Phosphatidylinositol-3 kinase signaling controls survival and stemness of hematopoietic stem and progenitor cells.
Hematopoietic stem and progenitor cells (HSPCs) are multipotent cells giving rise to all blood lineages during life. HSPCs emerge from the ventral wall of the dorsal aorta (VDA) during a specific timespan in embryonic development through endothelial hematopoietic transition (EHT). We investigated the ontogeny of HSPCs in mutant zebrafish embryos lacking functional pten, an important tumor suppressor with a central role in cell signaling. Through in vivo live imaging, we discovered that in pten mutant embryos a proportion of the HSPCs died upon emergence from the VDA, an effect rescued by inhibition of phosphatidylinositol-3 kinase (PI3K). Surprisingly, inhibition of PI3K in wild-type embryos also induced HSPC death. Surviving HSPCs colonized the caudal hematopoietic tissue (CHT) normally and committed to all blood lineages. Single-cell RNA sequencing indicated that inhibition of PI3K enhanced survival of multipotent progenitors, whereas the number of HSPCs with more stem-like properties was reduced. At the end of the definitive wave, loss of Pten caused a shift to more restricted progenitors at the expense of HSPCs. We conclude that PI3K signaling tightly controls HSPCs survival and both up- and downregulation of PI3K signaling reduces stemness of HSPCs.


Decreasing incidence rates of osteoporotic hip fractures in Ecuador during the COVID-19 pandemic.
We aimed at comparing the incidence of hip fractures in older adults from Ecuador before and during the COVID-19 pandemic. There was a significant reduction in the number of hip fractures, with no change in the length of hospital stay, mortality, and case-fatality rate, during the period of social isolation. The impact that the COVID-19 pandemic has had on fragility fractures is being recently evaluated in the literature. Despite this, data from Latin America in this regard is scarce. This study aims to compare the incidence rate of hip fractures before and during the COVID-19 pandemic in older adults who received care in the public and private health system of Ecuador. This was a descriptive and retrospective study that analyzed data of individuals aged 60 years and older who had hip fractures before and during the COVID-19 pandemic. The information was obtained from the National Hospital Discharge Yearbook. We calculated the incidence, average length of hospital stay, mortality, and case-fatality rate associated with hip fractures. There was a significant reduction in the incidence of hip fractures in adults 60 or older during the period of social isolation due to COVID-19. Between March and December 2019, there was an incidence of 152 hip fractures per 100,000 inhabitants, whereas during the same period but in 2020 in the incidence was 110 per 100,000 inhabitants (p < 0.0001). The main decrease was observed in women aged 80 or more. The average length of hospital stay did not show significant changes. Mortality displayed a non-significant decrease (p = 0.14), although this decrease was significant among women (p = 0.02). Case-fatality rate showed a non-significant increase for the whole group (p = 0.68) and for men (p = 0.09). Hip fracture rates decreased significantly in adults aged 60 and older in 2020 compared to 2019. This decrease of hip fracture incidence rates was mainly due to the reductions observed in older people and women. The average length of hospital stay, mortality, and case-fatality rate associated with hip fractures did not show significant changes during the pandemic.


High-sensitivity nanometer-scale infrared spectroscopy using a contact mode microcantilever with an internal resonator paddle.
Infrared (IR) spectroscopy is one of the most widely used techniques for identifying and characterizing materials, but is diffraction limited to a spatial resolution of no smaller than several micrometers. This paper reports IR spectroscopy with 100 nm spatial resolution, using a tunable laser whose absorption in an organic layer is measured via atomic force microscopy. Wavelength-dependent absorption in the sample results in local thermomechanical deformation, which is sensed using the sharp tip of a resonant atomic force microscope cantilever. We introduce a cantilever and system design capable of 100 nm spatial resolution and a 6 x sensitivity improvement over previous approaches.


Lung cinescintigraphy in the dynamic assessment of ventilation and mucociliary clearance of asbestos cement workers.
To verify in vivo whether lung cinescintigraphy confirms the effect of asbestos on the patency of the smallest airways and on the efficiency of mucociliary clearance in asbestos cement workers. 39 male subjects were examined: 30 asbestos cement workers and nine workers never exposed to occupational respiratory irritants. All subjects had a chest radiograph (International Labour Organisation (ILO) 1980); standard questionnaire on chronic bronchitis; spirometry; arterial blood gas analysis; carbon monoxide transfer factor (TLcosb); pulmonary O2 and CO2 ductances (DuO2, DuCO2); electrocardiogram; and lung cinescintigraphy after radioaerosol inhalation for the measurement of mucociliary clearance time in vivo in the smallest ciliated airways and for the assessment of radioaerosol deposition in alveoli (alveolar deposition index). Apart from nine non-exposed subjects, the 30 asbestos cement workers were so classified on the basis of chest radiography: nine of them as healthy exposed, 10 with pleural plaques, and 11 with asbestosis. The four groups had similar ages, work seniority, and smoking habits. Exercise dyspnoea was significantly more frequent in asbestos cement workers. Lung function variables of workers with effects related to asbestos were significantly lower than the other two groups. The PaO2, TLcOsb and DuO2 mean values were significantly lower in exposed workers than non-exposed. The mean PacO2 value was significantly higher in the asbestosis group than in the other three groups. Workers with effects related to asbestos showed a significantly lower alveolar deposition index and a significantly higher mucociliary clearance time than the other two groups. Subjects with asbestosis showed similar differences from those with pleural plaques. Lung cinescintigraphy confirms in vivo the effects of asbestos on bronchiolar and alveolar patency and on efficiency of mucociliary clearance in the smallest ciliated airways. Finally, lung cinescintigraphic variables are able to discriminate workers with asbestosis from those with pleural plaques.


Protocol for a randomized controlled trial comparing the Circle of Security-parenting (COS-P) with treatment as usual in child mental health services.
The quality of a child's attachment to its primary caregiver plays an important role for its long-term socioemotional development. While 'secure' attachment is associated with better outcomes, 'insecure' attachment is associated with a higher risk of externalizing and internalizing symptoms. Children referred to mental health services show much higher rates of insecure attachment than the general population, yet the parent-child relationship is rarely in treatment focus. Attachment quality is closely associated with parental sensitive responsiveness that is target of attachment-based interventions like Circle of Security (COS). COS has shown to improve attachment quality and the well-being of both children and parents. No randomized controlled trials have investigated the effect of COS on parental sensitivity and child psychiatric symptoms in child mental health services. To investigate whether COS-Parenting (COS-P) can increase observed maternal sensitivity and decrease children's psychiatric symptoms as an add on to treatment as usual (TAU). In a randomized controlled parallel superiority trial COS-P is compared with TAU for parents of children referred to child mental health services (n = 186). Families are randomized 2:1 to intervention or control group, if their child is between 3 and 8 years old and scores ≥ 93d percentile on both the CBCL total score and the oppositional defiant disorder or conduct disorder subscale. Primary outcome is maternal sensitivity, secondary and exploratory outcomes include, among others, child psychiatric symptoms, parental stress and coping with children's negative emotions. Outcomes and adverse events are assessed before (T0) and after 10 weeks of treatment (T1) and 6 months later (T2). Regression analysis and /or ANOVA will be used for all outcomes. Targeting the parent-child relation has the potential to reduce psychiatric symptoms in children. This trial will provide valuable information if attachment-based interventions like COS-P can enhance treatment as usual in child mental health services. ClinicalTrials.gov Identifier: NCT03578016.


Overview of Research on Germline Genetic Variation in Immune Genes and Cancer Outcomes.
Since the late 19th century, the immune system has been known to play a role in cancer risk, initiation, and progression. Genome-wide association studies (GWAS) have identified hundreds of genetic risk loci for autoimmune and inflammatory diseases, yet the connection between human genetic variation and immune-mediated response to cancer treatments remains less well-explored. Understanding inherited genetic variation, with respect to germline genetic polymorphisms that affect immune system pathways, could lead to greater insights about how these processes may best be harnessed to successfully treat cancer. Our goal in this manuscript was to understand progress and challenges in assessing the role of inherited genetic variation in response to cancer treatments. Overall, the 39 studies reviewed here suggest that germline genetic variation in immune system-related genes may potentially affect responses to cancer treatments. Although further research is needed, considering information on germline immune genetic variation may help, in some cases, to optimize cancer treatment.


Household characteristics associated with rodent presence and Leptospira infection in rural and urban communities from Southern Chile.
Rodents are well-recognized reservoirs of Leptospira, contributing to its maintenance in endemic areas and playing a role in the public health risk associated with the infection. This study sought to provide some insights into rodent populations from Chile and their Leptospira carriage. In total, 393 rodents were trapped in 177 households. Higher rodent counts were associated with year 2 of the study, rainfall, and number of rodent signs. There was an inverse correlation with the number of cats. The number of rodents was higher in villages compared with slums (rate ratio = 3.23) but modified by average household age. Eighty rodents (20.4%) tested positive for Leptospira: 19.7% on the farms, 25.9% in villages, and 12.3% in the slums. Prevalence was 22.5% in Mus musculus, 20.7% in Rattus rattus, 21.1% in wild rodents, and 10.3% in R. norvegicus. Seasonal and temporal effects were the major determinants of Leptospira infection in rodent populations.


Be careful with your principal components.
Principal components analysis (PCA) is a common method to summarize a larger set of correlated variables into a smaller and more easily interpretable axes of variation. However, the different components need to be distinct from each other to be interpretable otherwise they only represent random directions. This is a fundamental assumption of PCA and, thus, needs to be tested every time. Sample correlation matrices will always result in a pattern of decreasing eigenvalues even if there is no structure. Tests are, therefore, needed to discern real patterns from illusionary ones. Furthermore, the loadings of the vectors need to be larger than expected by random data to be useful in the calculation of PC-scores. PC-scores calculated from nondistinct PC's have very large standard errors and cannot be used for biological interpretations. I give a number of examples to illustrate the potential problems with PCA. Robustness of the PC's increases with increasing sample size but not with the number of traits. I review a few simple test statistics appropriate for testing PC's and use a real-world example to illustrate how this can be done using randomization tests. PCA can be very useful but great care is needed to avoid spurious results.


Psychological interventions that target sleep reduce pain catastrophizing in knee osteoarthritis.
Pain catastrophizing is a significant risk factor for patients with knee osteoarthritis (KOA) and thus is a target for many psychological interventions for pain. This study examined if interventions targeting sleep found to be effective in improving sleep in KOA also reduce pain catastrophizing measured as a trait through the pain catastrophizing scale and measured as a daytime and nocturnal state through daily diaries. Secondary analyses were conducted on data collected as part of a randomized controlled trial assessing the effectiveness of cognitive behavioral therapy for insomnia in patients with KOA at 5 different time points: pretreatment, midtreatment and posttreatment and at 3- and 6-month follow-up. One hundred patients diagnosed with KOA and insomnia were randomized to receive either 8 sessions of cognitive behavioral therapy for insomnia or a placebo intervention of behavioral desensitization. Multilevel modeling revealed that both intervention groups showed a significant reduction pretreatment to posttreatment in all 3 measures of pain catastrophizing and maintained stable levels through the 6-month follow-up. Increased sleep continuity early in treatment (pretreatment to midtreatment), but not reductions in pain, was associated with a reduction in trait and nocturnal catastrophizing later in treatment (midtreatment to posttreatment). These results suggest that short interventions focusing on sleep can significantly reduce pain catastrophizing even in a clinical population with low baseline levels of catastrophizing, possibly through improving sleep continuity.


Primary ovarian insufficiency: a toolkit for the busy clinician.
Primary ovarian insufficiency (sometimes known as premature ovarian insufficiency) is a result of loss of ovarian follicular activity before the age of 40 years. It is an endocrine deficiency state in women, characterized by premature estrogen deprivation. In the absence of estrogen replacement, women experience bothersome menopause symptoms and a predisposition to accelerated aging and multimorbidity accumulation. Unless a true contraindication exists, estrogen therapy is recommended at least until the age of natural menopause. This Practice Pearl summarizes the clinical manifestations, diagnostic evaluation, and management of primary ovarian insufficiency.


Resistance to peer influence and crime desistance in emerging adulthood: A moderated mediation analysis.
The current study sought to determine whether resistance to peer pressure contributes to crime desistance by inhibiting proactive criminal thinking (moral disengagement), a social-cognitive variable that has been found to mediate the peer influence effect. Using data from the Pathways to Desistance study (N = 1,354), resistance to peer influence and peer delinquency were tested as predictors of subsequent offending when participants were 18, 19, and 20 years of age. Whereas peer delinquency (PD) facilitated future offending by increasing moral disengagement (MD), peer resistance inhibited future offending by decreasing moral disengagement. There were no significant differences between the two indirect effects. although only PD achieved a direct effect on future offending above and beyond the indirect effect of the PD → MD → future offending pathway. Peer resistance and PD were found to interact positively with each other in support of a moderated mediation hypothesis in which the inhibitory effects of resistance on moral disengagement were strongest at lower levels of PD. This indicates that juvenile offenders have the greatest chance of desisting from crime when they have fewer delinquent friends and stronger peer resistance skills, for even nondelinquent friends can provide occasional opportunities for crime. These results suggest that peer resistance training may be an effective adjunct in the treatment of juvenile offenders. (PsycINFO Database Record (c) 2018 APA, all rights reserved).


[Use of antibiotics among Danish children in 1998. Consumption on a national basis].
The aim was to evaluate the use of antibiotics (AB) among Danish children in 1998. Patient specific data was collected on antibacterials for systemic use (ATC-group J01) prescribed for children aged 0-15 years in 1998. Data included a total of 585,491 prescriptions for 331,979 individuals. AB were prescribed for 33% of all Danish children on one or several occasions in 1998. Younger children were more likely to be treated with AB than older children. The one-year-prevalence of the use of AB was 63.2% for the 1-year-olds and 18.3% for 12-year-old children in 1998. The type of AB used varied with age, with younger children more prone to receive broad-spectrum AB. Eighty-three percent of all AB-prescriptions were prescribed by general practitioners. It is concluded, that AB, especially the broad-spectrum AB, are more often prescribed for younger than for older children and adults in Denmark in 1998.


"Nonspecific" increases in plasma immunoreactive calcitonin in healthy individuals: discrimination from medullary thyroid carcinoma by a new extraction technique.
Occasional seemingly healthy individuals have above-normal concentrations of calcitonin-like immunoreactivity in their plasma, which can lead to mistaken diagnosis of thyroidal or other cancer. We measured immunoreactive calcitonin (CT) before and after extracting the plasma on columns of silica (to improve sensitivity and specificity of the assay for monomeric calcitonin) in five "healthy high-CT" men (I), five patients with known medullary thyroid carcinoma (II), and 30 normal controls (III). Median (and range) values (pg/mL = ng/L) for whole-plasma immunoreactive CT in these groups were, respectively, 379 (157-526), 429 (174-563), and 33 (less than 25-92). Dose-dilution curves for plasma samples from group I did not parallel the standard curve, in contrast to samples from the other two groups. Values for extractable CT from plasma from groups I and III, however, were indistinguishable, but remained significantly increased in group II. Infusions of Ca, 2 mg/kg body wt. in 5 min, produced the expected (normal) increases in extractable CT in group I. The occasional factor (or factors) in plasma of healthy persons that interferes in assays for CT is eliminated by the silica extraction method, and in this way such cases can be distinguished from cases of medullary thyroid carcinoma.


[Deficiency of the fatty-acid oxidising enzyme medium-chain acyl-CoA dehydrogenase (MCAD) in an adult, detected during a neonatal screening programme].
In a trial running since October 2003 in the Dutch provinces of Friesland, Groningen, Drenthe and Overijssel neonatal screening for medium-chain acyl-CoA dehydrogenase (MCAD) deficiency has been added to the regular newborn screening programme for phenylketonuria, congenital hypothyroidism and adrenogenital syndrome. One of the questions to be answered by this trial is the cause of the strong variation in clinical expression of the disorder. Underdiagnosing is an important factor in this phenomenon, as shown by the data of a family of which the case histories of the two oldest children were discussed in this journal in 1965. Both children died at a very young age. Recently, MCAD deficiency was diagnosed in the youngest child of this family, now a 34-year-old woman. This family history illustrates the variable clinical expression of MCAD deficiency, which can cause death but can also run a milder or even subclinical course. Moreover, this family history shows that the underdiagnosis of MCAD deficiency in deceased children may be a cause of the apparently limited clinical detection rate of this disease, for which a simple treatment consisting of life-style and dietary measures is available after diagnosis.


The functional domains of the murine Thy-1 gene promoter.
The Thy-1 gene promoter resembles a "housekeeping" promoter in that it is located within a methylation-free island, lacks a canonical TATA box, and displays heterogeneity in the 5'-end termini of the mRNA. Using transgenic mice, we show that this promoter does not confer any tissue specificity and is active only in a position-dependent manner. It can only be activated in a tissue-specific manner by elements that lie downstream of the initiation site. We have analyzed the functional domains of the minimal Thy-1 promoter and show that the dominant promoter elements consist of multiple binding sites for the transcription factor Sp1, an inverted CCAAT box, and sequences proximal to the transcription start site. DNase I and gel mobility shift assays show the binding of a number of nuclear factors to these elements, including Sp1 and CP1. Our results show that the structure of this promoter only permits productive interactions of the two transcription factors Sp1 and CP1 with the basal transcription machinery in the presence of enhancer sequences.


Social coding of the multisensory space around us.
Multisensory integration of stimuli occurring in the area surrounding our bodies gives rise to the functional representation of peripersonal space (PPS). PPS extent is flexible according to the affective context and the target of an action, but little is known about how social context modulates it. We used an audiotactile interaction task to investigate PPS of individuals during social interaction. Participants had to detect as fast as possible a tactile stimulus while task-irrelevant looming sounds were presented, while they were paired as collaborative dyads and as competitive dyads. We also measured PPS in participants seated near an inactive individual. PPS boundaries were modulated only when participants collaborated with a partner, in the form of an extension on the right hemispace and independently of the location of the partner. This suggests that space processing is modified during collaborative tasks. During collective actions, a supra-individual representation of the space of action could be at stake in order to adapt our individual motor control to an interaction as a group with the external world. Reassessing multisensory integration in the light of its potential social sensitivity might reveal that low-level mechanisms are modified by the need to interact with others.


Push-pull monorail technique to overcome a difficult course through the left internal jugular vein.
We assessed the outcomes of a push-pull monorail technique to overcome a difficult anatomical course through the left internal jugular vein in implantable port insertions. From December 2018 to May 2021, a total of 5326 patients were referred for implantable port insertion in our interventional unit, among which 472 cases were requested for insertion on the left side. Our monorail technique was applied only when the catheter tip entered the azygos vein instead of the superior vena cava (n = 8). The technique consists of a puncture at the distal tip of the port catheter with a 21-gauge micropuncture needle, advancing a 0.018-inch hair-wire to the guide, providing support for the pre-assembled port, and advancing the microsheath over the hair-wire to prevent extrusion of the catheter. The push-pull monorail technique was performed in eight patients, and all cases were technically successful, exhibiting a technical success rate of 100%. There were no immediate or delayed complications. The push-pull monorail technique is helpful in overcoming the difficult anatomical course through the left internal jugular vein during implantable port insertion.


Can ischemia-modified albumin be a valuable indicator of tissue ischemia in polycythemia vera?
The red cell expansion in polycythemia vera (PV) causes hyperviscosity affecting blood flow, which plays a major role in the pathogenesis of both microcirculatory disturbances and ultimately thromboses. Ischemia-modified albumin (IMA) is produced during an ischemic states and is present in blood in early and easily detectable levels. This study investigated whether IMA is a useful adjunct in the determination of ischemia in patients with PV, prior to them exhibiting clinical evidence of thrombotic complications. Blood IMA levels were determined in 20 PV patients and in 20 healthy individuals using a method described by Bar-Or. Mean IMA levels in the PV group were significantly higher than those of the control group (P<0.05). At the optimum cutoff point (0.193 absorbance units), the sensitivity and specificity of IMA were 80 and 100% to ischemia, respectively. In conclusion, IMA may be a valuable biochemical marker in predicting tissue ischemia in PV before the signs of vascular disturbances occur.


A functional fibroblast growth factor-1 immunoglobulin fusion protein.
Proteins of the fibroblast growth factor (FGF) family play diverse roles in embryonic development, angiogenesis, and wound healing. The most well studied targets of FGF activity typically are cells of mesodermal and neuroectodermal origin; in addition, expression of FGF-1 (acidic FGF) is increased at several sites of chronic immunologic injury, and recent studies show that FGF-1 also may interact with cells of the immune system. In some human T cells, FGF-1 can induce signals necessary for production of interleukin-2, a key cytokine required for T cell proliferation. To better characterize the interaction of FGF-1 with FGF receptors on T cells, a fusion protein was constructed containing a portion of the constant region of human IgG1 (Fc) at the amino terminus of FGF-1. The Fc-FGF-1 fusion protein retained FGF function as determined by stimulation of tyrosine phosphorylation and DNA synthesis in NIH 3T3 cells. Binding of the intact fusion protein to FGF receptor 1 (FGFR1) on T cells was demonstrated by immunoprecipitation of the receptor bound to Fc-FGF-1 and by flow cytometry showing binding of fusion protein to T cells expressing FGFR1. This functional Fc-FGF-1 protein should prove useful in identifying FGFR-expressing cells.


The concordance between RNA-seq and microarray data depends on chemical treatment and transcript abundance.
The concordance of RNA-sequencing (RNA-seq) with microarrays for genome-wide analysis of differential gene expression has not been rigorously assessed using a range of chemical treatment conditions. Here we use a comprehensive study design to generate Illumina RNA-seq and Affymetrix microarray data from the same liver samples of rats exposed in triplicate to varying degrees of perturbation by 27 chemicals representing multiple modes of action (MOAs). The cross-platform concordance in terms of differentially expressed genes (DEGs) or enriched pathways is linearly correlated with treatment effect size (R(2)0.8). Furthermore, the concordance is also affected by transcript abundance and biological complexity of the MOA. RNA-seq outperforms microarray (93% versus 75%) in DEG verification as assessed by quantitative PCR, with the gain mainly due to its improved accuracy for low-abundance transcripts. Nonetheless, classifiers to predict MOAs perform similarly when developed using data from either platform. Therefore, the endpoint studied and its biological complexity, transcript abundance and the genomic application are important factors in transcriptomic research and for clinical and regulatory decision making.


Transfer and control of the orientation of 3D nanostructures fabricated by nanoskiving.
This communication describes the transfer and stacking of slabs of epoxy containing nanostructures fabricated by nanoskiving. Removal of the sacrificial layer and epoxy matrices produces 3D nanostructures via control over the position and alignment of each sequential layer. The process uses mild etchants and transfers the nanoskived features without wrinkling or damage. We demonstrate the utility of this method of transfer with aligned and intersecting nanowires and arrays of nano-holes and crescents.


Attenuated forearm vasodilator response to atrial natriuretic factor in the elderly.
The vasodilator potency of human atrial natriuretic factor-(99-126) was investigated in the forearm vascular bed of 10 young and 10 elderly normotensive volunteers with venous occlusion strain gauge plethysmography. Atrial natriuretic factor was infused at six increasing dose steps into the brachial artery from 0.001 up to 0.3 microgram/min/100 ml of forearm volume. This induced a mean +/- SEM increase in blood flow from 1.4 +/- 0.2 up to 6.0 +/- 1.0 ml/min/100 ml in the young and from 1.4 +/- 0.2 up to 3.9 +/- 0.6 ml/min/100 ml in the elderly. The dose-response curves of forearm blood flow and of forearm vascular resistance after increasing infusion rates of atrial natriuretic factor were shifted to the right in the elderly when compared with the young subjects. The mean percent decrease in forearm vascular resistance, induced by atrial natriuretic factor, during this dose-response curve averaged -31 +/- 3% in the elderly versus -56 +/- 3% in the young subjects (p = 0.0002). The calculated forearm spillover of the second messenger of atrial natriuretic factor, cyclic guanosine monophosphate, significantly increased from baseline values of 1.2 +/- 1.1 and 0.7 +/- 0.5 pmol/min/100 ml in young and elderly subjects, respectively, up to 23.2 +/- 5.0 and 30.5 +/- 7.0 pmol/min/100 ml during the highest dose of atrial natriuretic factor (both p less than 0.01 versus baseline). There were no significant differences in the increments of the forearm spillover of this second messenger between both age groups.(ABSTRACT TRUNCATED AT 250 WORDS)


Total internal reflection fluorescent microscopy.
This review discusses applications of fluorescence microscopy using totally internally reflected excitation light. When totally internally reflected in a transparent solid at its interface liquid, the excitation light beam penetrates only a short distance into the liquid. This surface electromagnetic field, called the 'evanescent wave', can selectively excite fluorescent molecules in the liquid near the interface. Total internal reflection fluorescence (TIRF) has been used to examine the cell/substrate contact regions of primary cultured rat myotubes with acetylcholine receptors labelled by fluorescent alpha-bungarotoxin and human skin fibroblasts labelled with a membrane-incorporated fluorescent lipid. TIRF examination of cell/surface contacts dramatically reduces background from cell autofluorescence and debris. TIRF has also been combined with fluorescence photobleaching recovery and correlation spectroscopy to measure the chemical kinetic binding rates and surface diffusion constant of fluorescent labelled serum protein binding (at equilibrium) to a surface.


Avulsion of globe following maxillofacial trauma.
Avulsion of the eye globe is a rare condition that is unexpected in maxillofacial trauma and results from severe trauma to the orbit and face. A search of the literature reveals very few reported cases. The present article reports a case of this singular, challenging condition and comments on several other related causes, pathophysiology mechanisms, risks, complications, and management.


Formulation Graphs for Mapping Structure-Composition of Battery Electrolytes to Device Performance.
Advanced computational methods are being actively sought to address the challenges associated with the discovery and development of new combinatorial materials, such as formulations. A widely adopted approach involves domain-informed high-throughput screening of individual components that can be combined together to form a formulation. This manages to accelerate the discovery of new compounds for a target application but still leaves the process of identifying the right "formulation" from the shortlisted chemical space largely a laboratory experiment-driven process. We report a deep learning model, the Formulation Graph Convolution Network (F-GCN), that can map the structure-composition relationship of the formulation constituents to the property of liquid formulation as a whole. Multiple GCNs are assembled in parallel that featurize formulation constituents domain-intuitively on the fly. The resulting molecular descriptors are scaled based on the respective constituent's molar percentage in the formulation, followed by integration into a combined formulation descriptor that represents the complete formulation to an external learning architecture. The use case of the proposed formulation learning model is demonstrated for battery electrolytes by training and testing it on two exemplary data sets representing electrolyte formulations vs battery performance: one data set is sourced from the literature about Li/Cu half-cells, while the other is obtained by lab experiments related to lithium-iodide full-cell chemistry. The model is shown to predict performance metrics such as Coulombic efficiency (CE) and specific capacity of new electrolyte formulations with the lowest reported errors. The best-performing F-GCN model uses molecular descriptors derived from molecular graphs (GCNs) that are informed with HOMO-LUMO and electric moment properties of the molecules using a knowledge transfer technique.


Synergistic coupling between 3D bioprinting and vascularization strategies.
Three-dimensional (3D) bioprinting offers promising solutions to the complex challenge of vascularization in biofabrication, thereby enhancing the prospects for clinical translation of engineered tissues and organs. While existing reviews have touched upon 3D bioprinting in vascularized tissue contexts, the current review offers a more holistic perspective, encompassing recent technical advancements and spanning the entire multistage bioprinting process, with a particular emphasis on vascularization. The synergy between 3D bioprinting and vascularization strategies is crucial, as 3D bioprinting can enable the creation of personalized, tissue-specific vascular network while the vascularization enhances tissue viability and function. The review starts by providing a comprehensive overview of the entire bioprinting process, spanning from pre-bioprinting stages to post-printing processing, including perfusion and maturation. Next, recent advancements in vascularization strategies that can be seamlessly integrated with bioprinting are discussed. Further, tissue-specific examples illustrating how these vascularization approaches are customized for diverse anatomical tissues towards enhancing clinical relevance are discussed. Finally, the underexplored intraoperative bioprinting (IOB) was highlighted, which enables the direct reconstruction of tissues within defect sites, stressing on the possible synergy shaped by combining IOB with vascularization strategies for improved regeneration.


Interleukin-24 protects against liver injury in mouse models.
Interleukin-24 (IL-24) binds to two kinds of receptor complexes, namely IL-20R1/IL-20R2 and IL-20R2/IL-22R1, which are also bound by IL-20. IL-20 plays a detrimental role in liver fibrosis. Due to the sharing of receptor complexes, we aimed to determine whether IL-24 also participates in liver fibrosis. Clinical biopsy specimens from various stages of liver fibrosis were used to analyze IL-24 expression. IL-24 protein was administered to mice with thioacetamide (TAA)-induced liver injury. The direct effects of IL-24 on mouse primary hepatocytes or hepatic stellate cells (HSCs) were analyzed. Wild-type, IL-20R1-, and IL20R2-deficient mice were used to establish a model of acute TAA-induced liver injury. Among patients with more severe liver fibrosis, there was a reduced IL-24/IL-20 ratio. Administration of IL-24 protein protected mice from TAA-induced liver injury and reduction of liver inflammation by antioxidant effects. IL-24 protected hepatocytes from TAA-induced apoptosis and prevented liver fibrosis through the inhibition of the HSCs activation. The protective role of IL-24 acted on liver cells were mainly IL-20R1-independent. IL-20R2-deficient mice exhibited more severe liver injury upon TAA treatment, thus confirming the protective role of IL-24. IL-24 plays a key protective role in the progression of liver injury and has therapeutic potential for treating liver injuries. This work was supported by the Ministry of Science and Technology of Taiwan (MOST 106-2320-B-006-024) and Taiwan Liver Disease Prevention & Treatment Research Foundation.


How did national life expectation related to school years in developing countries - an approach using panel data mining.
The purpose of the study was to probe into the changes in life expectancy associated with schooling years found by the Organization for Economic Co-operation and Development (OECD). The study was based on the OECD database from the period 2000 to 2006. The data of thirty countries were constructed to allow comparisons over time and across these countries. Panel data analysis was used to estimate the relationship of national education, as defined as school years, with life expectancy. The control factors considered were numbers of practicing physicians, practicing nurses, hospital beds, and GDP. We used fixed effects of both country and time through linear regression, the coefficient of school years in relation to life expectancy was statistically significant but negative. This finding is not in accord with the hypothesis that investing in human capital through education stimulates better health outcomes. Within developing countries, educational attainment is no longer keeping the same pace with life expectancy as before. Therefore, we suggest that an effective education policy should cover diverse topics, for example, balancing economic growth and mental hygiene, to improve national life expectancy.


Relevance of focal cerebral ischemia models. Experience with fibrinolytic agents.
Despite the limitations of individual ischemia models, experience with fibrinolytic agents suggests that 1) early intervention with rt-PA may result in rapid thrombolysis, functional recovery, and decreased mortality in small animal stroke thromboembolism models, 2) rt-PA has no general effect on clinical recovery following MCA occlusion and reperfusion in the nonhuman primate at dose rates capable of producing very high circulating rt-PA levels, while u-PA has an apparently salutary effect, and 3) intravenous infusion of rt-PA or u-PA early after ischemia/infarction in several model systems is not associated with significant intracerebral hemorrhage. The true clinical relevance of these general impressions must await the completion of human studies and studies in well-conceived models designed to define the vascular consequences to be expected from reperfusion achievable with thrombolytic agents.


A sister group contrast using untargeted global metabolomic analysis delineates the biochemical regulation underlying desiccation tolerance in Sporobolus stapfianus.
Understanding how plants tolerate dehydration is a prerequisite for developing novel strategies for improving drought tolerance. The desiccation-tolerant (DT) Sporobolus stapfianus and the desiccation-sensitive (DS) Sporobolus pyramidalis formed a sister group contrast to reveal adaptive metabolic responses to dehydration using untargeted global metabolomic analysis. Young leaves from both grasses at full hydration or at 60% relative water content (RWC) and from S. stapfianus at lower RWCs were analyzed using liquid and gas chromatography linked to mass spectrometry or tandem mass spectrometry. Comparison of the two species in the fully hydrated state revealed intrinsic differences between the two metabolomes. S. stapfianus had higher concentrations of osmolytes, lower concentrations of metabolites associated with energy metabolism, and higher concentrations of nitrogen metabolites, suggesting that it is primed metabolically for dehydration stress. Further reduction of the leaf RWC to 60% instigated a metabolic shift in S. stapfianus toward the production of protective compounds, whereas S. pyramidalis responded differently. The metabolomes of S. stapfianus leaves below 40% RWC were strongly directed toward antioxidant production, nitrogen remobilization, ammonia detoxification, and soluble sugar production. Collectively, the metabolic profiles obtained uncovered a cascade of biochemical regulation strategies critical to the survival of S. stapfianus under desiccation.


Ergonomics in the clinical laboratory.
Ergonomics can improve work quality, increase productivity, raise morale, reduce absenteeism, and reduce workers compensation. Laboratory managers are responsible for recognizing employees at risk of developing CTSs which represent 65 percent of all injuries reported. It is a wise investment to prevent injuries. It needs to be stressed to laboratorians that just because one is not suffering now, does not mean suffering will not occur in the future. All of these micro- (mini-) traumas add up over the years. Once there is pain, there are things that can be done to alleviate the discomfort or disability, but that part of the body is never the same again. CTDs are not inexpensive; the National Council on Compensation Insurance reports the average CTD victim compensation is $29,000. Policies and procedures are needed in every facility to address ergonomic safety issues. All employees should be educated about tasks or situations putting an individual at risk for CTDs. Employees should communicate with management and management should be open to information about potential or real risk situations. Safety is everyone's job.


Polarized electron probes of magnetic surfaces.
The magnetic properties of surfaces are now being explored with electron spectroscopies that use electron spin polarization techniques. The increased activity in surface magnetic measurements with polarized electron beams is spurred by new scientific and technological challenges and is made feasible by recent advances in the technology of sources and detectors of polarized electrons. The ability to grow thin films and to engineer artificial structures permits new phenomena to be investigated at magnetic surfaces and interfaces. For such investigations, spin-polarized electron techniques-such as polarized electron scattering, polarized photoemission, polarized Auger spectroscopy, and scanning electron microscopy with polarization analysis-have been and will probably continue to be used to great advantage.


Literature-aided meta-analysis of microarray data: a compendium study on muscle development and disease.
Comparative analysis of expression microarray studies is difficult due to the large influence of technical factors on experimental outcome. Still, the identified differentially expressed genes may hint at the same biological processes. However, manually curated assignment of genes to biological processes, such as pursued by the Gene Ontology (GO) consortium, is incomplete and limited. We hypothesised that automatic association of genes with biological processes through thesaurus-controlled mining of Medline abstracts would be more effective. Therefore, we developed a novel algorithm (LAMA: Literature-Aided Meta-Analysis) to quantify the similarity between transcriptomics studies. We evaluated our algorithm on a large compendium of 102 microarray studies published in the field of muscle development and disease, and compared it to similarity measures based on gene overlap and over-representation of biological processes assigned by GO. While the overlap in both genes and overrepresented GO-terms was poor, LAMA retrieved many more biologically meaningful links between studies, with substantially lower influence of technical factors. LAMA correctly grouped muscular dystrophy, regeneration and myositis studies, and linked patient and corresponding mouse model studies. LAMA also retrieves the connecting biological concepts. Among other new discoveries, we associated cullin proteins, a class of ubiquitinylation proteins, with genes down-regulated during muscle regeneration, whereas ubiquitinylation was previously reported to be activated during the inverse process: muscle atrophy. Our literature-based association analysis is capable of finding hidden common biological denominators in microarray studies, and circumvents the need for raw data analysis or curated gene annotation databases.


Exploring tutor perceptions and current practices in facilitating diagnostic reasoning in preclinical medical students: Implications for tutor professional development needs.
This article was migrated. The article was marked as recommended. Introduction: This study explores tutors' experience of teaching diagnostic reasoning (DR) - a key component of clinical reasoning - to build understanding into the use of explicit strategies in facilitating development of DR skills in preclinical medical students. Methods: A qualitative, interpretive study was undertaken with 14 preclinical problem-based learning tutors who participated in semi-structured interviews. A thematic analysis was conducted to identify key factors that influence students' learning of diagnostic reasoning. Results: Tutor dispositions towards facilitating learning of DR were variable in this study. Explicit strategies to teach DR were thought to exert positive influences on the development of DR skills, through improving student knowledge and reducing potential error. The advantages of using explicit strategies to teach DR outweighed the perceived difficulties identified in this context. Explicit strategies may need modification for preclinical students and the focus should be on building knowledge of classic presentations and developing metacognitive awareness. Conclusion: The use of explicit educational strategies will contribute to facilitating preclinical student learning of DR skills. Tutor professional development is a key component in the successful implementation of these strategies.


Lower levels of erythrocyte membrane fluidity in diabetic patients. A spin label study.
The dynamic properties of intact erythrocyte membrane in diabetic patients were investigated by means of electron spin resonance using three stearic acid spin labels (SAL): 5-, 12-, and 16-SAL. Significantly lower levels of erythrocyte membrane fluidity were revealed with 16-SAL as a probe in diabetic patients when compared with normal controls. However, there were no significant differences in fluidity values using 5- or 12-SAL between the two groups. Therefore, it became obvious that the decrease in fluidity was located in deeper sites (hydrophobic region) of the erythrocyte double membrane in diabetic patients. It was strongly suggested that changes in the membrane cholesterol to phospholipid molar ratios are not a principal factor contributing to the fluidity change. A significant increase of sphingomyelin and decrease of phosphatidylethanolamine were found in the erythrocyte membrane of diabetic patients and an alteration in membrane phospholipid classes and their acyl-chains could conceivably be related to the fluidity change. There were no significant correlations between change in membrane fluidity and most plasma lipids, plasma lecithin-cholesterol acyl-transferase activities, erythrocyte glycosylated hemoglobin, erythrocyte adenosine triphosphate, fasting blood glucose or duration of the disease. Plasma cholesterol of high-density lipoprotein showed a significant negative correlation with the membrane fluidity values. Some of the possible factors contributing to and the significance of the lower levels of erythrocyte membrane fluidity were discussed in conjunction with both metabolic and clinical aspects in diabetic patients.


Single-cell metabolite profiling enables information-rich classification of lymphocyte types and subtypes.
Lymphocytes play crucial roles in the human immune system; however, detailed metabolite characteristics need to be further investigated. Herein, we propose a lymphocyte classification method based on metabolite profiling at the single-cell level. The percentages of different lymphocyte types were calculated with a low margin of error, confirming that the metabolites could serve as a basis for lymphocyte classification. Furthermore, we analyzed the CD4/CD8 ratio in human peripheral blood to verify the feasibility of this method for the classification of lymphocyte subtypes. The proposed method is expected to be a potential tool for the clinical diagnosis of lymphocyte-related diseases.


Endothelial extraction of tracer water varies with extravascular water in dog lungs.
In multiple indicator-dilution experiments, transvascular passage of a permeating indicator is conventionally derived from the up-slope separation of the curve of the permeating indicator from that of a vascular reference and is expressed as the extraction (Ec). Extraction may be limited by the barrier (barrier-limited distribution). It may be limited by the volume of distribution accessible to it; in the time domain of an indicator-dilution experiment, the passage to and distribution in the extravascular volume are rapid relative to the velocity of blood in the exchange vessels. We examine here the relations of the extraction of tracer water as tritium oxide (THO) [Ec(THO)] and of the extraction of tracer sodium as 22Na [Ec(22Na)] to extravascular lung water, delta V wev, by adding isotonic fluid to the gas phase of the lungs. The net convective transvascular passage of water is negligible relative to the transendothelial molecular exchange. In 10 experiments in vivo and in 10 experiments in isolated perfused lungs, Ec(THO) increases as delta V wev increases. Ec(22Na) and the permeability-surface area product (PS) for 22Na do not change as delta V wev increases. We conclude that the extraction of THO is determined mainly by the volume accessible to it (flow- or volume-limited distribution) and that the extraction of 22Na is determined mainly by the resistance of the endothelium (barrier-limited distribution). A diffusion limitation in the added alveolar fluid rather than a barrier limitation at the endothelium may moderate Ec(THO).


Lysophosphatidic acid decreases epidermal growth factor receptor binding in airway epithelial cells.
We showed previously that treatment of human airway smooth muscle cells and lung fibroblasts with lysophosphatidic acid (LPA) increases the binding of epidermal growth factor (EGF) to EGF receptors (EGFRs). The purpose of this study was to determine whether LPA also regulates EGFR binding in airway epithelial cells. Airway epithelial cells were incubated in the absence or presence of 10 microM LPA for increasing times, and binding of 125I-EGF to intact cells on ice was measured. Exposure to LPA for only 15 min caused a 30 to 70% decrease in EGFR binding in a dose-dependent manner, depending on the cell line. This decrease in binding was sustained to at least 18 h in BEAS-2B and primary human bronchial epithelial cells. In contrast, the LPA-induced decrease in binding reversed rapidly in two lung cancer epithelial cell lines, H292 and A549, returning to control levels within 3 h. LPA increased phosphorylation of the EGFR in BEAS-2B cells, and this phosphorylation was inhibited by both 4-(3'-chloroanilino)-6,7-dimethoxy-quinazoline (AG1478; EGFR tyrosine kinase inhibitor) and N-[(2R)-2-(hydroxamidocarbonylmethyl)-4-methylpentanoyl]-l-tryptophan methylamide (GM6001; matrix metalloproteinase inhibitor) but not by CRM197 (heparin-binding EGF inhibitor). AG-1478 and GM6001 also inhibited the LPA-induced decrease in EGFR binding but only by 50%, suggesting only partial involvement of EGFR transactivation in the decrease in EGFR binding. In summary, LPA stimulates a decrease in EGFR binding in airway epithelial cells that is sustained in normal cells but that rapidly reverses in cancer cells. LPA-induced transactivation of EGFRs occurs and contributes to the decrease in EGFR binding, but additional pathway(s) may also be involved.


Evidence for immune activation against oxidized lipoproteins in inactive phases of juvenile chronic arthritis.
Oxidative stress contributes to joint inflammation and damage in rheumatoid arthritis. In a mobile inflamed joint, exercise induced multiple cycles of hypoxia-reperfusion injury may lead to the creation of a redox environment in which oxido-reductase systems, by NADPH mechanisms, produce highly reactive chemical species (i.e., oxygen free radicals). We investigated 2 endproducts of lipid peroxidation, malonildialdehyde (MDA) and diene conjugates (DC), and the formation of antibodies against oxidized low density lipoproteins (Ab oxLDL) in juvenile chronic arthritis (JCA), and assessed the role of oxidative phenomena in different phases and subsets of this disease. To assess the role of oxidative stress in JCA, we measured the endproducts of lipid peroxidation, MDA and DC, by the increase of absorbance at 586 nm and 234 nm, respectively, and the levels of Ab oxLDL by ELISA in the sera of 58 patients with JCA and 21 healthy controls. Due to crossreactivity between Ab oxLDL and anticardiolipin antibodies (aCL), the sera were also tested by a standard ELISA for IgG-aCL. The patients were divided into 3 subsets: 29 with pauciarticular (pauci), 15 with polyarticular (poly), and 14 with systemic (sys) onset disease, and then were subdivided, according to different variables appropriate to each subset, reflecting active and inactive disease, into 30 active (14 pauci, 8 poly, 8 sys) and 28 inactive (15 pauci, 7 poly, 6 sys). Levels of Ab oxLDL were significantly increased in the whole group of patients (566.6 +/- 263.0 vs 206.6 +/- 136.3 mU/ml; p < 0.001) and in each of the type of onset (pauci 660.8 +/- 272.1, p < 0.001; poly 341.3 +/- 134.7, p < 0.01; sys 497.8 +/- 114.8, p < 0.001) compared to controls. Ab oxLDL were higher in the inactive than in the active group (743.5 +/- 231.9 and 404.4 +/- 169.9; p < 0.001). MDA and DC levels were not increased significantly in patients' sera. No patient was positive for IgG-aCL. These findings suggest that MDA and DC cannot be considered major markers of oxidative stress in JCA and that the Ab oxLDL may represent a delayed sign of oxidative stress previously induced by the inflammatory process in patients with JCA.


A combination of derepression of the lac operator-repressor system with positive induction by glucocorticoid and metal ions provides a high-level-inducible gene expression system based on the human metallothionein-IIA promoter.
We and others have introduced the use of the lac operator-repressor system as a method for providing inducible gene expression for gene transfer experiments in animal cells (M. C.-T. Hu, and N. Davidson, Cell 48:555-566, 1987; J. Figge, C. Wright, C. J. Collins, T. M. Roberts, and D. M. Livingston, Cell 52:713-722, 1988). To improve the dynamic range of such an inducible system, we have investigated the effects of combining the relief by isopropyl-beta-D-thiogalactoside (IPTG) of negative control by the lac system with positive induction by the natural inducers glucocorticoids and cadmium ion for a system based on the human metallothionein-IIA gene promoter. We used the chloramphenicol acetyltransferase gene as a reporter gene and inserted a lacO sequence into the promoter between the GC box and metal-responsive element 1, between metal-responsive element 1 and the TATA box, or between the TATA box and the transcription start site. Surprisingly, all of these insertions had a significant inhibitory effect on promoter activity even in the absence of repressor. However, with these lacO-containing constructs, the levels of gene expression after induction by glucocorticoid, Cd2+, or both were considerably reduced in cells engineered to express the lac repressor. Derepression by IPTG, coupled with induction by both dexamethasone and Cd2+ ion, then provided a high level of induced expression, i.e., by a factor of approximately 100 over the basal level of expression. However, inserting the lacO sequence well upstream just before the glucocorticoid-responsive element had much smaller effects on expression levels in both repressor-negative and repressor-positive cells. This study describes a new, high-level-inducible promoter system for gene transfer experiments. The observed effects are discussed in terms of current models of the mechanisms by which transcription factors control gene expression.


Ellipsoidal shell subtraction model of right ventricular volume. Comparison with regional free wall dimensions as indexes of right ventricular function.
Pulse-transit sonomicrometry was used to measure the base-apex (a), anteroposterior (b), and septal-free wall (c) diameters of the left ventricle and the septal-free wall diameter of the right ventricle (d) in eight excised and three isolated, pump-perfused canine heart preparations, as well as in nine conscious dogs. In the three perfused hearts and in four of the excised hearts, right ventricular free wall regional segment lengths and segment area also were assessed. Biventricular volumes were measured directly with intracavitary balloons in all isolated hearts. When left ventricular balloon volume was held constant, relations between right ventricular free wall dimensions and right ventricular balloon volume were highly linear. With increments in left ventricular volume, however, these relations remained linear but shifted progressively upward, indicating an independent relation between right ventricular free wall dimensions and left ventricular cavitary volume. An ellipsoidal shell subtraction model (pi/6.abd minus right ventricular free wall volume) was developed to estimate right ventricular cavitary volume from cardiac dimensions. With this method, a highly linear relation was observed between calculated right ventricular volume and right ventricular balloon volume (mean r = 0.99 +/- 0.01). Moreover, this relation appeared to be independent of changes in left ventricular balloon volume. With the shell subtraction model, dynamic right ventricular volume was computed in nine conscious dogs, and in four, stroke volume derived from dimensions was compared with right ventricular stroke volume measured with ultrasonic flow probes. A highly linear relation was observed, suggesting the accuracy of the shell subtraction method in vivo. Right ventricular end-systolic pressure-volume and stroke work/end-diastolic volume relations then were evaluated, and both proved to be highly linear in the right ventricle (both mean r = 0.99 +/- 0.01). Thus, the shell subtraction model allows a simple estimate of dynamic right ventricular volume in the intact heart and facilitates assessment of right ventricular performance in vivo.


Efficient processing of an antigenic sequence for presentation by MHC class I molecules depends on its neighboring residues in the protein.
Processing of endogenously synthesized proteins generates short peptides that are presented by MHC class I molecules to CD8 T lymphocytes. Here it is documented that not only the sequence of the presented peptide but also the residues by which it is flanked in the protein determine the efficiency of processing and presentation. This became evident when a viral sequence of proven antigenicity was inserted at different positions into an unrelated carrier protein. Not different peptides, but different amounts of the antigenic insert itself were retrieved by isolation of naturally processed peptides from cells expressing the different chimeric proteins. Low yield of antigenic peptide from an unfavorable integration site could be overcome by flanking the insert with oligo-alanine to space it from disruptive neighboring sequences. Notably, the degree of protection against lethal virus disease related directly to the amount of naturally processed antigenic peptide.


Antisense Oligodeoxynucleotide-Mediated Gene Knockdown in Pollen Tubes.
Specific gene knockdown mediated by the antisense oligodeoxynucleotides (AODNs) strategy emerged as a rapid and effective tool for probing gene role in plant cells, particularly tip-growing pollen tubes. Here, we describe the protocol for the successful employment of AODN technique in growing tobacco pollen tubes, covering AODN design, application, and analysis of the results. We also discuss the advantages and drawbacks of this method.


Transepidermal water loss in young and aged healthy humans: a systematic review and meta-analysis.
Transepidermal water loss (TEWL) is regarded as one of the most important parameters for characterizing skin barrier function but an agreed upon definition of what a "normal" TEWL is does not exist. In order to determine generalizable TEWL values for healthy adults, a systematic review and meta-analysis was conducted. The databases MEDLINE and EMBASE and publication lists were screened. After full-text appraisal of 398 studies, 231 studies were excluded due to unclear or insufficient reporting. 167 studies providing data about 50 skin areas were included in the final data synthesis. Pooled sample sizes ranged from n = 5 for the left cheek and the left lower back to a maximum of n = 2,838 for the right midvolar forearm area. The lowest TEWL of 2.3 (95 % CI 1.9-2.7) g/m(2)/h was calculated for the breast skin, the highest TEWL of 44.0 (39.8-48.2) g/m(2)/h for the axilla. TEWL in individuals being 65 years and above was consistently lower compared to the group of 18- to 64-year-old individuals. The quality of reporting TEWL in humans should be increased in future studies.


Automated real-time monitoring of human pluripotent stem cell aggregation in stirred tank reactors.
The culture of human induced pluripotent stem cells (hiPSCs) at large scale becomes feasible with the aid of scalable suspension setups in continuously stirred tank reactors (CSTRs). Innovative monitoring options and emerging automated process control strategies allow for the necessary highly defined culture conditions. Next to standard process characteristics such as oxygen consumption, pH, and metabolite turnover, a reproducible and steady formation of hiPSC aggregates is vital for process scalability. In this regard, we developed a hiPSC-specific suspension culture unit consisting of a fully monitored CSTR system integrated into a custom-designed and fully automated incubator. As a step towards cost-effective hiPSC suspension culture and to pave the way for flexibility at a large scale, we constructed and utilized tailored miniature CSTRs that are largely made from three-dimensional (3D) printed polylactic acid (PLA) filament, which is a low-cost material used in fused deposition modelling. Further, the monitoring tool for hiPSC suspension cultures utilizes in situ microscopic imaging to visualize hiPSC aggregation in real-time to a statistically significant degree while omitting the need for time-intensive sampling. Suitability of our culture unit, especially concerning the developed hiPSC-specific CSTR system, was proven by demonstrating pluripotency of CSTR-cultured hiPSCs at RNA (including PluriTest) and protein level.


Allergic bronchopulmonary aspergillosis among asthmatics.
Allergic bronchopulmonary aspergillosis (ABPA), an immunologically mediated lung disease is now well recognised as a significant cause of chronic lung disease in the West. This study aimed at determining the extent of this entity in 540 patients with severe bronchial asthma and 96 patients with mild or moderate asthma. Of these 134 were subjected to detailed examinations in the form of a skin test, serological test, sputum examination and peripheral blood eosinophil count. Eight patients (1.2% of the total and 6% of selected patients) had evidence of ABPA. A further 50 patients (8% of total and 37% of selected patients) had evidence of allergy to the fungus as depicted by positive skin test reactivity. The latter patients could be either early ABPA or potential candidates for the development of ABPA later. Skin test sensitivity to both Aspergillus fumigatus and A niger were demonstrated in Sri-Lankan asthmatics. Skin test sensitivity could be considered a screening test for the diagnosis of ABPA. A scheme for the diagnosis of ABPA using simple criteria could be devised keeping in mind the cost and availability of tests.


Performance analysis of cooperative virtual MIMO systems for wireless sensor networks.
Multi-Input Multi-Output (MIMO) techniques can be used to increase the data rate for a given bit error rate (BER) and transmission power. Due to the small form factor, energy and processing constraints of wireless sensor nodes, a cooperative Virtual MIMO as opposed to True MIMO system architecture is considered more feasible for wireless sensor network (WSN) applications. Virtual MIMO with Vertical-Bell Labs Layered Space-Time (V-BLAST) multiplexing architecture has been recently established to enhance WSN performance. In this paper, we further investigate the impact of different modulation techniques, and analyze for the first time, the performance of a cooperative Virtual MIMO system based on V-BLAST architecture with multi-carrier modulation techniques. Through analytical models and simulations using real hardware and environment settings, both communication and processing energy consumptions, BER, spectral efficiency, and total time delay of multiple cooperative nodes each with single antenna are evaluated. The results show that cooperative Virtual-MIMO with Binary Phase Shift Keying-Wavelet based Orthogonal Frequency Division Multiplexing (BPSK-WOFDM) modulation is a promising solution for future high data-rate and energy-efficient WSNs.


ATP-sensitive potassium channels mediate contraction-induced attenuation of sympathetic vasoconstriction in rat skeletal muscle.
Sympathetic vasoconstriction is sensitive to inhibition by metabolic events in contracting rat and human skeletal muscle, but the underlying cellular mechanisms are unknown. In rats, this inhibition involves mainly alpha2-adrenergic vasoconstriction, which relies heavily on Ca2+ influx through voltage-dependent Ca2+ channels. We therefore hypothesized that contraction-induced inhibition of sympathetic vasoconstriction is mediated by ATP-sensitive potassium (KATP) channels, a hyperpolarizing vasodilator mechanism that could be activated by some metabolic product(s) of skeletal muscle contraction. We tested this hypothesis in anesthetized rats by measuring femoral artery blood flow responses to lumbar sympathetic nerve stimulation or intraarterial hindlimb infusion of the specific alpha2-adrenergic agonist UK 14,304 during KATP channel activation with diazoxide in resting hindlimb and during KATP channel block with glibenclamide in contracting hindlimb. The major new findings are twofold. First, like muscle contraction, pharmacologic activation of KATP channels with diazoxide in resting hindlimb dose dependently attenuated the vasoconstrictor responses to either sympathetic nerve stimulation or intraarterial UK 14,304. Second, the large contraction-induced attenuation in sympathetic vasoconstriction elicited by nerve stimulation or UK 14,304 was partially reversed when the physiologic activation of KATP channels produced by muscle contraction was prevented with glibenclamide. We conclude that contraction-induced activation of KATP channels is a major mechanism underlying metabolic inhibition of sympathetic vasoconstriction in exercising skeletal muscle.


Autosomal recessive cerebellar ataxia with oculomotor apraxia (ataxia-telangiectasia-like syndrome) is linked to chromosome 9q34.
Ataxia with oculomotor apraxia (ataxia-telangiectasia-like syndrome [AOA]; MIM 208920) is an autosomal recessive disorder characterized by ataxia, oculomotor apraxia, and choreoathetosis. These neurological features resemble those of ataxia-telangiectasia (AT), but in AOA there are none of the extraneurological features of AT, such as immunodeficiency, neoplasia, chromosomal instability, or sensitivity to ionizing radiation. It is unclear whether these patients have a true disorder of chromosomal instability or a primary neurodegenerative syndrome, and it has not been possible to identify the defective gene in AOA, since the families have been too small for linkage analysis. We have identified a new family with AOA, and we show that the patients have no evidence of chromosomal instability or sensitivity to ionizing radiation, suggesting that AOA in this family is a true primary cerebellar ataxia. We have localized the disease gene, by linkage analysis and homozygosity mapping, to a 15.9-cM interval on chromosome 9q34. This work will ultimately allow the disease gene to be identified and its relevance to other types of autosomal recessive cerebellar ataxias to be determined.


Venezuelan Equine Encephalitis Virus Capsid Implicated in Infection-Induced Cell Cycle Delay in vitro.
Venezuelan equine encephalitis virus (VEEV) is a positive sense, single-stranded RNA virus and member of the New World alphaviruses. It causes a biphasic febrile illness that can be accompanied by central nervous system involvement and moderate morbidity in humans and severe mortality in equines. The virus has a history of weaponization, lacks FDA-approved therapeutics and vaccines in humans, and is considered a select agent. Like other RNA viruses, VEEV replicates in the cytoplasm of infected cells and eventually induces apoptosis. The capsid protein, which contains a nuclear localization and a nuclear export sequence, induces a shutdown of host transcription and nucleocytoplasmic trafficking. Here we show that infection with VEEV causes a dysregulation of cell cycling and a delay in the G0/G1 phase in Vero cells and U87MG astrocytes. Cells infected with VEEV encoding a capsid NLS mutant or treated with the capsid-importin α interaction inhibitor G281-1485 were partially rescued from this cell cycle dysregulation. Pathway analysis of previously published RNA-sequencing data from VEEV infected U87MG astrocytes identified alterations of canonical pathways involving cell cycle, checkpoint regulation, and proliferation. Multiple cyclins including cyclin D1, cyclin A2 and cyclin E2 and other regulators of the cell cycle were downregulated in infected cells in a capsid NLS dependent manner. Loss of Rb phosphorylation, which is a substrate for cyclin/cdk complexes was also observed. These data demonstrate the importance of capsid nuclear localization and/or importin α binding for inducing cell cycle arrest and transcriptional downregulation of key cell cycle regulators.


Crack cocaine use in Barcelona: a reason of worry.
In Spain, crack cocaine use is silently increasing. In Barcelona, an intentional sample was selected to describe the general characteristics of this consumption. Participants were submitted to an interview and data were analyzed through qualitative research procedures. Users are young males and of low socioeconomic status and formal education. The major pattern of use is compulsive. Illegal income activities are the choice for crack cocaine or money acquisition, increasing individual and social health costs. Polydrug use is a matter of concern. Although these findings can not be generalized, they should be considered for the development of public policies to adequately address crack cocaine users' needs.


Pb induces the release of CXCL10 and CCL2 chemokines via mtROS/NF-κB activation in BV-2 cells.
Lead (Pb), a well-known environmental pollutant, could cause damage of microglia, the resident macrophages vitally regulating inflammation in brain. Previous studies have found that Pb exposure induces typical pro-inflammatory factors release, such as tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β), but what effects of Pb treatment below the dose causing these factors release are unknown. Thus, cytokines assay was performed to identify the factors released from Pb-treated BV-2 cells at 2.5μM, causing no effects on TNF-α, IL-1β, and IL-6 release and cell death. Cytokines assay identified low doses of Pb exposure mainly induce an increase in specific chemokines, including CXCL10, CCL2, and CXCL2, which were confirmed by ELISA. Subsequent assessment found Pb could damage mitochondria function and generate mitochondrial reactive oxygen species (mtROS), and Mito TEMPO, a specific inhibitor of mtROS, suppressed Pb-caused upregulation of CXCL10 and CCL2, but not CXCL2. Finally, we determined that mtROS mediated Pb-induced activation of NF-κB pathway, as Mito TEMPO treatment inhibited P-p65/p65 escalation during Pb treatment. Inhibition of NF-κB pathway by Bay11-7821 suppressed the release of CXCL10 and CCL2. Collectively, low dose of Pb induces the release of CXCL10 and CCL2 chemokines, but not TNF-α and IL-1β, via mtROS/NF-κB activation in BV-2 cells.


[Genotoxicity of doxorubicin assessed on Saccharomyces cerevisiae cell model].
The genotoxicity of doxorubicin was assessed by the induction of ribonucleotide reductase gene expression and by the oxidative stress induction in Saccharomyces cerevisiae as a eukaryote cell model. Doxorubicin induced dose-dependent inhibition of cell proliferation, increased the intracell concentration of GSH and GSSG, and did not significantly influence the malonic dialdehyde concentration. Doxorubicin activated (independently of the concentration) ribonucleotide reductase gene expression, thus increasing GSH concentration. The increase in GSSG concentration is probably indicative of the development of oxidative stress in Saccharomyces cerevisiae cells, although this is not accompanied by an increase the MDA concentration in the cells.


Endoscopic anti-reflux valve for post-esophagectomy reflux: an animal study.
Background and study aims: Gastric reflux into the remnant esophagus after subtotal or partial esophagectomy is associated with impairment in patient quality of life. We evaluated the feasibility, safety, and potential effectiveness of a novel procedure using a new endoscopic suturing device to create an anti-reflux valve (funnel) in pigs after esophagectomy with gastric tube reconstruction. Methods: The endoscopic procedure was performed in four pigs using a semi-full-thickness endoscopic suturing system (OverStitch; Apollo Endosurgery, Austin, Texas, USA). The operating time, funnel height, ratio of the height of the funnel to the diameter of the gastric tube, and adverse events associated with the procedure were evaluated. The "reflux angle" was measured morphologically and functionally during a reflux induction test, using contrast medium or artificial gastric fluid, by tilting the operating table gradually from a head up to a head down position. Reflux angles before and after funnel creation were compared. Results: The procedure was successful in all four animals. The median operating time was 43 minutes, and the median funnel height was 17 mm (56.7 % of gastric tube diameter). There were no adverse events associated with the procedure. The reflux angle was lower after the procedure compared with before in both morphological and functional assessments in all four cases. Conclusions: Endoscopic anti-reflux funnel creation using OverStitch was feasible, safe, and potentially effective.


Bioactive constituents in Prunus africana: geographical variation throughout Africa and associations with environmental and genetic parameters.
Prunus africana--an evergreen tree found in Afromontane forests--is used in traditional medicine to cure benign prostate hyperplasia. Different bioactive constituents derived from bark extracts from 20 tree populations sampled throughout the species' natural range in Africa were studied by means of GC-MSD. The average concentration [mg/kgw/w] in increasing order was: lauric acid (18), myristic acid (22), n-docosanol (25), ferulic acid (49), β-sitostenone (198), β-sitosterol (490), and ursolic acid (743). The concentrations of many bark constituents were significantly correlated and concentration of n-docosanol was highly significantly correlated with all other analytes. Estimates of variance components revealed the highest variation among populations for ursolic acid (66%) and the lowest for β-sitosterol (20%). In general, environmental parameters recorded (temperature, precipitation, altitude) for the samples sites were not correlated with the concentration of most constituents; however, concentration of ferulic acid was significantly correlated with annual precipitation. Because the concentration of compounds in bark extracts may be affected by tree size, the diameter of sampled plants at 1.3m tree height (as proxy of age) was recorded. The only relationship with tree diameter was a negative correlation with ursolic acid. Under the assumption that genetically less variable populations have less variable concentrations of bark compounds, correlations between variation parameters of the concentration and the respective genetic composition based on chloroplast and nuclear DNA markers were assessed. Only variation of β-sitosterol concentration was significantly correlated with haplotypic diversity. The fixation index (F(IS)) was positively correlated with the variation in concentration of ferulic acid. Principal Components Analysis (PCA) indicated a weak geographic pattern. Mantel tests, however, revealed associations between the geographic patterns of bioactive constituents and the phylogenetic relationship among the populations sampled. This suggests an independent evolution of bark metabolism within different phylogeographical lineages, and the molecular phylogeographic pattern is partly reflected in the variation in concentration of bark constituents. The results have important implications for the design of strategies for the sustainable use and conservation of this important African tree species.


The response of Escherichia coli to ciprofloxacin and norfloxacin.
The action of ciprofloxacin and norfloxacin on two strains of Escherichia coli was studied by diverse methods including electronmicroscopy, viable counting and continuous turbidimetric monitoring. During the first few hours of exposure to inhibitory concentrations of the drugs, the opacity of bacterial cultures continued to increase for a period that was inversely proportional to the drug concentration. This change corresponded to the appearance of filamentous bacteria, swollen forms and some lysis. There was subsequently a gradual drop in opacity during which extensive lysis occurred. As judged by viable counts of bacteria washed free of drug, cell death occurred within 30 min of first exposure to the drugs and continued over a 3-h period. Ultrastructure studies demonstrated that lysis was preceded by the formation of vacuoles, predominantly at the poles of the cells. At these sites, breaks in the cell walls eventually occurred, resulting in extrusion of the cytoplasmic contents.


EGF Contributes to Hypertrophy of Human Ligamentum Flavum via the TGF-β1/Smad3 Signaling Pathway.
Background: The most common spinal disorder in elderly is lumbar spinal canal stenosis (LSCS). Previous studies showed that ligamentum flavum hypertrophy (LFH) with fibrosis as the main pathological change is one of the pathogenic factors leading to LSCS. Epidermal Growth Factor (EGF) is known to have an intimate relationship with fibrosis in various tissues. Nevertheless, currently, there are few studies regarding EGF in LFH. The effect of EGF on the development of LFH is unknown, and the underlying pathomechanism remains unclear. In this study, we investigated the role of EGF in LFH and its potential molecular mechanism. Methods: First, the expression levels of EGF, phosphorylation of EGF receptor (pEGFR), Transforming growth factor-β1 (TGF-β1), Phosphorylated Smad3 (pSmad3), collagen I and collagen III were examined via immunohistochemistry and Western blot in LF tissues from patients with LSCS or Non-LSCS. Second, primary LF cells were isolated from adults with normal LF thickness and were cultured with different concentrations of exogenous EGF with or without erlotinib/TGF-β1-neutralizing antibody. Results: The results showed that EGF, pEGFR, TGF-β1, pSmad3, collagen I and collagen III protein expression in the LSCS group was significantly higher than that in the Non-LSCS group. Meanwhile, pEGFR, TGF-β1, pSmad3, collagen I and collagen III protein expression was significantly enhanced in LF cells after exogenous EGF exposure, which can be notably blocked by erlotinib. In addition, pSmad3, collagen I and collagen III protein expression was blocked by TGF-β1-neutralizing antibody. Conclusions: EGF promotes the synthesis of collagen I and collagen III via the TGF-β1/Smad3 signaling pathway, which eventually contributes to LFH.


Automatic segmentation of brain glioma based on XY-Net.
Glioma is a malignant primary brain tumor, which can easily lead to death if it is not detected in time. Magnetic resonance imaging is the most commonly used technique to diagnose gliomas, and precise outlining of tumor areas from magnetic resonance images (MRIs) is an important aid to physicians in understanding the patient's condition and formulating treatment plans. However, relying on radiologists to manually depict tumors is a tedious and laborious task, so it is clinically important to investigate an automated method for outlining glioma regions in MRIs. To liberate radiologists from the heavy task of outlining tumors, we propose a fully convolutional network, XY-Net, based on the most popular U-Net symmetric encoder-decoder structure to perform automatic segmentation of gliomas. We construct two symmetric sub-encoders for XY-Net and build interconnected X-shaped feature map transmission paths between the sub-encoders, while maintaining the feature map concatenation between each sub-encoder and the decoder. Moreover, a loss function composed of the balanced cross-entropy loss function and the dice loss function is used in the training task of XY-Net to solve the class unevenness problem of the medical image segmentation task. The experimental results show that the proposed XY-Net has a 2.16% improvement in dice coefficient (DC) compared to the network model with a single encoder structure, and compare with some state-of-the-art image segmentation methods, XY-Net achieves the best performance. The DC, HD, recall, and precision of our method on the test set are 74.49%, 10.89 mm, 78.06%, and 76.30%, respectively. The combination of sub-encoders and cross-transmission paths enables the model to perform better; based on this combination, the XY-Net achieves an end-to-end automatic segmentation of gliomas on 2D slices of MRIs, which can play a certain auxiliary role for doctors in grasping the state of illness.


Customer value & business success in the 21st century.
Many health care organizations are realizing the importance of understanding the needs and expectations of customers and prospective customers, coupled with developing services that best meet and exceed those expectations. These concepts represent an innovative yet fundamental approach to building success beyond the clinical aspects. To create, build on, and measure the "customer value factor" in its bottom line, health care organizations must (1) understand how meeting and exceeding the needs and expectations of customers will become the most significant differentiator of true business value, (2) learn how to develop realistic, measurable, and successful customer value processes within complex organizations, (3) understand how customer value can link with the power of strategic communications to become the new currency for the next millennium.


Gut microbiota links to serum ferritin and cognition.
Iron is required for the replication and growth of almost all bacterial species and in the production of myelin and neurotransmitters. Increasing clinical studies evidence that the gut microbiota plays a critical role in iron metabolism and cognition. However, the understanding of the complex iron-microbiome-cognition crosstalk remains elusive. In a recent study in the Aging Imageomics cohort (n = 1,030), we identified a positive association of serum ferritin (SF) with executive function (EF) as inferred from the semantic verbal fluency (SVF,) the total digit span (TDS) and the phonemic verbal fluency tests (PVF). Here, we explored the potential mechanisms by analyzing the gut microbiome and plasma metabolome using shotgun metagenomics and HPLC-ESI-MS/MS, respectively. Different bacterial species belonging to the Proteobacteria phylum (Klebsiella pneumoniae, Klebsiella michiganensis, Unclassified Escherichia) were negatively associated both with SF and executive function. At the functional level, an enrichment of microbial pathways involved in phenylalanine, arginine, and proline metabolism was identified. Consistently, phenylacetylglutamine, a metabolite derived from microbial catabolism of phenylalanine, was negatively associated with SF, EF, and semantic memory. Other metabolites such as ureidobutyric acid and 19,20-DiHDPA, a DHA-derived oxylipin, were also consistently and negatively associated with SF, EF, and semantic memory, while plasma eicosapentaenoic acid was positively associated. The associations of SF with cognition could be mediated by the gut microbiome through microbial-derived metabolites.


Dietary and biomarker estimates of fatty acids and risk of colorectal cancer.
The associations between intake of or circulating fatty acids and risk of colorectal cancer (CRC) are unclear. We examined prospectively the associations between dietary or biomarker fatty acids and CRC. For 41,514 men and women, aged 40-69 years, baseline (1990-94) dietary intakes of fatty acids were estimated using a food frequency questionnaire and plasma phospholipid (PPL) fatty acids were measured for 4,205 participants including 395 CRC cases, according to a case-cohort design. Hazard ratios were computed using Cox regression adjusting for education, alcohol intake, smoking status, physical activity and total energy intake; and stratified for gender, ethnicity and family history of cancer, with age as the time scale. We assessed the heterogeneity of associations with colon and rectal cancers. PPL saturated fatty acids (SFAs) were positively associated with CRC risk, while total n-3 polyunsaturated fatty acids (PUFA) and long chain marine n-3 PUFAs showed inverse associations, significant only for 22:5 n-3. No significant associations were observed for dietary fatty acid intakes but positive associations with CRC of borderline significance were seen for both dietary and PPL linoleic acid. Positive associations with dietary palmitic acid (16:0), MUFAs and n-6 PUFAs were seen for rectal but not colon cancers. PPL 22:6 n-3 was inversely associated with rectal cancer. Limiting intakes of SFAs and MUFAs could be assisted by following existing guidelines to limit red and processed meats which are important sources in the Australian diet. Our observations regarding linoleic acid should be examined further.


Surface plasmon resonance: a label-free tool for cellular analysis.
Surface plasmon resonance (SPR) is a popular technique that allows for sensitive, specific, label-free and real-time assessment of biomolecular interactions. SPR is a nondestructive, modular and flexible tool for various applications in biomedical sciences ranging from cell sorting, cell surface characterization and drug discovery. In this review, we will discuss more specifically how SPR is used to monitor the dynamics of various types of cellular binding events and morphological adherence changes in response to external stimuli.


The human cytomegalovirus glycoprotein B gene (ORF UL55) is expressed early in the infectious cycle.
Northern hybridizations were carried out using mRNA preparations of human cytomegalovirus (HCMV)-infected cultures and gene-specific antisense RNA probes for transcriptional analysis of the gene cluster composed of genes for DNA polymerase, glycoprotein B (gB), herpes simplex virus-infected cell protein 18.5 homologue p130 and a major DNA-binding protein corresponding to open reading frames (ORFs) UL54-UL57, respectively. Monocistronic transcripts of 5 kb and 3.7 kb were found for ORFs UL54 and UL55, respectively, and five additional high molecular mass overlapping transcripts of 14 kb, 10 kb, 10 kb, 8 kb and 6 kb were found. Mapping of 5' ends showed that transcription was initiated at the expected distance downstream of predicted TATA elements; in the case of a UL56-specific transcript two potential initiation sites were identified. Transcription was found to terminate at the expected distance downstream of either of two prominent polyadenylation consensus motifs in the region of UL54. All transcripts were identified early in the infectious cycle, except for the UL55 (gB)-specific transcript of 3.7 kb which was not synthesized until late post-infection. However, specific immunoreactions demonstrated the presence of a gB-specific polypeptide early after infection in the absence of viral DNA synthesis. It is suggested that a bicistronic transcript of 8 kb encoded by ORFs UL55 and UL54 is involved in biosynthesis of early HCMV gB.


Establishment and maintenance of repression by bacteriophage lambda: the role of the cI, cII, and c3 proteins.
To define the events necessary for the establishment and maintenance of repression in a lambda-infected cell, we have studied the requirements for efficient synthesis of the cI protein ("lambda-repressor"). Three classes of lambda mutants defective in the establishment of repression are also defective in the appearance of cI protein activity at the normal time. Two of these mutational classes (cII(-) and cIII(-)) probably result from inactivation of lambda-specified proteins, but the third class (cy(-)) may involve a structural defect. We conclude that at least three regulatory elements are likely to be required for the normal turn-on of cI protein synthesis in an infected nonlysogenic cell: cII and cIII proteins and an "active" y-region of lambda DNA. From these and other results, the complete role of cII and cIII proteins in the establishment of repression may involve a bifunctional regulatory activity: positive regulation of the cI gene and negative regulation of late genes. A possible molecular model for cII and cIII action is discussed. Since the cII and cIII genes are repressed by the cI protein under conditions of stable lysogeny, a separate mechanism is required for the maintenance of cI protein synthesis. After infection of a lysogen by cII(-) phage, the rate of increase of cI protein activity is substantially greater than after infection of a nonlysogen. From these and other results, the cI protein may also have a bifunctional regulatory activity: positive regulation of the cI gene and negative regulation of early lytic genes.


The impact of obesity on patient survival in liver transplant recipients: a meta-analysis.
It is controversial if obesity has an impact on overall survival after liver transplantation (LT). The goal of this study was to determine if obesity impacts liver transplant recipient survival. Through subgroup analysis, we also evaluated different body mass index (BMI) thresholds and the confounding effect of ascites on survival. A systematic literature search from 1990 until July 2013. The main outcome was to evaluate the impact of obesity on survival in adult LT recipients. Dochotomous outcomes were reported as relative risk (RR) with 95% confidence intervals (CI). Thirteen studies with a total 2275 obese and 72 212 non obese patients were included in the analysis. The combined analysis showed no difference in mortality between control and increased weight patients (RR = 0.97, 95% CI [0.82, 1.13], P = 0.66) at last follow-up. Moreover, no differences in mortality were noted in subgroup analysis comparing different BMI thresholds. There was also no differences in survival when BMI was adjusted for ascites or in studies where the liver disease severity was similar. Obese patients had worse survival than nonobese patients in pooled analysis of studies which had similar causes of liver disease (RR = 0.69, 95% CI [0.52, 0.92], P = 0.01). The results of our pools analysis suggest that BMI does not specifically impact patient survival. However, obese patients have worse survival when analysis was performed in studies whose cohorts of obese and nonobese patients had similar causes of liver disease.


Developmental profiles of the intrinsic properties and synaptic function of auditory neurons in preterm and term baboon neonates.
The human fetus starts to hear and undergoes major developmental changes in the auditory system during the third trimester of pregnancy. Although there are significant data regarding development of the auditory system in rodents, changes in intrinsic properties and synaptic function of auditory neurons in developing primate brain at hearing onset are poorly understood. We performed whole-cell patch-clamp recordings of principal neurons in the medial nucleus of trapezoid body (MNTB) in preterm and term baboon brainstem slices to study the structural and functional maturation of auditory synapses. Each MNTB principal neuron received an excitatory input from a single calyx of Held terminal, and this one-to-one pattern of innervation was already formed in preterm baboons delivered at 67% of normal gestation. There was no difference in frequency or amplitude of spontaneous excitatory postsynaptic synaptic currents between preterm and term MNTB neurons. In contrast, the frequency of spontaneous GABA(A)/glycine receptor-mediated inhibitory postsynaptic synaptic currents, which were prevalent in preterm MNTB neurons, was significantly reduced in term MNTB neurons. Preterm MNTB neurons had a higher input resistance than term neurons and fired in bursts, whereas term MNTB neurons fired a single action potential in response to suprathreshold current injection. The maturation of intrinsic properties and dominance of excitatory inputs in the primate MNTB allow it to take on its mature role as a fast and reliable relay synapse.


Lujan-Fryns syndrome (mental retardation, X-linked, marfanoid habitus).
The Lujan-Fryns syndrome or X-linked mental retardation with marfanoid habitus syndrome is a syndromal X-linked form of mental retardation, affecting predominantly males. The prevalence is not known for the general population. The syndrome is associated with mild to moderate mental retardation, distinct facial dysmorphism (long narrow face, maxillary hypoplasia, small mandible and prominent forehead), tall marfanoid stature and long slender extremities, and behavioural problems. The genetic defect is not known. The diagnosis is based on the presence of the clinical manifestations. Genetic counselling is according to X-linked recessive inheritance. Prenatal testing is not possible. There is no specific treatment for this condition. Patients need special education and psychological follow-up, and attention should be given to diagnose early psychiatric disorders.


Dual catheter and double-lumen cerebrospinal fluid shunt systems with backflow mechanisms.
We previously developed a novel functional benchtop apparatus to simulate catheter occlusion in vitro utilizing avian vitelline membrane and chalaza to test catheter designs and de-obstruction techniques. Here, we study the integration of double-lumen catheter-mediated backflow in the shunt system assembly and its potential for an in-line de-obstruction of an obstructed ventricular catheter. A double-lumen catheter was connected to a standard proximal shunt catheter for all trials. One limb of the double-lumen catheter was connected to the backflow mechanisms and allowed to loop back for fluid access. A micropump and a bi-corporal electromagnetic pump were utilized to provide various degrees of backflow at predetermined intervals. Flow rates were measured after initial occlusion and after implementation of the backflow mechanisms, and degrees of catheter blockage was calculated as a percentage of the unoccluded flow rate. Flow visualization was also used. In baseline blockage of less than 50%, the average occluding agent weighed 0.3-0.6 g with baseline flow rates of 8.5-11.9 mL/min. After 5 min of backflow using a micropump, the degree of blockage was reduced in 50% of trials. Additional backflow for 5 min did not provide further improvements in flow rate. In baseline blockage of greater than 50%, the average occluding agent weighed 0.8-1.3 g with baseline flow rates of 1.1-4.2 mL/min. After 5 min of backflow, the system demonstrated a decreased blockage in 20% of trials; additional backflow for 5 min further improved the flow rate in 40% of the total trials. Only magnetic plates provided enough force to provide pulsatile backflow in the bi-corporal electromagnetic system. The preliminary results of connecting a standard proximal catheter in series with a double-lumen catheter show a slight change in the percent occlusion from the baseline status several times when the retrograde flow occurred via one limb of the catheter. Additionally, the de-obstruction seems related to the length of the interval of the backflow and the initial percentage occlusion of the proximal catheter. The statistical analysis does not reveal a statistically significant reduction in occlusion in the proximal catheter with either backflow interval.


Follicle dominance and ovarian asymmetry after luteectomy in rhesus monkeys.
Previous work demonstrated that asymmetrical ovarian activity accompanies morphological asymmetry during the ovarian cycle in rhesus and cynomolgus macaques. This study was designed to determine whether functional ovarian asymmetry could be used to detect the upcoming dominant follicle (DF) even before it was grossly visible. Revealing a latent DF in this manner would permit a better estimate of the time when dominance of the follicle selected to ovulate is attained. To accomplish this, rhesus monkeys were luteectomized at midluteal phase to synchronize subsequent follicle growth, and 4 or 8 days later either the ipsilateral or contralateral ovary was removed. Unilateral ablation at day 4 (when no DF is grossly apparent) of either ovary produced symmetrical responses: the interval from luteectomy (CLX) to the next luteinizing hormone (LH) surge was extended by about 4 days in both groups (P less than 0.01), i.e., from about 12.5 days to 16.7 +/- 1.6 and 17.0 +/- 1.5 days (mean +/- SE). In contrast, hemiovariectomy at day 8 produced markedly divergent asymmetrical responses. Removal of the ipsilateral ovary 8 days after CLX did not affect the timing of the next LH surge (13.2 +/- 0.6 days), which ordinarily occurs about 12.5 days after CLX alone. However, ablation of the contralateral ovary (bearing the next DF) on day 8 extended the interval from CLX to the next LH surge from about 12.5 to 26.6 +/- 1.3 days. These findings indicate that, during the normal ovarian cycle when menses occurs 2--4 days after luteolysis, the follicle destined to ovulate becomes dominant between the 2nd and 6th day and that attainment of dominance signals the completion of a follicle selection process that begins or resumes promptly after luteolysis.