COBAS AMPLICOR: fully automated RNA and DNA amplification and detection system for routine diagnostic PCR.
The COBAS AMPLICOR system automates amplification and detection of target nucleic acids, making diagnostic PCR routine for a variety of infectious diseases. The system contains a single thermal cycler with two independently regulated heating/cooling blocks, an incubator, a magnetic particle washer, a pipettor, and a photometer. Amplified products are captured on oligonucleotide-coated paramagnetic microparticles and detected with use of an avidin-horseradish peroxidase (HRP) conjugate. Concentrated solutions of amplicon or HRP were pipetted without detectable carryover. Amplified DNA was detected with an intraassay CV of < 4.5%; the combined intraassay CV for amplification and detection was < 15%. No cross-reactivity was observed when three different target nucleic acids were amplified in a single reaction and detected with three target-specific capture probes. The initial COBAS AMPLICOR menu includes qualitative tests for diagnosing infections with Chlamydia trachomatis, Neisseria gonorrhoeae, Mycobacterium tuberculosis, and hepatitis C virus. All tests include an optional Internal Control to provide assurance that specimens are successfully amplified and detected.


[Violence in hospital psychiatric units].
The present report is based on a review of literature and shows that violence is a rare manifestation, which results from the intrications of numerous factors and which is difficult to predict even in the short term. Preventive work should be based on the knowledge of some personal as well as relational risk factors.


Mortality, teratogenicity and growth inhibition of three glyphosate formulations using Frog Embryo Teratogenesis Assay-Xenopus.
Ample evidence around the world exists suggesting a link between exposure to glyphosate, toxicity and perturbed physiological functions in non-target organisms. Although glyphosate formulations are widely used for weed and alien plant management, their ecotoxicological information remain scanty. Using the 96-hour Frog Embryo Teratogenesis Assay-Xenopus protocol, embryotoxicity and teratogenicity of three glyphosate-based formulations were assessed. Embryos of Xenopus laevis were exposed to Roundup, Kilo Max and Enviro Glyphosate at concentration of 0.3-1.3, 130-280 and 320-560 mg acid equivalent (a.e.)/L respectively. The results showed Roundup to be more toxic than the other formulations with a 96-hour LC50 of 1.05 mg a.e/L. compared with 207 mg a.e./L, and 466 mg a.e./L for Kilo Max and Enviro Glyphosate respectively. Although, both Roundup and Kilo Max formulations show inhibition on growth of the embryo-larva (P ˂ .05), the minimum concentration inhibiting growth ratios of the three formulations was >0.30 baseline, indicating no significant growth inhibiting effect in the formulations. For teratogenicity, Roundup and Enviro Glyphosate formulations exhibited increasing teratogenic traces, with the teratogenic index at 1.7 and 1.6 respectively. Kilo Max formulation shows low teratogenicity with the teratogenic index at 1.4. Characteristic malformation induced by these formulations included generalized edema, cardiac and abdominal edema, improper gut formation and axial malformations. This study confirms that these formulations could be a potential physiological and ecological health disruptor, particularly concerning teratogenicity and growth disruption. Further studies to characterize the contributions of their surfactants will be invaluable.


Long-chain polyunsaturated fatty acid supplementation in infants born at term.
This section is under preparation and will be included in the next issue. The n-3 and n-6 essential fatty acids alpha linolenic acid and linoleic acid are the precursors of the n-3 and n-6 longchain polyunsaturated fatty acids (LCPUFA). Controversy exists over whether LCPUFA are essential nutrients during infancy. The aim of this review is to assess whether supplementation of formula with LCPUFA is safe and of benefit to term infants. Trials were identified by MEDLINE, checking reference lists of articles and conference proceedings, and by personal communication. All randomised trials of formula supplemented with LCPUFA and with clinical endpoints were reviewed. Eight randomised studies were identified. One of these studies was excluded due to supplementation commencing after three weeks of age, and one is awaiting assessment. The remaining six trials were assessed to be of good quality. Visual acuity was assessed by visual evoked potentials at 4 and 7-8 months in the studies of Makrides et al (1995 & 1996). The supplemented infants had an improved rate of visual maturation at both time points in the first study, but no difference was found between the groups at 4 months in the second study. Visual acuity was assessed by Teller acuity cards in the study of Carlson et al (1996), Clausen et al (1996) and Austed et al (1997). Supplementation had no consistent effect on visual acuity over the first year of life. General development was measured in five studies, all with small numbers. Two suggested a benefit of supplementation. One reported a benefit with supplementation at 4 months ( developmental quotient, DQ measured with the Brunet Lezine test, Agostini et al 1995), but when the groups were reassessed with the same test at 1 and 2 years, no differences were found in DQ (Agostini et al 1997). The other study reported a benefit with supplementation at 9 months (novelty preference measured by Fagan Infantest, Clausen et al 1996). The Adelaide studies showed no effect of supplementation on DQ at one year (Bayley Scales of Infant Development (BSID), Makrides et al 1995 &1996). The Portland study showed no effect of supplementation on development at one year (BSID) or at three years (Stanford Binet IQ test). The Portland study showed no effect of supplementation on language development at one year (McArthur Communicative Development Inventory, Janowsky et al 1995) or at three years (Peabody Picture Vocabulary Test, Scott et al 1997). Growth was measured in a small number of infants in the studies of Makrides et al (1995 & 1996) and Austed et al (1997). There was no suggestion that LCPUFA supplementation reduced the growth of term infants. At present there is little evidence from randomised trials of LCPUFA supplementation to support the hypothesis that LCPUFA supplementation confers any benefit on visual or cognitive development. There are no data from the randomised trials presently available to indicate that LCPUFA supplements influence the growth of term infants.


Minimizing risk using prediction uncertainty in neural network estimation fusion and its application to papermaking.
The paper presents Bayesian information fusion theory in the context of neural-network model combination. It shows how confidence measures can be combined with individual model estimates to minimize risk through the fusion process. The theory is illustrated through application to the real task of quality prediction in the papermaking industry. Prediction uncertainty estimates are calculated using approximate Bayesian learning. These are incorporated into model combination as confidence measures. Cost functions in the fusion center are used to control the influence of the confidence measures and improve the performance of the resultant committee.


Use of the Behavioral Model of Health Care Use to identify correlates of use of treatment for panic attacks in the community.
The aim of this study was to identify predisposing, enabling, and need factors associated with health service use for panic attacks among adults in the United States using the Behavioral Model of Health Care Use. Data were drawn from the National Comorbidity Survey (n = 8098), a community-based household sample representative of the United States adult (age 15-54) population. Stepwise logistic regression models were used to compute odds ratios (with 95 % confidence intervals) measuring the association between predisposing, enabling, and need factors and the self-reported health service use for treatment of panic attacks. In addition to perceived (perception of poor mental health) and evaluated need (severity of panic attacks, psychiatric morbidity), predisposing (being older, married, more educated, white) factors were independently associated with use of treatment for panic attacks. Distinct correlates for use of primary care, specialized mental health treatment, and use of psychotropic medication were found. In addition to need factors, predisposing factors are independently associated with the use of services for panic attacks in the community. These results suggest that factors other than severity of panic and degree of psychiatric morbidity may influence use of services for panic. Future studies that examine whether these factors reflect barriers to care for those in need of mental health treatment may help to inform programs aimed at targeting those with unmet need in the community.


Spectroscopic properties of poly(9,9-dioctylfluorene) thin films possessing varied fractions of β-phase chain segments: enhanced photoluminescence efficiency via conformation structuring.
Poly(9,9-dioctylfluorene) (PFO) is a widely studied blue-emitting conjugated polymer, the optoelectronic properties of which are strongly affected by the presence of a well-defined chain-extended "β-phase" conformational isomer. In this study, optical and Raman spectroscopy are used to systematically investigate the properties of PFO thin films featuring a varied fraction of β-phase chain segments. Results show that the photoluminescence quantum efficiency (PLQE) of PFO films is highly sensitive to both the β-phase fraction and the method by which it was induced. Notably, a PLQE of ∼69% is measured for PFO films possessing a ∼6% β-phase fraction induced by immersion in solvent/nonsolvent mixtures; this value is substantially higher than the average PLQE of ∼55% recorded for other β-phase films. Furthermore, a linear relationship is observed between the intensity ratios of selected Raman peaks and the β-phase fraction determined by commonly used absorption calibrations, suggesting that Raman spectroscopy can be used as an alternative means to quantify the β-phase fraction. As a specific example, spatial Raman mapping is used to image a mm-scale β-phase stripe patterned in a glassy PFO film, with the extracted β-phase fraction showing excellent agreement with the results of optical spectroscopy. © 2016 The Authors. Journal of Polymer Science Part B: Polymer Physics Published by Wiley Periodicals, Inc. J. Polym. Sci., Part B: Polym. Phys. 2016, 54, 1995-2006.


Evolution of sagittal spinal shape for the development of thoracic ossification of ligamentum flavum.
Thoracic ossification of the ligamentum flavum (TOLF) is a rare pathology for which limited research exists. While it is known that mechanical factors play a role in the development of TOLF, little is currently understood about the sagittal alignment and related mechanical stress involved in its development. This study aims to describe the sagittal alignment of patients with TOLF based on the pathologic evolution of the Roussouly classification. The current study evaluated the preoperative Roussouly type in consecutive patients who underwent posterior decompressive laminectomy with or without posterior screw fixation for TOLF between January 2015 and December 2021. The post-evolution sagittal alignments were analyzed using the classic Roussouly classification based on sacral slope (SS). To determine the pre-evolution Roussouly type, the patients were retrospectively classified using their individual PI and PT values. Lumbopelvic parameters and morphological index including inflection point (IP), lumbar apex (LA), and lordosis distribution index (LDI) were also evaluated. Forty-three patients (21 women and 22 men) were included; their mean age was 64.21 ± 11.01 years (range 43-81). The most affected level was T10-11 (48.83%). The mean PI was 50.81 ± 9.56°, the mean SS was 33.11 ± 8.61°and the mean PT was 17.69 ± 7.89°. According to the post-evolution Roussouly classification, type 2 shape was the most frequently observed type (n = 23, 53.5%) in the post-evolution classification while type 3 was the most common type observed in the pre-evolution classification (n = 22, 51.5% and P = .00). The level of IP and LA in type 3 moved caudally (around L2 and L4/5 level, respectively) and the LDI increased (77.98 ± 8.08%) than the normal standard value. The authors found that the majority of the patients had a false type 2 spine, which had evolved pathologically from Roussouly type 3 and exhibited increased LDI, a lowered level of IP, and a lowered level of LA. These changes of spinal shape, including the transition to long hypolordosis and increased length of the thoracic kyphosis, may have accentuated tensile stress at the lower thoracic spine and contributed to the development of TOLF.


Tubal flushing for subfertility.
A possible therapeutic effect of diagnostic tubal patency testing has been debated in the literature for half a century. Further debate surrounds whether oil-soluble or water-soluble contrast media might have the bigger fertility-enhancing effect. Historically a variety of agents have been used to 'flush' the fallopian tubes, although tubal flushing does not currently form part of routine practice in the treatment of fertility delay. To evaluate the effect of flushing a woman's fallopian tubes with oil- or water-soluble contrast media on subsequent fertility outcomes in couples with infertility. We searched the Cochrane Menstrual Disorders and Subfertility Group's specialised register of trials (searched 2 March 2004), MEDLINE (1966 to Mar 2004), EMBASE (1980 to Mar 2004), Biological Abstract (1980 to Mar 2004) and reference lists of articles. All randomised trials where tubal flushing with oil-soluble contrast media or tubal flushing with water-soluble media was compared with one another or with no treatment were considered for inclusion in the review. Eleven randomised controlled trials were identified and included in this review. All trials were assessed for quality criteria. The studied primary outcome was live birth (and ongoing pregnancy), with secondary outcomes pregnancy, miscarriage, ectopic pregnancy, treatment complications including pain, intravasation of contrast medium, infection and haemorrhage, and image quality. Tubal flushing with oil-soluble media versus no intervention (three trials including 381 participants) was associated with a significant increase in the odds of live birth (Peto OR 2.98, 95%CI 1.40 to 6.37) and of pregnancy (Peto OR 3.30, 95%CI 2.00 to 5.43). There were no data from RCTs to assess tubal flushing with water-soluble media versus no intervention. For the comparison of tubal flushing with oil-soluble media versus tubal flushing with water-soluble media (six trials including 1,483 participants), the increase in the odds of live birth for tubal flushing with oil-soluble versus water-soluble media (Peto OR 1.49, 95%CI 1.05 to 2.11) was based on two trials where statistical heterogeneity was present and the higher quality trial showed no significant difference; there was no evidence of a significant difference in the odds of pregnancy (Peto OR 1.24, 95%CI 0.97 to 1.57). The addition of oil-soluble media to flushing with water-soluble media (water-soluble plus oil-soluble media versus water-soluble media alone; three trials including 555 participants) showed no evidence of a significant difference in the odds of pregnancy (Peto OR 1.18, 95%CI 0.82 to 1.70) or live birth (Peto OR 1.06, 95%CI 0.64 to 1.77). Adverse event data were only available for the comparison OSCM versus WSCM: OSCM carried a significantly higher chance of intravasation (Peto OR 5.41, 95%CI 2.57 to 11.37) but a significantly lower chance of immediate pain (Peto OR 0.53, 95%CI 0.34 to 0.84), prolonged pain (Peto OR 0.26, 95%CI 0.15 to 0.45) and post-procedure bleeding (Peto OR 0.22, 95%CI 0.15 to 0.31), and no serious adverse events were reported. There is evidence of effectiveness of tubal flushing with oil-soluble contrast media in increasing the odds of pregnancy and live birth versus no intervention. The limited evidence of an increase in the odds of live birth from tubal flushing with oil-soluble contrast media versus water-soluble contrast media must be interpreted cautiously. Further robust randomised trials, comparing oil-soluble versus water-soluble media and comparing water-soluble media versus no intervention, would be a useful further guide to clinical practice. Further research is merited to ascertain the mechanism of the fertility enhancing effect of oil-soluble media, as to whether this is a 'tubal flushing' phenomenon, an effect on the intraperitoneal environment, or an implantation enhancing effect on the endometrium.


Differential regulation of chemokine expression by estrogen in human periodontal ligament cells.
Estrogen modulates inflammatory responses, but the mechanisms involved have not yet been identified. Periodontal ligament (PDL) cells produce chemokines (a group of chemoattractant molecules that recruit leukocytes) and it has been suggested that estrogen modulates periodontal inflammation by regulating the expression of chemokines by PDL cells. Therefore, the objectives of this study were to investigate the regulation of chemokine ligand 2 [CCL2/monocyte chemoattractant protein 1 (MCP-1)], chemokine ligand 3 [CCL3/macrophage inflammatory protein-1α (MIP-1α)] and chemokine ligand 5 (CCL5/RANTES) by estrogen in human PDL cells. PDL cells were obtained from the PDL of premolars, extracted for orthodontic reasons, from two boys and two girls (16 and 17 years of age). PDL cell CCL2, CCL3 and CCL5 mRNA transcripts were determined by quantitative real-time PCR. The concentrations of CCL2, CCL3 and CCL5 proteins were determined by ELISAs. Treatment with 0.5 μg/mL of lipopolysaccharide (LPS, from Escherichia coli) + 100 nm 17β-estradiol (E(2) ) for 24 h reduced the expression of CCL3 mRNA by about 40% compared to PDL cells treated with LPS alone. Attenuation of CCL3 mRNA was not associated with a decrease in CCL3 protein within 48 h, suggesting a slow turnover of the CCL3 protein. Interindividual differences in the effects of E(2) on CCL5 mRNA expression were observed. E(2) (100 nm) increased the expression of CCL5 by 40-60% in PDL cells derived from two subjects but reduced the expression of CCL5 by about 30% in cells from another subject. CCL2 mRNA and CCL2 protein were highly expressed, but not regulated by E(2) . Similar data were observed in cells obtained from both boys and girls. Regulation, by estrogen, of chemokine expression in PDL cells shows a complex pattern involving the down-regulation as well as the up-regulation of chemokines, suggesting that estrogen exerts both anti-inflammatory and proinflammatory effects through these mechanisms.


A randomized controlled trial of the concrete biosand filter and its impact on diarrheal disease in Bonao, Dominican Republic.
A number of household water treatment and safe storage technologies, such as chlorine disinfection, solar disinfection, and ceramic filtration, have been documented for their ability to reduce diarrheal disease and improve microbial water quality. The biosand filter (BSF) is a promising household water treatment technology in use by > 500,000 people globally. The purpose of this research was to document the ability of BSFs to improve water quality and to reduce diarrheal disease in user compared with non-user households in a randomized controlled trial in Bonao, Dominican Republic, during 2005-2006. During the 6-month intervention period, 75 BSF households had significantly improved drinking water quality on average compared with 79 control households (P < 0.001). Based on random intercepts logistic regression, BSF households had 0.53 times the odds of diarrheal disease as control households, indicating a significant protective effect of the BSF against waterborne diarrheal disease.


Vascularized bone graft from the iliac crest for the treatment of nonunion of the proximal part of the scaphoid with an avascular fragment.
It was hypothesized that nonunion of the proximal third of the scaphoid associated with avascular necrosis could be treated successfully with a free vascularized bone graft obtained from the iliac crest. Fifteen patients who had a nonunion of the proximal part of the scaphoid that had been present for an average of two years and three months (range, nine months to seven years) were managed with use of a free vascularized bone graft obtained from the iliac crest. Avascularity of the scaphoid, as assessed on preoperative radiographs, was characterized by loss of trabecular structure, collapse of subchondral bone, and formation of bone cysts. The results of the procedure were assessed in terms of osseous union, pain, active motion of the wrist, and osteoarthritis. Postoperatively, vascularity of the scaphoid was evaluated with use of magnetic resonance imaging and color Doppler ultrasonography. The average duration of follow-up was six years and one month (range, two years and one month to eight years and one month). Preoperatively, one patient had had pain with any movement of the wrist and fourteen had had pain after strenuous manual labor or sports activity. The average pain score, derived with use of a 10-point visual analog scale, was 2.4 points (range, 1.0 to 6.7 points). Postoperatively, union was achieved in twelve patients; six were pain-free, and six had occasional pain during strenuous manual labor or sports activity, or both. The average pain score for these twelve patients was 1.1 points (range, 0.0 to 4.2 points) on the visual analog scale. Preoperatively, osteoarthritis was limited to the region between the radial styloid process and the distal part of the scaphoid in fourteen patients and to the radioscaphoid region in one patient. Postoperatively, the degree of osteoarthritis remained unchanged in seven of the twelve patients who had union and progressed to the radioscaphoid region in five. Vascularity, as seen on the imaging studies, was restored in all twelve patients who had union. The nonunion persisted in three patients, all of whom had progressive osteoarthritis leading to carpal collapse. The index procedure was successful in twelve of the fifteen patients who had a symptomatic nonunion of the proximal part of the scaphoid associated with avascular necrosis and osteoarthritis that was limited to the radioscaphoid joint.


Tracking the Eye Movement of Four Years Old Children Learning Chinese Words.
Storybook reading is the major source of literacy exposure for beginning readers. The present study tracked 4-year-old Chinese children's eye movements while they were reading simulated storybook pages. Their eye-movement patterns were examined in relation to their word learning gains. The same reading list, consisting of 20 two-character Chinese words, was used in the pretest, 5-min eye-tracking learning session, and posttest. Additionally, visual spatial skill and phonological awareness were assessed in the pretest as cognitive controls. The results showed that the children's attention was attracted quickly by pictures, on which their attention was focused most, with only 13% of the time looking at words. Moreover, significant learning gains in word reading were observed, from the pretest to posttest, from 5-min exposure to simulated storybook pages with words, picture and pronunciation of two-character words present. Furthermore, the children's attention to words significantly predicted posttest reading beyond socioeconomic status, age, visual spatial skill, phonological awareness and pretest reading performance. This eye-movement evidence of storybook reading by children as young as four years, reading a non-alphabetic script (i.e., Chinese), has demonstrated exciting findings that children can learn words effectively with minimal exposure and little instruction; these findings suggest that learning to read requires attention to the basic words itself. The study contributes to our understanding of early reading acquisition with eye-movement evidence from beginning readers.


Multimodality image fusion and planning and dose delivery for radiation therapy.
Image-guided radiation therapy (IGRT) relies on the quality of fused images to yield accurate and reproducible patient setup prior to dose delivery. The registration of 2 image datasets can be characterized as hardware-based or software-based image fusion. Hardware-based image fusion is performed by hybrid scanners that combine 2 distinct medical imaging modalities such as positron emission tomography (PET) and computed tomography (CT) into a single device. In hybrid scanners, the patient maintains the same position during both studies making the fusion of image data sets simple. However, it cannot perform temporal image registration where image datasets are acquired at different times. On the other hand, software-based image fusion technique can merge image datasets taken at different times or with different medical imaging modalities. Software-based image fusion can be performed either manually, using landmarks, or automatically. In the automatic image fusion method, the best fit is evaluated using mutual information coefficient. Manual image fusion is typically performed at dose planning and for patient setup prior to dose delivery for IGRT. The fusion of orthogonal live radiographic images taken prior to dose delivery to digitally reconstructed radiographs will be presented. Although manual image fusion has been routinely used, the use of fiducial markers has shortened the fusion time. Automated image fusion should be possible for IGRT because the image datasets are derived basically from the same imaging modality, resulting in further shortening the fusion time. The advantages and limitations of both hardware-based and software-based image fusion methodologies are discussed.


In vivo distribution of cerebrospinal fluid tracer in human upper spinal cord and brain stem.
BACKGROUNDIntrathecal injection is an attractive route through which drugs can be administered and directed to the spinal cord, restricted by the blood-spinal cord barrier. However, in vivo data on the distribution of cerebrospinal fluid (CSF) substances in the human spinal cord are lacking. We conducted this study to assess the enrichment of a CSF tracer in the upper cervical spinal cord and the brain stem.METHODSAfter lumbar intrathecal injection of a magnetic resonance imaging (MRI) contrast agent, gadobutrol, repeated blood samples and MRI of the upper cervical spinal cord, brain stem, and adjacent subarachnoid spaces (SAS) were obtained through 48 hours. The MRI scans were then analyzed for tracer distribution in the different regions and correlated to age, disease, and amounts of tracer in the blood to determine CSF-to-blood clearance.RESULTSThe study included 26 reference individuals and 35 patients with the dementia subtype idiopathic normal pressure hydrocephalus (iNPH). The tracer enriched all analyzed regions. Moreover, tracer enrichment in parenchyma was associated with tracer enrichment in the adjacent SAS and with CSF-to-blood clearance. Clearance from the CSF was delayed in patients with iNPH compared with younger reference patients.CONCLUSIONA CSF tracer substance administered to the lumbar thecal sac can access the parenchyma of the upper cervical spinal cord and brain stem. Since CSF-to-blood clearance is highly individual and is associated with tracer level in CSF, clearance assessment may be used to tailor intrathecal treatment regimes.FUNDINGSouth-Eastern Norway Regional Health and Østfold Hospital Trust supported the research and publication of this work.


The COSMIN study reached international consensus on taxonomy, terminology, and definitions of measurement properties for health-related patient-reported outcomes.
Lack of consensus on taxonomy, terminology, and definitions has led to confusion about which measurement properties are relevant and which concepts they represent. The aim was to clarify and standardize terminology and definitions of measurement properties by reaching consensus among a group of experts and to develop a taxonomy of measurement properties relevant for evaluating health instruments. An international Delphi study with four written rounds was performed. Participating experts had a background in epidemiology, statistics, psychology, and clinical medicine. The panel was asked to rate their (dis)agreement about proposals on a five-point scale. Consensus was considered to be reached when at least 67% of the panel agreed. Of 91 invited experts, 57 agreed to participate and 43 actually participated. Consensus was reached on positions of measurement properties in the taxonomy (68-84%), terminology (74-88%, except for structural validity [56%]), and definitions of measurement properties (68-88%). The panel extensively discussed the positions of internal consistency and responsiveness in the taxonomy, the terms "reliability" and "structural validity," and the definitions of internal consistency and reliability. Consensus on taxonomy, terminology, and definitions of measurement properties was reached. Hopefully, this will lead to a more uniform use of terms and definitions in the literature on measurement properties.


An Approach to In Vitro Manufacturing of Hypertrophic Cartilage Matrix for Bone Repair.
Devitalized hypertrophic cartilage matrix (DCM) is an attractive concept for an off-the-shelf bone graft substitute. Upon implantation, DCM can trigger the natural endochondral ossification process, but only when the hypertrophic cartilage matrix has been reconstituted correctly. In vivo hypertrophic differentiation has been reported for multiple cell types but up-scaling and in vivo devitalization remain a big challenge. To this end, we developed a micro tissue-engineered cartilage (MiTEC) model using the chondrogenic cell line ATDC5. Micro-aggregates of ATDC5 cells (approximately 1000 cells per aggregate) were cultured on a 3% agarose mold consisting of 1585 microwells, each measuring 400 µm in diameter. Chondrogenic differentiation was strongly enhanced using media supplemented with combinations of growth factors e.g., insulin, transforming growth factor beta and dexamethasone. Next, mineralization was induced by supplying the culture medium with beta-glycerophosphate, and finally we boosted the secretion of proangiogenic growth factors using the hypoxia mimetic phenanthroline in the final stage of in vivo culture. Then, ATDC5 aggregates were devitalized by freeze/thawing or sodium dodecyl sulfate treatment before co-culturing with human mesenchymal stromal cells (hMSCs). We observed a strong effect on chondrogenic differentiation of hMSCs. Using this MiTEC model, we were able to not only upscale the production of cartilage to a clinically relevant amount but were also able to vary the cartilage matrix composition in different ways, making MiTEC an ideal model to develop DCM as a bone graft substitute.


Particle-resolved lattice Boltzmann simulations of 3-dimensional active turbulence.
Collective behaviour in suspensions of microswimmers is often dominated by the impact of long-ranged hydrodynamic interactions. These phenomena include active turbulence, where suspensions of pusher bacteria at sufficient densities exhibit large-scale, chaotic flows. To study this collective phenomenon, we use large-scale (up to N = 3 × 106) particle-resolved lattice Boltzmann simulations of model microswimmers described by extended stresslets. Such system sizes enable us to obtain quantitative information about both the transition to active turbulence and characteristic features of the turbulent state itself. In the dilute limit, we test analytical predictions for a number of static and dynamic properties against our simulation results. For higher swimmer densities, where swimmer-swimmer interactions become significant, we numerically show that the length- and timescales of the turbulent flows increase steeply near the predicted finite-system transition density.


Ion solvation in liquid mixtures: effects of solvent reorganization.
Using field-theoretic techniques, we study the solvation of salt ions in liquid mixtures, accounting for the permanent and induced dipole moments, as well as the molecular volume of the species. With no adjustable parameters, we predict solvation energies in both single-component liquids and binary liquid mixtures that are in excellent agreement with experimental data. Our study shows that the solvation energy of an ion is largely determined by the local response of the permanent and induced dipoles, as well as the local solvent composition in the case of mixtures, and does not simply correlate with the bulk dielectric constant. In particular, we show that, in a binary mixture, it is possible for the component with the lower bulk dielectric constant but larger molecular polarizability to be enriched near the ion.


Experimental set-up for studies of microwave-induced hyperthermia in rats.
A low-cost microprocessor-based temperature controller for hyperthermia experiments on rats is described. The system directs a microwave generator, used for heating, by feedback power regulating signals in accordance with the temperature in the animal. The microwave power is pulsed for short on-and-off periods and the temperature recordings are carried out during the off periods. More than 300 hyperthermia runs have been carried out on rats using this fully automated unit. The controller can direct the hyperthermia to the predetermined level with a deviation of +/- 0.1 degree C for systemic hyperthermia. For local hyperthermia in the liver, individual recorded mean temperatures were up to -0.5 degree C from the preset temperature.


Multi-family psychoeducation groups in the treatment of children with mood disorders.
This study examined the impact of adjunctive multi-family psychoeducation groups (MFPG) on mood-disordered children aged 8 to 11 and their families. Participants were 35 children and 47 parents from families randomly assigned to either immediate MFPG plus treatment as usual (n = 18) or a 6-month wait-list condition plus treatment as usual (n = 17). At the 6 month follow up, immediate treatment families reported: Increased parental knowledge about childhood mood symptoms; increased positive family interactions as reported by the parent; increased perceptions of parental support as reported by children; and increased utilization of appropriate services by families. Expected impact on decreasing negative family interactions was not found. Results are largely consistent with hypothesized findings and support the need to further investigate the adjunctive role of psychoeducation in the treatment of childhood mood disorders.


[Regional features of the electrical activity of the neocortex in the dog over a wide range of frequencies].
Autospectral characteristics (ASC) of potentials of different neocortical areas of dogs in a state of quiet wakefulness were studied in a wide frequency band (1-100 Hz) by means of correlation-spectral analysis. A transition from evaluation of the mean power tendencies to a construction of distributions of different power levels for each of five arbitrarily chosen frequency bands permitted to reveal regional differences in ASC of each animal as well as interindividual EEG distinctions. The regional differences observed in all frequency bands (though not equally expressed) including the higher ones indicate not only the informativity of high-frequency components of electrical neocortical activity but also their neuronal origin.


The population genetics of ploidy change in unicellular fungi.
Changes in ploidy are a significant type of genetic variation, describing the number of chromosome sets per cell. Ploidy evolves in natural populations, clinical populations, and lab experiments, particularly in unicellular fungi. Predicting how ploidy will evolve has proven difficult, despite a long history of theoretical work on this topic, as it is often unclear why one ploidy state outperforms another. Here, we review what is known about contemporary ploidy evolution in diverse fungal species through the lens of population genetics. As with typical genetic variants, ploidy evolution depends on the rate that new ploidy states arise by mutation, natural selection on alternative ploidy states, and random genetic drift. However, ploidy variation also has unique impacts on evolution, with the potential to alter chromosomal stability, the rate and patterns of point mutation, and the nature of selection on all loci in the genome. We discuss how ploidy evolution depends on these general and unique factors and highlight areas where additional experimental evidence is required to comprehensively explain the ploidy transitions observed in the field, the clinic, and the lab.


The UCSF post-baccalaureate reapplication program: a preliminary report.
The University of California San Francisco School of Dentistry initiated a pilot project in 1998 to assist economically and/or educationally disadvantaged students to gain admission to dental school. Students entering the program have completed the requirements for admission to dental school, have applied, and been denied admission, and have expressed a desire to care for underserved populations. The program features a summer preparation course for the Dental Admission Test (DAT), learning skill workshops, seminars, and assistance in the reapplication process. The students participate in clinical clerkships and during the fall and spring semesters are enrolled in upper-division and graduate-level science courses at a local university. Five students participated in the pilot project in 1998; they were admitted in 1999 and have successfully completed the first year of dental school. The sixteen students enrolled in 1999 have now been admitted to one or more dental schools. Fourteen students are currently enrolled in the 2000/2001 academic year program.


Use and limitations of modular metabolic control analysis in medicine and biotechnology.
By Metabolic Control Analysis (MCA), it has been shown that control on flux is in most cases shared by several enzymes rather than concentrated on one "rate-limiting step." This analysis also allows the quantification of the control exerted by groups (modules) of enzymes. The modules may correspond to spatial compartments or to functional units. A brief outline of the modular approach to MCA is given. The criteria by which the system can be modularized and the concept of monofunctional unit are explained. Various studies in which control analysis was applied to biotechnological and medical issues are reviewed. In particular, MCA has turned out to be helpful in the assessment of the severity of enzyme deficiencies. Another application is the search for target enzymes or enzyme groups where pharmaceuticals can suppress the metabolism of pathogenic microorganisms most. In biotechnology, modular and "traditional" control analyses are valuable tools for choosing the most promising targets for genetic manipulation so as to increase a biosynthetic flux. As control coefficients are linear approximations, the effect of enhancing the activities of enzymes to a larger extent is often overestimated. Further limitations such as the restriction to stationary states, uncertainties due to spatial heterogeneities and the impact of experimental error are discussed.


[Genomic of aging and predictive medicine].
New molecular-genetic methods stimulate substantial advances in complex diseases studies, speed up identification of new candidate genes participating in functional genetic modules (gene nets) associated with many common diseases. Decisive impact of predictive genetic studies in efficient implementation of genomic technology advances into presymptomatic identification of the subjects of high risk groups prone to various common complex diseases is reviewed. Substantial progress of genomic studies in genetic of aging processes, including complex metabolic processes and gene regulation is outlined. Advances of predictive medicine in pharmacogenomic, nutrigenomic, sport genomic as well as in genomic of aging substantiate real and soon progress in promotion active healthy longevity.


Sick building syndrome. III. Stachybotrys chartarum.
Increasingly, physicians are being asked to evaluate patients with putative environmentally associated illnesses. These can include a variety of problems, including infectious illnesses (Legionnaire's disease), chemical exposure in the workplace, and sick building syndromes. The latter has been an issue particularly in asthma because of the association of mold and increased bronchial responsiveness. Recently, attention has been focused on the mold Stachybotrys in human disease. Stachybotrys was first identified more than 60 years ago following an epidemic of stomatitis, rhinitis, conjunctivitis, pancytopenia, neurologic disorders, and death in horses. Since then, Stachybotrys has been identified in several outbreaks of disease in animals. It has also attracted attention as a possible agent in idiopathic pulmonary hemorrhage in infants. Stachybotrys is a relatively uncommon fungus but has been isolated from a variety of sources, including contaminated grains, tobacco, indoor air, insulator foams, and water-damaged buildings with high humidity. This fungus is particularly important because it is one of a series of fungi that produces trichothecenes mycotoxins; these mycotoxins are biologically active and can produce a variety of physiological and pathologic changes in humans and animals, including modulation of inflammation and altered alveolar surfactant phospholipid concentrations. The presence of Stachybotrys in a building does not necessarily imply a cause-and-effect relationship with illness, but should alert physicians and healthcare professionals to do more vigorous environmental testing. Guidelines are presented herein for intervention measures in the maintenance of heating, ventilation, and air-conditioning systems.


Intermolecular Interactions in 3-Aminopropyltrimethoxysilane, N-Methyl-3-aminopropyltrimethoxysilane and 3-Aminopropyltriethoxysilane: Insights from Computational Spectroscopy.
In this work, a computational spectroscopy approach was used to provide a complete assignment of the inelastic neutron scattering spectra of three title alkoxysilane derivatives-3-aminopropyltrimethoxysilane (APTS), N-methyl-3-aminopropyltrimethoxysilane (MAPTS), and 3-aminopropyltriethoxysilane (APTES). The simulated spectra obtained from density functional theory (DFT) calculations exhibit a remarkable match with the experimental spectra. The description of the experimental band profiles improves as the number of molecules considered in the theoretical model increases, from monomers to trimers. This highlights the significance of incorporating non-covalent interactions, encompassing classical NH···N, N-H···O, as well as C-H···N and C-H···O hydrogen bond contacts, to achieve a comprehensive understanding of the system. A distinct scenario emerges when considering optical vibrational techniques, infrared and Raman spectroscopy. In these instances, the monomer model provides a reasonable description of the experimental spectra, and no substantial alterations are observed in the simulated spectra when employing dimer and trimer models. This observation underscores the distinctive ability of neutron spectroscopy in combination with DFT calculations in assessing the structure and dynamics of molecular materials.


Experience with pancreas islets separation, immunoisolation and cryopreservation.
Experience of Warsaw Pancreas Laboratory is presented. Some improvement in the methods of rat, human and pig pancreases digestion, and in identification of Langerhans islets by means of intravenous injection of I-DTZ was achieved. For immunoisolation of islets, 2 methods were elaborated: capsules containing alginate/polyethyleneimine/protamine/heparin membrane prepared by modified Sun method, and microcapsules based on Zekorn method. Biocompability of hollow fibers, prepared with polypropylene (PP), surface modified PP (PPS) and polysulphone (PS) was assessed in vitro. Only PS fibers were fully compatible. It was shown, that the mixture of exocrine tissue did not influence in vitro insulin secretion, providing that alginate in which islets are embedded remain gelled. The efficacy of 3 methods of islets cryopreservation was compared: freezing in "semicontrolable" conditions, in programmable Kriomedpol machine, and vitrification. The highest percentage of frozen/thawed living cells, and the most reliable results were obtained with Kriomedpol method.


[An autopsy case of amyotrophic lateral sclerosis with prominent muscle cramps, fasciculation, and high titer of anti-voltage gated potassium channel (VGKC) complex antibody].
The patient was a 55-year-old male who had prominent fasciculation and muscle cramps. Muscle weakness and atrophy of the trunk, respiratory system, and extremities gradually progressed. On the basis of these features, we diagnosed this patient as having amyotrophic lateral sclerosis (ALS), however, the upper motor neuron signs were not significant. Following the detection of the anti-voltage gated potassium channel (VGKC) complex antibody at 907.5 pM (normal < 100 pM) and repetitive discharge in a nerve conduction study, immunotherapy with intravenous immunoglobulin, methylprednisolone (mPSL), double filtration plasmapheresis (DFPP), ciclosporin, and rituximab was introduced. mPSL and DFPP showed only tentative effectiveness for fasciculation and muscle cramps, respectively. Thereafter, muscle weakness progressed. The patient died of type II respiratory failure at the age of 57 years, about 2 years after the onset of the disease. At autopsy, a histopathological diagnosis of ALS with lower-motor-predominant degeneration was made. Characteristic cellular features, including Bunina bodies in the remaining lower motor neurons and phosphorylated TAR DNA-binding protein 43-kDa (pTDP-43)-immunopositive inclusions in both upper and lower motor neuron systems, were evident. At present, an immunological role of the anti-VGKC complex antibody in the development of cramp-fasciculation syndrome has been speculated. In this ALS patient, the antibodies might be associated with pathomechanisms underlying the characteristic symptoms.


Tinospora cordifolia protects from skeletal muscle atrophy by alleviating oxidative stress and inflammation induced by sciatic denervation.
Tinospora cordifolia (TC) is widely being used as immunomodulatory and re-juvenile drug and well described in Indian Ayurveda system of medicine. Rejuvenation also means the fine tuning of the skeletal muscles. Skeletal muscle related disorder, i.e. atrophy is major problem which arise due to cachexia, sarcopenia and immobilization. However, despite of the great efforts, there is scarcity of FDA approved drugs in the market to treat skeletal muscle atrophy. The current study was aimed to explore the in-vitro and in-vivo efficacy and mechanism of TC in myogenic differentiation and skeletal muscle atrophy to establish the possibility of its usage to counteract skeletal muscle atrophy. C2C12 cell lines were used to determine myogenic potential and anti-atrophic effects of T. cordifolia water extract (TCE). Its in-vitro efficacy was re-validated in vivo by supplementation of TCE at a dose of 200 mg/kg/p.o. for 30 days in denervated mice model of skeletal muscle atrophy. Effects of TCE administration on levels of oxidative stress, inflammatory markers and proteolysis were determined. TCE supplementation displayed increased lymphocyte proliferation and induced myogenic differentiation of C2C12 myoblasts by significantly increasing myocytes length and thickness, in comparison to control (p < 0.05). TCE supplementation decreased oxidative stress and inflammatory response by significantly modulating activities of catalase, glutathione peroxidase, lipid peroxidase, superoxide dismutase and β-glucuronidase (p < 0.05). It increased MF-20c expression and ameliorated degradation of muscle protein by down-regulating MuRF-1 and calpain activity. TCE supplementation promotes myogenic differentiation in C2C12 cell lines and prevents denervation induced skeletal muscle atrophy by antagonizing the proteolytic systems (calpain and UPS) and maintaining the oxidative defense mechanism of the cell. Hence, TCE can be used as a protective agent against muscle atrophy.


Cortical Excitability, Synaptic Plasticity, and Cognition in Benign Epilepsy With Centrotemporal Spikes: A Pilot TMS-EMG-EEG Study.
Children with benign epilepsy with centrotemporal spikes have rare seizures emerging from the motor cortex, which they outgrow in adolescence, and additionally may have language deficits of unclear etiology. We piloted the use of transcranial magnetic stimulation paired with EMG and EEG (TMS-EMG, TMS-EEG) to test the hypotheses that net cortical excitability decreases with age and that use-dependent plasticity predicts learning. We assessed language and motor learning in 14 right-handed children with benign epilepsy with centrotemporal spikes. We quantified two TMS metrics of left motor cortex excitability: the resting motor threshold (measure of neuronal membrane excitability) and amplitude of the N100-evoked potential (an EEG measure of GABAergic tone). To test plasticity, we applied 1 Hz repetitive TMS to the motor cortex to induce long-term depression-like changes in EMG- and EEG-evoked potentials. Children with benign epilepsy with centrotemporal spikes tolerate TMS; no seizures were provoked. Resting motor threshold decreases with age but is elevated above maximal stimulator output for half the group. N100 amplitude decreases with age after controlling for resting motor threshold. Motor cortex plasticity correlates significantly with language learning and at a trend level with motor learning. Transcranial magnetic stimulation is safe and feasible for children with benign epilepsy with centrotemporal spikes, and TMS-EEG provides more reliable outcome measures than TMS-EMG in this group because many children have unmeasurably high resting motor thresholds. Net cortical excitability decreases with age, and motor cortex plasticity predicts not only motor learning but also language learning, suggesting a mechanism by which motor cortex seizures may interact with language development.


Formulating injectable pastes of porous calcium phosphate glass microspheres for bone regeneration applications.
Current trends in regenerative medicine treatments for bone repair applications focus on cell-based therapies. These aim to deliver the treatment via a minimally invasive injection to reduce patient trauma and to improve efficacy. This paper describes the injectability of porous calcium phosphate glass microspheres to be used for bone repair based on their formulation, rheology and flow behavior. The use of excipients (xanthan gum, methyl cellulose and carboxyl methyl cellulose) were investigated to improve flow performance. Based on our results, the flow characteristics of the glass microsphere pastes vary according to particle size, surface area, and solid to liquid ratio, as well as the concentration of viscosity modifiers used. The optimal flow characteristics of calcium phosphate glass microsphere pastes was found to contain 40 mg/mL of xanthan gum which increased viscosity whilst providing elastic properties (∼29,000 Pa) at shear rates that mirror the injection process and the resting period post injection, preventing the glass microspheres from both damage and dispersion. It was established that a base formulation must contain 1 g of glass microspheres (60-125 μm in size) per 1 mL of cell culture media, or 0.48 g of glass microspheres of sizes between 125 and 200 μm. Furthermore, the glass microsphere formulations with xanthan gum were readily injectable via a syringe-needle system (3-20 mL, 18G and 14G needles), and have the potential to be utilized as a cell (or other biologics) delivery vehicle for bone regeneration applications.


Electroacupuncture Ameliorates Cognitive Impairment by Inhibiting the JNK Signaling Pathway in a Mouse Model of Alzheimer's Disease.
Electroacupuncture (EA) has become popular for its adjustable strength and frequency and easy quantification in the clinic and has demonstrated therapeutic potential for Alzheimer's disease (AD). However, the mechanism remains unknown. Abnormally activated c-Jun N-terminal kinase (JNK) has been closely related to the pathological process of AD. The aim of this study was to investigate the effect of EA on cognitive impairment and the role of the JNK signaling pathway in AD model amyloid precursor protein (APP)/presenilin 1 (PS1) mice. The memory and learning ability of each group was assessed using the Morris Water Maze (MWM). Immunofluorescence, immunohistochemistry and Western blot were performed to measure the expression of APP, JNK, phosphorylated (P-)JNK, mitogen-activated protein kinase 4 (MKK4), MKK7, c-Jun and caspase-3 in hippocampal tissue samples in APP/PS1 mice after EA intervention. Obvious cognitive deficits were observed in the AD model APP/PS1 mice in the MWM test and were associated with JNK signaling pathway activation and APP upregulation. Four weeks of EA significantly ameliorated the cognitive impairments and inhibited JNK signaling pathway activation and APP upregulation. Taken together, the findings demonstrated that EA can reverse cognitive deficits and substantially lower the burden of APP in AD model APP/PS1 mice, at least partially through inhibiting the JNK signaling pathway and regulating apoptosis signals. Therefore, EA may offer an effective alternative therapeutic approach for AD.


Left ventricular epicardial admittance measurement for detection of acute LV dilation.
There are two implanted heart failure warning systems incorporated into biventricular pacemakers/automatic implantable cardiac defibrillators and tested in clinical trials: right heart pressures, and lung conductance measurements. However, both warning systems postdate measures of the earliest indicator of impending heart failure: left ventricular (LV) volume. There are currently no proposed implanted technologies that can perform LV blood volume measurements in humans. We propose to solve this problem by incorporating an admittance measurement system onto currently deployed biventricular and automatic implantable cardiac defibrillator leads. This study will demonstrate that an admittance measurement system can detect LV blood conductance from the epicardial position, despite the current generating and sensing electrodes being in constant motion with the heart, and with dynamic removal of the myocardial component of the returning voltage signal. Specifically, in 11 pigs, it will be demonstrated that 1) a physiological LV blood conductance signal can be derived; 2) LV dilation in response to dose-response intravenous neosynephrine can be detected by blood conductance in a similar fashion to the standard of endocardial crystals when admittance is used, but not when only traditional conductance is used; 3) the physiological impact of acute left anterior descending coronary artery occlusion and resultant LV dilation can be detected by blood conductance, before the anticipated secondary rise in right ventricular systolic pressure; and 4) a pleural effusion simulated by placing saline outside the pericardium does not serve as a source of artifact for blood conductance measurements.


Pharmacocavernometry with prostaglandin E1.
The aim of this study was to define the normal response of cavernometry parameters with high-dose pharmacostimulation, to investigate repeatability of the method and to find out the incidence of cavernovenous leakage in impotence. 82 consecutive impotent patients and 10 potent control subjects underwent pharmacocavernometry with 40 micrograms of prostaglandin E1. In control subjects, the maintenance flow at 150 mm Hg (MF150) was 3-22 ml/min with an average of 6 ml/min (SD = 6 ml/min). Cavernometry was repeated in 17 patients, but the repeatability of quantitative parameters was disappointing. Cavernovenous leakage was estimated to be present in 55-67% of all impotent patients. Cavernometry with a high-dose prostaglandin E1 pharmacostimulation is a useful method in selected patients, but the consistency of the method should be improved.


Adolescent Health: Communication With Adolescent Patients.
Communication is an essential component in providing health care for adolescent patients. A critical part of adolescence is identity development. Affirmation of identity and emphasis on individual strengths are important for this age group. The concept of resilience is at the center of adolescent care and shapes how care is defined and delivered. A primary goal is to protect patients from harm through a combination of promotion of protective factors, including resilience, and risk factor reduction. In adolescents, use of motivational interviewing has been shown to decrease risky sexual behaviors, help prevent unplanned pregnancy, increase physical activity levels, and decrease substance use. Confidentiality is another essential component of care. The American Academy of Pediatrics (AAP) recommends use of the Strengths, School, Home, Activities, Drugs/substance use, Emotions/eating/depression, Sexuality, Safety (SSHADESS) screen for psychosocial assessment. Several other standardized, validated screening tools also may be valuable in guiding discussions and identifying risky behaviors. Sexual orientation, gender identity, religious, racial, and ethnic components of identity development should be addressed. The empowerment of adolescent patients to achieve personal independence in the health care setting is part of the transition from an adolescent approach to health care to an adult approach.


Using Machine Learning to Derive Just-In-Time and Personalized Predictors of Stress: Observational Study Bridging the Gap Between Nomothetic and Ideographic Approaches.
Investigations into person-specific predictors of stress have typically taken either a population-level nomothetic approach or an individualized ideographic approach. Nomothetic approaches can quickly identify predictors but can be hindered by the heterogeneity of these predictors across individuals and time. Ideographic approaches may result in more predictive models at the individual level but require a longer period of data collection to identify robust predictors. Our objectives were to compare predictors of stress identified through nomothetic and ideographic models and to assess whether sequentially combining nomothetic and ideographic models could yield more accurate and actionable predictions of stress than relying on either model. At the same time, we sought to maintain the interpretability necessary to retrieve individual predictors of stress despite using nomothetic models. Data collected in a 1-year observational study of 79 participants performing low levels of exercise were used. Physical activity was continuously and objectively monitored by actigraphy. Perceived stress was recorded by participants via daily ecological momentary assessments on a mobile app. Environmental variables including daylight time, temperature, and precipitation were retrieved from the public archives. Using these environmental, actigraphy, and mobile assessment data, we built machine learning models to predict individual stress ratings using linear, decision tree, and neural network techniques employing nomothetic and ideographic approaches. The accuracy of the approaches for predicting individual stress ratings was compared based on classification errors. Across the group of patients, an individual's recent history of stress ratings was most heavily weighted in predicting a future stress rating in the nomothetic recurrent neural network model, whereas environmental factors such as temperature and daylight, as well as duration and frequency of bouts of exercise, were more heavily weighted in the ideographic models. The nomothetic recurrent neural network model was the highest performing nomothetic model and yielded 72% accuracy for an 80%/20% train/test split. Using the same 80/20 split, the ideographic models yielded 75% accuracy. However, restricting ideographic models to participants with more than 50 valid days in the training set, with the same 80/20 split, yielded 85% accuracy. We conclude that for some applications, nomothetic models may be useful for yielding higher initial performance while still surfacing personalized predictors of stress, before switching to ideographic models upon sufficient data collection.


Evaluation of immunohistochemical markers in non-small cell lung cancer by unsupervised hierarchical clustering analysis: a tissue microarray study of 284 cases and 18 markers.
This study has investigated a panel of immunomarkers in non-small cell lung carcinoma (NSCLC). Unsupervised hierarchical clustering analysis was used to investigate the possibility of identifying different subgroups in NSCLC based on their molecular expression profile rather than morphological features. A tissue microarray consisting of 284 cases of NSCLC was constructed. Immunohistochemistry was used to detect the presence of 18 biomarkers including synaptophysin, chromogranin, bombesin, NSE, GFI1, ASH-1, p53, p63, p21, p27, E2F-1, cyclin D1, Bcl-2, TTF-1, CEA, HER2/neu, cytokeratin 5/6, and pancytokeratin. Univariate analysis of all 18 markers for prognostic significance was performed. Immunohistochemical scoring data for NSCLC were analysed by unsupervised hierarchical clustering analysis. Kaplan-Meier survival curves were plotted for the different cluster groups of lung tumours identified by this method. Analysis of the three different World Health Organization (WHO) subtypes (adenocarcinoma, squamous cell carcinoma, large cell carcinoma) of NSCLC individually showed that different markers were significant in different subtypes. For example, p53 and p63 were significant for squamous cell carcinoma (p = 0.007 and p = 0.03, respectively), whereas cyclin D1 and HER2/neu were significant prognostic markers for adenocarcinoma (p = 0.025 and p = 0.015, respectively). These markers were not significant prognostic predictors for NSCLC as a group. Hierarchical clustering analysis of NSCLC produced four separate cluster groups, although the vast majority of cases were found in two cluster groups, one dominated by squamous cell carcinoma and the other by adenocarcinoma. The clinical outcomes of cases from the four cluster groups were not significantly different. Prognostic indicators vary between different morphological subtypes of NSCLC. Unsupervised hierarchical clustering analysis, based on an extended immunoprofile, identifies two main cluster groups corresponding to adenocarcinoma and squamous cell carcinoma; cases of large cell carcinomas are assigned to one of these two groups based on their molecular phenotype.


Detecting Critical Functional Ingredients Group and Mechanism of Xuebijing Injection in Treating Sepsis.
Sepsis is a systemic inflammatory reaction caused by various infectious or noninfectious factors, which can lead to shock, multiple organ dysfunction syndrome, and death. It is one of the common complications and a main cause of death in critically ill patients. At present, the treatments of sepsis are mainly focused on the controlling of inflammatory response and reduction of various organ function damage, including anti-infection, hormones, mechanical ventilation, nutritional support, and traditional Chinese medicine (TCM). Among them, Xuebijing injection (XBJI) is an important derivative of TCM, which is widely used in clinical research. However, the molecular mechanism of XBJI on sepsis is still not clear. The mechanism of treatment of "bacteria, poison and inflammation" and the effects of multi-ingredient, multi-target, and multi-pathway have still not been clarified. For solving this issue, we designed a new systems pharmacology strategy which combines target genes of XBJI and the pathogenetic genes of sepsis to construct functional response space (FRS). The key response proteins in the FRS were determined by using a novel node importance calculation method and were condensed by a dynamic programming strategy to conduct the critical functional ingredients group (CFIG). The results showed that enriched pathways of key response proteins selected from FRS could cover 95.83% of the enriched pathways of reference targets, which were defined as the intersections of ingredient targets and pathogenetic genes. The targets of the optimized CFIG with 60 ingredients could be enriched into 182 pathways which covered 81.58% of 152 pathways of 1,606 pathogenetic genes. The prediction of CFIG targets showed that the CFIG of XBJI could affect sepsis synergistically through genes such as TAK1, TNF-α, IL-1β, and MEK1 in the pathways of MAPK, NF-κB, PI3K-AKT, Toll-like receptor, and tumor necrosis factor signaling. Finally, the effects of apigenin, baicalein, and luteolin were evaluated by in vitro experiments and were proved to be effective in reducing the production of intracellular reactive oxygen species in lipopolysaccharide-stimulated RAW264.7 cells, significantly. These results indicate that the novel integrative model can promote reliability and accuracy on depicting the CFIGs in XBJI and figure out a methodological coordinate for simplicity, mechanism analysis, and secondary development of formulas in TCM.


Gene expression analysis of the Streptococcus pneumoniae competence regulons by use of DNA microarrays.
Competence for genetic transformation in Streptococcus pneumoniae is coordinated by the competence-stimulating peptide (CSP), which induces a sudden and transient appearance of competence during exponential growth in vitro. Models of this quorum-sensing mechanism have proposed sequential expression of several regulatory genes followed by induction of target genes encoding DNA-processing-pathway proteins. Although many genes required for transformation are known to be expressed only in response to CSP, the relative timing of their expression has not been established. Overlapping expression patterns for the genes cinA and comD (G. Alloing, B. Martin, C. Granadel, and J. P. Claverys, Mol. Microbiol. 29:75-83, 1998) suggest that at least two distinct regulatory mechanisms may underlie the competence cycle. DNA microarrays were used to estimate mRNA levels for all known competence operons during induction of competence by CSP. The known competence regulatory operons, comAB, comCDE, and comX, exhibited a low or zero initial (uninduced) signal, strongly increased expression during the period between 5 and 12 min after CSP addition, and a decrease nearly to original values by 15 min after initiation of exposure to CSP. The remaining competence genes displayed a similar expression pattern, but with an additional delay of approximately 5 min. In a mutant defective in ComX, which may act as an alternate sigma factor to allow expression of the target competence genes, the same regulatory genes were induced, but the other competence genes were not. Finally, examination of the expression of 60 candidate sites not previously associated with competence identified eight additional loci that could be induced by CSP.


Oxygen and carbon dioxide dual gas-responsive and switchable microgels prepared from emulsion copolymerization of fluoro- and amino-containing monomers.
We report herein the design and preparation of microgels that are responsive to both O2 and CO2 gases. The microgels were synthesized through soap-free emulsion copolymerization of O2-responsive monomer 2,3,4,5,6-pentafluorostyrene (FS) and CO2-responsive monomer 2-(diethylamino)ethyl methacrylate (DEA) with N,N'-methylenebis(acrylamide) (BisAM) as the cross-linker. The P(DEA-co-FS) microgels dispersed in aqueous solution could undergo volume phase transitions triggered by O2 and/or CO2 aeration. The particles were very responsive to CO2, while their responsivity to O2 was moderate. Microgels having different levels of the responsivity could be designed and prepared by varying the FS content in the copolymer. The phase transitions were also highly reversible, and the initial states of microgels could be easily recovered by "washing off" the trigger gases with N2. Multicycle O2, CO2, and N2 aerations were applied, and no loss in the dual gas responsivity and switchability was observed.


WiFi-Based Real-Time Calibration-Free Passive Human Motion Detection.
With the rapid development of WLAN technology, wireless device-free passive human detection becomes a newly-developing technique and holds more potential to worldwide and ubiquitous smart applications. Recently, indoor fine-grained device-free passive human motion detection based on the PHY layer information is rapidly developed. Previous wireless device-free passive human detection systems either rely on deploying specialized systems with dense transmitter-receiver links or elaborate off-line training process, which blocks rapid deployment and weakens system robustness. In the paper, we explore to research a novel fine-grained real-time calibration-free device-free passive human motion via physical layer information, which is independent of indoor scenarios and needs no prior-calibration and normal profile. We investigate sensitivities of amplitude and phase to human motion, and discover that phase feature is more sensitive to human motion, especially to slow human motion. Aiming at lightweight and robust device-free passive human motion detection, we develop two novel and practical schemes: short-term averaged variance ratio (SVR) and long-term averaged variance ratio (LVR). We realize system design with commercial WiFi devices and evaluate it in typical multipath-rich indoor scenarios. As demonstrated in the experiments, our approach can achieve a high detection rate and low false positive rate.


Molecular Cloning and Motif Identification of the Sheep Musclin Gene Promoter.
Musclin is a bioactive factor that functions in regulating the muscle growth and metabolism. To investigate the transcriptional regulatory mechanism of the gene, the 1.4 kb musclin promoter in sheep was cloned (GenBank accession: JX966391) and the sequence was analyzed to predict the motifs associated with muscle growth. Next the enhanced green fluorescent protein (EGFP) was selected as the reporter gene and various wild-type and motif-mutant vectors were constructed. The transcriptional regulatory activities were compared by observing the fluorescence strength and detecting the EGFP mRNA expression in C2C12 myoblasts transfected with the vectors. The results showed that the different lengths of promoters could drive the transcription of EGFP and the mutation of some motifs up- or downregulated the activity of the promoter. Furthermore, the electrophoresis mobility shift assay showed that these motifs regulated the musclin gene transcription through binding to the corresponding transcriptional factors in sheep muscle tissue.


Essential thrombocythemia with portal vein thrombosis and splenic infarction successfully treated with platelet apheresis.
A 63-year-old diabetic woman presented to the outpatient clinic with a 1-week history of abdominal pain. On complete evaluation, she was diagnosed to have essential thrombocythemia. Abdominal imaging revealed portal vein thrombosis with a large splenic infarct. The patient was started on anticoagulant, antiplatelet and cytoreductive therapy. In view of persistent high platelet count, plasma apheresis was done, following which the patient's platelet counts were reduced. Essential thrombocythemia has a high rate of complications, resulting in significant morbidity and mortality. Few cases of this disease and its treatment have been described in the literature, especially pertaining to the Indian scenario. Further studies are needed to establish a multidisciplinary algorithm for its diagnosis and to elucidate the guidelines for the successful treatment of the condition.


Australian dental practitioners experience of burnout.
The aim of this study was to investigate factors associated with burnout in Australian dental practitioners. A cross-sectional survey of 1483 Australian dental practitioners was conducted from Oct to Dec 2021. Participants reported burnout using the Sydney Burnout Measure, and aspects of mental health including psychological distress, depression, and anxiety disorders. Predictors of burnout were explored using a generalized linear model with a stepped approach including demographic, health and risk factor variables. One in four participants were classified as likely to be experiencing burnout. Burnout was associated with working in a regional/rural location (β = 2.82, p < 0.001), an academic/non-clinical role (β = 5.01, p = 0.037), more years of experience as a dental practitioner (β = 0.08, p = 0.022), a current diagnosis of depression (β = 2.38, p = 0.049), moderate/severe psychological distress (β = 7.16, p < 0.001), poor self-rated physical health (β = 5.84, p < 0.001) and increasing alcohol consumption (β = 0.17, p = 0.020). Participants who scored high on resilience (β = -0.23, p = 0.002) or perfectionism (β = -0.24, p < 0.001) had lower burnout scores. Australian dental practitioners appear to be at high risk of burnout which may impact on their health and wellbeing and ability to deliver patient care. There is a need for support services to manage burnout and education to address workplace and environmental factors that may contribute to burnout.


Myometrial constitutive nitric oxide synthase expression is increased during human pregnancy.
Nitric oxide (NO), derived from L-arginine by the action of nitric oxide synthase (NOS), is a mediator of many diverse biological activities, including vasodilation, neurotransmission and inhibition of platelet adhesion. A role for NO in the maintenance of rat and rabbit pregnancy is supported by a variety of studies. A recent study in women demonstrated that myometrial inducible NOS (iNOS) expression was greater in the early third trimester than either the late third trimester or in the non-pregnant condition, suggesting that increased iNOS expression is involved in the maintenance of human pregnancy. Constitutive NOS (cNOS) expression was not determined. The aim of this study was to compare constitutive NOS (both eNOS and bNOS) expression in the human non-pregnant uterus, preterm pregnant uterus (25-34 weeks gestation) and term pregnant uterus (>37 weeks gestation) using immunohistochemistry and Western blotting. Preterm pregnant samples were taken from women with a variety of pathologies necessitating early delivery. We found that eNOS and bNOS protein concentrations were greater in the preterm pregnant myometrium than non-pregnant myometrium. eNOS, but not bNOS, protein concentration was lower in myometrial samples obtained at term compared with those obtained preterm. We conclude that the constitutive isoforms of NOS are also up-regulated in human pregnancy and may play a role in the maintenance of myometrial quiescence.


Health Data Privacy in the COVID-19 Pandemic Context: Discourses on HIPAA.
Considering the impacts of the COVID-19 pandemic on health service delivery, the US Office for Civil Rights (OCR) updated the policies on health data processing, and Health Insurance Portability and Accountability Act (HIPAA). In this study, we investigated discourses on HIPAA in relation to COVID-19. Through a search of media sources in the Factiva database, relevant texts were identified. We applied a text mining approach to identify concepts and themes in these texts. Our analysis revealed six central themes, namely, Health, HIPAA, Privacy, Security, Patients, and Need, as well as their associated concepts. Among these, Health was the most frequently discussed theme. It comprised concepts such as public, care, emergency, providers, telehealth, entity, use, discretion, OCR, Health and Human Services (HHS), enforcement, business, and services. Our discourse analysis of media outlets highlights the role of health data privacy law in the response to global public health emergencies and demonstrates how discourse analysis and computational methods can inform health data protection policymaking in the digital health era.


Consecutive influenza surveillance of neuraminidase mutations and neuraminidase inhibitor resistance in Japan.
The large consumption of neuraminidase inhibitors (NAIs) for the treatment of influenza virus infections places Japan at risk of becoming the epicenter of the global spread of NAI-resistant viruses. To clarify NA amino acid mutations of epidemic influenza viruses in Japan and their related NAI resistance. A total of 1791 samples, including 396 A/H1N1pdm09, 1117 A/H3N2, and 278 B isolates, were collected to determine of their 50% inhibitory concentration (IC50 ) values by NAIs (oseltamivir, zanamivir, peramivir, and laninamivir) during the Japanese seasons from 2011-2012 to 2016-2017. Then, 380 samples including 49 A/H1N1pdm09, 251 A/H3N2, and 80 B isolates were sequenced for the entire NA genes. Neuraminidase inhibitor-resistant A/H1N1pdm09 viruses were detected at a frequency of 1.3% (5/396 isolates) in the epidemic seasons. None of the A/H3N2 and B viruses developed resistance to any of the four NAIs during the six seasons. Only five and 13 AA mutations were detected in the NA catalytic sites of A/H1N1pdm09 and A/H3N2 viruses, respectively. No mutations were observed in the catalytic sites of B viruses. Four of the five mutations in the catalytic sites of A/H1N1pdm09 consisted of H275Y, which was related to high resistance to oseltamivir and peramivir. Most (10/13) of the catalytic site mutations in A/H3N2 were associated with MDCK-passaged induction (D151G/N). Finally, no mutations related to substantial NAI resistance were detected in the A/H3N2 and B viruses examined. These findings suggest that the NA catalytic sites of influenza viruses are well preserved. Even in Japan, no spread of NAI-resistant viruses has been observed, and A/H1N1pdm09 viruses carrying H275Y remain limited.


Assessing Concussion Knowledge Among Recreational Surfers and Comparing Results to Concussion Knowledge Among Soccer Players: A Pilot Study.
Concussions are caused by physical trauma to the head, face, or neck and can be sustained while surfing, increasing the risk of drowning. The purpose of this pilot study was to establish a preliminary assessment of concussion knowledge in a group of adult recreational surfers. Using the standardized Concussion Knowledge Index, an anonymous survey was conducted with 55 surfers. The Concussion Knowledge Index and similar statistical measures were used in a previous study of adult soccer players in England. Data from these 2 groups were compared. The preliminary data suggests that the group of adult surfers demonstrate more concussion knowledge than the group of adult soccer players. Further study into surfers' knowledge of concussion with a larger sample size could increase the clinical utility and generalizability of this study.


Changes in corticospinal and spinal reflex excitability through functional electrical stimulation with and without observation and imagination of walking.
Functional electrical stimulation (FES), a method for inducing muscle contraction, has been successfully used in gait rehabilitation for patients with deficits after neurological disorders and several clinical studies have found that it can improve gait function after stroke and spinal cord injury. However, FES gait training is not suitable for patients with walking difficulty, such as those with severe motor paralysis of the lower limbs. We have previously shown that action observation combined with motor imagery (AO + MI) of walking induces walking-related cortical activity. Therefore, we combined FES, which alternately generates dorsiflexion and plantar flexion, with AO + MI as an alternative to gait training. The present study investigates the transient effects of 20-min of FES simultaneously with and without AO + MI of walking on corticospinal and spinal reflex excitability in able-bodied participants. We measured motor evoked potentials and Hoffmann-reflexes to assess corticospinal and spinal reflex excitability at rest before and after the 20-min FES with and without the AO + MI. Our results show that FES without AO + MI did not change excitability (p > 0.05), while FES with AO + MI facilitated corticospinal excitability (p < 0.05). This facilitation likely occurred due to the synchronization of sensory inputs from FES and cortical activity during AO + MI. Facilitation was observed only in the dorsiflexor but not the plantar flexor muscle (p < 0.05), suggesting muscle specificity of the facilitation. These results demonstrate the effectiveness of combining FES with AO + MI and pave the way for novel neurorehabilitation strategies for patients with neurological gait deficits.


Mutants with base changes at the 3'-end of the 16S RNA from Escherichia coli. Construction, expression and functional analysis.
The functionally important 3' domain of the ribosomal 16S RNA was altered by in vitro DNA manipulations of a plasmid-encoded 16S RNA gene. By in vitro DNA manipulations two double mutants were constructed in which C1399 was converted to A and G1401 was changed to either U or C and a single point mutant was made wherein G1416 was changed to U. Only one of the mutated rRNA genes could be cloned in a plasmid under the control of the natural rrnB promoters (U1416) whereas all three mutations were cloned in a plasmid under the control of the lambda PL promoter. In a strain coding for the temperature-sensitive lambda repressor cI857 the mutant RNAs could be expressed conditionally. We could show that all three mutant rRNAs were efficiently incorporated into 30S ribosomes. However, all three mutants inhibited the formation of stable 70S particles to various degrees. The amounts of mutated rRNAs were quantified by primer extension analysis which enabled us to assess the proportion of the mutated ribosomes which are actively engaged in in vivo protein biosynthesis. While ribosomes carrying the U1416 mutation in the 16S RNA were active in vivo a strong selection against ribosomes with the A1399/U1401 mutation in the 16S RNA from the polysome fraction is apparent. Ribosomes with 16S RNA bearing the A1399/C1401 mutation did not show a measurable protein biosynthesis activity in vivo. The growth rate of cells harbouring the different mutations reflected the in vivo translation capacities of the mutant ribosomes. The results underline the importance of the highly conserved nucleotides in the 3' domain of the 16S RNA for ribosomal function.


Routine preoperative blood testing in pediatric neurosurgery.
The frequency with which routine preoperative blood test results predict perioperative or postoperative complications is insignificant. The unnecessary ordering of routine tests increases the financial costs and patients' distress. The authors evaluated the effects of routine preoperative testing on patient management and the overall financial costs. The authors retrospectively reviewed the medical records and laboratory data for 355 children admitted to the neurosurgical department for elective procedures over a 5-year period (January 2008-December 2012). They excluded all patients admitted for imaging or surgical procedures requiring local anesthesia, and they recorded the results of preoperative and previous (up to 6 months before surgery) blood tests and any abnormalities noted. As a result of the 3489 blood tests ordered preoperatively for 328 (94.6%) of the 355 patients, 29 abnormalities (9%) were detected. Most of these abnormal values were near the reference range, and none significantly affected the progression of scheduled procedures. For only 1 patient (0.28%) was the procedure cancelled because of an abnormality (preoperative partial thromboplastin time), which further testing showed to be a false-positive result. The cost of these tests over 5 years was 5205-10,410 euros ($6766-$13,533 US). Preoperative tests should be selectively requested on the basis of clinical indication.


Thrombotic disorders.
The causes and consequences of thrombosis are of interest and concern to a wide range of clinicians and scientists. Accordingly, the Royal College of Physicians joined with the Haemostasis and Thrombosis Task Force of the British Society for Haematology to hold a conference on thrombotic disorders in October 1995. The aim of the conference was to highlight and discuss recent advances in identifying patients at risk of thrombosis and ways of inhibiting the coagulation cascade and platelet function.


Dyspepsia and IBS symptoms in patients with NERD, ERD and Barrett's esophagus.
Irritable bowel syndrome (IBS) and functional dyspepsia (FD) are highly prevalent in the general population as does gastroesophageal reflux disease (GERD). Therefore, it is expected that these conditions may frequently overlap. We aimed at evaluating the presence ofFD and IBS symptoms in patients with erosive (ERD), non-erosive reflux disease (NERD) and Barrett's esophagus (BE). 71 patients presenting at the reflux disease outpatient clinic were prospectively included in this study. 33 patients had NERD, 25 ERD and 13 BE according to the Montreal classification. All patients with ERD and NERD had typical reflux symptoms, as assessed by a validated GERD questionnaire (RDQ). The diagnosis of functional dyspepsia and IBS symptoms was assessed according to the Rome III criteria. IBS symptoms (bloating, abdominal pain, constipation and diarrhea) were slightly more prevalent in NERD (54.6, 63.6, 21.20, 24.2%, respectively) than in ERD (48.0, 44.0, 12.0, 20.0%, respectively) and in BE (53.9, 23.10, 15,4, 23.1%, respectively), but none of these differences reached statistical significance. NERD patients had more FD symptoms than patients with ERD or BE, but again this difference did not reach statistical significance. Our data show that IBS and FD are common in the entire spectrum of GERD. The presence of these disorders might explain why many patients with GERD are deemed as treatment failures if they have no complete symptom relief with proton pump inhibitors.


The role of small molecule Flt3 receptor protein-tyrosine kinase inhibitors in the treatment of Flt3-positive acute myelogenous leukemias.
Flt3 is expressed by early myeloid and lymphoid progenitor cells and it regulates the proliferation and differentiation of hematopoietic cells. Flt3 is activated by the Flt3 ligand, the monomeric form of which is a polypeptide of about 200 amino acid residues. Both membrane-associated and soluble Flt3 ligands, which are a product of the same gene, function as noncovalent dimers. FLT3 mutations occur in about one-third of newly diagnosed acute myelogenous leukemia (AML) patients. This disease is a malignancy of hematopoietic progenitor cells with a variable clinical course; the incidence of this disorder is more than twice that of patients with chronic myelogenous leukemias (20,000 vs. 8500 new patients per year, respectively, in the United States). FLT3 internal tandem duplication (ITD) results from the head-to-tail duplication of from one to more than 100 amino acids within the juxtamembrane domain and such duplication occurs in about 20-25 % of patients with acute myelogenous leukemias. FLT3 tyrosine kinase (FLT3 TK) mutations, usually within the activation segment, occur in 5-10 % of these patients. The mainstay for the care of acute myelogenous leukemias include daunorubicin or idarubicin and cytarabine. Older patients who are not candidates for such traditional therapy are usually given 5-azacitidine, decitabine, or clofarabine. The addition of orally effective small molecule Flt3 inhibitors to these therapies may prolong event-free and overall survival, a subject of ongoing clinical studies. Midostaurin is US FDA-approved in combination with standard cytarabine and daunorubicin for first-line induction chemotherapy and in combination with cytarabine for second-line consolidation chemotherapy in the treatment of acute myelogenous leukemias with FLT3-postive mutations. Moreover, gilteritinib is a Flt3 multikinase inhibitor that is also FDA approved for the care of adult patients with relapsed or refractory acute myelogenous leukemias with FLT3 mutations. Quizartinib is a Flt3 multikinase inhibitor that was approved by the Ministry of Health, Labor and Welfare (MHLW) of Japan for the treatment of adult patients with relapsed/refractory Flt3-positive acute myelogenous leukemias. Gilteritinib and quizartinib bind to Flt3 with the inactive DFG-Dout structure and are classified as type II inhibitors. Furthermore, ponatinib is a multikinase inhibitor that is approved as therapy for Philadelphia chromosome-positive acute lymphoblastic and chronic myelogenous leukemias; it is used off label for the treatment of patients with acute myelogenous leukemias. Moreover, sorafenib is FDA-approved for the treatment of hepatocellular, renal cell, and differentiated thyroid cancers and it is used off label as maintenance therapy following allogeneic hematopoietic stem cell transplantation in the treatment of acute myelogenous leukemias. Other drugs that are in clinical trials for the treatment of this disorder include sunitinib, crenolanib, FF10101, and lestaurtinib. Unlike chronic myelogenous leukemias, which result solely from the formation of the BCR-Abl chimeric protein kinase, acute myelogenous leukemias result from multi-factorial causes and are prone to be resistant to both cytotoxic and targeted therapies. Consequently, there is a pressing need for better understanding the etiologies of acute myelogenous leukemias and for the development of more effective therapies.


Cytokine expression profiling in human leukocytes after exposure to hypertonic and isotonic fluids.
Resuscitation from hemorrhagic shock causes profound immunologic changes. The tonicity of fluids used for resuscitation clearly influences the immune response. Our study was designed to determine whether isotonic and hypertonic fluids exert their differential effects on immune response by altering the cytokine gene profile of human leukocytes. The cDNA array method was used to profile transcriptional responses after exposure to hypertonic and isotonic fluids. Blood from seven healthy volunteers was incubated for 30 minutes with isotonic (10% dextran-40 and lactated Ringer's [LR] solution) and hypertonic (7.5% hypertonic saline and hypertonic dextran [HTD]) fluids. The volumes of isotonic fluids used were equal to the volume of blood, whereas the volumes of hypertonic fluids were adjusted to keep the salt load identical to the LR group. The cDNA array technique was used to measure the gene expression of 23 common cytokines. Increased gene transcription of proinflammatory cytokines (interleukin [IL]-1alpha, IL-6, IL-10, and tumor necrosis factor-alpha) as well as others (IL-5, IL-7, and IL-16) was found after incubation with resuscitation fluids. Variances were noted depending on the type of fluid: HTD and LR solution did not induce expression of IL-5, and HTD also did not induce IL-1beta expression. Genes encoding IL-1alpha, IL-6, IL-9, and tumor necrosis factor-alpha had low level baseline expression in leukocytes isolated from unstimulated blood, and their expression increased markedly after exposure to resuscitation fluids. The inducible transcripts included IL-1beta, IL-7, IL-10, and IL-16. However, there was no difference in cytokine expression profile between isotonic and hypertonic fluids. Exposure of human leukocytes to resuscitation fluids causes an increase in cytokine gene expressions compared with undiluted blood. This expression profile is largely independent of the type of fluid used.


The crystal structure of plant-specific calcium-binding protein AtCBL2 in complex with the regulatory domain of AtCIPK14.
Calcium signals mediate a multitude of plant responses to external stimuli. Calcineurin B-like (CBL) proteins and their target kinases, CBL-interacting protein kinases (CIPKs), represent important relays in plant calcium signaling. CBL interacts with CIPK through a conserved motif (NAF/FISL motif) within the C-terminal regulatory domain. To better understand the functional role of the CBL-CIPK system, we determined the crystal structure of AtCBL2 in complex with the regulatory domain of AtCIPK14 at 1.2 A resolution. The NAF/FISL motif is inserted into a hydrophobic crevice within AtCBL2, accompanied by a large displacement of the helices and loop on the opposite side of the NAF/FISL motif from the C-terminal region, which shields the hydrophobic crevice in free form. Ca(2+) are coordinated within four EF hands in AtCBL2 in bound form. This calcium coordination pattern differs from that in the structure of the SOS3-SOS2 complex previously reported. Structural comparison of the two structures shows that the recognition of CBL by CIPK is performed in a similar manner, but inherent interactions confer binding affinity and specificity.


[Research methods in dentistry 1. Methods to quantify retention of complete dentures].
Retention is not an unambiguous variable. The degree of retention is dependent on biologic and physiologic properties of a complete denture and the denture-bearing and surrounding tissues. Quantifying retention forces of a complete denture can be carried out with three types of methods: subjective methods, methods with clinical more or less objective criteria, and nearly entire objective methods using measurement equipment. The subjective and most of the methods with clinical criteria are not or little reliable. However, for epidemiological research the methods with clinical criteria are very pragmatic. Of all objective methods using measurement equipment, the gnathodynamometer is the only apparatus with proven reliability.


SLC39A1 contribute to malignant progression and have clinical prognostic impact in gliomas.
Gliomas are one of the most common primary tumors of the central nervous system, and have an unfavorable prognosis. SLC39A1 is a zinc ion transport protein which inhibits the progression of prostate cancer. By studying the role and mechanism of SLC39A1 in the progression of gliomas, perhaps a new therapeutic target can be provided for their treatment. The TCGA, CCGA, GSE16011, GSE44971 and GSE11260 data sets were employed to evaluate the expression level of SLC39A1 in paracancerous and glioma tissues. In addition, Kaplan-Meier analysis, Cox analysis, and the ESTIMATE and CIBERSORT algorithms were used to analyze its prognostic value and immune infiltration correlation. A CCK-8 and flow cytometer were used to measure the effects of SLC39A1 on U87 cell proliferation or apoptosis; RT-qPCR and western blot were used to detect its effects on the expression of MMP2\MMP9. SLC39A1 has up-regulated expression in glioma tissues. High SLC39A1 expression predicted significantly worse survival. Univariate and multivariate analysis show that SLC39A1 independently indicated poor prognosis in patients with gliomas. The expression of SLC39A1 is significantly correlated with clinical pathological parameters such as Grade, IDH mutation status, and 1p19q codeletion status. In vitro experimental results show that SLC39A1 promotes proliferation of glioma cells, inhibits their apoptosis, and promotes expression of MMP2\MMP9. In addition, it may affect infiltration of immune cells into the glioma microenvironment. SLC39A1 may serve as a new prognostic biomarker and potential target for treatment of gliomas.


Computational Evaluation of Amorphous Carbon Coating for Durable Silicon Anodes for Lithium-Ion Batteries.
We investigate the structural, mechanical, and electronic properties of graphite-like amorphous carbon coating on bulky silicon to examine whether it can improve the durability of the silicon anodes of lithium-ion batteries using molecular dynamics simulations and ab-initio electronic structure calculations. Structural models of carbon coating are constructed using molecular dynamics simulations of atomic carbon deposition with low incident energies (1-16 eV). As the incident energy decreases, the ratio of sp² carbons increases, that of sp³ decreases, and the carbon films become more porous. The films prepared with very low incident energy contain lithium-ion conducting channels. Also, those films are electrically conductive to supplement the poor conductivity of silicon and can restore their structure after large deformation to accommodate the volume change during the operations. As a result of this study, we suggest that graphite-like porous carbon coating on silicon will extend the lifetime of the silicon anodes of lithium-ion batteries.


The history of Dupuytren's contracture.
In 1833 Dupuytren described what he referred to as an "affectation" of the palmar aponeurosis. This disorder continues to bear his name a century and a half later despite the fact that Dupuytren was not the first to describe it. The author contemplates the contributions of Felix Platter, Henry Cline, and Sir Astley Cooper in the management of this "maladie."


Construction, replication, and chromatin structure of TRP1 RI circle, a multiple-copy synthetic plasmid derived from Saccharomyces cerevisiae chromosomal DNA.
Transformation studies with Saccharomyces cerevisiae (bakers' yeast) have identified DNA sequences which permit extrachromosomal maintenance of recombinant DNA plasmids in transformed cells. It has been hypothesized that such sequences (called ARS for autonomously replicating sequence) serve as initiation sites for DNA replication in recombinant DNA plasmids and that they represent the normal sites for initiation of replication in yeast chromosomal DNA. We have constructed a novel plasmid called TRP1 R1 Circle which consists solely of 1,453 base pairs of yeast chromosomal DNA. TRP1 RI Circle contains both the TRP1 gene and a sequence called ARS1. This plasmid is found in 100 to 200 copies per cell and is relatively stable during both mitotic and meiotic cell cycles. Replication of TRP1 RI Circle requires the products of the same genes (CDC28, CDC4, CDC7, and CDC8) required for replication of chromosomaL DNA. Like chromosomal DNA, its replication does not occur in cells arrested in the B1 phase of the cell cycle by incubation with the yeast pheromone alpha-factor. In addition, TRP1 RI Circle DNA is organized into nucleosomes whose size and spacing are indistinguishable from that of bulk yeast chromatin. These results indicate that TRP1 RI Circle has the replicative and structural properties expected for an origin of replication from yeast chromosomal DNA. Thus, this plasmid is a suitable model for further studies of yeast DNA replication in both cells and cell-free extracts.