences as well as defense-related genes associated with anthracnose resistance. We summarized current knowledge about C. sublineola populations and its virulence. Illustration of the sorghum-C. sublineola interaction model based on the current understanding is also provided. We highlighted the importance of genomic resources of both organisms for integrated omics research to unravel the key molecular components underpinning compatible and incompatible sorghum-anthracnose interactions. Furthermore, sorghum-breeding strategy employing rapid sorghum germplasm screening, systems biology, and molecular tools is presented.


Niche construction, biological evolution, and cultural change.
We propose a conceptual model that maps the causal pathways relating biological evolution to cultural change. It builds on conventional evolutionary theory by placing emphasis on the capacity of organisms to modify sources of natural selection in their environment (niche construction) and by broadening the evolutionary dynamic to incorporate ontogenetic and cultural processes. In this model, phenotypes have a much more active role in evolution than generally conceived. This sheds light on hominid evolution, on the evolution of culture, and on altruism and cooperation. Culture amplifies the capacity of human beings to modify sources of natural selection in their environments to the point where that capacity raises some new questions about the processes of human adaptation.


Systematic of alcohol-induced simple coacervation in aqueous gelatin solutions.
Turbidity measurements performed at 450 nm were used to follow the process of simple coacervation when 1% (w/v) aqueous alkali processed gelatin (type-B) solutions were titrated with methanol, ethanol, propanol, and tert-butyl alcohol at various pHs of the solution ranging from pH = 5 to 8 and ionic strengths varying from I = 0.01 to 0.1 M NaCl. The titration profiles clearly established the transition points in terms of the percentage of volume of alcohol added relative to that of solvent corresponding to the first occurrence of turbidity (Vt) and a point of turbidity maximum (Vp). Addition of more alcohol drove the system toward precipitation. The values of Vt and Vp characterized the initiation of intramolecular folding and intermolecular aggregate formation of the charge neutralized gelatin molecules and the subsequent micro coacervate droplet formation. The state of intermolecular aggregates and that of folded gelatin molecules could be characterized by dynamic laser light scattering experiments, which implied spontaneous segregation of particle sizes preceding coacervation. The aggregates constitute the coacervate phase while the folded gelatin molecules mostly stay dispersed in the supernatant. The data taken together reveal the role played by solution entropy in addition to that of electrostatic and solute-solvent interactions, which had been overlooked hitherto.


Automated classification of severity of age-related macular degeneration from fundus photographs.
To evaluate an automated analysis of retinal fundus photographs to detect and classify severity of age-related macular degeneration compared with grading by the Age-Related Eye Disease Study (AREDS) protocol. Following approval by the Johns Hopkins University School of Medicine's Institution Review Board, digitized images (downloaded AT http://www.ncbi.nlm.nih.gov/gap/) of field 2 (macular) fundus photographs from AREDS obtained over a 12-year longitudinal study were classified automatically using a visual words method to compare with severity by expert graders. Sensitivities and specificities, respectively, of automated imaging, when compared with expert fundus grading of 468 patients and 2145 fundus images are: 98.6% and 96.3% when classifying categories 1 and 2 versus categories 3 and 4; 96.1% and 96.1% when classifying categories 1 and 2 versus category 3; 98.6% and 95.7% when classifying category 1 versus category 3; and 96.0% and 94.7% when classifying category 1 versus categories 3 and 4; Development of an automated analysis for classification of age-related macular degeneration from digitized fundus photographs has high sensitivity and specificity when compared with expert graders and may have a role in screening or monitoring.


Measuring the coherence function of continuous-wave lasers by a photorefractive grating method.
We demonstrate a simple, adjustment-insensitive technique for measuring the temporal coherence function of cw multimode and monomode semiconductor lasers, using two-beam coupling in photorefractive InP and CdTe crystals. The emission spectra of the diodes are measured independently. The coherence functions are also calculated from these spectra and agree with the photorefractive measurements. Coupling of two partially coherent waves in low-speed photorefractive media is described theoretically. The range of the experimental parameters in which the method of coherence measurement is correct is given.


The use of bone marrow culture for the diagnosis of melioidosis.
We have evaluated prospectively the contribution of bone marrow culture to the diagnosis of melioidosis. Bone marrow (BMC) and blood cultures (BC) were collected concurrently from 105 patients with suspected acute, severe melioidosis. 67 patients were subsequently proved to have the disease whilst other significant organisms were isolated from these specimens in 5 cases. Overall, 67.2% of BC and 64.2% of BMC from melioidosis patients grew Pseudomonas pseudomallei. Time to positivity did not differ significantly in paired BC and BMC specimens. These results do not support the routine use of BMC in the diagnosis of acute, severe melioidosis. In one patient with pulmonary melioidosis, however, blood cultures were repeatedly negative, whilst bone marrow grew P. pseudomallei, and this preceded the development of a distant focus of infection. This suggests that culture of bone-marrow may be of value in certain blood culture-negative patients with melioidosis.


Efficacy of a modified improved technique for detecting red cell haemoglobin H inclusions.
Alpha-thalassaemia is a common disease in Taiwan. A feature useful in diagnosis is the excess of of beta-chains that result from impaired alpha-chain production. These excess chains assemble into beta 4 tetramers (i.e. Hb H) which can be detected by its rapid anodal migration on alkaline electrophoretic media as seen in Hb H disease. However, this technique cannot, and conventional Hb H inclusion staining rarely can visualize alpha-thalassaemic traits due to small quantities of Hb H formed in these patients. The staining for Hb H inclusion bodies uses brilliant cresyl blue (BCB) or methylene blue (MB) as an oxidant to denature Hb H as intracellular inclusions. We have improved the technique of Jones (who modified the original method in order to obtain enriched young red cells) by prolonging the incubation time from 30 min to 3 h. The sensitivity of this modified improved method was 91% for detecting obligatory alpha-thalassaemic traits as shown in table. No false positive results were seen in beta-thalassaemia or in others. We believe this can be used as a confirmatory test in heterozygous alpha 1-thalassaemia and homozygous alpha 2-thalassaemia.


LRRC8A promotes Glaesserella parasuis cytolethal distending toxin-induced p53-dependent apoptosis in NPTr cells.
Glaesserella parasuis is an early colonizer of the swine upper respiratory tract and can break through the respiratory barrier for further invasion. However, the mechanisms underlying G. parasuis increases epithelial barrier permeability remain unclear. This study demonstrates that G. parasuis cytolethal distending toxin (CDT) induces p53-dependent apoptosis in new-born piglet tracheal (NPTr) cells. Moreover, we report for the first time that leucine-rich repeat-containing protein 8A (LRRC8A), an essential subunit of the volume-regulated anion channel (VRAC), involves in apoptosis of NPTr cells mediated by G. parasuis CDT. Pharmacological inhibition of VRAC with either PPQ-102 or NS3728 largely attenuated CDT-induced apoptosis in NPTr cells. Additionally, experiments with cells knocked down for LRRC8A using small interfering ribonucleic acid (siRNA) or knocked out LRRC8A using CRISPR/Cas9 technology showed a significant reduction in CDT-induced apoptosis. Conversely, re-expression of Sus scrofa LRRC8A in LRRC8A-/- NPTr cells efficiently complemented the CDT-induced apoptosis. In summary, these findings suggest that LRRC8A is pivotal for G. parasuis CDT-induced apoptosis, providing novel insights into the mechanism of apoptosis caused by CDT.


Sensorimotor Processing in the Basal Ganglia Leads to Transient Beta Oscillations during Behavior.
Brief epochs of beta oscillations have been implicated in sensorimotor control in the basal ganglia of task-performing healthy animals. However, which neural processes underlie their generation and how they are affected by sensorimotor processing remains unclear. To determine the mechanisms underlying transient beta oscillations in the LFP, we combined computational modeling of the subthalamo-pallidal network for the generation of beta oscillations with realistic stimulation patterns derived from single-unit data recorded from different basal ganglia subregions in rats performing a cued choice task. In the recordings, we found distinct firing patterns in the striatum, globus pallidus, and subthalamic nucleus related to sensory and motor events during the behavioral task. Using these firing patterns to generate realistic inputs to our network model led to transient beta oscillations with the same time course as the rat LFP data. In addition, our model can account for further nonintuitive aspects of beta modulation, including beta phase resets after sensory cues and correlations with reaction time. Overall, our model can explain how the combination of temporally regulated sensory responses of the subthalamic nucleus, ramping activity of the subthalamic nucleus, and movement-related activity of the globus pallidus leads to transient beta oscillations during behavior.SIGNIFICANCE STATEMENT Transient beta oscillations emerge in the normal functioning cortico-basal ganglia loop during behavior. Here, we used a unique approach connecting a computational model closely with experimental data. In this way, we achieved a simulation environment for our model that mimics natural input patterns in awake, behaving animals. We demonstrate that a computational model for beta oscillations in Parkinson's disease (PD) can also account for complex patterns of transient beta oscillations in healthy animals. Therefore, we propose that transient beta oscillations in healthy animals share the same mechanism with pathological beta oscillations in PD. This important result connects functional and pathological roles of beta oscillations in the basal ganglia.


Reversing the Effects of a Peripheral Nerve Block with Normal Saline: A Randomised Controlled Trial.
The objective of the present study was to determine whether or not the effects of peripheral nerve block can be reversed by flushing normal saline down a peripheral nerve block catheter following the completion of arteriovenous (AV) fistula surgery. In the present study, 38 patients undergoing AV fistula surgery were recruited, and a brachial plexus block with a peripheral nerve catheter was established. Following surgery, the patients were randomised to either the control group or the washout group, where 10 mL of normal saline was flushed down the peripheral nerve catheter at 15-minute intervals for 1 h while the patients were in the postoperative recovery room. An observer blinded to the patient group allocation assessed motor and sensory functions in all patients at 15-minute intervals for 1 h, and pain scores were recorded. There was no difference in time to resolution of motor or sensory block in the two groups. The median changes in the motor score were 1.5 out of 10 for the control group and 2 for the washout group (p=0.95). The median changes in the sensory score were 3 out of 10 for the control group and 1 for the washout group (p=0.14). There were no differences in pain scores over the study period in either group (p=0.44). We were unable to show any useful improvement in block resolution with normal saline washout of supraclavicular or infraclavicular brachial plexus blocks following AV fistula surgery.


Photodynamic therapy-induced killing is enhanced in depigmented metastatic melanoma cells.
The resistance of pigmented human melanomas over their unpigmented counterparts to a number of therapies has suggested that the presence of intracellular melanin plays a role in rendering these cells less susceptible to cell death, probably through the ability of this pigment to act as an intracellular antioxidant, thus neutralizing chemotherapeutic-induced ROS (reactive oxygen species). PDT (photodynamic therapy) was recently suggested as an attractive, adjunctive therapy owing to its cellular specificity and limited side effects. In the present study, we propose that first depigmenting melanomas with a reversible TYR (tyrosinase) inhibitor such as PTU (phenylthiourea) increases their susceptibility to HYP-PDT (hypericin-mediated PDT). Pigmented [UCT Mel-1 (University of Cape Town melanoma cell line 1)] and unpigmented (A375) melanomas were first characterized with respect to their TYR activities and melanin quantities and then treated with a TYR inhibitor for 48 h. Cell viability assays after treatment with 3 μM HYP-PDT showed a significant increase in cell death in depigmented melanomas compared with untreated melanomas that returned to the level of untreated melanoma cells on removing the TYR inhibitor. The present study supports the hypothesis that combining the inhibition of melanogenesis with PDT should be explored as a valid therapeutic target for the management of advanced melanoma.


Toxicity of chemical components of fine particles inhaled by aged rats: effects of concentration.
This study tested the hypothesis that exposure to mixtures containing fine particles and ozone (O3) would cause pulmonary injury and decrements in functions of immunological cells in exposed rats (22-24 months old) in a dose-dependent manner. Rats were exposed to high and low concentrations of ammonium bisulfate and elemental carbon and to 0.2 ppm O3. Control groups were exposed to purified air or O3 alone. The biological end points measured included histopathological markers of lung injury, bronchoalveolar lung fluid proteins, and measures of the function of the lung's innate immunological defenses (macrophage antigen-directed phagocytosis and respiratory burst activity). Exposure to O3 alone at 0.2 ppm did not result in significant changes in any of the measured end points. Exposures to the particle mixtures plus O3 produced statistically significant changes consistent with adverse effects. The low-concentration mixture produced effects that were statistically significant compared to purified air but, with the exception of macrophage Fc receptor binding, exposure to the high-concentration mixture did not. The effects of the low- and high-concentration mixtures were not significantly different. The study supports previous work that indicated that particle + O3 mixtures were more toxic than O3 alone.


Keeping safe. Continuous glucose monitoring (CGM) in persons with Type 1 diabetes and impaired awareness of hypoglycaemia: a qualitative study.
To further our understanding of individual use and experience of continuous glucose monitoring (CGM) in adults with Type 1 diabetes and impaired awareness of hypoglycaemia, we conducted a qualitative study supplementary to a randomized controlled trial, using semi-structured interviews. Twenty-three participants of the IN CONTROL trial were interviewed within 4 weeks after the last study visit. The interview centred around experiences of CGM, taking into account the person's expectations prior to the trial. The interview was semi-structured, using open-ended questions and, if needed, prompts were offered to elicit further responses. Using thematic analysis, the interview transcripts were coded independently by three members of the research team. The consolidated criteria for reporting qualitative research (COREQ) were followed. Overall, CGM was experienced as helpful in gaining more insight into glucose variability, and temporarily improved sense of control, reduced distress and made participants less dependent on others. However, some participants experienced confrontation with CGM output as intrusive, while some reported frustration due to failing technique and difficulty trusting the device. Participants reported active and passive self-management behaviours mirroring individual differences in attitudes and coping styles. In adults with Type 1 diabetes at risk of recurrent hypoglycaemia due to impaired awareness of hypoglycaemia, CGM use enhances a sense of control and safety for most, but not all. Future studies should further explore differential use of CGM in this population in the context of active and passive self-management styles.


A flexible nonlinear mixed effects model for HIV viral load rebound after treatment interruption.
Characterization of HIV viral rebound after the discontinuation of antiretroviral therapy is central to HIV cure research. We propose a parametric nonlinear mixed effects model for the viral rebound trajectory, which often has a rapid rise to a peak value followed by a decrease to a viral load set point. We choose a flexible functional form that captures the shapes of viral rebound trajectories and can also provide biological insights regarding the rebound process. Each parameter can incorporate a random effect to allow for variation in parameters across individuals. Key features of viral rebound trajectories such as viral set points are represented by the parameters in the model, which facilitates assessment of intervention effects and identification of important pretreatment interruption predictors for these features. We employ a stochastic expectation-maximization (StEM) algorithm to incorporate HIV-1 RNA values that are below the lower limit of assay quantification. We evaluate the performance of our model in simulation studies and apply the proposed model to longitudinal HIV-1 viral load data from five AIDS Clinical Trials Group treatment interruption studies.


A study design to assess the safety and efficacy of on-pump versus off-pump coronary bypass grafting: the ROOBY trial.
Since the late 1960s, coronary artery bypass graft (CABG-only) procedures were traditionally performed using a heart-lung machine on an arrested heart (on-pump). Over the past decade, an increasing number CABG-only procedures were performed on a beating heart (off-pump). Advocates of the off-pump approach expect to reduce many of the adverse side effects related to using the heart-lung machine, while advocates for the on-pump procedure raise concerns related to graft patency rates and long-term event-free survival for the off-pump technique. The U.S. Department of Veteran Affairs (VA) Cooperative Studies Program funded a randomized, multicenter clinical trial comparing the clinical and resource-related outcomes following on-pump versus off-pump techniques for veterans undergoing a non-emergent CABG-only procedure. The planning committee was faced with several critically important challenges to assure feasibility of study costs and required sample size; generalizability to non-VA surgical practices; and comparability of clinically meaningful results. These challenges are discussed. This study is a prospective, randomized, multicenter, single blinded (patient) clinical trial that compares on-pump and off-pump techniques for veterans requiring non-emergent CABG-only procedures. There will be 2200 patients randomized at 17 VA Medical Centers when the five-year recruitment period ends on 15 April 2007. There are two primary objectives: a short-term objective to assess the immediate impact of the two techniques on 30-day mortality/morbidity and a long-term objective to assess one-year mortality/morbidity. Major secondary outcomes are one-year graft patency rates and change in neuropsychological assessments from baseline to one year. All patients are assessed at 30 days post-surgery or discharge from the hospital, whichever is latest, and at one-year post-surgery. During planning, several key issues had to be decided. These included 1) choosing primary objectives: a short-term (30-day) and a long-term (one-year) objective were chosen; 2) choosing primary outcome measures: composite measures were selected to ensure sufficient end-points; 3) standardization of surgical techniques: minimal standardization required but guidelines and continuing discussions on both techniques provided; 4) establishing criteria for surgeons and residents for participation: surgeons required to have completed 20 off-pump procedures prior to doing study procedures and residents, in presence of study surgeon, capable of doing either procedure; 5) identifying metrics of cognitive dysfunction sensitive to treatment: a neuropshychologist hired who centrally monitors cognitive functioning testing; and 6) blinding participants of surgical procedure: attempt to blind participants. Areas of concern are whether all surgeons sufficiently experienced on the off-pump procedure, should residents have been allowed to do study surgeries, should techniques have been standardized more and were the best neurocognitive tests selected. The study design presented allows for a balanced and fair assessment of the on-pump and off-pump CABG procedures across a diversity of clinical outcomes and resource use metrics. Its results have the potential to influence clinical cardiac surgical practice in the future.


Aureococcus anophagefferens (Pelagophyceae) genomes improve evaluation of nutrient acquisition strategies involved in brown tide dynamics.
The pelagophyte Aureococcus anophagefferens causes harmful brown tide blooms in marine embayments on three continents. Aureococcus anophagefferens was the first harmful algal bloom species to have its genome sequenced, an advance that evidenced genes important for adaptation to environmental conditions that prevail during brown tides. To expand the genomic tools available for this species, genomes for four strains were assembled, including three newly sequenced strains and one assembled from publicly available data. These genomes ranged from 57.11 to 73.62 Mb, encoding 13,191-17,404 potential proteins. All strains shared ~90% of their encoded proteins as determined by homology searches and shared most functional orthologs as determined by KEGG, although each strain also possessed coding sequences with unique functions. Like the original reference genome, the genomes assembled in this study possessed genes hypothesized to be important in bloom proliferation, including genes involved in organic compound metabolism and growth at low light. Cross-strain informatics and culture experiments suggest that the utilization of purines is a potentially important source of organic nitrogen for brown tides. Analyses of metatranscriptomes from a brown tide event demonstrated that use of a single genome yielded a lower read mapping percentage (~30% of library reads) as compared to a database generated from all available genomes (~43%), suggesting novel information about bloom ecology can be gained from expanding genomic space. This work demonstrates the continued need to sequence ecologically relevant algae to understand the genomic potential and their ecology in the environment.


Development of surgical skill with singular neurectomy using human cadaveric temporal bones.
Profound anatomical knowledge and surgical experience are essential for safe otological surgery. The surgeon's learning curve is evaluated in performing Gacek's singular neurectomy on cadaveric specimens. One otological surgeon performed Gacek's approach on 96 halves of human heads embalmed according to Thiel's method, divided into four groups (24 halves per group) and evaluated them concurrent to the evaluation of an anatomist after a first surgical attempt. Successful operations were subdivided into "direct hits" of the osseous canal of the posterior ampullary nerve also known as the singular nerve and "indirect hits" with access to the posterior ampullary recess. Unsuccessful operations showed "no hit" of the nerve without lesion of the membranous labyrinth. "Indirect" or "no hits" were reinvestigated in a second attempt to evaluate possible reclassifications due to a learning process of the surgeon. The order of dissection, the rate of success and the changes of results in correlation with the numbers of dissected specimens were documented. The success rate significantly increased from 54.2% direct hits after the first group to 87.36% in the fourth group after the first attempt. Successful operations were performed in 86.5% after completion of the first attempt and 97.9% after the second attempt. The number of new allocations decreased from 11 cases in the first group of dissected specimens to zero in the fourth group. This paper strengthens the value of cadaveric training for surgeons and the crucial role of dissection of a large number of specimens in improvement of the surgeon's experience and success rate.


Epitopes of adhesion-perturbing monoclonal antibodies map within a predicted alpha-helical domain of the integrin beta 1 subunit.
Several recent studies have demonstrated the involvement of various domains of the beta 1 integrin subunit in ligand binding. Thus, specific amino acids have been shown to be important in divalent cation binding, and others have been implicated by peptide crosslinking to play an intimate role in integrin-ligand interactions. Added to these data are previous observations that a group of adhesion-blocking anti-chicken beta 1 antibodies mapped within the first 160 amino acid residues of the subunit. These observations suggested that this region plays a critical role in integrin ligand recognition. In order to further define the domain in which the epitopes for these antibodies are clustered, a series of mouse/chicken chimeric beta 1 constructs were examined for their reactivity with each of these antibodies. Most of the antibodies recognize a region between residues 124 to 160 of the chicken beta 1 subunit. Computer modeling predicted a possible amphipathic alpha-helical configuration for the region between residues 141 to 160. Consistent with this prediction, circular dichroism and NMR analysis revealed a tendency for a synthetic peptide containing these residues to form an alpha-helix. The significance of this structural characteristic was demonstrated by a mutation at residue 149 that disrupted the alpha-helix formation and resulted in a loss of the ability to form heterodimers with alpha subunits, localize to focal contacts, or be transported to the cell surface. The direct involvement of residues 141 to 160 in ligand binding was supported by the ability of a peptide with this sequence to elute integrins from a fibronectin affinity column. Thus, our data suggest that residues 141 to 160 of the integrin beta 1 subunit, when arranged in an alpha-helix configuration, participate in ligand binding.


Kinetic modelling and simulation of laccase catalyzed degradation of reactive textile dyes.
A kinetic model based on Michaelis-Menten equation was developed to simulate the dye decolourisation of Reactive Black 5 (RB5), Reactive Blue 114 (RB114), Reactive Yellow 15 (RY15), Reactive Red 239 (RR239) and Reactive Red 180 (RR180) dyes by commercial laccase. The unusual kinetic behavior of some of these reactions suggests that the kinetic model must consider the activation of the laccase-mediator system. Several reactions at different concentrations of each dye were performed in batch reactors and time courses were obtained. A LSODE code to solve the differential equation obtained from the batch reactor was combined with an optimization Fortran program to obtain the theoretical time courses. The time courses obtained from the developed program were compared with the experimentally obtained ones to estimate the kinetic constants that minimized the difference between them. The close correlation between the predicted and the experimental results seems to support the reliability of the established models.


Emx2 and Foxg1 inhibit gliogenesis and promote neuronogenesis.
Neural stem cells (NSCs) give rise to all cell types forming the cortex: neurons, astrocytes, and oligodendrocytes. The transition from the former to the latter ones takes place via lineage-restricted progenitors in a highly regulated way. This process is mastered by large sets of genes, among which some implicated in central nervous system pattern formation. The aim of this study was to disentangle the kinetic and histogenetic roles exerted by two of these genes, Emx2 and Foxg1, in cortico-cerebral precursors. For this purpose, we set up a new integrated in vitro assay design. Embryonic cortical progenitors were transduced with lentiviral vectors driving overexpression of Emx2 and Foxg1 in NSCs and neuronal progenitors. Cells belonging to different neuronogenic and gliogenic compartments were labeled by spectrally distinguishable fluoroproteins driven by cell type-specific promoters and by cell type-specific antibodies and were scored via multiplex cytofluorometry and immunocytofluorescence. A detailed picture of Emx2 and Foxg1 activities in cortico-cerebral histogenesis resulted from this study. Unexpectedly, we found that both genes inhibit gliogenesis and promote neuronogenesis, through distinct mechanisms, and Foxg1 also dramatically stimulates neurite outgrowth. Remarkably, such activities, alone or combined, may be exploited to ameliorate the neuronal output obtainable from neural cultures, for purposes of cell-based brain repair.


HIV-1 restriction by endogenous APOBEC3G in the myeloid cell line THP-1.
HIV-1 replication in CD4-positive T lymphocytes requires counteraction of multiple different innate antiviral mechanisms. Macrophage cells are also thought to provide a reservoir for HIV-1 replication but less is known in this cell type about virus restriction and counteraction mechanisms. Many studies have combined to demonstrate roles for APOBEC3D, APOBEC3F, APOBEC3G and APOBEC3H in HIV-1 restriction and mutation in CD4-positive T lymphocytes, whereas the APOBEC enzymes involved in HIV-1 restriction in macrophages have yet to be delineated fully. We show that multiple APOBEC3 genes including APOBEC3G are expressed in myeloid cell lines such as THP-1. Vif-deficient HIV-1 produced from THP-1 is less infectious than Vif-proficient virus, and proviral DNA resulting from such Vif-deficient infections shows strong G to A mutation biases in the dinucleotide motif preferred by APOBEC3G. Moreover, Vif mutant viruses with selective sensitivity to APOBEC3G show Vif null-like infectivity levels and similarly strong APOBEC3G-biased mutation spectra. Importantly, APOBEC3G-null THP-1 cells yield Vif-deficient particles with significantly improved infectivities and proviral DNA with background levels of G to A hypermutation. These studies combine to indicate that APOBEC3G is the main HIV-1 restricting APOBEC3 family member in THP-1 cells.


Effects of ginger (Zingiber officinale) on plasma glucose level, HbA1c and insulin sensitivity in type 2 diabetic patients.
The present study was aimed to evaluate the effects of Zingiber officinale on some biochemical parameters in type 2 diabetic (DM2) patients. In a randomized double-blind placebo controlled trial, 64 patients with DM2 were assigned to ginger or placebo groups (receiving 2 g/d of each). A 3 d diet record, anthropometric measurements and concentrations of fasting blood glucose (FPG), HbA1c, lipid profile (including total cholesterol, triglyceride, low density lipoprotein and high density lipoprotein) and also the homeostasis model assessment (HOMA) and quantitative insulin-sensitivity check index (QUICKI) were determined before and after 2 months of intervention. Ginger supplementation significantly lowered the levels of insulin (11.0 ± 2.3 versus 12.1 ± 3.3; p = 0.001), LDL-C (67.8 ± 27.2 versus 89.2 ± 24.9; p = 0.04), TG (127.7 ± 43.7 versus 128.2 ± 37.7; p = 0.03) and the HOMA index (3.9 ± 1.09 versus 4.5 ± 1.8; p = 0.002) and increased the QUICKI index (0.313 ± 0.012 versus 0.308 ± 0.012; p = 0.005) in comparison to the control group; while, there were no significant changes in FPG, TC, HDL-C and HbA1c (p > 0.05). In summary, ginger supplementation improved insulin sensitivity and some fractions of lipid profile in DM2 patients. Therefore it may be considered as a useful remedy to reduce the secondary complications of DM2.


Role of regulatory T cells in a new mouse model of experimental autoimmune myositis.
Polymyositis is a rare and severe inflammatory muscle disorder. Treatments are partially efficacious but have many side effects. New therapeutic approaches must be first tested in a relevant animal model. Regulatory CD4+CD25+ T cells (Tregs) have been rediscovered as a pivotal cell population in the control of autoimmunity, but the connection between polymyositis and Tregs is currently unknown. To develop a reproducible experimental autoimmune myositis model of polymyositis, mice were immunized once a week for 3 weeks with 1 mg of partially purified myosin emulsified in complete Freund's adjuvant. All mice injected with myosin and complete Freund's adjuvant developed myositis. The infiltrates were composed of CD4(+) and CD8(+) cells, as well as macrophages, but did not contain B lymphocytes. In mice that were depleted of Tregs, the myositis was more severe, as determined by quantitative scoring of muscle inflammation (2.36 +/- 0.9 vs. 1.64 +/- 0.8, P = 0.019). In contrast, injection of in vitro expanded polyclonal Tregs at the time of immunization significantly improved the disease (quantitative score of inflammation 0.87 +/- 1.06 vs. 2.4 +/- 0.67, P = 0.047). Transfer of sensitized or CD4(+)-sorted cells from the lymph nodes of experimental autoimmune myositis mice induced myositis in naïve, irradiated, recipient mice. Thus, experimental autoimmune myositis is a reproducible, transferable disease in mice, both aggravated by Treg depletion and improved by polyclonal Treg injection.


A genetic analysis of the pteridine biosynthetic enzyme, guanosine triphosphate cyclohydrolase, in Drosophila melanogaster.
Strains with mutant eye color were surveyed for levels of GTP cyclohydrolase (GTP CH), the first enzyme acting in the biosynthesis of pteridines, the pigments causing red eye color in Drosophila. Six strains were found to have reduced GTP CH activity. In five of the six strains, the reduction of activity is apparent only in the adult head of homozygous mutants. We show that mutations in Punch (2-97, Pu) have severe effects on GTP CH activity. In most cases, the reduction of activity is apparent in all tissues and stages that express the enzyme. The activity of GTP CH is shown to be closely correlated with the number of Pu+ genes in the genome. One ethyl methanesulfonate (EMS)-induced Pu mutant has a GTP CH enzyme that is unstable when compared with the wild-type enzyme. Mutations in Pu fall into three general classes. The largest class has a recessive lethal and eye color phenotype, 50% or higher GTP CH activity in heterozygotes, and equivalent defects in all tissues. A second class is dominant in eye color phenotype and recessive lethal, with less than 50% GTP CH activity in heterozygotes. The third class is homozygous viable and has severe reduction of activity in the adult head, but no or less severe loss in other tissues.


Outbreaks of healthcare-associated infections linked to water-containing hospital equipment: a literature review.
Healthcare-associated infections (HAIs) are a significant cause of morbidity and mortality in hospitalized patients. Water in the environment can be a source of infection linked to outbreaks and environmental transmission in hospitals. Water safety in hospitals remains a challenge. This article has summarized available scientific literature to obtain an overview of outbreaks linked to water-containing hospital equipment and strategies to prevent such outbreaks. We made a list of water-containing hospital equipment and devices in which water is being used in a semi-closed circuit. A literature search was performed in PubMed with a search strategy containing the names of these medical devices and one or more of the following words: outbreak, environmental contamination, transmission, infection. For each medical device, we summarized the following information: the function of the medical device, causes of contamination, the described outbreaks and possible prevention strategies. The following water-containing medical equipment or devices were identified: heater-cooler units, hemodialysis equipment, neonatal incubators, dental unit waterlines, fluid warmers, nebulizers, water traps, water baths, blanketrol, scalp cooling, and thermic stimulators. Of the latter three, no literature could be found. Of all other devices, one or more outbreaks associated with these devices were reported in the literature. The water reservoirs in water-containing medical devices can be a source of microbial growth and transmissions to patients, despite the semi-closed water circuit. Proper handling and proper cleaning and disinfection can help to reduce the microbial burden and, consequently, transmission to patients. However, these devices are often difficult to clean and disinfect because they cannot be adequately opened or disassembled, and the manufacturer's cleaning guidelines are often not feasible to execute. The development of equipment without water or fluid containers should be stimulated. Precise cleaning and disinfection guidelines and instructions are essential for instructing healthcare workers and hospital cleaning staff to prevent potential transmission to patients.


Occurrence of and referral to specialists for pain-related diagnoses in First Nations and non-First Nations children and youth.
Indigenous youth have higher rates of chronic health conditions interfering with healthy development, including high rates of ear, dental, chest and musculoskeletal pain, as well as headache, arthritis and mental health issues. This study explores differences in pain-related diagnoses in First Nations and non-First Nations children. Data from a study population of age- and sex-matched First Nations and non-First Nations children and youth were accessed from a specific region of Atlantic Canada. The primary objective of the study was to compare diagnosis rates of painful conditions and specialist visits between cohorts. The secondary objective was to determine whether there were correlations between early physical pain exposure and pain in adolescence (physical and mental health). Although ear- and throat-related diagnoses were more likely in the First Nations group than in the non-First Nations group (ear 67.3% v. 56.8%, p < 0.001; throat 89.3% v. 78.8%, p < 0.001, respectively), children in the First Nations group were less likely to see a relevant specialist (ear 11.8% v. 15.5%, p < 0.001; throat 12.7% v. 16.1%, p < 0.001, respectively). First Nations newborns were more likely to experience an admission to the neonatal intensive care unit (NICU) than non-First Nations newborns (24.4% v. 18.4%, p < 0.001, respectively). Non-First Nations newborns experiencing an NICU admission were more likely to receive a mental health diagnosis in adolescence, but the same was not found with the First Nations group (3.4% v. 5.7%, p < 0.03, respectively). First Nations children with a diagnosis of an ear or urinary tract infection in early childhood were almost twice as likely to have a diagnosis of headache or abdominal pain as adolescents (odds ratio [OR] 1.9, 95% confidence interval [CI] 1.1-3.0, and OR 1.7, 95% CI 1.2-2.3, respectively). First Nations children were diagnosed with more pain than non-First Nations children, but did not access specific specialists or mental health services, and were not diagnosed with mental health conditions, at the same rate as their non-First Nations counterparts. Discrepancies in pain-related diagnoses and treatment are evident in these specific comparative cohorts. Community-based health care access and treatment inquiries are required to determine ways to improve care delivery for common childhood conditions that affect health and development.


Diffusion tensor imaging in the human spinal cord: development, limitations, and clinical applications.
Diffusion tensor imaging (DTI) is currently the only non-invasive in vivo assessment of white matter tract integrity. Capitalizing on the diffusion properties of water within an axon, DTI enables the visualization of tissue structure at a microscopic scale. Furthermore, measurements of anisotropy and diffusivity enable the detection of subtle details of the effects of injury that cannot be detected using conventional magnetic resonance techniques. Recently, DTI has been applied to the spinal cord, and results have demonstrated it to be a valuable tool for assessing the extent of white matter damage in numerous spinal cord-related conditions including multiple sclerosis, spinal cord injury, amyotrophic lateral sclerosis, myelitis, and spinal cord tumors. The purpose of this review is to discuss the technical limitations of the imaging method within the spinal cord, review possible solutions, and highlight the current uses and the potential clinical application of this technique.


Spinal Sagittal Imbalance is Associated with Vertebral Fracture without a Definite History of Falls: Cross-Sectional, Comparative Study of Cohort with and without a Distal Radius Fracture.
Treating osteoporosis in patients with a distal radius fracture (DRF) became paramount at the Fracture Liaison Service. Spinal sagittal imbalance emerged as a risk factor for subsequent fractures. Therefore, here we investigated the spinal profile of patients with DRF to investigate its association with a history of falls and prevalent vertebral fractures. We reviewed the cases of 162 women presenting with DRF and 162 age-matched women without fracture who underwent an osteoporosis evaluation including bone mineral density (BMD) and lateral spine imaging. We compared the incidence of prevalent vertebral fracture and sagittal vertical axis (SVA) to measure spinal sagittal imbalance. We also performed a regression analysis of the risks of prevalent vertebral fracture, such as age, body mass index (BMI), BMD, and SVA. The SVA was significantly smaller (indicating more stable sagittal balance) in patients with a DRF versus controls (16 mm vs. 34 mm, respectively; p<0.001). The incidence of a prevalent vertebral fracture was similar between groups (12% vs. 15%, respectively; p=0.332). In both groups, the SVA was significantly greater in those with versus without a vertebral fracture. The vertebral fracture was significantly associated with age and SVA but not BMI or spinal BMD. Spinal sagittal balance was superior in DRF patients, yet the frequency of prevalent vertebral fractures was similar. The identification of this unique spinal profile in patients with DRF may increase our understanding of osteoporotic fractures.


Detergent-mediated protein aggregation.
Because detergents are commonly used to solvate membrane proteins for structural evaluation, much attention has been devoted to assessing the conformational bias imparted by detergent micelles in comparison to the native environment of the lipid bilayer. Here, we conduct six 500-ns simulations of a system with >600,000 atoms to investigate the spontaneous self assembly of dodecylphosphocholine detergent around multiple molecules of the integral membrane protein PagP. This detergent formed equatorial micelles in which acyl chains surround the protein's hydrophobic belt, confirming existing models of the detergent solvation of membrane proteins. In addition, unexpectedly, the extracellular and periplasmic apical surfaces of PagP interacted with the headgroups of detergents in other micelles 85 and 60% of the time, respectively, forming complexes that were stable for hundreds of nanoseconds. In some cases, an apical surface of one molecule of PagP interacted with an equatorial micelle surrounding another molecule of PagP. In other cases, the apical surfaces of two molecules of PagP simultaneously bound a neat detergent micelle. In these ways, detergents mediated the non-specific aggregation of folded PagP. These simulation results are consistent with dynamic light scattering experiments, which show that, at detergent concentrations ≥600 mM, PagP induces the formation of large scattering species that are likely to contain many copies of the PagP protein. Together, these simulation and experimental results point to a potentially generic mechanism of detergent-mediated protein aggregation.


A retrospective radiographic review of hemophilic shoulder arthropathy.
Bleeding into large joints is the most common orthopaedic manifestation of hemophilia. Involvement of the shoulder rarely is reported, lending to its relative neglect in the orthopaedic literature. Through retrospective chart and radiographic reviews, the incidence and progression of radiographic and clinical changes that occur in the shoulder of patients with hemophilia is described. We retrospectively reviewed 822 patient charts and found 93 patients with symptomatic shoulders. Seventy-nine of these 93 patients were rated according to Pettersson's scoring method. The clinical charts were reviewed for the presence of shoulder symptoms and dysfunction. A spectrum of radiographic changes was seen beginning with mild subchondral irregularity and greater tuberosity cyst formation progressing to joint space narrowing, osteophyte formation, marginal erosion, and deformity. Of the 54 patients for whom an adequate shoulder history was available, the severity of symptoms correlated with the Pettersson score. The current study is the largest to date examining the type and progression of radiographic changes caused by hemophilic shoulder arthropathy. We observed a consistent pattern of radiographic changes, the severity of which correlated directly with shoulder symptoms.


Lewinella litorea sp. nov., isolated from marine sand.
A Gram-stain-negative, aerobic, non-motile and ovoid- or rod-shaped bacterial strain, designated HSMS-39T, was isolated from marine sand sampled at Hongsung, Republic of Korea. Strain HSMS-39T grew optimally at 30 °C and in the presence of 1.0-2.0 % (w/v) NaCl. Phylogenetic trees based on 16S rRNA gene sequences showed that strain HSMS-39T fell within the clade comprising the type strains of Lewinella species, clustering with the type strain of Lewinella marina showing 16S rRNA gene sequence similarity of 99.1 %. It exhibited 16S rRNA gene sequence similarities of less than 95.5 % to the type strains of the other Lewinella species. Strain HSMS-39T contained MK-7 as the predominant menaquinone and iso-C17 : 1ω9c, iso-C15 : 0 and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) as the major fatty acids. The major polar lipids of strain HSMS-39T were phosphatidylethanolamine and one unidentified phospholipid. The DNA G+C content of strain HSMS-39T was 60.0 mol%. The mean DNA-DNA relatedness value between strain HSMS-39T and the type strain of L. marina was 15 %. The average nucleotide identity value between strain HSMS-39Tand the type strain of L. marina was 81.87 %. The phylogenetic and genetic data and differential phenotypic properties indicated that strain HSMS-39T is separated from other recognized species of the genus Lewinella. On the basis of the polyphasic data, strain HSMS-39T is considered to represent a novel species of the genus Lewinella, for which the name Lewinella litorea sp. nov. is proposed. The type strain is HSMS-39T (=KACC 19866T=NBRC 113585T).


Determination of standardized ileal crude protein and amino acid digestibilities in protein supplements for piglets.
The objective of the study was to determine standardized ileal digestibilities (SID) of crude protein (CP) and amino acids (AA) in six ingredients commonly used in weanling pigs diets. The ingredients consisted of extruded soybeans (SBe), high-protein soybean meal (SBMhp), soy protein concentrate (SPC), hydrolyzed wheat gluten (WGh), conventional fish meal 1 (FM1) and extracted fish meal 2 (FM2e). Each assay feed ingredient was added to a purified cornstarch-based basal diet supplemented with casein and crystalline AA, and SID of CP and AA of assay ingredients were estimated using the difference method. A total of 12 3-week-old barrows were surgically fitted with T-cannulas at the distal ileum. The piglets were allocated to four simultaneous 3 × 3 Latin-square designs with three animals and three periods each, resulting in six observations per assay diet. After 3 to 5 days recovery from surgery, each assay diet was fed at a level of 30 g/kg body weight. Each experimental period consisted of 5 days for adaptation to the assay diets and 2 days for ileal digesta collection. The SID of CP, Lys, Met, Thr and Trp were 73%, 78%, 76%, 66% and 71% in SBe; 80%, 84%, 87%, 75% and 81% in SBMhp; 86%, 89%, 89%, 78% and 83% in SPC; 87%, 60%, 88%, 76% and 79% in WGh; 87%, 92%, 94%, 86% and 86% in FM1; and 79%, 86%, 89%, 80% and 74% in FM2e, respectively. The greatest SID of indispensable AA were observed for FM1 and SPC (SID 85% for most indispensable AA), followed by WGh, FM2e and SBMhp (SID 80% for most indispensable AA) and the smallest SID were obtained for SBe (SID < 80% for most indispensable AA). The SID of CP and indispensable AA in both types of fish meal, SBMhp and WGh were similar to SPC (P = 0.063 to 0.855), except for Arg (P = 0.010) and Lys (P = 0.001) in WGh, and Phe (P = 0.044) and Trp (P = 0.037) in FM2e. The SID of CP and indispensable AA were smaller in SBe compared with SPC (P < 0.001 to P = 0.017), except for Lys (P = 0.136). The SID of CP and indispensable AA were consistently smaller in FM2e compared with FM1 with significant differences for CP (P = 0.035), Phe (P = 0.028) and Trp (P = 0.008). Digestibility values measured in the present study can be used to formulate diets for piglets based on standardized ileal digestible contents of CP and AA.


MicroRNA-26a induces osteosarcoma cell growth and metastasis via the Wnt/β-catenin pathway.
MicroRNAs (miRNAs/miRs) are a type of highly conserved, small non-coding RNA that are vital to the post-transcriptional regulation of gene expression via base pairing with target mRNA 3'-untranslated regions (3'-UTRs). Several studies have indicated that the abnormal expression of miRNAs occurs frequently in human osteosarcoma (OS). In the present study, the role of miR-26a in the progression and metastasis of OS was investigated using reverse transcription-quantitative polymerase chain reaction, a luciferase activity assay, cell viability assay, in vitro migration and invasion assays, transfection and western blot analysis. miR-26a was upregulated in OS tissues and cell lines, and the expression of miR-26a was indicated to affect the proliferation, migration and invasion of OS Saos-2 cells. At the molecular level, the results showed that glycogen synthase kinase-3β (GSK-3β) was identified as a target of miR-26a, and the ectopic expression of miR-26a inhibited GSK-3β by directly binding to the 3'-UTR. Therefore, the expression of miR-26a was negatively correlated with GSK-3β in the OS tissues. These data suggest that miR-26a is significant in the proliferation of human OS cells due to the direct regulation of Wnt/β-catenin signaling.


Sudden infant death with periventricular leukomalacia.
Periventricular leukomalacia (PVL) is a form of cerebral infarction occurring in neonates, particularly in low-weight and premature infants. PVL is well-known to neonatologists, but generally considered nonfatal. Many infants with PVL die in the hospital with multiple medical problems. Those infants with PVL who survive because of intensive care will have serious motor and sensory deficits, but these problems are rarely recognized before one year of age. When infants with PVL die at home, death seems sudden and unexpected. However, it is important to distinguish death caused by PVL from the Sudden Infant Death Syndrome because the implications for the family are quite different. This case report emphasizes that PVL may be fatal.


Think fungus NOT just a crypto-meningitis in AIDS!
Extrapulmonary cryptococcosis has been defined as AIDS defining illness in HIV infected people. Cryptococcal meningitis is the commonest meningitis with advanced immune deficiency. Therefore clinicians ask for tests only for detection of cryptococci which may be misleading. A prospective study of suspected fungal meningitis with CSF fungal culture is carried out. 70 ART naive cases of suspected fungal meningitis in HIV cases were subjected to CSF cytochemistry, smear exam and CSF fungal culture. The CSF culture was positive in 75.6% cases of these 21 were C. Neoformans as against 28 of Rhodotorula. In addition candida, aspergillus, geotrichum, trichosporon were isolated. Apart from c. neoformans, other fungi also cause meningitis. Each case of suspected fungal meningitis, may be subjected for CSF fungal culture for proper and adequate management. If facility for fungal culture is not available and if CSF smear shows evidence of fungal infection then standard therapy with Amphotericin may be instituted earlier to reduce mortality. This is the largest series isolating Rhodotorula from CSF in AIDS patients.


Randomised controlled trial to determine the appropriate time to initiate peritoneal dialysis after insertion of catheter to minimise complications (Timely PD study).
The most appropriate time to initiate dialysis after surgical insertion of Tenckhoff catheters is not clear in the literature. There is the possibility of peritoneal dialysis (PD) complications such as leakage and infection if dialysis is started too soon after insertion. However, much morbidity and expense could be saved by reducing dependency on haemodialysis (HD) by earlier initiation of PD post catheter insertion. Previous studies are observational and mostly compare immediate with delayed use. The primary objective is to determine the safest and shortest time interval between surgical placement of a Tenckhoff catheter and starting PD. This is a randomised controlled trial of patients who will start PD after insertion of Tenckhoff catheter at Royal Brisbane and Women's Hospital (RBWH) or Rockhampton Base Hospital (RBH) who meet the inclusion criteria. Patients will be stratified by site and diabetic status. The patients will be randomised to one of three treatment groups. Group 1 will start PD one week after Tenckhoff catheter insertion, group 2 at two weeks and group 3 at four weeks. Nurses and physicians will be blinded to the randomised allocation. The primary end point is the complication rate (leaks and infection) after initiation of PD. The study will determine the most appropriate time to initiate PD after placement of a Tenckhoff catheter. ACTRN12610000076077.


Topically applied anaesthetics for treating perineal pain after childbirth.
Perineal trauma is a major problem affecting millions of women around the world each year. The degree of perineal pain and discomfort associated with perineal trauma is often underestimated. Pain often interferes with basic daily activities for the woman such as walking, sitting and passing urine and also negatively impacts on motherhood experiences. To assess the effects of topically applied anaesthetics for relief of perineal pain following childbirth whilst in hospital and following discharge. We searched the Cochrane Pregnancy and Childbirth Group Trials Register (February 2004), CINAHL (1982 to December 2002) and MIDIRS (last searched February 2003). We checked reference lists of trials and review articles. Randomised controlled trials comparing topically applied anaesthetic with no treatment, placebo or alternative treatment. Two authors independently assessed trial eligibility and quality and double-entered the data. We contacted study authors for additional information. Eight trials made up of 976 women were included in the review. Five of these trials measured pain experienced up to 24 hours after birth but different methods to assess pain were used in each of the studies. All five trials showed no difference in pain relief when the topical anaesthetic was compared with placebo. One of these studies looked at topical anaesthetics compared with indomethacin vaginal suppositories but there was no significant difference in mean pain scores. All trials reported only short-term follow up (up to four days). Two trials looked at additional analgesia taken for perineal pain, with one trial finding that less additional analgesia was required with epifoam use in comparison with placebo (relative risk (RR) 0.58, 95% confidence interval (CI) 0.40 to 0.84, one trial, 97 women). However, lignocaine/lidocaine showed no difference with regard to additional analgesia use. Adverse effects were not formally measured in the studies; however, some studies commented that there were no side-effects severe enough to discontinue treatment. One study found that the women in the treatment group were more satisfied than the placebo group (RR 0.09, 95% CI 0.01 to 0.65, one trial, 103 women). Evidence for the effectiveness of topically applied local anaesthetics for treating perineal pain is not compelling. There has been no evaluation for the long-term effects of topically applied local anaesthetics.


Alzheimer disease identification using amyloid imaging and reserve variables: proof of concept.
Several factors may influence the relationship between Alzheimer disease (AD) lesions and the expression of dementia, including those related to brain and cognitive reserve. Other factors may confound the association between AD pathology and dementia. We tested whether factors thought to influence the association of AD pathology and dementia help to accurately identify dementia of the Alzheimer type (DAT) when considered together with amyloid imaging. Participants with normal cognition (n = 180) and with DAT (n = 25), aged 50 years or older, took part in clinical, neurologic, and psychometric assessments. PET with the Pittsburgh compound B (PiB) tracer was used to measure brain amyloid, yielding a mean cortical binding potential (MCBP) reflecting PiB uptake. Logistic regression was used to generate receiver operating characteristic curves, and the areas under those curves (AUC), to compare the predictive accuracy of using MCBP alone vs MCBP together with other variables selected using a stepwise selection procedure to identify participants with DAT vs normal cognition. The AUC resulting from MCBP alone was 0.84 (95% confidence interval [CI] = 0.73-0.94; cross-validated AUC = 0.80, 95% CI = 0.68-0.92). The AUC for the predictive equation generated by a stepwise model including education, normalized whole brain volume, physical health rating, gender, and use of medications that may interfere with cognition was 0.94 (95% CI = 0.90-0.98; cross-validated AUC = 0.91, 95% CI = 0.85-0.96), an improvement (p = 0.025) over that yielded using MCBP alone. Results suggest that factors reported to influence associations between AD pathology and dementia can improve the predictive accuracy of amyloid imaging for the identification of symptomatic AD.


Predicting everyday functional abilities of dementia patients with the Mini-Mental State Examination.
The Mini-Mental State Examination is a widely used cognitive screening measure. The purpose of the present study was to assess how 5 specific clusters of Mini-Mental State Examination items (ie, subscores) correlate with and predict specific areas of daily functioning in dementia patients, 61 patients with varied forms of dementia were administered the Mini-Mental State Examination and an observation-based daily functional test (the Direct Assessment of Functional Status). The results revealed that the orientation and attention subscores of the Mini-Mental State Examination correlated most significantly with most functional domains. The Mini-Mental State Examination language items correlated with all but the shopping and time orientation tasks, while the Mini-Mental State Examination recall items correlated with the Direct Assessment of Functional Status time orientation and shopping tasks. Stepwise regression analyses found that among the Mini-Mental State Examination subscores, orientation was the single, best independent predictor of daily functioning.


National Trends in the Prevalence, Treatment, and Associated Spinal Diagnoses Among Pediatric Spondylolysis Patients.
Spondylolysis is an increasingly common diagnoses for young individuals and presents with a wide range of pathological and clinical findings. Most patients are treated conservatively, and surgery is reserved for severe cases. This is a populations study defining the incidence of spondylolysis in the Kids' Inpatient Database (KID) and assess trends in diagnoses, causes, and treatments. Retrospective analysis of the prospectively collected information in KID was performed for the years 2003 through 2012. Patients with a diagnosis of spondylolysis (ICD-9-CM 756.11) between the ages of 0 and 20 years in the KID were identified. Incidence of spondylolysis was established using KID-supplied hospital- and year-adjusted trend weights. Demographics including age, race, gender, and Charlson Comorbidity Index were assessed for all spondylolysis patients. Primary outcome measures were yearadjusted and hospital-adjusted incidence of spondylolysis. Secondary outcome measures were concurrent diagnoses and surgical details. Six hundred and sixteen patients with a diagnosis of spondylolysis (329 with primary diagnosis) were identified (female: 53.8%; age: 15.27 ± 3.32 years). The incidence of spondylolysis is 7 per 100,000 patients nationally. Spondylolysis incidence has increased over time (p < 0.001) though the operative rate for spondylolysis has remained the same in the last decade (70% average, p = 0.52). The average CCI is 0.234, the average length of stay is 3.76 days and 92.4% of patients were discharged home. The etiology of the spondylolysis was trauma in 8.6% of patients (3.2% car crash, 1.9% pedestrian, 1.3% fall, 1.3% assault, 1.1% other transport, 1.0% sports, 0.3% motorcycle, 0.2% firearm, 0.2% bicycle; 1.9% reported multiple trauma etiologies). The most common concurrent diagnoses for all spondylolysis patients were spondylolisthesis (28%), idiopathic scoliosis (4.4%), cerebral palsy (1.9%), and spina bifida (1.8%). Four hundred and thirty patients with spondylolysis underwent surgical treatment and 40% of the surgically treated patients had spondylolisthesis. The rate of fusions was 54.9% fusions and 21% decompression, though the rate of fusions or decompressions being performed for spondylolysis has remained the same in the last decade (average fusion rate: 55%; average decompression rate: 18%; both p > 0.05). Levels fused and complications did not differ depending on whether or not decompression was performed (p > 0.05). The posterior-only approach was used in 62.2% of surgeries and were mostly 2 to 3 level procedures (63.5%). Perioperative complications occurred in 8.1% of patients, with the most common complications being device-related (2.3%), respiratory (1.5%), and digestive (1.5%). The national incidence of spondylolysis has increased over time, and the surgical rate and treatment techniques have remained constant. The most common concurrent diagnoses were idiopathic scoliosis, cerebral palsy, and spina bifida. Further work is required to determine the significance of these trends and associations.


Caregiving Time of Unpaid Family Caregivers Assisting Older Adults With Hearing Difficulty.
Objectives: We aimed to investigate the association of older adults' hearing difficulty status with caregiving time. Methods: We used data from two linked surveys of Medicare beneficiaries and family caregivers. Hearing difficulty was defined by hearing aid use and hearing capacity in functional settings. Weighted multivariable linear regression examined the association between hearing difficulty and caregiving time. Stratified analyses were conducted to investigate the moderation effects of caregiving networks and care recipient's dementia status. Results: Among 3003 caregivers, those who assisted older adults with hearing difficulty were observed to spend greater time providing care (β = 1.18, 95% Confidence Interval [CI]: 1.04, 1.32). Stronger associations in magnitude were observed among caregivers without caregiving networks (β = 1.35, 95% CI: 1.13, 1.56) and assisting older adults without dementia (β = 1.22; 95% CI = 1.06, 1.37). Discussion: Hearing difficulty may contribute to greater caregiving time and remains a potential target for caregiver support programs.


[An unexpected stage of alkalosis in the dynamics of the early posthemorrhagic period].
A study was made on acid-base metabolism in early posthemorrhagic period as exemplified by examination of patients presenting with gastrointestinal hemorrhage. It has been ascertained that hemorrhage is accompanied by a mixed variant of the acid-base state (ABS) deviation, namely metabolic lactate-acidosis and respiratory alkalosis. In the time-related course of posthemorrhagic period such deviations persist in patients with lethal outcome; with the disease running a favourable course the above deviations are found to return to normal quite soon. The development of complications leads to staging in ABC, its stages being as follows: stage I--the initial stage, stage II--persisting metabolic acidosis and respiratory alkalosis, stage III--alkalosis, stage IV--normalization, with stage III of ABS being encouraged by hypocapnia caused by function disorders of the lungs in early posthemorrhagic period, normalization of cell metabolism, increase in the rate of urination as a reflection of the third earlier identified stage of water metabolism, with the H+ excretion in the urine at the previous level. The identified ABS stage III threatens coming trouble, being accompanied by metabolic deviations together with a risk of function disorder of the myocardium.


Accuracy of radiologic and endoscopic diagnosis of gastric ulcer.
Patients with upper abdominal pain are often examined with both double contrast study of the stomach and endoscopy. On the basis of the results of the two examinations four diagnostic criteria of an ulcer can be formed: 1) radiography reveals an ulcer, 2) endoscopy reveals an ulcer, 3) both radiography and endoscopy reveal an ulcer, and 4) radiography and/or endoscopy reveals an ulcer. In a prospective study the accuracy of each of the four diagnostic criteria was examined. Eighty-two randomly selected outpatients had a double contrast barium examination and an upper gastrointestinal endoscopy performed by staff personnel. The diagnosis of a specialist in upper gastrointestinal endoscopy was used as the standard. For the four diagnostic criteria the overall accuracy ranged from 0.80 to 0.88. The predictive value of a positive test result was around 0.70 and the predictive value of a negative test result ranged from 0.81 to 0.96. The specificity ranged from 0.87 to 0.95, and the sensitivity from 0.38 to 0.90. It is concluded that from a clinical point of view, the accuracy of the four diagnostic criteria does not differ to an extent that justifies recommendation of one diagnostic criterion of gastric ulcer rather than the other.


Effects of compressive lesions on intraoperative human spinal cord elasticity.
Although evaluating tissue elasticity has various clinical applications, spinal cord elasticity (SCE) in humans has never been well documented. In this study, the authors aimed to evaluate the impact of compression on human SCE in vivo. The authors prospectively assessed SCE using intraoperative shear wave elastography (SWE). All consecutive patients undergoing spinal cord (SC) decompression (laminectomy or corpectomy) between June 2018 and June 2019 were included. After intraoperative exposure of the patient's dura mater, at least three SWE measurements of the SC and its coverings were performed. Intraoperative neurological monitoring in the form of motor and somatosensory evoked potentials was utilized. Cases were divided into two groups based on the state of SC compression following bone removal (laminectomy or corpectomy): patients with adequate decompression (the decompressed SC group [DCG]) following bone removal and patients with remining compression, e.g., compressing tumor or instability (the compressed SC group [COG]). A total of 25 patients were included (8 females and 17 males) with a mean age of 48.28 ± 21.47 years. Most cases were degenerative diseases (10 cases) followed by tumors (6 cases), and the compression was observed at cervical (n = 14), thoracic (n = 9), and conus medullaris (n = 2) levels. The COG (6 cases) expressed significantly higher elasticity values, i.e., greater stiffness (median 93.84, IQR 75.27-121.75 kPa) than the decompressed SC in DCG (median 9.35, IQR 6.95-11.22 kPa, p < 0.001). Similarly, the compressed dura mater in the COG was significantly stiffer (mean ± SD 121.83 ± 70.63 kPa) than that in the DCG (29.78 ± 18.31 kPa, p = 0.042). Following SC decompression in COG, SCE values were significantly reduced (p = 0.006; adjusted for multiple comparisons). Intraoperative monitoring demonstrated no worsening from the baseline. The current study is to the authors' knowledge the first to quantitatively demonstrate increased stiffness (i.e., elasticity value) of the human SC and dura mater in response to external compression in vivo. It appears that SCE is a dynamic phenomenon and is reduced following decompression. Moreover, the evaluation of human SCE using the SWE technique is feasible and safe. Information from future studies aiming to further define SCE could be valuable in the early and accurate diagnosis of the compressed SC.


The utility of cardiac biomarkers and echocardiography for the early detection of bevacizumab- and sunitinib-mediated cardiotoxicity.
The recent introduction of novel anticancer therapies, including bevacizumab (BVZ) and sunitinib (SNT), is associated with an increased risk of cardiotoxicity. However, early identification of left ventricular (LV) systolic dysfunction may facilitate dose modification and avoid the development of advanced heart failure. Using a murine model of BVZ- and SNT-mediated cardiotoxicity, we investigated whether cardiac biomarkers and/or tissue velocity imaging (TVI) using echocardiography can detect early changes in cardiac function, before a decrease in LV ejection fraction is identified. A total of 75 wild-type C57Bl/6 male mice were treated with either 0.9% saline, BVZ, or SNT. Serial monitoring of blood pressure, high-sensitivity troponin I, and echocardiographic indexes were performed over a 14-day study period, after which the mice were euthanized for histological and biochemical analyses. Mice treated with either BVZ or SNT developed systemic hypertension as early as day 7, which increased by day 14. Cardiac biomarkers, specifically high-sensitivity troponin I, were not predictive of early LV systolic dysfunction. Although conventional LV ejection fraction values decreased at day 13 in mice treated with either BVZ or SNT, TVI confirmed early LV systolic dysfunction at day 8. Histological and biochemical analysis demonstrated loss of cellular integrity, increased oxidative stress, and increased cardiac apoptosis in mice treated with BVZ or SNT therapy at day 14. In a murine model of BVZ- or SNT-mediated cardiomyopathy, noninvasive assessment by TVI detected early LV systolic dysfunction before alterations in conventional echocardiographic indexes.


Primary immunodeficiencies.
Primary immunodeficiencies include a variety of disorders that render patients more susceptible to infections. If left untreated, these infections may be fatal. The disorders constitute a spectrum of more than 80 innate defects in the body's immune system. Primary immunodeficiencies generally are considered to be relatively uncommon. There may be as many as 500,000 cases in the United States, of which about 50,000 cases are diagnosed each year. Common primary immunodeficiencies include disorders of humoral immunity (affecting B-cell differentiation or antibody production), T-cell defects and combined B- and T-cell defects, phagocytic disorders, and complement deficiencies. Major indications of these disorders include multiple infections despite aggressive treatment, infections with unusual or opportunistic organisms, failure to thrive or poor growth, and a positive family history. Early recognition and diagnosis can alter the course of primary immunodeficiencies significantly and have a positive effect on patient outcome.


Effect of growth environment on spatial expression of involucrin by human epidermal keratinocytes.
Involucrin, the major protein precursor of the cornified envelope, is expressed during terminal differentiation of human keratinocytes, both in vivo and in vitro. In epidermis, the onset of synthesis is several layers above the basal layer, but in stratified cultures of keratinocytes on tissue culture plastic involucrin synthesis begins in the first suprabasal layer. To investigate the reason for this premature expression, the distribution of involucrin was studied in epidermis from different body sites, in organotypic cultures and in transplants of keratinocytes onto nude mice. We found that premature expression was not associated with poor morphological differentiation, because involucrin synthesis began immediately above the basal layer even when distinct basal, spinous, granular and cornified layers were formed in organotypic cultures recombined with dermis. The site of involucrin expression in culture did not depend on the number of cornified layers present. The only conditions which resulted in an upward shift in the site of synthesis were in 3-week old transplants on nude mice. We conclude that the site of onset of involucrin synthesis is not determined by the degree of morphological differentiation of the tissue, and discuss other factors which may be involved.


The hrp genes of Pseudomonas cichorii are essential for pathogenicity on eggplant but not on lettuce.
Pseudomonas cichorii causes necrotic lesions in eggplant and rot in lettuce. Through transposon insertion into P. cichorii strain SPC9018 we produced two mutants, 4-57 and 2-99, that lost virulence on eggplant but not lettuce. Analyses showed that a transposon was inserted into the hrpG gene in 4-57 and the hrcT gene in 2-99. Nucleotide sequences of the hrp genes of SPC9018 are homologous to those of Pseudomonas viridiflava BS group strains. The pathogenicity of 4-57 on eggplant was restored by transformation with an hrpF operon, originating from either SPC9018 or the BS group member P. viridiflava strain 9504 (Pv9504). These data suggested the involvement of hrp genes in the pathogenicity of SPC9018 on eggplant, and functional conservation of hrpF operons between SPC9018 and Pv9504. Both the hrpS mutant and the hrpL mutant were unable to cause necrotic lesions on eggplant leaves but retained their pathogenicity against lettuce. These results suggest that the pathogenicity of P. cichorii is hrp-dependent in eggplant, but not in lettuce.


Understanding impacts of zoo visitors: Quantifying behavioural changes of two popular zoo species during COVID-19 closures.
Visitors are normally a prominent and constant feature in a zoo animals' environment with more than 700 million people visiting zoos and aquariums worldwide, annually. Animal-visitor interactions can be enriching and stimulating and are now considered within the Five Domains of animal welfare assessment. Zoo closures as a result of COVID-19 provided a unique opportunity to monitor the impact of abrupt and prolonged removal of visitors on two popular zoo species. Data were collected at four facilities (n = 3 slender-tailed meerkats, n = 1 African penguin) during COVID-19 zoo closures and up to one month following reopening to the public. Meerkats showed increased positive social interactions, increased alert behaviours, and reduced environmental interactions in the first month post-opening, as compared to closure periods. They also used more of their enclosures during periods of closure and spent longer than would be expected in zones furthest from visitor viewing areas when facilities reopened. African penguins showed no behavioural change between open and closure periods. Enclosure usage during both observation periods was relatively even and no differences were observed in enclosure use between open and closure periods. These results will enable an advanced understanding of the impact that people have on the behaviour of zoo animals, which has ramifications for animals used in close encounters and other 'visitor experiences' in the future. Understanding relationships between animals and people is applicable in all managed animal settings. The results from this study are of practical use in managing visitor access to animals moving forwards, including enclosure location and design, to ensure a positive visitor experience that does not negatively impact animal behaviour.


FIGO good practice recommendations on the use of prenatal corticosteroids to improve outcomes and minimize harm in babies born preterm.
For women with a singleton or a multiple pregnancy in situations where active neonatal care is appropriate, and for whom preterm birth is anticipated between 24 and 34 weeks of gestation, one course of prenatal corticosteroids should ideally be offered 18 to 72 h before preterm birth is expected to improve outcomes for the baby. However, if preterm birth is expected within 18 h, prenatal corticosteroids should still be administered. One course of corticosteroids includes two doses of betamethasone acetate/phosphate 12 mg IM 24 h apart, or two doses of dexamethasone phosphate 12 mg IM 24 h apart. In women in whom preterm birth is expected within 72 h and who have had one course of corticosteroids more than a week previously, one single additional course of prenatal corticosteroids could be given at risk of imminent delivery. Prenatal corticosteroids should not be offered routinely to women in whom late preterm birth between 34 and 36 weeks is anticipated. In addition, prenatal corticosteroids should not be given routinely before cesarean delivery at term. Neither should prenatal corticosteroids be given "just in case". Instead, prenatal steroid administration should be reserved for women for whom preterm birth is expected within no more than 7 days, based on the woman's symptoms or an accurate predictive test.


miR-31-NUMB Cascade Modulates Monocarboxylate Transporters to Increase Oncogenicity and Lactate Production of Oral Carcinoma Cells.
Oral squamous cell carcinoma (OSCC) is among the leading causes of cancer-associated death worldwide. miR-31 is an oncogenic miRNA in OSCC. NUMB is an adaptor protein capable of suppressing malignant transformation. Disruption of the miR-31-NUMB regulatory axis has been demonstrated in malignancies. Mitochondrial dysfunction and adaptation to glycolytic respiration are frequent events in malignancies. Monocarboxylate transporters (MCTs) function to facilitate lactate flux in highly glycolytic cells. Upregulation of MCT1 and MCT4 has been shown to be a prognostic factor of OSCC. Here, we reported that miR-31-NUMB can modulate glycolysis in OSCC. Using the CRISPR/Cas9 gene editing strategy, we identified increases in oncogenic phenotypes, MCT1 and MCT4 expression, lactate production, and glycolytic respiration in NUMB-deleted OSCC subclones. Transfection of the Numb1 or Numb4 isoform reversed the oncogenic induction elicited by NUMB deletion. This study also showed, for the first time, that NUMB4 binds MCT1 and MCT4 and that this binding increases their ubiquitination, which may decrease their abundance in cell lysates. The disruptions in oncogenicity and metabolism associated with miR-31 deletion and NUMB deletion were partially rescued by MCT1/MCT4 expression or knockdown. This study demonstrated that NUMB is a novel binding partner of MCT1 and MCT4 and that the miR-31-NUMB-MCT1/MCT4 regulatory cascade is present in oral carcinoma.


Simulation of tremor on 3-dimentional musculoskeletal model of wrist joint and experimental verification?
A musculoskeletal model that allows to simulate the tremor of wrist joint with three degrees of freedom (DoFs) is developed. The model includes five muscles, extensor carpi radialis brevis, extensor carpi radialis longus, extensor carpi ulnaris, flexor carpi radialis and flexor carpi ulnaris. Simulation of tremor generation based on the 3-DoF model is performed. The tremor disorder can be generated in two directions: flexion-extension and radia-ulnar deviation. Accordingly, experiment is conducted on healthy subjects to verify the feasibility of artificial tremor generation via functional electrical stimulation (FES). Simulation results have shown qualitative agreement with the experimental results.


Circumferential tensile stiffness of glaucomatous trabecular meshwork.
Our previous work indicated that a larger circumferential Young's modulus (E) of trabecular meshwork (TM) correlated with a higher outflow facility (C) in normal human donor eyes. The current study investigated the influence of glaucomatous TM stiffness and cellularity on C. Eight left eyes from glaucomatous human donors were perfused within 48 hours post mortem. Values of C were determined at pressures of 10, 20, 30, and 40 mm Hg. The TM was then dissected and imaged with optical coherence tomography to determine its cross-sectional area. Uniaxial tensile stress was applied longitudinally to TM segments to determine stress-strain curves. E was calculated at zero strain, representing the circumferential stiffness of the TM at a relaxed state. Confocal images of DAPI-stained TM segments were used to determine cellularity after mechanical stretching. C (μL/min/mm Hg) of glaucomatous eyes was 0.18 ± 0.02 (mean ± SE) at 10 mm Hg and decreased to 0.11 ± 0.02 when the pressure was increased to 40 mm Hg. C measured at 30 and 40 mm Hg correlated with TM cellularity. E was 12.5 MPa and 1.4 (geometric mean and SE) and did not statistically correlate with postmortem time, age, C, or cellularity. Compared with data of normal eyes observed in a previous study, C in glaucomatous eyes was reduced significantly, and the amount of reduction increased with increasing the pressure. E of glaucomatous TM was approximately one-fifth that of normal TM. Prospective studies are needed to further investigate the influence of TM tensile stiffness on outflow regulation.


Role of IL28B gene polymorphism and cell-mediated immunity in spontaneous resolution of acute hepatitis C.
A single-nucleotide polymorphism (SNP; rs12979860) near the IL28B gene has been associated with spontaneous and treatment-induced hepatitis C virus clearance. We investigated predictors of spontaneous disease resolution in a cohort of patients with acute hepatitis C (AHC), analyzing epidemiological, clinical and virological parameters together with IL28B.rs12979860 genotypes and cell-mediated immunity (CMI). Fifty-six symptomatic AHC patients were enrolled and followed prospectively. CMI was measured in 31 patients at multiple time points by interferon-γ enzyme-linked immunospot assay and was correlated to the IL28B.rs12979860 SNP. Eighteen patients had a self-limiting AHC that was associated with female sex (P = .028), older age (P = .018), alanine aminotransferase level >1000 U/L (P = .027), total bilirubin level >7 mg/dL (P = .036), and IL28B.rs12979860 genotype CC (P = .030). In multivariate analysis, only CC genotype was independently associated with self-limiting AHC (odds ratio, 5.3; 95% confidence interval, 1.1-26.5). Patients with the CC genotype with self-limiting AHC had a stronger (P = .02) and broader (P = .013) CMI than patients with the CT genotype with chronically evolving AHC. In patients with chronically evolving disease, CC genotype was associated with a broader CMI compared to CT genotype (P = .028). A negative CMI was more frequently associated with CT genotype among persistently infected patients (P = .043) and with persistent infection among CT patients (P = .033). . Self-limiting AHC was independently associated with CC genotype. The correlation between IL28B.rs12979860 genotypes and CMI is suggestive of a possible important role of CMI in favoring hepatitis C virus clearance in CC patients.


Mice doubly deficient for the Polycomb Group genes Mel18 and Bmi1 reveal synergy and requirement for maintenance but not initiation of Hox gene expression.
Polycomb group genes were identified as a conserved group of genes whose products are required in multimeric complexes to maintain spatially restricted expression of Hox cluster genes. Unlike in Drosophila, in mammals Polycomb group (PcG) genes are represented as highly related gene pairs, indicative of duplication during metazoan evolution. Mel18 and Bmi1 are mammalian homologs of Drosophila Posterior sex combs. Mice deficient for Mel18 or Bmi1 exhibit similar posterior transformations of the axial skeleton and display severe immune deficiency, suggesting that their gene products act on overlapping pathways/target genes. However unique phenotypes upon loss of either Mel18 or Bmi1 are also observed. We show using embryos doubly deficient for Mel18 and Bmi1 that Mel18 and Bmi1 act in synergy and in a dose-dependent and cell type-specific manner to repress Hox cluster genes and mediate cell survival of embryos during development. In addition, we demonstrate that Mel18 and Bmi1, although essential for maintenance of the appropriate expression domains of Hox cluster genes, are not required for the initial establishment of Hox gene expression. Furthermore, we show an unexpected requirement for Mel18 and Bmi1 gene products to maintain stable expression of Hox cluster genes in regions caudal to the prospective anterior expression boundaries during subsequent development.


[Delayed diagnosis of HIV infection in the Spanish VACH cohort [1997-2002]].
To study the prevalence of delayed diagnosis of HIV infection and associated factors. A cross sectional study of patients included in the Spanish VACH cohort who had been diagnosed with HIV infection between 1997 and 2002 was performed. Delayed diagnosis was defined as patients diagnosed with HIV infection and AIDS simultaneously or within the first month after the first positive serologic test, or those with a first CD4+ cell count below 200/ml. The epidemiological characteristics of these patients were compared with those of the remaining patients Of 2,820 new cases of HIV infection, delayed diagnosis was found in 506 (18%). These patients differed from the remaining patients in their lower mean age and higher HIV viral load, as well as in their distribution by sex (higher proportion of males), occupational status, history of incarceration in prison, and HIV-risk transmission group. The median survival during follow-up was significantly lower among AIDS patients with a delayed diagnosis. Delayed diagnosis remains a cause for concern in our environment, due to its magnitude and its association with mortality. Some epidemiological characteristics provide clues to guide future programs directed at increasing information and improving prevention.


Genetic Mapping by Bulk Segregant Analysis in Drosophila: Experimental Design and Simulation-Based Inference.
Identifying the genomic regions that underlie complex phenotypic variation is a key challenge in modern biology. Many approaches to quantitative trait locus mapping in animal and plant species suffer from limited power and genomic resolution. Here, I investigate whether bulk segregant analysis (BSA), which has been successfully applied for yeast, may have utility in the genomic era for trait mapping in Drosophila (and other organisms that can be experimentally bred in similar numbers). I perform simulations to investigate the statistical signal of a quantitative trait locus (QTL) in a wide range of BSA and introgression mapping (IM) experiments. BSA consistently provides more accurate mapping signals than IM (in addition to allowing the mapping of multiple traits from the same experimental population). The performance of BSA and IM is maximized by having multiple independent crosses, more generations of interbreeding, larger numbers of breeding individuals, and greater genotyping effort, but is less affected by the proportion of individuals selected for phenotypic extreme pools. I also introduce a prototype analysis method for simulation-based inference for BSA mapping (SIBSAM). This method identifies significant QTL and estimates their genomic confidence intervals and relative effect sizes. Importantly, it also tests whether overlapping peaks should be considered as two distinct QTL. This approach will facilitate improved trait mapping in Drosophila and other species for which hundreds or thousands of offspring (but not millions) can be studied.


The automatic implantable defibrillator. New Modality for treatment of life-threatening ventricular arrhythmias.
The automatic implantable defibrillator continuously monitors cardiac rhythm, identifies ventricular fibrillation and then delivers corrective defibrillatory discharges when indicated; it weighs 250 grams and has a volume of 145 cc. When a suitable arrhythmia is detected, a 25 Joule pulse is delivered through a superior vena cava catheter electrode and another electrode placed over the cardiac apex. As of March 1981, sixteen survivors of multiple cardiac arrests refractory to antiarrhythmic therapy had undergone implantation of the automatic defibrillator. There was no operative mortality and the morbidity was minimal. Electrophysiologic studies were performed before and after surgery to confirm failure of drug therapy and to ensure the device's ability to terminate malignant arrhythmias. Eight spontaneous and fourteen of the seventeen induced malignant arrhythmias were properly recognized and corrected by the device. The discharges were well tolerated by awake patients. A number of problems including recycling delays and spurious discharges have been identified and corrected. There were three late deaths with pulmonary edema noted in two patients, and asystole in one. The autopsies revealed no myocardial damage attributable to the automatic defibrillator. Although the ultimate role of this approach to prevention of sudden arrhythmic death has yet to be determined, the results obtained to date are encouraging and indicate that a useful modality for treating malignant ventricular arrhythmias has been added to our armamentarium.