Glaucoma and optical coherence tomography changes in migraine: A comparative cross-sectional study.
To study the prevalence of glaucoma among adults with migraine and the effect of migraine on peripapillary retinal nerve fiber layer (pRNFL) and central macular thickness (CMT) using optical coherence tomography (OCT) compared to those without migraine headache, i.e. in tension-type headache (TTH) and normal group. One hundred and eleven patients (222 eyes) were recruited in three groups. migraine, TTH, and normal subjects visiting hospital outpatient services. After noting demographic details and pertinent history, ophthalmological evaluation including optic disc for glaucomatous changes along with computerized visual field testing and OCT for pRNFL thickness and CMT was performed in all eyes. Continuous variables were compared using ANOVA or Kruskal-Wallis test, while categorical variables including the association of glaucoma with migraine were analyzed using Chi-square or Fisher's exact test. Prevalence of glaucoma in migraine group (12.2%) was more than in comparison groups (6.8% in TTH, 4.1% in normal) which was however not significant (Fisher's exact P = 0.207). Average pRNFL thickness (103.59 ± 12.82 μm) and thickness in nasal (90.49 ± 19.19 μm) and temporal quadrants (70.58 ± 16.13 μm) and CMT (213.78 ± 19.81 μm) were significantly reduced (ANOVA P < 0.05) in migraine patients when compared to the other groups and this was independent of the presence of glaucoma. Prevalence of glaucoma is not significantly higher in migraine patients. However, migraine causes thinning of retinal layers on OCT that is statistically significant.


Bayesian predictive probabilities: a good way to monitor clinical trials.
Bayesian methods, with the predictive probability (PredP), allow multiple interim analyses with interim posterior probability (PostP) computation, without the need to correct for multiple looks at the data. The objective of this paper was to illustrate the use of PredP by simulating a sequential analysis of a clinical trial. We used data from the Laryngobloc trial that planned to include 480 patients to demonstrate the equivalence of success between a laryngoscopy performed with the Laryngobloc® device and a control device. A crossover Bayesian design was used. The success rates of the two laryngoscopy devices were compared. Interim analyses, computed from random numbers of subjects, were simulated. The PostP of equivalence rapidly reached the predefined bound of 0.95. The PredP computed with an equivalence margin of 10% reached the efficacy bound between 352 and 409 of the 480 included patients. If a frequentist analysis had been made on the basis of 217 out of 480 subjects, the study would have been prematurely stopped for equivalence. The PredP indicated that this result was nonetheless unstable and that the equivalence was, thus far, not guaranteed. Based on these interim analyses, we can conclude with a sufficiently high probability that the equivalence would have been met on the primary outcome before the predetermined end of this particular trial. If a Bayesian approach using PredP had been used, it would have allowed an early termination of the trial by reducing the calculated sample size by 15-20%.


Volunteer faculty: what rewards or incentives do they prefer?
Clerkship directors and college administrators have concerns about recruitment and retention of practicing physicians for volunteer teaching. There is a paucity of data regarding the rewards and incentives offered to, or desired by, the nonsalaried community-based practicing physicians who volunteer their time to teach. This study was designed to gain information about rewards and incentives from volunteer teachers in pediatric, family practice, and internal medicine clerkships. We surveyed nonsalaried physician teachers of internal medicine, family medicine, and pediatrics across the United States. The survey focused on teachers' evaluation of the rewards or incentives offered by the programs in the following categories: educational opportunities, services or gifts, recognition bestowed by the school, academic appointments, and monetary payments. Respondents rated each item from 1 (not appreciated) to 5 (very much appreciated). They also were asked to rank order the rewards or incentives (with the addition of a category of personal satisfaction) from 1 (least appreciated) to 6 (most appreciated). Educational opportunities received high ratings, especially when the school bore the cost of providing a service. Payment for teaching was offered to 37% of the respondents, and those who were paid rated it higher. Overall, payment for teaching had a mean appreciation score of 3.94, second only to travel and meeting registration reimbursement (4.27). However, in the rank order listing, personal satisfaction had the highest rank (5.16). In contrast, payment for teaching (2.92) and gifts or services from the college (2.53) were at the bottom of the rank order. The survey asked practicing physicians the value they placed on awards and incentives provided to them by the college. This information should help administrators and clerkship directors in recruiting and retaining community-based practicing physicians for teaching.


The role of long distance migration in the rural renaissance, in gentrification, and in growth of the Sunbelt.
The author "has examined trends in interstate migration during the 1970s for the four regions of the United States and, in particular, explored the relationship of this migration with nonmetropolitan expansion and the return to the city movement. The data show that the former trend is a very real one which has persisted throughout the decade and that population growth through interstate migration in nonmetropolitan America is, on balance, the result of nonblacks moving to the South from elsewhere in the nation. Except in the West, central cities continue to experience substantial losses of population through interstate migration. Interstate migration continues to be directed toward suburbs, and is especially vigorous in the West." Data are from the March 1975 and March 1979 Current Population Surveys.


SPATIAL VARIABILITY OF INDOOR RADON CONCENTRATION IN SCHOOLS: IMPLICATIONS ON RADON MEASUREMENT PROTOCOLS.
The requirements about radon measurements in schools and public buildings included in most of the national and international legislations are generally restricted to all the rooms located at the ground floor and basement, assuming the soil beneath the building as the main source of indoor radon. In order to verify such an assumption for small buildings having at maximum two floors, a preliminary study was performed in 50 schools located in 15 municipalities of the Republic of Srpska. Results of this study suggest that a protocol requiring measurements at the ground floor only may be considered adequate. Due to the high radon spatial variability for rooms at the ground floor, it is preferable to require measurements in a high number of rooms (preferably in all of them) in order to assess the compliance with the reference level established by the legislation.


Structure and stability of xenon insertion compounds of hypohalous acids, HXeOX [X=F, Cl, and Br]: an ab initio investigation.
The structure and stability of xenon-inserted hypohalous acids HXeOX (X=F, Cl, and Br) have been investigated theoretically using ab initio molecular orbital calculations. All these molecules are found to consist of a nearly linear HXeO moiety and a bend XeOX fragment. Geometrical parameters of HXeOX are comparable with that of experimentally observed HXeOH species. The dissociation energies corresponding to the lowest-energy fragmentation products, HOX+Xe have been computed to be -398.1, -385.5, and -386.7 kJmol for HXeOF, HXeOCl, and HXeOBr, respectively, at the MP2 level of theory. The respective barrier heights corresponding to the bent transition states (H-Xe-O bending mode) have been calculated to be 138.1, 138.4, and 138.2 kJmol with respect to HXeOX minimum. These species are found to be metastable in their respective potential-energy surface, and the dissociation energies corresponding to the H+Xe+OX products are found to be 56.8, 66.0, and 80.8 kJmol for HXeOF, HXeOCl, and HXeOBr, respectively. The energies corresponding to the H+Xe+O+X dissociation channel have been computed to be 272.0, 309.3, and 299.7 kJmol for HXeOF, HXeOCl, and HXeOBr, respectively, at the same level of theory. Energetics as well as geometrical considerations suggests that it may be possible to prepare these species experimentally similar to that of HXeOH species at low-temperature laser photolysis experiments.


[Generation of transgenic mouse of dentin sialoprotein and transgene expression analysis].
To generate the transgenic mouse model of DSP and perform transgene expression analysis by RT-PCR. Plasmid pcDNA3.1-CX was constructed by substituting promoter cbeta-actin for CMV promoter of pcDNA3.1, and the ultimate transgenic vector, pcDNA3.1-CX-dsp, was constructed by cloning DSP coding sequence into pcDNA3.1-CX. The pcDNA3.1-CX-dsp plasmid was linearized and microinjected into the male pronucleus of the zygotes. The tail DNA of pups was tested by PCR and Southern blot. A member of F1 generation of one positive mouse was used to perform transgene expression analysis by RT-PCR. 717 embryos were implanted to 29 recipient pseudopregnant mice, 4 of the 67 pups carrying the transgene. Expression of DSP was detected in a member of F1 generation of one positive mouse by RT-PCR. Founders of the DSP transgenic mouse were obtained successfully, and the expression of DSP was primarily confirmed.


Comparing the outcome of two surgical treatments for snoring using a patient-orientated measure of benefit.
Many different surgical techniques are currently used to treat simple snoring and their outcomes are difficult to compare. We used the Glasgow Benefit Inventory (GBI) to examine the outcome of two different surgical procedures. Thirty patients had been treated by uvulopalatopharyngoplasty (UPPP) and fifty-six by laser palatoplasty. The GBI scores showed close agreement with other subjective outcome measures. The two groups gave similar scores at 15 months postoperatively, and there was no deterioration in score with time. The GBI could be used to evaluate new surgical techniques for snoring, and could be useful in research and audit in this field.


Spontaneous liver rupture in hypereosinophilic syndrome: a rare but fatal complication.
We report a rare case of spontaneous liver rupture in a patient with hypereosinophilic syndrome (HES), of which the diagnosis was delayed, resulting in a fatal outcome. The diagnostic criteria and treatment of HES with hepatic involvement were reviewed. The possible cause of spontaneous liver rupture in HES and its management were also discussed. To our knowledge, this is the first case report of spontaneous liver rupture in HES. We emphasized the need of a high index of suspicion in diagnosing HES, so that early treatment could be initiated.


Comparison of tools for the assessment of inappropriate prescribing in hospitalized older people.
RATIONAL, AIMS AND OBJECTIVE: The aim of the present study was to compare the ease of use and the capability of four approaches [Medication Appropriateness Index (MAI), the Beers' criteria 2003, the Improved Prescribing in the Elderly Tool (IPET) and Health Plan Employer Data and Information Set (HEDIS)] in assessing changes in medication appropriateness in elderly patients over a period of hospitalization. A retrospective observational study in two hospitals in Northern Ireland using the four measures was undertaken, involving a cohort of 192 patients (aged > 65 years). Medication appropriateness assessments were made at three stages during the patients' hospital 'journey', that is, at admission, during their inpatient stay and at discharge. The identifying rates of inappropriate prescribing in elderly patients in hospital used validated screening tools: MAI, the Beers' criteria 2003, the IPET and HEDIS. The MAI was the most comprehensive approach but was also the most time consuming to apply. Data derived using the MAI indicated clearly that there was improved medication appropriateness over the three hospital stay stages (P < 0.001). Although this trend was also significant for the Beers' criteria 2003 (P < 0.05) and the IPET (P < 0.05) approaches, the HEDIS was unable to differentiate changes in appropriateness over time. There was a good correlation between data derived using the MAI and the Beers' criteria 2003 and the IPET approaches; this correlation was not evident for the HEDIS. The MAI is the most convincing tool in evaluating medication appropriateness, but is very time consuming to apply. Beers' criteria 2003 and the IPET perform to an acceptable standard within the clinical setting and are more practical in their application. The HEDIS, although simplest to apply, does not have the sensitivity to measure change in appropriateness over time.


CCL19-IgG prevents allograft rejection by impairment of immune cell trafficking.
An adaptive immune response is initiated in the T cell area of secondary lymphoid organs, where antigen-presenting dendritic cells may induce proliferation and differentiation in co-localized T cells after T cell receptor engagement. The chemokines CCL19 and CCL21 and their receptor CCR7 are essential in establishing dendritic cell and T cell recruitment and co-localization within this unique microenvironment. It is shown that systemic application of a fusion protein that consists of CCL19 fused to the Fc part of human IgG1 induces effects similar to the phenotype of CCR7-/- animals, like disturbed accumulation of T cells and dendritic cells in secondary lymphoid organs. CCL19-IgG further inhibited their co-localization, which resulted in a marked inhibition of antigen-specific T cell proliferation. The immunosuppressive potency of CCL19-IgG was tested in vivo using murine models for TH1-mediated immune responses (delayed-type hypersensitivity) and for transplantation of different solid organs. In allogeneic kidney transplantation as well as heterotopic allogeneic heart transplantation in different strain combinations, allograft rejection was reduced and organ survival was significantly prolonged by treatment with CCL19-IgG compared with controls. This shows that in contrast to only limited prolongation of graft survival in CCR7 knockout models, the therapeutic application of a CCR7 ligand in a wild-type environment provides a benefit in terms of immunosuppression.


Electrochemical dissolution of fractured nickel-titanium instruments in human extracted teeth.
To assess the effect of sodium chloride concentration in fluoridated solutions during the electrochemical dissolution of fractured rotary endodontic instruments. Two solutions were assessed (solution 1: NaF 12 g L-1 + NaCl 1 g L-1 , pH = 5.0; and solution 2: NaF 12 g L-1 + NaCl 180 g L-1 , pH = 5.0) using two tests: the ProTaper Universal F1 (PTU F1) instrument polarization test and the polarization test for intracanal PTU F1 fragments fractured in mandibular incisors. In the first test, two sets of five instruments were separately and partially immersed in each solution, and the electrical current was evaluated over 30 min. In the second test, 45 PTU F1 instruments were fractured within the root canals of mandibular incisors and subjected to potentiodynamic polarization for 30 min. The electrical current and the variations in the length of PTU F1 fragments were measured. The data were analysed statistically (anova and Wilcoxon and Mann-Whitney tests, respectively). Solution 2 was associated with more corrosive effects in both tests. In the first test, the PTU F1 instruments immersed in solution 2 had a higher electrical current (P < 0.001) and had a total dissolution time of approximately 540 s. In the second test, a larger difference between the baseline and final lengths of the fragments was noted in solution 2 (P = 0.011). Saturation of fluoridated solution with sodium chloride led to an increase in electrical current and microscopic reductions in the length of fractured instrument fragments subjected to electrochemical dissolution.


Relationship between self-reported intensity of headache and magnitude of surface EMG.
The study was designed to examine the relationship between self-reported intensity of headache and surface EMG. 98 patients, diagnosed by their neurologists with "muscle-contraction headaches" (tension-type headaches) were referred to evaluate their suitability for biofeedback therapy. At the time of examination, they were asked to rate their average headache intensity on a 10-point scale. Surface EMG data were collected to assess actual muscle contraction. Analysis indicated that among patients diagnosed with muscle contraction headache, there is a positive significant correlation between self-reported intensity of headache and actual muscle-contraction. The current data lend support to the hypothesis that the tension in the headaches currently described as "tension-type" may in fact refer to actual muscular tension or contraction.


Investing in the future of inpatient dermatology: the evolution and impact of specialized dermatologic consultation in hospitalized patients.
Inpatient dermatology has transitioned from units that admitted and cared for patients with chronic dermatoses to consultative services that provide a wide breadth of care, leading to a paradigm shift in the role and impact of dermatologists in the inpatient setting. Consultative dermatology provides a distinct and essential service in the care of hospitalized patients, leading to improved care quality along with reductions in inappropriate health care spending.


PAI-1 promotes the accumulation of exudate macrophages and worsens pulmonary fibrosis following type II alveolar epithelial cell injury.
Fibrotic disorders of the lung are associated with perturbations in the plasminogen activation system. Specifically, plasminogen activator inhibitor-1 (PAI-1) expression is increased relative to the plasminogen activators. A direct role for this imbalance in modulating the severity of lung scarring following injury has been substantiated in the bleomycin model of pulmonary fibrosis. However, it remains unclear whether derangements in the plasminogen activation system contribute more generally to the pathogenesis of lung fibrosis beyond bleomycin injury. To answer this question, we employed an alternative model of lung scarring, in which type II alveolar epithelial cells (AECs) are specifically injured by administering diphtheria toxin (DT) to mice genetically engineered to express the human DT receptor (DTR) off the surfactant protein C promoter. This targeted AEC injury results in the diffuse accumulation of interstitial collagen. In the present study, we found that this targeted type II cell insult also increases PAI-1 expression in the alveolar compartment. We identified AECs and lung macrophages to be sources of PAI-1 production. To determine whether this elevated PAI-1 concentration was directly related to the severity of fibrosis, DTR(+) mice were crossed into a PAI-1-deficient background (DTR(+) : PAI-1(-/-) ). DT administration to DTR(+) : PAI-1(-/-) animals caused significantly less fibrosis than was measured in DTR(+) mice with intact PAI-1 production. PAI-1 deficiency also abrogated the accumulation of CD11b(+) exudate macrophages that were found to express PAI-1 and type-1 collagen. These observations substantiate the critical function of PAI-1 in pulmonary fibrosis pathogenesis and provide new insight into a potential mechanism by which this pro-fibrotic molecule influences collagen accumulation. Copyright © 2012 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.


[Prognostic markers of multiple sclerosis].
Multiple sclerosis (MS) is a chronic autoimmune disease with a high degree of heterogeneity due to course and prognosis. More than half of MS patients do not discuss their long-term prognosis with the doctor. At the same time, most patients consider the importance of personalized prognosis for decisions in family planning, career, and medical interventions. Clinical markers are used to determine the prognosis in routine clinical practice. However, it is nominally divided into positive and negative factors. It allows making a general conclusion about the MS prognosis. Neuroimaging and biological markers are used mostly for research purposes. But also they are already actively used in clinical trials for endpoint`s investigation. The review describes studies investigating prognostic clinical, neuroimaging, and biological markers.


Comparing evolvability and variability of quantitative traits.
There are two distinct reasons for making comparisons of genetic variation for quantitative characters. The first is to compare evolvabilities, or ability to respond to selection, and the second is to make inferences about the forces that maintain genetic variability. Measures of variation that are standardized by the trait mean, such as the additive genetic coefficient of variation, are appropriate for both purposes. Variation has usually been compared as narrow sense heritabilities, but this is almost always an inappropriate comparative measure of evolvability and variability. Coefficients of variation were calculated from 842 estimates of trait means, variances and heritabilities in the literature. Traits closely related to fitness have higher additive genetic and nongenetic variability by the coefficient of variation criterion than characters under weak selection. This is the reverse of the accepted conclusion based on comparisons of heritability. The low heritability of fitness components is best explained by their high residual variation. The high additive genetic and residual variability of fitness traits might be explained by the great number of genetic and environmental events they are affected by, or by a lack of stabilizing selection to reduce their phenotypic variance. Over one-third of the quantitative genetics papers reviewed did not report trait means or variances. Researchers should always report these statistics, so that measures of variation appropriate to a variety of situations may be calculated.


Lack of bone neoplasms and persistence of bone efficacy in cynomolgus macaques after long-term treatment with teriparatide [rhPTH(1-34)].
In rats, teriparatide [rhPTH(1-34)] causes marked increases in bone mass and osteosarcoma. In primates, teriparatide causes lesser increases in bone mass, and osteosarcomas have not been reported. Previous studies in primates were not designed to detect bone tumors and did not include a prolonged post-treatment observation period to determine whether tumors would arise after cessation of treatment. Ovariectomized (OVX), skeletally mature, cynomolgus monkeys (n = 30 per group) were given teriparatide for 18 mo at either 0 or 5 microg/kg/d subcutaneously. After 18 mo of treatment, subgroups of six monkeys from both groups were killed and evaluated, whereas all remaining monkeys entered a 3-yr observation period in which they did not receive teriparatide. Surveillance for bone tumors was accomplished with plain film radiographs, visual examination of the skeleton at necropsy, and histologic evaluation of multiple skeletal sites. Quantitative assessments of bone mass, architecture, and strength were also performed. After the 18-mo treatment period, vertebral BMD, BMC, and strength (ultimate load) were increased by 29%, 36%, and 52%, respectively, compared with OVX controls. Proximal femur BMD, BMC, and strength were also increased by 15%, 28% and 33%, respectively. After 3 yr without treatment, no differences in bone mass or strength at the vertebra were observed relative to OVX controls; however, the femoral neck showed significant persistence in stiffness (20%), BMC (14%), and trabecular BV/TV (53%), indicating a retention of teriparatide efficacy at the hip. Radiographs and histology did not identify any bone proliferative lesions or microscopic lesions of osteosarcoma at the end of the treatment or observation period. These data indicate that teriparatide did not induce bone proliferative lesions over a 4.5-yr interval of observation, including 18 mo of treatment and 3 yr of follow-up observation. Bone analyses confirmed that teriparatide caused increases in bone mass and strength, consistent with previous studies. During the withdrawal phase, beneficial effects of teriparatide treatment on the vertebra were lost; however, some of the beneficial effects on the proximal femur persisted for 3 yr after cessation of treatment. Although the lack of bone tumors in this study provides some additional reassurance regarding the safety of teriparatide for the primate skeleton, the small group size and other limitations of this, or any other animal study, limit the ability to draw definitive conclusions regarding the risk of bone tumor developments in patients.


Downregulation of CYP1A2, CYP2B6, and CYP3A4 in Human Hepatocytes by Prolyl Hydroxylase Domain 2 Inhibitors via Hypoxia-Inducible Factor-α Stabilization.
Hypoxia-inducible factor (HIF) is associated with the expression of CYP, but the underlying mechanism remains uncertain. In this study, we investigated the effect of HIF-α stabilization caused by novel prolyl hydroxylase domain (PHD) 2 inhibitors, which are HIF-α stabilizers that mimic hypoxia, on the expressions of CYP1A2, CYP2B6, and CYP3A4 in human hepatocytes. An mRNA expression analysis of human hepatocytes treated with PHD2 inhibitors for 72 hours showed the downregulation of genes encoding CYP1A2, CYP2B6, and CYP3A4. The mRNA repressions were accompanied with an increase in erythropoietin protein, a marker of HIF-α stabilization, indicating that HIF-α stabilization was involved in the downregulation of the CYP isoforms. To understand the underlying mechanisms, we assessed the relationship between the expressions of the CYP isoforms and those of their regulating transcription factors [aryl hydrocarbon receptor (AhR), AhR nuclear translocator (ARNT), constitutive androstane receptor (CAR), pregnane X receptor (PXR), and retinoid X receptor (RXR)] in human hepatocytes treated with the HIF-α stabilizers. As a result, the mRNA level of AhR did not decrease, although ARNT expression was repressed. On the other hand, the mRNA expression levels of CAR, PXR, and RXR were repressed and closely associated with those of CYP2B6 and CYP3A4. Although the underlying mechanism of the downregulation for CYP1A2 remains unclear, the presently reported results suggest that the downregulation of CYP2B6 and CYP3A4 via HIF-α stabilization is caused by a decrease in the expressions of CAR, PXR, and RXR. SIGNIFICANCE STATEMENT: We showed that hypoxia-inducible factor (HIF)-α stabilization downregulates CYP1A2, CYP2B6, and CYP3A4 using prolyl hydroxylase domain 2 inhibitors, which are HIF-α stabilizers, as a new tool to mimic hypoxia in human hepatocytes. To understand the underlying mechanisms, we assessed the relationship between the expressions of the CYP isoforms and those of their regulating transcription factors. Our findings would contribute to a better understanding of the hypoxia-triggered regulatory mechanism of drug-metabolizing enzymes in human hepatocytes.


Clinical Application of Circulating Tumor Cells in Gastric Cancer.
Early detection and accurate monitoring of cancer is important for improving clinical outcomes. Endoscopic biopsy and/or surgical resection specimens are the gold standard for diagnosing gastric cancer and are also useful for selecting therapeutic strategies based on the analysis of genomic/immune parameters. However, these approaches cannot be easily performed because of their invasiveness and because these specimens do not always reflect tumor dynamics and drug sensitivities during therapeutic processes, especially chemotherapy. Accordingly, many researchers have tried to develop noninvasive novel biomarkers that can monitor real-time tumor dynamics for early diagnosis, prognostic evaluation, and prediction of recurrence and therapeutic efficacy. Circulating tumor cells (CTCs) are metastatic cells that are released from the primary tumors into the blood stream and comprise a crucial step in hematogenous metastasis. CTCs, as a liquid biopsy, have received a considerable amount of attention from researchers since they are easily accessible in peripheral blood, avoiding the invasiveness associated with traditional biopsy techniques; they can also be used to derive clinical information for monitoring disease status. In this review, with respect to CTCs, we summarize the metastatic cascade, detection methods, clinical applications, and prospects for patients with gastric cancer.


Aetiology and management of the secondary brow.
The finding that a brow presenting within the pelvic cavity in the mento-anterior position during the second stage of labour could be easily and safely delivered by means of Kielland's forceps rotation and extraction led to a search for the cause of this rare phenomenon since normally brows are associated with non-engagement of the head. A personal series of six cases, collected over 39 years, were studied. Particular attention was paid to pelvic size and shape with detailed X-ray pelvimetry being obtained in the puerperium from the latter three patients. In all cases the pelvises were found to be commodious and, in the three where X-ray pelvimetries were taken, they were found to be anthropoid in shape, which is known to favour the occipito-posterior position. Past and subsequent pregnancies conducted by the author were in every instance associated with an occipito-posterior position, and a mechanism of labour for secondary brow presentations until impasse occurred was suggested. It was concluded that the condition only occurred when the pelvis was roomy and anthropoid in shape, and, for those experienced in their use, Kielland's forceps rotation and extraction was preferable to caesarean section. Furthermore, aficionados of the ventouse were warned to avoid attempting delivery with that instrument for such would be doomed to failure.


Flash Brillouin Scattering: A Confocal Technique for Measuring Glass Transitions at High Scan Rates.
Glass transition temperatures T g are most commonly measured by differential scanning calorimetry, a method that has been extended to the flash scanning calorimetry (FSC) regime by reducing sample volumes. However, significant manual preparation effort can render FSC impractical for, e.g., local probing of spatially heterogeneous specimens. Another strategy can be to select a small volume by focusing down a laser beam, where Brillouin Light Scattering (BLS) is a proven method for confocal T g measurement. Here, we introduce Flash Brillouin Scattering, which extends BLS to fast scan rates, achieved by periodically heating the probed region with an infrared laser. For comparison with conventional BLS, we first characterize T g of pure glycerol, and show how rapid quenching produces a less packed glass with downshifted sound velocity. We then turn toward its aqueous solutions, which crystallize too fast for a nonflash approach, and demonstrate scan rates in excess of 105 K/s. These results are of interest not only because glycerol is a model system for hydrogen-bonded glass formers, but also because of its applications as a cryoprotectant for frozen biological samples. Light scattering studies of the latter, currently limited to cryo-Raman spectroscopy, are likely to be complemented by the technique introduced here.


An epidemiologic overview of acute kidney injury in intensive care units.
Acute kidney injury (AKI) is a frequent event among critically ill patients hospitalized in intensive care units (ICU) and represents a global public health problem, being imperative an interdisciplinary approach. To investigate, through literature review, the AKI epidemiology in ICUs. METHODS: Online research in Medline, Scientific Electronic Library Online, and Latin American and Caribbean Literature in Health Sciences databases, with analysis of the most relevant 47 studies published between 2010 and 2017. Data of the 67,033 patients from more than 300 ICUs from different regions of the world were analyzed. The overall incidence of AKI ranged from 2.5% to 92.2%, and the mortality from 5% to 80%. The length of ICU stay ranged from five to twenty-one days, and the need for renal replacement therapy from 0.8% to 59.2%. AKI patients had substantially higher mortality rates and longer hospital stays than patients without AKI. AKI incidence presented high variability among the studies. One of the reasons for that were the different criteria used to define the cases. Availability of local resources, renal replacement therapy needs, serum creatinine at ICU admission, volume overload, and sepsis, among others, influence mortality rates in AKI patients.


Prioritization of putatively detrimental variants in euploid miscarriages.
Miscarriage is the spontaneous termination of a pregnancy before 24 weeks of gestation. We studied the genome of euploid miscarried embryos from mothers in the range of healthy adult individuals to understand genetic susceptibility to miscarriage not caused by chromosomal aneuploidies. We developed GP , a pipeline that we used to prioritize 439 unique variants in 399 genes, including genes known to be associated with miscarriages. Among the prioritized genes we found STAG2 coding for the cohesin complex subunit, for which inactivation in mouse is lethal, and TLE4 a target of Notch and Wnt, physically interacting with a region on chromosome 9 associated to miscarriages.


Cycloaddition of benzyne to SWCNT: towards CNT-based paddle wheels.
The cycloaddition of benzyne to SWCNT has been carried out for the first time. Raman spectroscopy, TGA, HR-TEM, UV-vis-NIR as well as XPS have been used for products characterization.


Incorporation of Coumarin-Based Fluorescent Monomers into Co-Oligomeric Molecules.
With the purpose of modifying organic fluorescent dyes based on the coumarin scaffold, and developing and evaluating a route to its incorporation into a polymeric backbone, a study was conducted on the co-polymerization of 3-vinylcoumarins with styrene and methyl acrylate using 2,2-azobis(isobutyronitrile) (AIBN) as the radical initiator. The structural and photophysical characterization proved the incorporation of the coumarin monomers into the polymeric chain and further showed a decrease in the fluorescence quantum yields in the co-oligomers.


Ewing's sarcoma of the orbit with intracranial extension: A rare cause of unilateral proptosis.
Ewing's sarcoma causing unilateral proptosis along with bifrontal extradural infiltration in a child is an unusual presentation. A female patient presented with features of painless proptosis of the left eye with visual deterioration. Her radiology revealed an infiltrating intraorbital, extraconal tumor with intracranial bifrontal extradural extension causing mass effect. Total excision of the intraorbital and intracranial part of the tumor along with postoperative chemo- and radiotherapy brought about a substantial relief. The clinicoradiological presentation and management of this rare entity are discussed.


Low level laser therapy on experimental myopathy.
The aim of the present work was to study the effect of Helium-Neon (HeNe) and Gallium Arsenide (GaAs) laser upon nitric oxide (NO) plasma levels, an inflammatory biomarker associated with oxidative stress, in rats with experimental myopathy. These were evaluated through histological assessment. The groups studied were: (A) control (intact rats that received LLLT sham exposures), (B) rats with myopathy and sacrificed at 24 h later, (C) rats with myopathy and sacrificed 8 days later, (D) rats with myopathy and treated with HeNe laser, (E) rats with myopathy and treated with GaAs laser, (F) intact rats treated with HeNe laser and (G) intact rats treated with GaAs laser. Myopathy was induced by injecting 50μl of 1% carrageenan λ (type IV) in the left gastrocnemius muscle. Low Level Laser Therapy (LLLT) was applied with 9.5 J.cm(-2) daily for 10 consecutive days with each laser. The determination of the NO was made by spectrophotometry. The muscles were stained with Hematoxylin-Eosin and examined by optic microscopy. Quantitative variables were statistically analyzed by the Fisher test, and categorical by applying Pearson's Chi Squared test at p <0.05 for all cases. In groups B and C, NO was significantly increased compared to groups A, D, E, F and G (p<0.05). In group C, the percentage of area with inflammatory infiltration was significantly increased compared to the other groups (p<0.001). LLLT decreased plasma levels of NO in rats with experimental myopathies and significant muscle recovery.


Characterization of stationary phases by a linear solvation energy relationship utilizing supercritical fluid chromatography.
Supercritical fluid chromatography was utilized in combination with the Abraham model of linear solvation energy relationship to characterize 11 different HPLC stationary phases. System constants were determined at one supercritical fluid chromatography condition for each stationary phase. The results indicate that several types of silica columns, including type B silica, type C silica, and fused core silica, are very similar in their retention behavior. Several aromatic stationary phases were characterized and it was found that, in contrast to the other phases studied, all of the aromatic stationary phases had positive contributions from the dispersion/cavity (v) term of the linear solvation energy relationship. Several aliphatic phases were characterized and there were several linear solvation energy relationship constants that differentiated the phases from each other, mainly the polar terms (dipolarity and hydrogen bonding). One stationary phase, a fused core pentafluorophenyl (PFP) phase, had very poor regression quality. The column volume of this phase was lower than the others in the study, which may have had some impact on the results of the regression.


Sciatic nerve entrapment in a child. Case report.
A child was brought for evaluation of signs of unilateral chronic progressive sciatic nerve dysfunction found to be due to nerve entrapment in the thigh by a fibrovascular band. Sectioning of the band was followed by marked improvement in the nerve function. Compression by a band is a rare but treatable cause of sciatic neuropathy.


Influence of iron on plasma interleukin-2 and gamma interferon level in iron deficiency anemia.
to find out the the influence of iron on the levels of plasma IL-2 and IFNgamma in iron deficiency anemia patient. the study subjects were patients with IDA (intended samples) selected from the accessible population that met the inclusion and exclusion criteria using consecutive sampling technique and by signing an informed consent. Blood samples were taken for measurement of IL-2 and IFNgamma concentrations as well as list of questionnaires for obtaining data on age, gender, body weight, infections suffered, use of immunosuppressive medicines, malnutrition, malignancy and genetic disorder, and IDA diagnosis. The levels of IL-2 and IFNgamma were measured before and after 8 weeks of iron tablet adminitration using immunoassay solid phase ELISA. Descriptive statistics was used to illustrate patient's characteristics and frequency distribution of various variables. The average difference of IL-2 and IFNgamma concentrations was also examined pre and post administration of iron tablets for 8 weeks using Wilkoxon sign rank test. in the study, 26 IDA patients were eligible, chosen from 64 IDA patients who were then given iron tablets for 8 weeks. By the end of the treatment, they were tested for complete blood analysis, serum ferritin, and cytokines of plasma. Sign rank test of Wilcoxon was used to find out direction and degree of difference between the two paired groups which showed a significant difference between pre and post iron tablet administration of hemoglobin (Z= -4.561; p<0.001), MCV (Z= -4.276; p<0.001), MCH (Z= -3.616; p<0.001) and feritin (Z= -3.556; p<0.001). After 8 weeks of iron tablets, the study also noted, increasing of plasma IL-2 and IFNgamma. Before treatment level of plasma IL-2 was 7.65 pg/l then became 29.3 pg/l after treatment (Z=- 3.508, p<0.001) and plasma IFNg before treatment was 10.15 pg/l became 46.7 pg/l after treatment (Z= -4.241, p<0.001). concentration of IL2 dan IFNgamma in plasma significantly increased after administration of iron tablets for 8 weeks as compared to that before iron treatment.


Planned Parenthood Federation of America v. Bowen, 25 February 1988.
The plaintiffs were family planning (FP) services and physicians who sought a preliminary injunction against enforcement of regulations promulgated by the Secretary of Health and Human Services of the US. The regulations prohibited counseling and referral for abortion services by FP services that receive funds under Title X of the Public Health Services Act. The Court held that the regulations exceeded the Secretary's authority, since nothing in Title X or legislative history relating to its enactment indicated that Congress intended to prohibit FP services from offering information with respect to abortions. The Court also ruled that the regulations violated a woman's Fifth Amendment liberty interest in choosing to have an abortion, her First Amendment right to receive necessary information to exercise her liberty interest, and a physician's First Amendment right to disseminate necessary information to patients. The Court granted the request for a preliminary injunction. On 15 June 1988, the same court granted the plaintiffs' request for a permanent injunction against enforcement of the regulations. On 3 March 1988, the US District Court, D. Massachusetts, also enjoined enforcement of the regulations (Commonwealth of Massachusetts vs. Bowen [679 F.Supp. 137]). On 30 June 1988, however, the US District Court, S.D. New York, rejected a challenge to the regulations, ruling that the Secretary did not exceed his authority and that the regulations were not unconstitutional (State of New York vs. Bowen [690 F.Supp. 1261]).


Ultrasound-Induced DNA Damage and Cellular Response: Historical Review, Mechanisms Analysis, and Therapeutic Implications.
The biological effects of ultrasound may be classified into thermal and nonthermal mechanisms. The nonthermal effects may be further classified into cavitational and noncavitational mechanisms. DNA damage induced by ultrasound is considered to be related to nonthermal cavitations. For this aspect, many in vitro studies on DNA have been conducted for evaluating the safety of diagnostic ultrasound, particularly in fetal imaging. Technological advancement in detecting DNA damage both in vitro and in vivo have elucidated the mechanism of DNA damage formation and their cellular response. Damage to DNA, and the residual damages after DNA repair are implicated in the biological effects. Here, we discuss the historical evidence of ultrasound on DNA damage and the mechanism of DNA damage formation both in vitro and in vivo, compared with those induced by ionizing radiation. We also offer a commentary on the safety of ultrasound over X-ray-based imaging. Also, understanding the various mechanisms involved in the bioeffects of ultrasound will lead us to alternative strategies for use of ultrasound for therapy.


Effects of low concentrations of o(3) on net photosynthesis, dark respiration, and chlorophyll contents in aging hybrid poplar leaves.
Chronic exposure of hybrid poplar (Populus deltoides x trichocarpa) plants to low concentrations of ozone had negative impact upon net photosynthetic capacity, dark respiration, and leaf chlorophyll contents. Exposure to as much as 0.20 microliters per liter O(3) had no immediate effect on net photosynthesis (P(n)), but chronic exposure to 0.125 or 0.085 microliters per liter had a number of gradual effects on CO(2) exchange. These included increased dark respiration and consequently increased light compensation points in very young leaves (4-6 days old); and decreased P(n), leaf chlorophyll a and b contents, light saturation points, and apparent quantum yields in leaves 10 to 70 days old. Decreased P(n) was partially due to accelerated aging in leaves exposed to O(3), and lightsaturated P(n) was linearly related to chlorophyll a + b contents. Differences in light-saturated P(n) between control and O(3)-treated leaves of the same age were mostly due to photosaturation in O(3)-treated leaves and to a much lesser extent to lowered apparent quantum yields. Also, since P(n) and dark respiration were most affected by O(3) at different leaf ages, distinct modes of action are suggested. The effects of leaf aging on CO(2) exchange were considerable, but typical of other species. However, careful monitoring of the interacting effects of leaf age and pollutant exposure was needed in order to characterize the impact of chronic O(3) exposure upon CO(2) exchange.


Borderline ovarian tumors in Finland: epidemiology and familial occurrence.
A total of 1,197 borderline ovarian tumors were reported to the Finish Cancer Registry in 1973-1992. The mean age of the patients was 52 years, while the mean age of the 7,060 patients with an invasive epithelial ovarian carcinoma reported in the same time period was 62 years. The incidence of borderline ovarian tumors did not increase with age after patients became 35 years old and older. The overall age-adjusted incidence of borderline ovarian tumors was 1.8 per 100,000 women-years. Familial cancer occurrence during 1967-1992 was studied among relatives of 144 index patients diagnosed in 1980-1982. No borderline ovarian tumors were detected in the relatives, and only one of the 446 female first-degree relatives had an epithelial ovarian cancer. The expected number (borderline and invasive combined) was 1.9. The mothers of the index patients had an increased risk for pancreatic cancer (standardized incidence ratio 4.9, 95% confidence interval 1.0-14.3) and for cancer of the uterine cervix (standardized incidence ratio 7.8, 95% confidence interval 1.6-22.8). No significant increase in cancer risk was observed for fathers, brothers, or sisters of the patients with borderline ovarian tumors.


Microscopic localization of active gelatinases in equine osteochondritis dissecans (OCD) cartilage.
To investigate the relationship between matrix metalloproteinase (MMP) activity and osteochondritis dissecans (OCD) in the equine joint. Equine articular cartilage was obtained from normal (N = 8) and osteochondrotic (OCD) (N = 6) femoropatellar joints from horses at necropsy. The activity of gelatinase MMPs was determined in sections of cartilage by in situ gelatin zymography. Gelatinase activity was markedly increased in articular cartilage obtained from OCD samples and was particularly prominent in the deep cartilage zone. Activity was only seen in the pericellular area of chondrocytes. In addition, in OCD cartilage there were vertical lines of activity, starting from the deep zone and radiating towards the articular surface. In contrast, normal cartilage showed only a very small amount of gelatinolytic activity, which was not restricted to specific cartilage zones. Gelatin zymography of culture supernatants from isolated chondrocytes demonstrated increased production of MMP-2 and MMP-9 from OCD chondrocytes. Sections of articular cartilage from OCD lesions revealed MMP activity, especially in the deep zone adjacent to the calcified subchondral bone. This MMP activity could account for the loss of cartilage integrity in the deep cartilage zone and the vertical lines of activity could represent areas of mechanical weakness, likely to result in fissures and the release of cartilage fragments into the joint space.


Structure-Dependent Effects of Phthalates on Intercellular and Intracellular Communication in Liver Oval Cells.
Humans are exposed to phthalates released from plastics, cosmetics, or food on a daily basis. Phthalates have low acute liver toxicity, but their chronic exposures could induce molecular and cellular effects linked to adverse health outcomes, such as liver tumor promotion or chronic liver diseases. The alternation of gap junctional intercellular communication (GJIC) and MAPK-Erk1/2 pathways in liver progenitor or oval cells can disrupt liver tissue homeostatic mechanisms and affect the development and severity of these adverse outcomes. Our study with 20 different phthalates revealed their structurally dependent effects on liver GJIC and MAPK-Erk1/2 signaling in rat liver WB-F344 cell line with characteristics of liver oval cells. The phthalates with a medium-length side chain (3-6 C) were the most potent dysregulators of GJIC and activators of MAPK-Erk1/2. The effects occurred rapidly, suggesting the activation of non-genomic (non-transcriptional) mechanisms directly by the parental compounds. Short-chain phthalates (1-2 C) did not dysregulate GJIC even after longer exposures and did not activate MAPK-Erk1/2. Longer chain (≥7 C) phthalates, such as DEHP or DINP, moderately activated MAPK-Erk1/2, but inhibited GJIC only after prolonged exposures (>12 h), suggesting that GJIC dysregulation occurs via genomic mechanisms, or (bio)transformation. Overall, medium-chain phthalates rapidly affected the key tissue homeostatic mechanisms in the liver oval cell population via non-genomic pathways, which might contribute to the development of chronic liver toxicity and diseases.


[Surgical treatment of carcinoma of the stomach invading to the esophago-gastric junction].
In order to elucidate the clinico-pathologic features of gastric cancer invading to esophago-gastric junction and to discuss about rational surgical treatment, 111 cases during the 23-year period between 1957 to 1983 were selected. Of these patients, a total gastrectomy with lower esophagectomy or subtotal esophagectomy on 82 (74%), a cardiofundectomy on 26 (26%) and caudal hemipancreatectomy with splenectomy on 99 (89%) were carried out. It is severely impossible to eradicate primary tumor with an adequate margin of the esophagus and intrathoracic lymph nodes only by abdominal approach in resecting gastric carcinoma invading to esophago-gastric junction. Our experience emphasizes that a laparotomy with median sternotomy is ideal one, in addition, blunt dissection of the thoracic esophagus or right thoracotomy may be done if indicated.


Form and Function of the Vertebrate and Invertebrate Blood-Brain Barriers.
The need to protect neural tissue from toxins or other substances is as old as neural tissue itself. Early recognition of this need has led to more than a century of investigation of the blood-brain barrier (BBB). Many aspects of this important neuroprotective barrier have now been well established, including its cellular architecture and barrier and transport functions. Unsurprisingly, most research has had a human orientation, using mammalian and other animal models to develop translational research findings. However, cell layers forming a barrier between vascular spaces and neural tissues are found broadly throughout the invertebrates as well as in all vertebrates. Unfortunately, previous scenarios for the evolution of the BBB typically adopt a classic, now discredited 'scala naturae' approach, which inaccurately describes a putative evolutionary progression of the mammalian BBB from simple invertebrates to mammals. In fact, BBB-like structures have evolved independently numerous times, complicating simplistic views of the evolution of the BBB as a linear process. Here, we review BBBs in their various forms in both invertebrates and vertebrates, with an emphasis on the function, evolution, and conditional relevance of popular animal models such as the fruit fly and the zebrafish to mammalian BBB research.


Immuno-PET Imaging and Radioimmunotherapy of Lymphomas.
Monoclonal antibodies (Ab) have revolutionized the management of lymphomas, the most common hematologic malignancy in adults. Indeed, incorporation of rituximab into the regimen for indolent non-Hodgkin's lymphomas (NHLs) has dramatically improved treatment response and disease outcome. Yet, newer Ab therapeutics against promising antigen targets need to be developed to treat refractory or relapsed patients. Treatment efficacy can be further enhanced by conjugating toxic molecules to the Abs. Radioimmunotherapy (RIT) harnesses Abs as vehicles for targeted delivery of therapeutic radionuclide payloads for direct killing of targeted tumor cells. Positron emission tomography (PET) with radiolabeled Abs (called immuno-PET) can facilitate the development of new Ab therapeutics and RIT by providing pharmacokinetic and pharmacodynamic information and by quantifying tumor antigen level relevant for treatment decision. Immuno-PET has recently gravitated toward labeling Abs with 89Zr, a radiometal with a 3.3 day half-life that is trapped following Ab internalization and thus provides high-resolution PET images with excellent contrast. Immuno-PET methods against major lymphoma antigens including CD20 and other promising targets are currently under development. With continued improvements, immuno-PET has the potential to be used in lymphoma management as an imaging biomarker for patient selection and assessment of treatment response.


Distinct spatial activation of intrinsic and extrinsic apoptosis pathways in natural scrapie: association with prion-related lesions.
Neurodegeneration and gliosis are the main neuropathological features of prion diseases. However, the molecular mechanisms involved in these processes remain unclear. Several studies have demonstrated changes in the expression of apoptotic factors and inflammatory cytokines in animals with experimental infection. Here we present the expression profiles of 15 genes implicated in the intrinsic and extrinsic apoptotic pathways in the central nervous systems of sheep naturally infected with scrapie. Expression changes obtained by real-time RT-PCR were also compared with the extent of classical scrapie lesions, such as prion deposition, neuronal vacuolisation, spongiosis, and astrogliosis as well as with the activation of caspase-3, using a stepwise regression. The results suggest that the factors assessed participate in apoptotic or inflammatory functions, depending on the affected area. The mitochondrial apoptosis pathway was associated with prion deposition in the prefrontal cortex (the less affected area), and with activation of caspase-3-mediated cell death via over-expression of BAK. In addition to its known association with astroglial activation, the extrinsic apoptosis pathway was also related to cell death and neuronal vacuolisation.


Pathogenesis of Anti-melanoma Differentiation-Associated Gene 5 Antibody-Positive Dermatomyositis: A Concise Review With an Emphasis on Type I Interferon System.
Anti-melanoma differentiation-associated gene 5 antibody-positive dermatomyositis (MDA5+ DM) is typically characterized by cutaneous manifestations, amyopathic or hypomyopathic muscle involvement, and a high incidence of rapid progressive interstitial lung disease (RP-ILD). However, the exact etiology and pathogenesis of this condition has yet to be fully elucidated. Melanoma differentiation-associated gene 5 (MDA5), as the autoantigen target, is a member of the retinoic acid-inducible gene-I (RIG-I) family. The MDA5 protein can function as a cytosolic sensor that recognizes viral double-strand RNA and then triggers the transcription of genes encoding type I interferon (IFN). Therefore, it was presumed that viruses might trigger the overproduction of type I IFN, thus contributing to the development of MDA5+ DM. Emerging evidence provides further support to this hypothesis: the increased serum IFNα level was detected in the patients with MDA5+ DM, and the type I IFN gene signature was upregulated in both the peripheral blood mononuclear cells (PBMCs) and the skin tissues from these patients. In particular, RNA sequencing revealed the over-expression of the type I IFN genes in blood vessels from MDA5+ DM patients. In addition, Janus kinase (JAK) inhibitors achieved the promising therapeutic effects in cases with interstitial lung disease (ILD) associated with MDA5+ DM. In this review, we discuss the role of the type I IFN system in the pathogenesis of MDA5+ DM.


Translation and evaluation of a comprehensive educational program for cardiac rehabilitation patients in Latin America: A multi-national, longitudinal study.
To translate, cross-culturally adapt and validate a comprehensive evidence- and theoretically-based CR education intervention in Latin America. First, best practices in translation and cross-cultural adaptation were applied through 6 steps. Then, the Spanish version was delivered to CR participants from programs in Colombia, Costa Rica and Peru for validation, such that the evaluation was pre-post, uncontrolled, pragmatic, observational, and prospective in design. Participants completed surveys assessing knowledge, health literacy, self-efficacy, and health behaviours. All outcomes were assessed pre-, and post-CR, as well as 6 months after CR completion. After translation of the patient guide from English to Spanish, 5 of the 9 booklets were culturally adapted. Two-hundred and forty-nine patients consented to participate, of which 184 (74 %) completed post-CR, and 121 (48 %) completed final assessments. There was a significant improvement in disease-related knowledge pre- to post-CR, as well as in health literacy, self-efficacy, and health behaviours (all p < 0.05). These gains were sustained 6 months post-program. With adjustment, CR attendance (i.e., exposure to the education) was associated with greater post-CR knowledge (ß = 0.026; p = 0.01). A patient education intervention for CR patients in Latin America has been validated, and wider implementation is warranted. Application of this first-ever validated CR education program for Spanish-speaking settings may result in secondary prevention.


HDAC6 and ovarian cancer.
The special class IIb histone deacetylase, HDAC6, plays a prominent role in many cellular processes related to cancer, including oncogenesis, the cell stress response, motility, and myriad signaling pathways. Many of the lessons learned from other cancers can be applied to ovarian cancer as well. HDAC6 interacts with diverse proteins such as HSP90, cortactin, tubulin, dynein, p300, Bax, and GRK2 in both the nucleus and cytoplasm to carry out these cancerous functions. Not all pro-cancer interactions of HDAC6 involve deacetylation. The idea of using HDAC6 as a target for cancer treatment continues to expand in recent years, and more potent and specific HDAC6 inhibitors are required to effectively down-regulate the tumor-prone cell signaling pathways responsible for ovarian cancer.


Impact of diet, exercise end diet combined with exercise programs on plasma lipoprotein and adiponectin levels in obese girls.
We studied the effect of three programs, diet restriction (D), individualized exercise training (E) at the maximal lipid oxidation point (LIPOXmax) and diet combined with exercise (D+E), on body mass, plasma lipoprotein and adiponectin levels in obese girls. Eighteen obese adolescents girls aged 12-14 years were studied. A longitudinal intervention was carried out, consisting of a two-month diet (D; -500 kcal·day-1), of individualized exercise (E; 4 days/week, 90 min·day-1) and of diet combined with exercise (D+E). Body mass, body mass index (BMI), body fat mass, waist circumference, substrate crossover point, LIPOXmax point, homeostasis model assessment (HOMA-IR) index, fasting levels of lipids and circulatory adiponectin, were measured in all subjects before and after the program. In subjects of the D+E group, body mass, BMI, body fat mass, waist circumference, HOMA-IR, low-density lipoprotein cholesterol (LDL-C) and total cholesterol / high-density lipoprotein cholesterol (TC/HDL-C) ratio were significantly lower, and HDL-C and adiponectin were higher after the program than that of subjects in the D or E groups. Diet/exercise improved the ability to oxidize lipids during exercise (crossover point: + 18.5 ± 3.4 of % Wmax; p < 0.01 and fat oxidation rate at LIPOXmax: + 89.7 ± 19.7 mg·min(-1); p < 0.01). In the D+E group, significant correlations were found between changes in body mass and adiponectin and between changes in the TC/HDL-C ratio and LIPOXmax. These findings show that the combined program of diet restriction and individualized exercise training at the LIPOXmax point is necessary to simultaneously improve body mass loss, adiponectin levels, as well as metabolic parameters, in obese girls. Key pointsDiet combined with exercise training improved body composition, adiponectin levels and metabolic parameters in obese girls.Diet only decreases body mass and LDL-C without improving fat oxidation and HDL- C.Individualized exercise training at LIPOXmax point improved the HDL-C and the circulatory adiponectin levels with any change of LDL-C and body composition.


Demographic, Husbandry, and Biosecurity Factors Associated with the Presence of Campylobacter spp. in Small Poultry Flocks in Ontario, Canada.
This study is part of a 2 year disease surveillance project conducted to establish the prevalence of poultry and zoonotic pathogens, including Campylobacter spp., among small poultry flocks in Ontario, Canada. For each post-mortem submission to the Animal Health Laboratory, a pooled sample of cecal tissue was cultured for Campylobacter spp., and a husbandry and biosecurity questionnaire was completed by the flock owner (n = 153). Using both laboratory and questionnaire data, our objective was to investigate demographic, husbandry, and biosecurity factors associated with the presence of Campylobacter spp. in small flocks. Two multivariable logistic regression models were built. In the farm model, the odds of Campylobacter spp. were higher in turkeys, and when birds were housed in a mixed group with different species and/or types of birds. The odds were lower when antibiotics were used within the last 12 months, and when birds had at least some free-range access. The effect of pest control depended on the number of birds at risk. In the coop model, the odds of Campylobacter spp. were lower when owners wore dedicated clothing when entering the coop. These results can be used to limit the transmission of Campylobacter spp. from small poultry flocks to humans.


Impact of Helium-Oxygen Administered via High Velocity Nasal Insufflation on Delivery of Inhaled Nitric Oxide.
Inhaled nitric oxide (iNO) may be continued during the transition from invasive to noninvasive respiratory support. Upper airway obstruction from laryngeal edema following extubation and lower airway obstruction from asthma and bronchiolitis may be managed with inhaled helium. The coadministration of helium with iNO and the impact on delivered amounts of iNO have not been extensively studied. A bench model simulating a spontaneously breathing infant received iNO at varying preset doses delivered with either helium-oxygen or nitrogen-oxygen via a Vapotherm unit. iNO levels were measured at the simulated trachea. Results from the two conditions were compared using t-tests. When nitrogen-oxygen was used, there was no difference between preset and measured iNO levels. A significant difference was present when helium-oxygen was used, with a 10-fold increase in measured iNO levels compared with preset values. The use of helium resulted in a significant increase in measured iNO at the level of the simulated trachea. Clinicians must be aware that iNO will not be delivered at prescribed doses when used with helium under the conditions used in this study.


Targeting integrins for the control of tumour angiogenesis.
The crucial role of cell extracellular matrix communication in angiogenesis is well established; thus, it is not surprising that integrins have gained considerable attention as targets for the treatment of neovascular disease. Given the diversity of ligands and complexity of integrin signalling, a new appreciation for the divergent roles of integrins in angiogenesis is emerging. It is becoming clear that integrins regulate angiogenesis in both a positive and negative manner. New studies have provided a better understanding of integrin structure as it relates to ligand binding and signalling. This new insight has opened exciting possibilities for the design of novel inhibitors for clinical applications. In this review, studies concerning the cooperative interactions between integrins and regulatory molecules and possible new strategies for controlling angiogenesis will be discussed.


Cytosolic abnormally hyperphosphorylated tau but not paired helical filaments sequester normal MAPs and inhibit microtubule assembly.
Neurofibrillary degeneration of abnormally hyperphosphorylated tau, a hallmark of Alzheimer's disease (AD) and related tauopathies, occurs both as cytosolic aggregated/oligomeric protein (AD P-tau) and as neurofibrillary tangles. The abnormal hyperphosphorylation not only results in the loss of tau function of promoting assembly and stabilizing microtubules but, in the case of the cytosolic AD P-tau, also in a gain of a toxic function whereby the pathological tau sequesters not only normal tau, but also the other two neuronal microtubule associated proteins (MAPs), MAP1A / MAP1B and MAP2, and causes inhibition and disruption of microtubules. The sequestration of normal MAPs leads to a slow but progressive degeneration of the affected neurons. The affected neurons defend against the toxic tau by continually synthesizing new normal tau as well as by packaging the abnormally hyperphosphorylated tau into polymers, i.e., neurofibrillary tangles of paired helical filaments, twisted ribbons and straight filaments. The filamentous tau is inert; it neither interacts with tubulin and stimulates it assembly, nor binds to normal MAPs and causes disruption of microtubules. These findings suggest the inhibition of tau abnormal hyperphosphorylation and not the aggregation of tau as the preferred therapeutic target for AD and related tauopathies.


Identification of influenza endonuclease inhibitors using a novel fluorescence polarization assay.
Influenza viruses have been responsible for the largest pandemics in the previous century. Although vaccination and prophylactic antiviral therapeutics are the primary defense against influenza virus, there is a pressing need to develop new antiviral agents to circumvent the limitations of current therapies. The endonuclease activity of the influenza virus PA(N) protein is essential for virus replication and is a promising target for novel anti-influenza drugs. To facilitate the discovery of endonuclease inhibitors, we have developed a high-throughput fluorescence polarization (FP) assay, utilizing a novel fluorescein-labeled compound (K(d) = 0.378 μM) and a PA(N) construct, to identify small molecules that bind to the PA(N) endonuclease active site. Several known 4-substituted 2,4-dioxobutanoic acid inhibitors with high and low affinities have been evaluated in this FP-based competitive binding assay, and there was a general correlation between binding and the reported inhibition of endonuclease activity. Additionally, we have demonstrated the utility of this assay for identifying endonuclease inhibitors in a small diverse targeted fragment library. These fragment hits were used to build a follow-up library that that led to new active compounds that demonstrate FP binding and anti-influenza activities in plaque inhibition assays. The assay offers significant advantages over previously reported assays and is suitable for high-throughput and fragment-based screening studies. Additionally the demonstration of the applicability of a mechanism-based "targeted fragment" library supports the general potential of this novel approach for other enzyme targets. These results serve as a sound foundation for the development of new therapeutic leads targeting influenza endonuclease.


Mutant p62P392L stimulation of osteoclast differentiation in Paget's disease of bone.
Paget's disease of the bone (PDB) is an autosomal dominant trait with genetic heterogeneity, characterized by abnormal osteoclastogenesis. Sequestosome 1 (p62) is a scaffold protein that plays an important role in receptor activator of nuclear factor κB (RANK) signaling essential for osteoclast (OCL) differentiation. p62P392L mutation in the ubiquitin-associated (UBA) domain is widely associated with PDB; however, the mechanisms by which p62P392L stimulate OCL differentiation in PDB are not completely understood. Deubiquitinating enzyme cylindromatosis (CYLD) has been shown to negatively regulate RANK ligand-RANK signaling essential for OCL differentiation. Here, we report that CYLD binds with the p62 wild-type (p62WT), non-UBA mutant (p62A381V) but not with the UBA mutant (p62P392L) in OCL progenitor cells. Also, p62P392L induces expression of c-Fos (2.8-fold) and nuclear factor of activated T cells c1 (6.0-fold) transcription factors critical for OCL differentiation. Furthermore, p62P392L expression results in accumulation of polyubiquitinated TNF receptor-associated factor (TRAF)6 and elevated levels of phospho-IκB during OCL differentiation. Retroviral transduction of p62P392L/CYLD short hairpin RNA significantly increased TRAP positive multinucleated OCL formation/bone resorption activity in mouse bone marrow cultures. Thus, the p62P392L mutation abolished CYLD interaction and enhanced OCL development/bone resorption activity in PDB.


Mucoid and nonmucoid Burkholderia cepacia complex bacteria in cystic fibrosis infections.
infection with Burkholderia cepacia complex (BCC) bacteria in cystic fibrosis (CF) is associated with an unpredictable rate of pulmonary decline. Some BCC, but not others, elaborate copious mucoid exopolysaccharide, endowing them with a gross mucoid phenotype, the clinical significance of which has not been described. to determine whether there was a correlation between bacterial mucoid phenotype, as assessed in a semiquantitative manner from plate culture, and severity of disease as assessed by the rate of decline in lung function. we performed a retrospective clinical review of 100 patients with CF attending the Vancouver clinics between 1981 and 2007 and analyzed the rate of lung function decline (% predicted FEV(1)). patients infected exclusively with nonmucoid BCC had a more rapid decline in lung function (annual FEV(1) change, -8.51 ± 2.41%) than those infected with mucoid bacteria (-3.01 ± 1.09%; P < 0.05). Linear mixed-effects data modeling revealed a statistically significant inverse association between semiquantitative mucoid exopolysaccharide production and rate of decline of lung function. In vitro incubation of BCC with ceftazidime and ciprofloxacin but not meropenem caused conversion of BCC from mucoid to nonmucoid. our data suggest an inverse correlation between the quantity of mucoid exopolysaccharide production by BCC bacteria and rate of decline in CF lung function. Certain antibiotics may induce a change in bacterial morphology that enhances their virulence. A simple in vitro test of bacterial mucoidy may be useful in predicting the rate of decline of respiratory function in CF.


Adrenaline-induced mobilization of T cells in HIV-infected patients.
The present study aimed to investigate lymphocyte mobilization from peripheral cell reservoirs in HIV-infected patients. Nine HIV-infected patients on stable highly active anti-retroviral therapy (HAART), eight treatment-naive HIV-infected patients and eight HIV- controls received a 1-h adrenaline infusion. The adrenaline infusion induced a three-fold increase in the concentration of lymphocytes in all three groups. All HIV-infected patients mobilized significantly higher numbers of CD8+ cells but less CD4+ cells. All subjects mobilized CD45RA+CD62L+ and CD8+CD28+ cells to a lesser extent than CD45RO+CD45RA- and CD8+CD28-cells. Furthermore, high numbers of CD8+CD38+ cells were mobilized only in the HIV-infected patients. It was therefore predominantly T cells with an activated phenotype which were mobilized after adrenaline stimulation. It is concluded that the HIV-associated immune defect induced an impaired ability to mobilize immune-competent cells in response to stress stimuli. Furthermore, the study does not support the idea that CD4+ T cells are trapped in lymph nodes by HIV antigens, because untreated and HAART-treated HIV-infected patients mobilized similar numbers of CD4+ T cells. Finally, no evidence was found for the existence of a HAART-induced non-circulating pool of CD4+ T cells.


Informed consent Hong Kong style: an instance of moderate familism.
This paper examines the practice of informed consent in Hong Kong by drawing on structured interviews conducted with eleven physicians, three patients, and four family members primarily at a well-established public hospital in Hong Kong. The findings of this study show that the Hong Kong approach to medical decision-making lies somewhere between that of America on the one hand, and mainland China on the other. It is argued that the practice of medical decision-making in Hong Kong can be modeled by a moderate familism that is directed towards achieving the best interests of the patient (1) as understood by the physician, (2) in consultation with the family, (3) under the prima facie presumption that consent is not required for disclosure of information to the family, (4) while aiming at an eventual albeit frequently partial and vague disclosure to the patient.


Optimized Silica-Binding Peptide-Mediated Delivery of Bactericidal Lysin Efficiently Prevents Staphylococcus aureus from Adhering to Device Surfaces.
Staphylococcal-associated device-related infections (DRIs) represent a significant clinical challenge causing major medical and economic sequelae. Bacterial colonization, proliferation, and biofilm formation after adherence to surfaces of the indwelling device are probably the primary cause of DRIs. To address this issue, we incorporated constructs of silica-binding peptide (SiBP) with ClyF, an anti-staphylococcal lysin, into functionalized coatings to impart bactericidal activity against planktonic and sessile Staphylococcus aureus. An optimized construct, SiBP1-ClyF, exhibited improved thermostability and staphylolytic activity compared to its parental lysin ClyF. SiBP1-ClyF-functionalized coatings were efficient in killing MRSA strain N315 (>99.999% within 1 h) and preventing the growth of static and dynamic S. aureus biofilms on various surfaces, including siliconized glass, silicone-coated latex catheter, and silicone catheter. Additionally, SiBP1-ClyF-immobilized surfaces supported normal attachment and growth of mammalian cells. Although the recycling potential and long-term stability of lysin-immobilized surfaces are still affected by the fragility of biological protein molecules, the present study provides a generic strategy for efficient delivery of bactericidal lysin to solid surfaces, which serves as a new approach to prevent the growth of antibiotic-resistant microorganisms on surfaces in hospital settings and could be adapted for other target pathogens as well.


Alcoholization of pyogenichepatic abscess with absolute alcohol in Bama minipigs.
Pyogenic hepatic abscess (PHA) is a rare, but potentially serious disease. At present, ultrasound-guided or computed tomography-guided percutaneous needle aspiration or catheter drainage is appropriate as a first-line treatment. However, it is difficult to aspirate or drain pus and to select the appropriate antibiotic therapy if the abscess consists of thick pus and polymicrobial confections, or its pathogenic bacterium is multidrug resistant and cryptogenic. Case studies of alcoholization provide a novel method to manage PHA. However, the efficacy and safety of this treatment should be further evaluated. In the present study, the therapeutic efficacy and complications of alcoholization for PHAs in Bama minipigs were investigated. PHAs were prospectively treated by ultrasound-guided percutaneous instillation of absolute alcohol in the abscess cavity. The criteria for considering a successful intervention were met in all minipigs subsequent to alcoholization twice within 14 days. The procedures were well tolerated in all animals, and there were no alcoholic adverse effects or procedure-associated complications. In conclusion, ultrasound-guided percutaneous alcoholization is a safe and effective procedure to manage PHA. The problems of thick pus aspiration and selection of an appropriate antibiotic observed in other treatments were resolved effectively using alcoholization. This technique may reduce the treatment period and possibly become a novel strategy for the management of PHA.


Beta-adrenergic stimulation restores oxygen extraction reserve during acute normovolemic hemodilution.
Compensatory increases in oxygen extraction (EO(2)) during acute normovolemic hemodilution (ANH) have the effect of decreasing tissue oxygen tension values, thus increasing the threat of tissue hypoxia. We hypothesized that if the beta-adrenergic agonist isoproterenol (ISOP) could augment cardiac output (CO) during ANH, it could reverse the increases in EO(2) and restore the margin of safety for tissue oxygenation. Studies were performed in seven anesthetized (isoflurane) dogs. CO was measured by using thermodilution, and regional blood flow (RBF) was measured by using radioactive microspheres. Systemic oxygen delivery (DO(2)), oxygen consumption (OV0312;O(2)), and EO(2), as well as regional DO(2), were calculated. Measurements were obtained under the following conditions in each dog: 1) baseline-1, 2) ISOP (0.1 micro g. kg(-1). min(-1) IV), 3) baseline-2, 4) ANH, and 5) ISOP during ANH. Hematocrit was 45% +/- 3% under baseline conditions and 18% +/- 3% during ANH. Before ANH, ISOP caused parallel increases in CO and systemic DO(2), which, in the presence of an unchanged OV0312;O(2), reduced EO(2). RBF increased in myocardium and spleen, decreased in pancreas, and did not change in brain, spinal cord, or other tissues. ANH caused increases in CO, which were insufficient to offset the decrease in arterial oxygen content, and thus systemic DO(2) declined; systemic OV0312;O(2) was maintained by an increase in EO(2). ANH-related increases in RBF maintained DO(2) in myocardium, brain, duodenum, and pancreas, whereas DO(2) declined in kidney and spleen. ISOP during ANH increased CO and systemic DO(2), which returned systemic EO(2) to baseline, and it increased RBF in myocardium, kidney, duodenum, and spleen. We conclude that 1) beta-adrenergic stimulation with ISOP restored the systemic EO(2) reserve during ANH, without apparent adverse effects in the individual body tissues, and that 2) the use of inotropic drugs, such as ISOP, may extend the limit to which hematocrit can be reduced safely during ANH. By restoring the oxygen extraction reserve, isoproterenol and other inotropic drugs can enhance the margin of safety and extend the limit to which hematocrit can be reduced safely during acute normovolemic hemodilution. The use of this approach will depend on the degree of hemodilution, the extent of mixed venous oxygen desaturation, and whether increases in cardiac output are possible or desirable.


Icariin-loaded porous scaffolds for bone regeneration through the regulation of the coupling process of osteogenesis and osteoclastic activity.
Icariin (IC) promotes osteogenic differentiation, and it may be a potential small molecule drug for local application in bone regeneration. Icariin-loaded hydroxyapatite/alginate (IC/HAA) porous composite scaffolds were designed in this study for the potential application of the sustainable release of icariin and subsequent bone regeneration. An icariin-loaded hydroxyapatite/alginate porous composite scaffold was prepared and characterized by SEM and HPLC for morphology and release behavior, respectively. The mechanical properties, degradation in PBS and cytotoxicity on BMSCs were also evaluated by MTT assay, compression strength and calculation of weight remaining ratio, respectively. Rabbit BMSCs were cocultured with IC/HAA scaffolds, and ALP activity and Alizarin Red staining were performed to evaluate osteogenic differentiation induction. The mRNA and protein expression level of an osteogenic gene was detected by RT-PCR and Western blotting, respectively. In vivo animal models of critical bone defects in the radius of rabbit were used. Four and 12 weeks after the implantation of IC/HAA scaffolds in the bone defect, radiographic images of the radius were obtained and scored by using the Lane and Sandhu X-ray scoring system. Tissue samples were also evaluated using H&E and Masson staining, and an osteogenic gene and Wnt signaling pathway genes were detected. A hydroxyapatite/alginate (HAA) porous composite scaffold-loaded icariin was fabricated using a freeze-drying method. Our data indicated that the icariin was loaded in alginate scaffold without compromising the macro/microstructure or mechanical properties of the scaffold. Notably, the IC/HAA promoted the proliferation of rBMSCs without exerting cytotoxicity on rBMSCs. In vivo, rabbit radius bone defect experiments demonstrated that the IC/HAA scaffold exhibited better capacity for bone regeneration than HAA, and IC/HAA upregulated the relative expression levels of an osteogenic gene and the Wnt signaling pathway genes. Most notably, the IC/HAA scaffold also inhibited osteoclast activity in vivo. Our data suggests a promising application for the use of HAA scaffolds to load icariin and promote bone regeneration in situ through mediation of the coupling processes of osteogenesis induction and osteoclast activity inhibition.


Therapy for acute otitis media. Preference of parents for oral or parenteral antibiotic.
To determine if parents prefer single-dose intramuscular (IM) therapy or standard 10-day oral therapy for treatment of acute otitis media (AOM). Parents were asked their preference at the time their child was enrolled in a randomized controlled trial comparing the clinical efficacy of single-dose IM ceftriaxone sodium with 10 days of oral amoxicillin and clavulanate potassium for AOM. Additional information was collected at days 3 to 5 and 14 to 16 after the initiation of the therapy. Primarily private practices; 15 sites. For this study, 648 children aged 3 months to 6 years were randomly assigned to receive IM (n=327) or oral (n=321) therapy. The groups were equivalent in all measured sociodemographic factors. At the time of enrollment, 85% of parents expressed a preference for single-dose IM therapy. At days 3 to 5, no differences were reported in days children were absent from school or day care, parental absence from work, or loss of sleep by children. However, more parents with children in the IM therapy group than in the oral therapy group reported loss of sleep by the parent (35% vs 26%, P=.02, chi(2)). At days 14 to 16, more parents with children in the IM group reported being "very satisfied" with the antibiotic (65%) compared with parents whose children were assigned to the oral therapy group (38%, P<.001). In comparing current therapy to past oral therapy for AOM, 71% of the parents with children in the IM therapy group reported more satisfaction with current therapy, in contrast to 21% of parents with children in the oral therapy group (P<.001). Of the parents, 83% indicated they would prefer single-dose IM therapy for AOM in the future. All of the parents prefer single-dose IM therapy for AOM over standard 10-day oral therapy.


A reproductive phase-dependent effect of dietary L-tryptophan on pineal gland and gonad of a nocturnal bird, Indian spotted owlet Athene brama.
Unlike other temperate owls, Indian spotted owlet Athene brama possesses a well-developed pineal gland that secrets moderate amount of hydroxy- (serotonin) and methoxy- (melatonin) indoles in circulation. However, in this study, we have reported the response of this endocrine gland to exogenous L-Tryptophan (precursor of the above indoles), and also its effect on gonads of this nocturnal bird. During breeding phase or pineal inactive phase (March), oral treatment of L-Trp (0.5 mg/100 g Bwt/day) significantly increased the pineal gland wt and plasma melatonin (MEL) level, while decreased the gonadal wt and plasma sex steroids levels (estradiol and progesterone in female and testosterone in male). Interestingly, during reproductively quiescent phase or pineal active phase (August), similar amount of L-Trp significantly decreased the plasma MEL level, while increased the above sex steroid levels in plasma. Finally, the results show a clear reproductive phase-dependent inverse effect of L-Trp on pineal gland and gonads for both sexes of the spotted owlets, and suggest that the therapeutic use of this amino acid would be a great advantage for controlling the reproduction of these economically important birds.


Characterization of a monoclonal antibody specific for the Ras-related GTP-binding protein Rho A.
The Rho family of small GTP-binding proteins is one of the three subgroups which, together with the Ras and Rab families, constitute the Ras-related superfamily. The Rho subgroup contains at least seven highly homologous members including 4 Rho proteins (RhoA, RhoC, RhoB, and RhoG), the Rac1 and Rac2 proteins, and CDC42Hs, which are involved in various aspects of cytoskeleton organisation and cell polarity. We have raised antibodies to individual members of the Rho family, and we report here the characterization of a monoclonal antibody (26C4) specific for RhoA. When used in western blot experiments, the 26C4 antibody recognizes the recombinant RhoA protein but not the almost identical RhoC or the RhoG, Rac and CDC42Hs proteins. Furthermore the 26C4 antibody identifies the natural RhoA protein in human lymphocyte cell extracts and was used to study the level of RhoA expression in several lymphoblastoid cell lines, and its association with the cell membrane.


Effectiveness of transforaminal epidural steroid injections in low back pain: a one year experience.
Transforaminal epidural steroid injections under fluoroscopy are an alternative treatment for lower back pain with radiculopathy. We followed 82 patients with a standardized telephone questionnaire at 2, 6, and 12 months after the first injection, in order to assess their effectiveness. Ninety-two patients with radiculopathic back pain due to spinal stenosis, herniated discs, spondylolisthesis, and degenerated discs, underwent transforaminal epidural steroid injections under fluoroscopy. Eighty-two patients were followed with a standardized telephone questionnaire. The population was divided into four groups: Group I, previous back surgery (16%); Group II, discogenic abnormalities: herniations, bulges, or degeneration, (42%); Group III, spinal stenosis (32%); Group IV, those without MRI (11%). Age ranged between 24 to 99 years, mean 64.5. Forty-seven were female, 35 male. Thirteen patients (16%) underwent one procedure, 27 patients (33%) two, 37 patients (45%) three, and five patients (6%) four, an average 2.4 procedures per patient. The pain scores for all patients improved significantly at all three time points (2, 6 and 12 months) compared to the initial mean pain score of 7.3 to mean pain scores of 3.4, 4.5 and 3.9 respectively. After one year, 36 patients did not take any pain medications. Greater than 50% improvement after one year was seen in 23% of Group I; 59% in Group II; 35% in Group III and 67% in Group IV. Transforaminal epidural steroid injections can offer significant pain reduction up to one year after initiation of treatment in patients with discogenic pain and possibly in patients with spinal stenosis.


Holliday junctions are formed in concentrated solutions of purified products of DNA amplification.
Previously, using concentrated solutions of PCR products of five different genes, we described the appearance in these solutions of DNA structures with molecular weights approximately twice greater than that of double-strand (ds) fragments and with even higher molecular weight. Since this phenomenon was shown to be not dependent on the size or sequence of the DNA fragments, we suggested that it is due to interaction of DNA duplexes. The double-sized dsDNA complex containing four polynucleotide strands of two DNA fragments was named a "tetramer". Our present work is devoted to elucidation of peculiarities of tetramer formation and its structure in solutions of a purified PCR product of p53 cDNA. We found that the intensity of tetramer formation depends on the concentration of the PCR product in solution. Three subsequent purifications of the PCR product were performed using DNA-binding matrix, but the tetramers appeared again after every procedure. After purification of PCR product preliminarily treated with S1-nuclease, tetramers appeared again, indicating that these structures are formed from dsDNA fragments. Purification of the tetramers on DNA-binding matrix led to the appearance of the initial dsDNA fragments as the main DNA structure. When electroelution and column filtration by centrifugation were used, the purification procedure was speeded up, and a solution with a higher amount of the tetramer was obtained. Electron microscopy revealed the presence of four-stranded symmetrical structures with crossing chains known as Holliday junctions. Thus, for the first time the ability of homologous dsDNA fragments to interact with the formation of Holliday junctions without participation of cell proteins has been demonstrated.


RNA tetraloop folding reveals tension between backbone restraints and molecular interactions.
In RNA, A-form helices are commonly terminated by tetraloops or 3' dangling ends. Aside from helices themselves, these helix-breaking motifs appear to be among the most frequent and repetitive structural elements of large folded RNAs. We show here that within a frequent type of tetraloop, cGNRAg (G is guanine, N is any base, R is purine, A is adenine), a tension exists between the backbone torsional energy of the loop and the energy contributed by molecular interactions (stacking and pairing). A model in which favorable bond rotamers are opposed by favorable stacking and pairing interactions is consistent with our observation that release of torsional restraints upon conversion of one or more loop riboses to more flexible trimethylene phosphate(s) contributes favorably to the enthalpy of folding. This effect presumably results from improved stacking and hydrogen-bonding interactions upon release of torsional restraints. The most obvious possibility for improving molecular interactions is a repositioning of A, which is proximal to the unfavorable torsion angles in native cGNRAg tetraloops, and which is unstacked on the 3' side and unpaired (it forms a single hydrogen bond with the opposing G). This tension between favorable bond rotamers and favorable molecular interactions may be representative of a general evolutionary strategy to prevent achievement of deep and irreversible thermodynamic wells in folded RNAs. Finally, we observe a simple stacking substructure with conserved geometry and sequence that forms a scaffold for both tetraloops and 3' dangling ends. It seems that simple substructures can build RNA motifs, which combine to establish the fundamental architecture of RNA.


Interpretive reporting to improve the effectiveness of clinical laboratory test results. An ECRI technology assessment.
Interpretive reporting encompasses a range of efforts by the clinical laboratory community to develop improved methods of transferring information from laboratories to physicians in a form that they can understand and use. The purpose of laboratory tests is to provide clinically useful information that can be used to answer a specific question for the clinician. However, growing physician reliance on laboratory and other diagnostic tests and the increased testing volumes made possible by laboratory automation have yielded some unintended negative effects. Laboratories can now provide more information than can be effectively assimilated. The resulting "information overload" can cause clinicians to misinterpret tests, ignore significant results, or fail to act appropriately when results indicate a treatable condition. Research indicates, however, that interpretive reporting improves information transfer and thus increases clinicians' understanding of the significance of laboratory test results.


Isolation of a plant glycoprotein involved with control of intercellular recognition.
A recognition molecule was isolated from stigmas of S-allele genotype S(2)S(2) of Brassica oleracea var. capitata L. After Sephadex chromatography, it eluted as a single symmetrical peak during diethylaminoethane-cellulose chromatography. A high degree of purity was affirmed by: sedimentation as a single peak during ultracentrifugation through 5 to 20% sucrose gradients; elution as a single peak from Sephadex G-100; visualization as a single band which stains with Coomassie blue and periodic acid Schiff reagent after electrophoresis on polyacrylamide gels. Other criteria supporting the conclusion that it is a glycoprotein are: (a) the highly purified preparation is anthrone-positive and has a Lowry protein to anthrone-positive carbohydrate ratio of 1.3; (b) the preparation contains arabinose, galactose, glucose, and mannose, although it is not precipitated by concanavalin A; (c) the immunological properties of the molecule are lost following protease treatment, and it has a molecular weight of 90,000 by Sephadex gel-filtration analysis and 54,500 by velocity sedimentation analysis.In vitro pretreatment of S(2)S(2) pollen with the post-diethylaminoethane-purified S(2) glycoprotein prevented the S(2)S(2) pollen from germinating on three classes of compatible stigmas: (a) mature stigmas of genotypes S(3)S(3) and S(8)S(8), which are non-self genotypes; (b) immature stigmas of genotype S(2)S(2), where incompatibility is not expressed; and (c) mature stigmas with a recessive S(2) allele. Pretreatment of S(3)S(3) and S(8)S(8) pollen with the S(2) glycoprotein did not interfere with their germination.


Discovery of the "RNA continent" through a contrarian's research strategy.
The International Human Genome Sequencing Consortium completed the decoding of the human genome sequence in 2003. Readers will be aware of the paradigm shift which has occurred since then in the field of life science research. At last, mankind has been able to focus on a complete picture of the full extent of the genome, on which is recorded the basic information that controls all life. Meanwhile, another genome project, centered on Japan and known as the mouse genome encyclopedia project, was progressing with participation from around the world. Led by our research group at RIKEN, it was a full-length cDNA project which aimed to decode the whole RNA (transcriptome) using the mouse as a model. The basic information that controls all life is recorded on the genome, but in order to obtain a complete picture of this extensive information, the decoding of the genome alone is far from sufficient. These two genome projects established that the number of letters in the genome, which is the blueprint of life, is finite, that the number of RNA molecules derived from it is also finite, and that the number of protein molecules derived from the RNA is probably finite too. A massive number of combinations is still involved, but we are now able to understand one section of the network formed by these data. Once an object of study has been understood to be finite, establishing an image of the whole is certain to lead us to an understanding of the whole. Omics is an approach that views the information controlling life as finite and seeks to assemble and analyze it as a whole. Here, I would like to present our transcriptome research while making reference to our unique research strategy.


Experimental transmission of cardiomyopathy syndrome (CMS) in Atlantic salmon Salmo salar.
Cardiomyopathy syndrome (CMS) is a disease of unknown aetiology, having significant economic impact as it primarily affects large, farmed Atlantic salmon Salmo salar L. in seawater, close to harvest. In the present study, we have demonstrated that CMS is a transmissible disease under experimental conditions. Histopathological lesions consistent with CMS were induced in Atlantic salmon post-smolts after injection of tissue homogenate from farmed fish diagnosed with CMS. Six weeks post-injection (p.i.), experimental fish started developing focal to multi-focal lesions in the atrial endo- and myocardium, with subsequent progression to the ventricle. This proceeded into severe endocarditis and subsequent myocarditis with mononuclear cell infiltration of the atrium and, to a lesser degree, the spongy layer of the ventricle. These lesions were consistent with histopathological findings in field outbreaks of CMS. From Week 33 p.i., lesions also appeared in the compact myocardium, with focal epicarditis adjacent to focal myocardial lesions. In conclusion, these results indicate that CMS has an infectious aetiology and should be treated as a potentially contagious disease.


A mathematical model of biomedical interventions for HIV prevention among men who have sex with men in China.
The new HIV treatment guidelines in China recommend antiretroviral therapy (ART) for all people living with HIV, but significant gaps in implementation still exist. Pre-exposure prophylaxis (PrEP) can effectively reduce the risk of HIV transmission among men who have sex with men (MSM). This study assessed the epidemiological impact and cost effectiveness of PrEP, enhanced biomedical interventions and their combination among MSM in China. A deterministic mathematical model was developed and projected over 20 years to assess the impact of the PrEP, biomedical interventions and their combinations. Incidence and prevalence of HIV were measured, and cost-effectiveness was assessed using incremental cost (international dollars, Int.$) per quality-adjusted life year (QALY) gained. A total of 0.78 million new HIV infections were estimated to occur over the next 20 years if no additional interventions are implemented among MSM. The PrEP-only strategy covering 25-75% of HIV-negative high-risk MSM can prevent 0.09-0.20 million (12.1-25.7%) new infections, at a cost of 17,277-18,452 Int.$/QALY. The optimal cost-effectiveness path is from test-and-treat to the combination strategy of test-and-treat and PrEP. Some strategies could almost eliminate new HIV infections over the next 20 years. PrEP, test-and-treat, and their combinations among MSM are effective and cost-effective relative to current policy. PrEP is an important and cost-effective addition to current policy in China.