[Endoprosthesis of the hip joint in progressive and chronic destructive tuberculous arthritis: problems and prospects].
The paper presents the results of endoprosthesis of the hip joint in 6 patients with destructive tuberculous coxitis. In 2 patients, the process was bilateral. All the patients received preoperative antituberculous therapy with 3-4 drugs (isoniazid, rifampicin, pyrazinamide, streptomycin) for 14-12 weeks. Within the first 6 weeks, isoniazid and rifampicin were given intravenously. A clinical stabilization was achieved in all cases, which permitted total endoprosthesis of the hip joint. In postoperative period, the patients began to be active on days 10-12 of antituberculous therapy that lasted as long as 6 months in their local dispensary. The follow-ups of the patients ranged from 1 to 5 years. The support capacity and function of the joint operated on were recovered in all cases. There was no recurrence of a tuberculous process at the site of endoprosthesis. Thus, the developed tactics of preoperative preparation and post-operative management permits endoprosthesis of the hip joint in patients with destructive tuberculous coxitis and recovery of articular support capacity and function. At the same time it is necessary to define clinical indications for this operation in each specific case.


Oxidative stress triggers aggregation of GFP-tagged Hsp31p, the budding yeast environmental stress response chaperone, and glyoxalase III.
The Saccharomyces cerevisiae Hsp31p protein belongs to the ubiquitous DJ-1/ThiJ/PfpI family. The most prominent member of this family is human DJ-1; defects of this protein are associated with Parkinson's disease pathogenesis. Numerous recent findings reported by our group and others have revealed the importance of Hsp31p for survival in the post-diauxic phase of cell growth and under diverse environmental stresses. Hsp31p was shown to possess glutathione-independent glyoxalase III activity and to function as a protein chaperone, suggesting that it has multiple cellular roles. Our previous work also revealed that HSP31 gene expression was controlled by multiple stress-related transcription factors, which mediated HSP31 promoter responses to oxidative, osmotic, and thermal stresses, toxic products of glycolysis, and the diauxic shift. Nevertheless, the exact role of Hsp31p within budding yeast cells remains elusive. Here, we aimed to obtain insights into the function of Hsp31p based on its intracellular localization. We have demonstrated that the Hsp31p-GFP fusion protein is localized to the cytosol under most environmental conditions and that it becomes particulate in response to oxidative stress. However, the particles do not colocalize with other granular subcellular structures present in budding yeast cells. The observed particulate localization does not seem to be important for Hsp31p functionality. Instead, it is likely the result of oxidative damage, as the particle abundance increases when Hsp31p is nonfunctional, when the cellular oxidative stress response is affected, or when cellular maintenance systems that optimize the state of the proteome are compromised.


The utility of the growth hormone (GH) releasing hormone-arginine test for diagnosing GH deficiency in adults with childhood acute lymphoblastic leukemia treated with cranial irradiation.
The insulin tolerance test (ITT) is the current standard diagnostic test for the diagnosis of adult GH deficiency (GHD), but alternative tests, such as the GHRH-arginine test, have been proposed. We investigated the sensitivity and specificity of the GHRH-arginine test using ITT as the gold standard in diagnosing GHD in a group of young adults treated with cranial irradiation (CRT) for childhood acute lymphoblastic leukemia (ALL). We estimated the positive and negative predictive values of the GHRH-arginine test among patients as well as a number of individual characteristics and therapy-related factors during both the GHRH-arginine test and ITT. Forty-three young adults, treated for childhood ALL with 18-30 Gy CRT and chemotherapy, were studied, and comparison was made with matched controls. We evaluated four different cutoff levels for GHD in the GHRH-arginine test: 5, 7.5, 9, and 16.5 microg/liter. Using 7.5 microg/liter as the cutoff yielded high specificity (94%), but at the same time the sensitivity was only 66%, which leads to a low negative predictive value (27%). In contrast, a failed GH response to the GHRH-arginine test accurately reflects the presence of radiation-induced GHD, illustrated by a high positive predictive value (95% at 7.5 microg/liter). Only age at CRT and body mass index remained significant predictors of the peak GH during the GHRH-arginine test. Because a high proportion of GHD patients show a normal response to the GHRH-arginine test, it cannot be used reliably to exclude GHD in these patients. Complementary ITT is also warranted to confirm GHD in obese patients.


Ni-Catalyzed Enantioselective Intermolecular Hydroamination of Branched 1,3-Dienes Using Primary Aliphatic Amines.
A Ni-catalyzed intermolecular enantioselective hydroamination of branched 1,3-dienes is reported. The method is broadly applicable, highly regio-, chemo-, and enantioselective, and provides direct access to valuable chiral allylic amines starting from linear or α-branched aliphatic primary amines or secondary amines. Mechanistic studies have been conducted using 31P NMR spectroscopy for reaction progress monitoring, isotopic labeling experiments (2H), and kinetic analysis. The resting state of the catalyst is a Ni-π-allyl complex, and the outer-sphere nucleophilic attack of H-bonded amine aggregates is proposed to be the rate-determining step. This hypothesis guided the identification of an improved set of reaction conditions for the enantioselective hydroamination of branched 1,3-dienes.


Protective effects of compounds from Garcinia mangostana L. (mangosteen) against UVB damage in HaCaT cells and hairless mice.
Ultraviolet B (UVB) radiation causes alterations in the skin, such as epidermal thickening, wrinkle formation and inflammation. Therefore, preventing UVB-induced skin damage can promote general health among the human population. Garcinia mangostana L. (mangosteen) is a fruit that has become a popular botanical dietary supplement because of its perceived role in promoting overall health. The present study investigated the photoprotective effects of α-, β-, γ-mangostins and gartanin against UVB radiation using the HaCaT immortalized human keratinocyte cell line as an in vitro model and hairless mice as an in vivo model. UVB radiation increased the expression of matrix metalloproteinase (MMP)‑1 and ‑9 and decreased the mRNA expression levels of involucrin, filaggrin and loricrin in HaCaT cells; however, these changes were attenuated by pretreating the cells with α-, β-, γ-mangostins and gartanin. Among these compounds, α-mangostin exhibited the greatest effects in reducing UVB-induced skin wrinkles, inhibited epidermal thickening in hairless mice in vivo. Exposure to UVB radiation increased the expression of MMPs and pro-inflammatory cytokines and activated mitogen-activated protein kinases in hairless mice, but these changes were attenuated by α-mangostin. The authors suggested that α-mangostin exerts anti-wrinkle and anti-aging properties.


Peripheral Blood Smears Distinguish Infective Fever after CAR-T Therapy.
Chimeric antigen receptor (CAR) T-cell therapy carries the risk of inducing severe and life-threatening toxicities such as cytokine release syndrome (CRS), neurotoxicity, and infection. Although CRS and infections have similar symptoms, their treatment strategies differ, and early diagnosis is very important. For CRS and infections, the fastest detection time currently takes more than 24 h, so a quick and simple method to identify a fever after CAR T-cell infusion is urgently needed. We enrolled 27 patients with recurrent fever treated with different types of CAR T-cells, including cluster of differentiation (CD) 7, CD19, CD22, and CD19-CD22 bicistronic CAR T-cells, and evaluated the infection events occurring in these patients. We detailed the morphology of CAR T-cells in peripheral blood smears (PBS) and reported the infection events, CAR transgene copy number, and inflammatory indicators within the first month after treatment. Similar morphological characteristics were observed in the PBS of different CAR T-cells, namely, enlarged cell bodies, deep outside and shallow inside basophilic blue cytoplasm, and natural killer (NK) cell-like purplish red granules. There were ten infections in nine of the twenty-seven patients (33%). The percentage of atypical lymphocytes in PBS was significantly associated with CAR transgene copy number and absolute lymphocyte count in all patients. The atypical lymphocyte percentage was significantly higher in the non-infection group. In conclusion, the unique morphology of CAR T-cells in PBS can be used to evaluate CAR T-cell kinetics and provide reliable evidence for the rapid early identification of fever after CAR T-cell infusion. ChiCTR-OPN-16008526; ChiCTR-OPN-16009847; ChiCTR2000038641; NCT05618041; NCT05388695.


A comparative study on the experimentally derived electron densities of three protease inhibitor model compounds.
In order to contribute to a rational design of optimised protease inhibitors which can covalently block the nucleophilic amino acids of the proteases' active sites, we have chosen three model compounds (aziridine , oxirane and acceptor-substituted olefin ) for the examination of their electron-density distribution. Therefore, high-resolution low temperature (9, 27 and 100 K) X-ray diffraction experiments on single-crystals were carried out with synchrotron and conventional X-radiation. It could be shown by the analysis of the electron density using mainly Bader's Theory of Atoms in Molecules, Volkov's EPMM method for interaction energies, electrostatic potentials and Gatti's Source Function that aziridine is most suitable for drug design in this field. A regioselective nucleophilic attack at carbon atom C1 could be predicted and even hints about the reaction's stereoselectivity could be obtained. Moreover, the comparison between two data sets of aziridine (conventional X-ray source vs. synchrotron radiation) gave an estimate concerning the reproducibility of the quantitative results.


Effect of socio-demographic factors on the association between multimorbidity and healthcare costs: a population-based, retrospective cohort study.
To estimate the attributable costs of multimorbidity and assess whether the association between the level of multimorbidity and health system costs varies by socio-demographic factors in young (<65 years) and older (≥65 years) adults living in Ontario, Canada. A population-based, retrospective cohort study SETTING: The province of Ontario, Canada PARTICIPANTS: 6 639 089 Ontarians who were diagnosed with at least one of 16 selected medical conditions on 1 April 2009. From the perspective of the publicly funded healthcare system, total annual healthcare costs were derived from linked provincial health administrative databases using a person-level costing method. We used generalised linear models to examine the association between the level of multimorbidity and healthcare costs and the extent to which socio-demographic variables modified this association. Attributable total costs of multimorbidity ranged from C$377 to C$2073 for young individuals and C$1026 to C$3831 for older adults. The association between the degree of multimorbidity and healthcare costs was significantly modified by age (p<0.001), sex (p<0.001) and neighbourhood income (p<0.001) in both age groups, and the positive association between healthcare costs and levels of multimorbidity was statistically stronger for older than younger adults. For individuals aged 65 years or younger, the increase in healthcare costs was more gradual in women than in their male counterparts, however, for those aged 65 years or older, the increase in healthcare costs was significantly greater among women than men. Lastly, we also observed that the positive association between the level of multimorbidity and healthcare costs was significantly greater at higher levels of marginalisation. Socio-demographic factors are important effect modifiers of the relationship between multimorbidity and healthcare costs and should therefore be considered in any discussion of the implementation of healthcare policies and the organisation of healthcare services aimed at controlling healthcare costs associated with multimorbidity.


[Early stages of myogenesis as seen through the action of the myf-5 gene].
Skeletal muscles in the vertebrate body are derived from the somites, epithelial spheres of cells which segment from the paraxial mesoderm in a rostral-caudal developmental gradient on either side of the neural tube. Initially, cells in the somite are multipotent and their fate depends on the environmental influences exerted by neighbouring tissues, notably the axial structures (neural tube and notochord), and the dorsal ectoderm. The ventralizing influence exerted by the notochord and floor plate of the neural tube through the action of sonic hedgehog, results in the differentiation of sclerotome which will give rise to cartilage and bone of the vertebral column and ribs. The dorsal derivatives of the somite, formed from cells in the dermomyotome, are derm and skeletal muscle. The onset of skeletal myogenesis is characterized by expression of myogenic factors, notably myf-5 and MyoD, members of the superfamily of helix-loop-helix transcription factors. Another member of the myogenic factor family, myogenin, is subsequently expressed and leads to muscle cell differentiation with activation of the downstream muscle-specific genes. Dorsalization of the somite and subsequent myogenesis depends on the presence of axial structures and dorsal ectoderm. The Wnt family of signalling molecules are potentially implicated in this process. Muscle progenitor cells present in the medial part of the dermomyotome activate myf-5 first and explant experiments have shown that the axial structures lead to the activation of this myogenic factor and subsequent myogenesis which results in the formation of the dorsal myotome in the central region of the somite. This contributes to the formation of axial muscles. Muscle progenitor cells in the lateral part of the dermomyotome preferentially activate MyoD and this depends on the presence of dorsal ectoderm. These cells will form the ventral aspect of the myotome, and later contribute to body wall muscles, for example. Part of the lateral progenitor population migrates away from the somite to form peripheral body muscles and the muscles of the limb. In this case myogenic factors are not initially expressed and these migratory cells are characterized by the expression of the paired-box gene Pax3. In explant experiments lateral mesoderm retards the induction of MyoD expression by dorsal ectoderm; in vivo this may be important to permit cell migration prior to differentiation. In mice carrying mutations in both MyoD and myf-5 no skeletal muscle forms, whereas myogenesis can take place in the absence of either MyoD or myf-5. Normally, cells in which one gene is activated first, subsequently co-express the other, so that there rapidly cease to be distinct MyoD+ or myf-5+ populations in the embryo. In myf-5-/- mice no myotome forms initially, but MyoD is subsequently activated. This takes place medially, as well as laterally, under the influence of the more mature neural tube and notochord. By targetting the myf-5 gene with an nlacZ reporter gene it has been possible to follow the fate of the early muscle progenitor cell population in which the myf-5 gene has been activated but no myf-5 protein is present. These beta-galactosidase positive cells delaminate from the dermomyotome, but instead of migrating under this epithelium to form the myotome, they migrate aberrantly. Some cells localize dorsally under the epiderm and begin to express the dermal marker, Dermo-1. Other muscle progenitor cells migrate ventrally into the sclerotomal compartment where they express an early sclerotomal marker, scleraxis. Later in the mutant mice, when cells from this compartment have condensed to form the cartilage of the ribs, beta-galactosidase positive cells are detectable within the ribs. These observations indicate that the early myogenic factor myf-5 is necessary to ensure the correct positioning of myogenic progenitor cells within the embryo. (ABSTRACT TRUNCATED)


Time since start of first-line therapy as a predictive clinical marker for nintedanib in patients with previously treated non-small cell lung cancer.
No predictive clinical or genetic markers have been identified or validated for antiangiogenic agents in lung cancer. We aimed to identify a predictive clinical marker of benefit for nintedanib, an angiokinase inhibitor, using data from two large second-line non-small cell lung cancer Phase III trials (LUME-Lung 1 ([LL1] and LUME-Lung 2). Predictive marker identification was conducted in a multi-step process using data from both trials; a hypothesis was generated, confirmed and validated. Statistical analyses included a stepwise selection approach, a recursive partitioning method and the evaluation of HRs, including treatment-by-covariate interactions. The marker was finally validated using a prospectively defined hierarchical testing procedure and treatment-by-covariate interaction for overall survival (OS) based on LL1. Time since start of first-line therapy (TSFLT) was identified as the only predictive clinical marker. A cut-off of 9 months was chosen for further analysis, based on HRs and recursive partitioning. The prospectively defined final validation using OS data from LL1 established the strong relationship between TSFLT and treatment with nintedanib. Patients with adenocarcinoma with TSFLT <9 months showed a greater survival benefit (median OS 10.9 vs 7.9 months, HR 0.75 [95% CI 0.60-0.92]; p=0.0073) compared with patients in the TSFLT >9 months group (median OS 17.0 vs 15.1 months, HR 0.89 [95% CI 0.66-1.19]). Patients with shorter TSFLT derive a greater progression-free survival and OS benefit from nintedanib. This clinical marker could be used for patient selection and further investigation is warranted regarding pathways promoting aggressive tumour growth and antiangiogenic tyrosine kinase inhibitor benefit.


Recent advances in the investigation of curcuminoids.
More than 30 Curcuma species (Zingiberaceae) are found in Asia, where the rhizomes of these plants are used as both food and medicine, such as in traditional Chinese medicine. The plants are usually aromatic and carminative, and are used to treat indigestion, hepatitis, jaundice, diabetes, atherosclerosis and bacterial infections. Among the Curcuma species, C. longa, C. aromatica and C. xanthorrhiza are popular. The main constituents of Curcuma species are curcuminoids and bisabolane-type sesquiterpenes. Curcumin is the most important constituent among natural curcuminoids found in these plants. Published research has described the biological effects and chemistry of curcumin. Curcumin derivatives have been evaluated for bioactivity and structure-activity relationships (SAR). In this article, we review the literature between 1976 and mid-2008 on the anti-inflammatory, anti-oxidant, anti-HIV, chemopreventive and anti-prostate cancer effects of curcuminoids. Recent studies on curcuminoids, particularly on curcumin, have discovered not only much on the therapeutic activities, but also on mechanisms of molecular biological action and major genomic effects.


Efficient solvent boundary potential for hybrid potential simulations.
A common challenge in computational biophysics is to obtain statistical properties similar to those of an infinite bulk system from simulations of a system of finite size. In this work we describe a computationally efficient algorithm for performing hybrid quantum chemical/molecular mechanical (QC/MM) calculations with a solvent boundary potential. The system is partitioned into a QC region within which catalytic reactions occur, a spherical region with explicit solvent that envelops the quantum region and is treated with a MM model, and the surrounding bulk solvent that is treated implicitly by the boundary potential. The latter is constructed to reproduce the solvation free energy of a finite number of atoms embedded inside a low-dielectric sphere with variable radius, and takes into account electrostatic and van der Waals interactions between the implicit solvent and the QC and MM atoms in the central region. The method was implemented in the simulation program pDynamo and tested by examining elementary steps in the reaction mechanisms of two enzymes, citrate synthase and lactate dehydrogenase. Good agreement is found for the energies and geometries of the species along the reaction profiles calculated with the method and those obtained by previous experimental and computational studies. Directions in which the utility of the method can be further improved are discussed.


A comparative genomics approach to prediction of new members of regulons.
Identifying the complete transcriptional regulatory network for an organism is a major challenge. For each regulatory protein, we want to know all the genes it regulates, that is, its regulon. Examples of known binding sites can be used to estimate the binding specificity of the protein and to predict other binding sites. However, binding site predictions can be unreliable because determining the true specificity of the protein is difficult because of the considerable variability of binding sites. Because regulatory systems tend to be conserved through evolution, we can use comparisons between species to increase the reliability of binding site predictions. In this article, an approach is presented to evaluate the computational predictions of regulatory sites. We combine the prediction of transcription units having orthologous genes with the prediction of transcription factor binding sites based on probabilistic models. We augment the sets of genes in Escherichia coli that are expected to be regulated by two transcription factors, the cAMP receptor protein and the fumarate and nitrate reduction regulatory protein, through a comparison with the Haemophilus influenzae genome. At the same time, we learned more about the regulatory networks of H. influenzae, a species with much less experimental knowledge than E. coli. By studying orthologous genes subject to regulation by the same transcription factor, we also gained understanding of the evolution of the entire regulatory systems.


Molecular differential diagnosis of renal carcinoma: from microscopes to microsatellites.
In the last decade, specific chromosomal alterations have been associated with different tumor types. These aberrations were originally detected by karyotyping and then by more sophisticated cytogenetic analysis. A few karyotypic alterations can be directly linked to distinct malignancies, such as the Philadelphia chromosome in acute lymphoblastic leukemia, loss of distal chromosome 3p 14 in small-cell lung cancer, the loss of distal chromosome 11p13 in Wilms' tumor, and loss or rearrangement of the short arm of chromosome 3 in clear and chromophobe RCC. The relative specificity of the latter findings enabled investigators to diagnose an occult renal clear-cell carcinoma from a supraclavicular lymph node metastasis by analysis of G-banded metaphase chromosomes obtained from this mass. A similar report based also on cytogenetic findings was published earlier. Karyotypic changes, however, detect only gross alterations visible to an observer. With more refined diagnostic tools, such as microsatellite analysis, other, even smaller, well defined lesions can be analyzed. A summary of the known frequencies of chromosomal losses is given in Table 1. The combination of certain LOH patterns has shown great promise in the differential diagnosis of renal tumors. The transfer of molecular genetics from the laboratory to surgical pathology and other clinical departments is a meaningful event and a challenging task. Molecular pathology is certain to become important in the diagnosis of tumors with unclear histology. Diagnosis based widely upon staining techniques and determination of a patient's prognosis by staging and grading alone will be increasingly accompanied by molecular genetic methods. Pathology may be on the verge of the greatest change since the introduction of the microscope.


Interleukin-1 mediates hemodynamic dysfunction and release of eicosanoids and tumor necrosis factor during graded bacteremia.
The pathophysiologic events of sepsis mediated by interleukin-1 (IL-1) remain ill-defined. The purpose of this study was to identify the circulatory derangements of which IL-1 was a necessary mediator and evaluate its interactions with tumor necrosis factor (TNF) and the eicosanoids during graded bacteremia. Eleven adult female swine were anesthetized, mechanically ventilated, and monitored with pulmonary artery catheters and arterial lines; they received intravenously either saline vehicle (septic control, n = 6) or human recombinant IL-1 receptor antagonist (IL-1ra, n = 5). The animals were then infused with Aeromonas hydrophila (10(9)/mL) for 4 h at rates gradually increased from .2 mL/kg/h to 4 mL/kg/h over 3 h, then sacrificed after 4 h. Mean arterial pressure (MAP), left ventricular stroke work index (LVSWI), and systemic vascular resistance index (SVRI) were recorded at baseline and hourly thereafter, and plasma 6-keto-PGF1alpha (6-KETO), tumor necrosis factor-alpha (TNF) and leukotrienes B4(LTB4) and C4D4E4 (LTCDE), pg/mL, were measured by ELISA. MAP, LVSWI, arterial P(O2) all decreased in the septic control group to levels significantly below those of the IL-1 antagonist animals. Circulating 6-KETO, LTCDE, and TNF increased significantly in all septic animals. Plasma LTB, and TNF were reduced by IL-1 blockade, compared with septic controls. TxB2 was not affected by IL-1 inhibition. There were no intergroup differences in platelet aggregation, but the in vitro aggregation response decreased from baseline in septic controls to 54+/-27% (p < .05). IL-1 is necessary to the development of systemic hypotension impaired LVSWI, and increased intravascular platelet aggregation during graded bacteremia. Conversely, IL-1 helps to maintain stroke volume and low SVRI in graded bacteremia, possibly through increased prostacyclin release. It may contribute to impaired pulmonary gas exchange and increased tissue oxygen demands. TNF release is stimulated in the presence of unopposed IL-1 and may be synergistic with it in the adverse hemodynamic effects of endogenous IL-1. IL-1 is required for increased leukotriene and prostacyclin levels in this model, but it is not involved in thromboxane release. Whether the lack of survival benefit from IL-1ra in human sepsis is due to these mixed cardiopulmonary and mediator effects, to species differences, or to timing of IL-1ra administration is not clear from the data.


Radiometric imaging by double exposure and gain calibration.
Due to low cost and small size, uncooled microbolometer-based thermal focal plane arrays are very attractive for radiometry. However, being non-cooled, they suffer from temporally and spatially dependent changes that require constant calibration. While the gain calibration can be reasonably realized by two-point correction, the offset due to internal radiation loads poses a complicated calibration scheme. We present a new computational optics approach that simplifies the essential calibration for temperature offset. Using two successive images of the object taken with different known blur levels, one can eliminate the object term from the image-formation equation, resulting in an equation for the unknown sensor offset. A general algebraic model is presented for the space-variant case followed by solutions using both direct inverse method and iterative solver. The new scheme allows restoration of the radiometric value within 1% error with the direct method, and 0.2% error with the iterative scheme. Account of the influence of realistic lens positioning error on restoration accuracy was given. Results using direct inverse methods for restoring the radiometric values yield restoration error with a good average error of 3.7% and less.


Regional vs. national renal sharing organizations: pros and cons.
Delayed graft function, defined as the need of dialysis in the first week after transplantation, neither due to immunological nor technical causes, determines a poor outcome of renal grafts. Delayed graft function is related to the cold ischemia time, which is shorter in local allocation programs. These, however, do not assure an optimal HLA-A,B,DR matching that can be provided by national allocation organizations. We reviewed 160 cadaveric kidney grafts performed in our local transplant network. Owing to the long waiting list caused by organ shortage, we were able to ensure both a high-grade histocompatibility and short cold ischemia times. The mean HLA-B,DR mismatch was 1.17. Cold ischemia time was < 24 h in 85% of cases. The incidence of DGF was 23.1%. In our experience a regional sharing program in the case of organ shortage provides good graft outcome (86.9% graft survival at 1 yr) with low incidence of delayed graft function.


Extraskeletal myxoid chondrosarcoma arising from the retroperitoneum.
A case of extraskeletal myxoid chondrosarcoma is presented. The tumor occurred in the retroperitoneum and systemic metastases were found at autopsy. The primary and metastatic tumors were soft and strikingly myxoid on gross appearance. Microscopic observation revealed undifferentiated malignant tumor having large amounts of myxoid substance and a small amount of well-differentiated chondrosarcoma element in the primary lesions. The authors obtained an immunohistochemical result that the tumor cells showed positivity for alpha-1-antitrypsin and alpha-1-antichymotrypsin. Regarding S-100 protein, the well-differentiated chondrosarcoma element revealed intense positivity, whereas the poorly differentiated myxoid areas were not positive except for a few tumor cells. This is the first case, to our knowledge, of extraskeletal myxoid chondrosarcoma arising from the retroperitoneum, and immunohistologic findings suggest that alpha-1-antitrypsin and alpha-1-antichymotrypsin may be available markers in poorly differentiated chondrosarcomas showing a negative reaction for S-100 protein.


The impact of community and provider-driven social accountability interventions on contraceptive use: findings from a cohort study of new users in Ghana and Tanzania.
Although contraceptive use has increased over 15 years, discontinuation rates remain high. Contraceptive use is becoming more important when addressing unmet need for family planning. Social accountability, defined here as collective processes for holding duty bearers to account for their actions, is a rights-based participatory process that supports service provision and person-centred care, as well as, informed decision-making among community members regarding their health. A study implemented in Ghana and Tanzania was designed to understand and evaluate how social accountability and participatory processes influences quality of care and client satisfaction and whether this results in increased contraceptive uptake and use. We report here on the relationship between social accountability and the use of modern contraceptives, i.e., contraceptive method discontinuation, contraceptive method switching, and contraceptive discontinuation. As part of Community and Provider driven Social Accountability Intervention (CaPSAI) Project, a cohort of women aged 15 to 49 years who were new users of contraception and accessing family planning and contraceptives services at the study facilities across both intervention and control groups were followed-up over a 12-month period to measure changes contraceptive use. In this cohort study over a one-year duration, we did not find a statistically significant difference in Ghana and Tanzania in overall method discontinuation, switching, and contraceptive discontinuation after exposure to a social accountability intervention. In Ghana but not in Tanzania, when stratified by the type of facility (district level vs. health centre), there were significantly less method and contraceptive discontinuation in the district level facility and significantly more method and contraceptive discontinuation in the health centres in the intervention group. In Ghana, the most important reasons reported for stopping a method were fear of side-effects, health concerns and wanting to become pregnant in the control group and fear of side-effects wanting a more effective method and infrequent sex in the intervention group. In Tanzania, the most important reasons reported for stopping a method were fear of side-effects, wanting a more effective method, and method not available in the control group compared to wanting a more effective method, fear of side-effects and health concerns in the intervention group. We did not demonstrate a statistically significant impact of a six-month CaPSAI intervention on contraceptives use among new users in Tanzania and Ghana. However, since social accountability have important impacts beyond contraceptive use it is important consider results of the intermediate outcomes, cases of change, and process evaluation to fully understand the impact of this intervention. The CaPSAI Project has been registered at Australian New Zealand Clinical Trials Registry (ACTRN12619000378123, 11/03/2019).


[Analgesia and sedation to supplement incomplete regional anesthesia].
Pharmacological praemedication. In patients receiving regional anaesthetics induction of deep sedation prior to the performance of the block should be avoided because during the installation of the nerve block it is an advantage to have a cooperative patient. Adequate anxiolytic effects are achieved by oral administration of chloracepate (0.3-0.5 mg/kg body weight). Intraoperative sedation. Once regional anaesthesia is established deep sedation or even a light sleep might be appropriate to improve the patient's comfort. Short acting i.v. substances are the agents of choice. Propofol (1.5-5 mg/kg per h) and midazolam (0.03-0.09 mg/kg per h) are recommended. Both substances should be titrated as needed. Since respiratory depression or loss of airway patency may occur, close observation and pulse oxymetric monitoring are mandatory. Intraoperative analgesia. Restlessness due to pain is not an indication for sedatives and/or hypnotics. Pain can be caused not only by incomplete regional anaesthesia, but also by a tourniquet or uncomfortable body positions, for example, and it should be treated in different ways according to its cause. In the case of an incomplete block, a catheter technique makes a top-up dose for augmentation possible; additional peripheral nerve blocks can also be used to complete the analgesia. If these attempts are unsuccessful, systemic analgesics (preferable narcotics) or even anaesthetics must be given. Opioids are recommended only in mild to moderate pain or discomfort. The risk of respiratory depression should be considered. The administration of oxygen by mask and pulse oxymetric monitoring are useful. Ketamine is a common drug with a potent analgesic effect, which possesses the advantages of good support for the cardiovascular system, because of its sympathomimetic action, and minimal depression of the ventilatory drive. However, with the exception of a few specific indications, Ketamine is not a drug that is initially an integral part of planned regional anaesthetic procedures. In case of incomplete regional blocks administration of ketamine is more frequently the "ultima ratio" following a number of previous, unsuccessful attempts-primarily with sedatives and/or opioids-to achieve a condition that will permit surgical procedures; as a result, the hypnotic and respiratory depressant effects of subsequently administered drugs are enhanced and potentiated. An important consequence of this complex pharmacodynamic interaction scenario is a potential loss of the advantages that would otherwise be gained by using "subanaesthetic" ketamine doses (< 0.5 mg/kg), namely: a cooperative patient who is breathing spontaneously and has an intact laryngopharyngeal reflex response and, therefore, an uncompromised airway competence. Pulse oxymetric monitoring of the potentially endangered respiratory function is obligatory. The individual transition to general anaesthesia is not easy to determine. Therefore, it is essential that, whenever the need arises, intubation and mechanical ventilation intervention procedures be carried out immediately.


Molecular phylogeny of Pemphiginae (Hemiptera: Aphididae) inferred from nuclear gene EF-1alpha sequences.
Three traditional tribes of Fordini, Pemphigini and Eriosomatini comprise Pemphiginae, and there are two subtribes in Fordini and Pemphigini, respectively. Most of the species in this subfamily live heteroecious holocyclic lives with distinct primary host specificity. The three tribes of Pemphigini (except Prociphilina), Eriosomatini and Fordini use three families of plants, Salicaceae (Populus), Ulmaceae (Ulums) and Anacardiaceae (Pistacia and Rhus), as primary hosts, respectively, and form galls on them. Therefore, the Pemphigids are well known as gall makers, and their galls can be divided into true galls and pseudo-galls in type. We performed the first molecular phylogenetic study of Pemphiginae based on molecular data (EF-1alpha sequences). Results show that Pemphiginae is probably not a monophylum, but the monophyly of Fordini is supported robustly. The monophyly of Pemphigini is not supported, and two subtribes in it, Pemphigina and Prociphilina, are suggested to be raised to tribal level, equal with Fordini and Eriosomatini. The molecular phylogenetic analysis does not show definite relationships among the four tribes of Pemphiginae, as in the previous phylogenetic study based on morphology. It seems that the four tribes radiated at nearly the same time and then evolved independently. Based on this, we can speculate that galls originated independently four times in the four tribes, and there is no evidence to support that true galls are preceded by pseudo-galls, as in the case of thrips and willow sawflies.


Geographical Variations of the Minimum Mortality Temperature at a Global Scale: A Multicountry Study.
Minimum mortality temperature (MMT) is an important indicator to assess the temperature-mortality association, indicating long-term adaptation to local climate. Limited evidence about the geographical variability of the MMT is available at a global scale. We collected data from 658 communities in 43 countries under different climates. We estimated temperature-mortality associations to derive the MMT for each community using Poisson regression with distributed lag nonlinear models. We investigated the variation in MMT by climatic zone using a mixed-effects meta-analysis and explored the association with climatic and socioeconomic indicators. The geographical distribution of MMTs varied considerably by country between 14.2 and 31.1 °C decreasing by latitude. For climatic zones, the MMTs increased from alpine (13.0 °C) to continental (19.3 °C), temperate (21.7 °C), arid (24.5 °C), and tropical (26.5 °C). The MMT percentiles (MMTPs) corresponding to the MMTs decreased from temperate (79.5th) to continental (75.4th), arid (68.0th), tropical (58.5th), and alpine (41.4th). The MMTs indreased by 0.8 °C for a 1 °C rise in a community's annual mean temperature, and by 1 °C for a 1 °C rise in its SD. While the MMTP decreased by 0.3 centile points for a 1 °C rise in a community's annual mean temperature and by 1.3 for a 1 °C rise in its SD. The geographical distribution of the MMTs and MMTPs is driven mainly by the mean annual temperature, which seems to be a valuable indicator of overall adaptation across populations. Our results suggest that populations have adapted to the average temperature, although there is still more room for adaptation.


[Income and smoking behavior in Germany--a secondary analysis of data from the 1995 microcensus].
We investigated to what extent two major parameters of the economic situation, namely the household income and the type of the major source of income, are related to smoking behaviour. Study variables are current cigarette smoking and ex-smoking. Databasis is the Public Use File of the "Microcensus 1995", which is a representative survey for Germany, conducted under the auspices of the German Federal Statistical Bureau. Included in the analysis were 185,822 persons aged 18 years and older. 30.9% of males and 18.2% of females are current smokers. 39% of males and 31.2% of females, who had been smokers at any time, stopped smoking. For persons aged 18 to 64 years, we observed strong income-related differences in the smoking prevalence. The higher the income the lower was the rate of smokers. Higher rates for ex-smokers were observed for persons older than 30 years, who had a high household income. For study subjects with a primary income based on unemployment or social welfare payments, we found for both sexes a remarkably higher smoking prevalence. Due to the fact that the underlying causes for the higher prevalence of smoking in the poorer segments of the population are not very well understood, it is rather difficult to establish a comprehensive prevention strategy for this high-risk population group.


Real-Time Pathophysiologic Correlates of Left Atrial Appendage Thrombus in Patients Who Underwent Transesophageal-Guided Electrical Cardioversion for Atrial Fibrillation.
Although current guidelines advocate using the CHA2DS2-VASc score to assess the risk of stroke in patients with atrial fibrillation (AF), compared with transesophageal echocardiography (TEE), its ability to predict left atrial appendage thrombus (LAAT) is limited. We studied 3,324 consecutive patients with sustained AF from our prospective registry of patients who underwent first-time TEE-guided electrical cardioversion (ECV) from May 2000 through March 2012. The association of CHA2DS2-VASc score or TEE risk factors with the occurrence of LAAT was analyzed. The mean (SD) age was 69 (12.5) years and 67% were men. LAAT was identified in 49 (1.5%) during pre-ECV TEE. Compared with patients without LAAT, those with LAAT had lower peak left atrial appendage emptying velocity (LAAEV) (17.2 ± 8.5 vs 36.6 ± 20.8; p <0.001) and left ventricular ejection fraction (LVEF) (39.9 ± 17.6 vs 51.4 ± 13.7; p <0.001); their CHA2DS2-VASc score also was higher, but the difference was not statistically significant (3.6 ± 1.4 vs 3.2 ± 1.6; p = 0.06). Multivariate logistic regression analysis identified an LVEF ≤40% (adjusted odds ratio 2.48, 95% confidence interval 1.38 to 4.46), LAAEV 20.3 to 33.9 cm/s (odds ratio 12.19, 95% confidence interval 1.53 to 96.86), and LAAEV ≤20.2 cm/s as independent predictors of LAAT. An LAAEV cut-point of 20 cm/s and an LVEF ≤40% were optimal for detecting LAAT (sensitivity 75% and 62%; specificity 77% and 75%; area under the curve 0.822 and 0.776, respectively). On follow-up, LAAT was an independent risk factor of subsequent ischemic stroke but did not influence survival. In conclusion, reduced LVEF and reduced LAAEV are important pathophysiologic correlates of left atrial appendage thrombogenesis and subsequent ischemic stroke in patients who underwent TEE-guided ECV for AF.


Deciphering cellular morphology and biocompatibility using polymer microarrays.
A quantitative and qualitative analysis of cellular adhesion, morphology and viability is essential in understanding and designing biomaterials such as those involved in implant surfaces or as tissue-engineering scaffolds. As a means to simultaneously perform these studies in a high-throughput (HT) manner, we report a normalized protocol which allows the rapid analysis of a large number of potential cell binding substrates using polymer microarrays and high-content fluorescence microscopy. The method was successfully applied to the discovery of optimal polymer substrates from a 214-member polyurethane library with mouse fibroblast cells (L929), as well as simultaneous evaluation of cell viability and cellular morphology. Analysis demonstrated high biocompatibility of the binding polymers and permitted the identification of several different cellular morphologies, showing that specific polymer interactions may provoke changes in cell shape. In addition, SAR studies showed a clear correspondence between cellular adhesion and polymer structure. The approach can be utilized to perform multiple experiments (up to 1024 single experiments per slide) in a highly reproducible manner, leading to the generation of vast amounts of data in a short time period (48-72 h) while reducing dramatically the quantities of polymers, reagents and cells used.


Isolation of clonal axenic strains of the symbiotic dinoflagellate Symbiodinium and their growth and host specificity(1).
The cnidarian-dinoflagellate mutualism is integral to the survival of the coral-reef ecosystem. Despite the enormous ecological and economic importance of corals, their cellular and molecular biology and the ways in which they respond to environmental change are still poorly understood. We have been developing a proxy system for examining the coral mutualism in which the dinoflagellate symbiont Symbiodinium is introduced into a clonal population of the host Aiptasia, a small sea anemone closely related to corals. To further develop the tools for this system, we generated five clonal, axenic strains of Symbiodinium and verified the lack of contaminants by growth on rich medium, microscopic examination, and PCR analysis. These strains were assigned to clades A (two strains), B, E, and F based on their chloroplast 23S rDNA sequences. Growth studies in liquid cultures showed that the clade B strain and one of the clade A strains were able to grow photoautotrophically (in light with no fixed carbon), mixotrophically (in light with fixed carbon), or heterotrophically (in dark with fixed carbon). The clade E strain, thought to be free-living, was able to grow photoautotrophically but not heterotrophically. Infection of an aposymbiotic Aiptasia host with the axenic strains showed consistent patterns of specificity, with only the clade B and one of the clade A strains able to successfully establish symbiosis. Overall, the Aiptasia-Symbiodinium association represents an important model system for dissecting aspects of the physiology and cellular and molecular biology of cnidarian-dinoflagellate mutualism and exploring issues that bear directly on coral bleaching.


Why sequence the genome of Mycobacterium tuberculosis?
Radical measures are required to prevent the grim predictions of the World Health Organisation for the deterioration of the global tuberculosis epidemic in the next century from becoming reality. Study of the nerve centre of the tubercle bacillus, its genome, by means of systematic deoxyribonucleic acid sequence analysis will provide a wealth of information about Mycobacterium tuberculosis that will undoubtedly fuel the next generation of research. In the coming year, this highly cost-effective approach will deliver an unprecedented amount of knowledge to catalyse the development of new, more efficient diagnostic tools and therapeutic interventions to detect, control and, ultimately, eliminate tuberculosis.


[Prion infections: most important medical and veterinary aspects].
The paper reviews the most important and topical information on prionic diseases in the medical and veterinary context. It gives data on bovine sponge-like encephalopathy and other new prionic infections in animals, as well as on a new type of the Greutzfeldt-jakob disease. The common pathology, pathogenesis, and transmission of the infections of this group, a prionic concept of their etiology, constituents of present-day epidemiological survey are generalized. Some most tasks of medical and veterinary importance, which are most essential to Russian science, are worded in the conclusion.


Conformational stability and membrane interaction of the full-length ectodomain of HIV-1 gp41: implication for mode of action.
Membrane fusion between the human immunodeficiency virus (HIV) and the target cell plasma membrane is correlated with conformational changes in the HIV gp41 glycoprotein, which include an early exposed conformation (prehairpin) and a late low energy six helix bundle (SHB) conformation also termed hairpin. Peptides resembling regions from the exposed prehairpin have been previously studied for their interaction with membranes. Here we report on the expression, purification, SHB stability, and membrane interaction of the full-length ectodomain of the HIV gp41 and its two deletion mutants, all in their SHB-folded state. The interaction of the proteins with zwitterionic and negatively charged membranes was examined by using various biophysical methods including circular dichroism spectroscopy, differential scanning calorimetry, lipid mixing of large unilamellar vesicles, and atomic force microscopy (AFM). All experiments were done in an acidic environment in which the protein remains in its soluble trimeric state. The data reveal that all three proteins fold into a stable coiled-coil core in aqueous solution and retain a stable helical fold with reduced coiled-coil characteristics in a zwitterionic and negatively charged membrane mimetic environment. Furthermore, in contrast with the extended exposed N-terminal domain, the folded gp41 ectodomain does not induce lipid mixing of zwitterionic membranes. However, it disrupts and induces lipid mixing of negatively charged phospholipid membranes (approximately 100-fold more effective than fusion peptide alone), which are known to be expressed more in HIV-1-infected T cells or macrophages. The results support the emerging model in which one of the roles of gp41 folding into the SHB conformation is to slow down membrane disruption effects induced by early exposed gp41. However, it can further affect membrane morphology once exposed to negatively charged membranes during late stages.


The anomalous health status of Texas border counties.
The health status of Texas border counties is anomalous in that the combination of poor health care services, low socioeconomic status, and low education rate does not result in the expected increase in age-adjusted mortality rate and shortened life expectancy. The anomaly appears to be due largely to the combination of a high Hispanic population density in this region, specifically Mexican-Americans, and to the phenomenon of Hispanic longevity. The latter is considered to be based on a variety of possible cultural factors. In the face of continuing escalations in the cost of health care, further definition of what those factors are and how they could be incorporated into the delivery of health care in our state should be a priority for state health policy.


Suspected carpal tunnel syndrome: Do nerve conduction study results and symptoms match?
To evaluate the diagnostic utility of nerve conduction studies (NCSs) by examining a population with a high pretest probability of carpal tunnel syndrome (CTS), including bilaterally and unilaterally symptomatic patients. Comparison of the results of NCSs with the results of prospective, pre-NCS, self-administered questionnaires of patients with clinical diagnoses of CTS referred for confirmatory NCSs. A tertiary care hospital neuro-electrophysiology laboratory. The study population consisted of 211 patients, 156 (73.9%) of whom were female. Population mean (range) age was 46.7 (21 to 88) years. Mean (range) symptom duration was 29.3 (1 to 300) months. Patient-reported symptom localization and NCS results. Results of NCSs were normal in 83 (39.3%) patients, were consistent with CTS in 121 (57.3%) patients, and suggested non-CTS abnormalities in 7 (3.3%) patients. Bilateral symptoms were reported by 139 (65.9%) patients, and isolated unilateral symptoms were reported by 72 (34.1%) patients. Those reporting bilateral symptoms had the highest agreement with NCS results at 38.8%. Unilateral NCS abnormalities were seen in 18.0% of those reporting bilateral hand symptoms. Discordant findings, either bilateral or opposite-side neuropathies, were seen in 25.6% of those reporting isolated right-sided symptoms, and in 55.2% of those reporting isolated left-sided symptoms. Based on these data the sensitivity of the NCS results was 49.1%, with a specificity of 62.5%. The positive predictive value was 86.4%, and the negative predictive value was 20.2%. Overall accuracy was 51.4%. Likelihood ratios were 1.3 with positive results, and 0.8 with negative results. In our study population NCS results did little to alter the pretest probability of CTS. Physicians using NCSs for verification of clinical diagnosis should be aware of potential limitations.


Social Isolation Modulates CLOCK Protein and Beta-Catenin Expression Pattern in Gonadotropin-Inhibitory Hormone Neurons in Male Rats.
Postweaning social isolation reduces the amplitude of the daily variation of CLOCK protein in the brain and induces lower reproductive activity. Gonadotropin-inhibitory hormone (GnIH) acts as an inhibitor in the reproductive system and has been linked to stress. Social isolation has been shown to lower neuronal activity of GnIH-expressing neurons in the dorsomedial hypothalamus (DMH). The exact mechanism by which social isolation may affect GnIH is still unclear. We investigated the impact of social isolation on regulatory cellular mechanisms in GnIH neurons. We examined via immunohistochemistry the expression of CLOCK protein at four different times throughout the day in GnIH cells tagged with enhanced fluorescent green protein (EGFP-GnIH) in 9-week-old adult male rats that have been raised for 6 weeks under postweaning social isolation and compared them with group-raised control rats of the same age. We also studied the expression of β-catenin-which has been shown to be affected by circadian proteins such as Bmal1-in EGFP-GnIH neurons to determine whether it could play a role in linking CLOCK in GnIH neurons. We found that social isolation modifies the pattern of CLOCK expression in GnIH neurons in the DMH. Socially isolated rats displayed greater CLOCK expression in the dark phase, while control rats displayed increased CLOCK expression in the light phase. Furthermore, β-catenin expression pattern in GnIH cells was disrupted by social isolation. This suggests that social isolation triggers changes in CLOCK and GnIH expression, which may be associated with an increase in nuclear β-catenin during the dark phase.


Association of Docosahexaenoic Acid Supplementation With Alzheimer Disease Stage in Apolipoprotein E ε4 Carriers: A Review.
The apolipoprotein E ε4 (APOE4) allele identifies a unique population that is at significant risk for developing Alzheimer disease (AD). Docosahexaenoic acid (DHA) is an essential ω-3 fatty acid that is critical to the formation of neuronal synapses and membrane fluidity. Observational studies have associated ω-3 intake, including DHA, with a reduced risk for incident AD. In contrast, randomized clinical trials of ω-3 fatty acids have yielded mixed and inconsistent results. Interactions among DHA, APOE genotype, and stage of AD pathologic changes may explain the mixed results of DHA supplementation reported in the literature. Although randomized clinical trials of ω-3 in symptomatic AD have had negative findings, several observational and clinical trials of ω-3 in the predementia stage of AD suggest that ω-3 supplementation may slow early memory decline in APOE4 carriers. Several mechanisms by which the APOE4 allele could alter the delivery of DHA to the brain may be amenable to DHA supplementation in predementia stages of AD. Evidence of accelerated DHA catabolism (eg, activation of phospholipases and oxidation pathways) could explain the lack of efficacy of ω-3 supplementation in AD dementia. The association of cognitive benefit with DHA supplementation in predementia but not AD dementia suggests that early ω-3 supplementation may reduce the risk for or delay the onset of AD symptoms in APOE4 carriers. Recent advances in brain imaging may help to identify the optimal timing for future DHA clinical trials. High-dose DHA supplementation in APOE4 carriers before the onset of AD dementia can be a promising approach to decrease the incidence of AD. Given the safety profile, availability, and affordability of DHA supplements, refining an ω-3 intervention in APOE4 carriers is warranted.


Potato, Solanum tuberosum, defense against Colorado potato beetle, Leptinotarsa decemlineata (Say): microarray gene expression profiling of potato by Colorado potato beetle regurgitant treatment of wounded leaves.
Colorado potato beetle (CPB) is a leading pest of solanaceous plants. Despite the economic importance of this pest, surprisingly few studies have been carried out to characterize its molecular interaction with the potato plant. In particular, little is known about the effect of CPB elicitors on gene expression associated with the plant's defense response. In order to discover putative CPB elicitor-responsive genes, the TIGR 11,421 EST Solanaceae microarray was used to identify genes that are differentially expressed in response to the addition of CPB regurgitant to wounded potato leaves. By applying a cutoff corresponding to an adjusted P-value of <0.01 and a fold change of >1.5 or <0.67, we found that 73 of these genes are induced by regurgitant treatment of wounded leaves when compared to wounding alone, whereas 54 genes are repressed by this treatment. This gene set likely includes regurgitant-responsive genes as well as wounding-responsive genes whose expression patterns are further enhanced by the presence of regurgitant. Real-time polymerase chain reaction was used to validate differential expression by regurgitant treatment for five of these genes. In general, genes that encoded proteins involved in secondary metabolism and stress were induced by regurgitant; genes associated with photosynthesis were repressed. One induced gene that encodes aromatic amino acid decarboxylase is responsible for synthesis of the precursor of 2-phenylethanol. This is significant because 2-phenylethanol is recognized by the CPB predator Perillus bioculatis. In addition, three of the 16 type 1 and type 2 proteinase inhibitor clones present on the potato microarray were repressed by application of CPB regurgitant to wounded leaves. Given that proteinase inhibitors are known to interfere with digestion of proteins in the insect midgut, repression of these proteinase inhibitors by CPB may inhibit this component of the plant's defense arsenal. These data suggest that beyond the wound response, CPB elicitors play a role in mediating the plant/insect interaction.


Heat acclimation: cardiac performance of isolated rat heart.
Cardiac performance was studied in the isolated perfused hearts of rats heat acclimated at 34 degrees C (AC) and their age-matched controls (C). The pressure-volume curves during isovolumetric conditions showed a shift to the right in AC compared with C hearts. At similar left ventricular (LV) volumes end-diastolic and peak systolic pressures of AC hearts were lower, but no difference was observed in the maximal pressure developed at the highest LV volumes measured. In both C and AC hearts the developed force decreased as pacing rate increased. AC and C heart responses were the same up to 250 pulses/min. At higher frequencies the amplitude of the developed force of AC hearts was smaller than that of the controls. In accordance the tension produced by very early premature beat reduced in AC compared with C hearts. Since no hypertrophy was observed in AC hearts, it is concluded that heat acclimation results in a change in the intrinsic properties of the AC hearts exhibited by increased compliance, reduced chamber stiffness, and a decrease in the tension developed for each volume load. It is also suggested that at a high beating rate AC hearts fail to restitute its contractility as quickly as C hearts.


A method improving the accuracy of fluorescence recovery after photobleaching analysis.
Fluorescence recovery after photobleaching has been an established technique of quantifying the mobility of molecular species in cells and cell membranes for more than 30 years. However, under nonideal experimental conditions, the current methods of analysis still suffer from occasional problems; for example, when the signal/noise ratio is low, when there are temporal fluctuations in the illumination, or when there is bleaching during the recovery process. We here present a method of analysis that overcomes these problems, yielding accurate results even under nonideal experimental conditions. The method is based on circular averaging of each image, followed by spatial frequency analysis of the averaged radial data, and requires no prior knowledge of the shape of the bleached area. The method was validated using both simulated and experimental fluorescence recovery after photobleaching data, illustrating that the diffusion coefficient of a single diffusing component can be determined to within approximately 1%, even for small signal levels (100 photon counts), and that at typical signal levels (5000 photon counts) a system with two diffusion coefficients can be analyzed with <10% error.


Blood pressure and hypertension in athletes: a systematic review.
Hypertension is reported to be the most prevalent risk factor for cardiovascular disease in elite athletes. We aimed to review blood pressure (BP) and prevalence of hypertension in different elite athletes, and study whether there was an association between high BP and left ventricular hypertrophy (LVH). A systematic review of studies reporting BP in athletes using search strategies developed for PubMed and EMBASE, including only studies with ≥100 participants. We collected data on BP, prevalence of hypertension, LVH and methods of BP measurement. Of 3723 records identified, 51 met the inclusion criteria. These included men and women (n=138,390), aged mostly between 18 and 40 years, from varied sports disciplines. Mean systolic BP varied from 109±11 to 138±7 mm Hg and mean diastolic BP from 57±12 to 92±10 mm Hg. Strength-trained athletes had higher BP than endurance-trained athletes (131.3±5.3/77.3±1.4 vs 118.6±2.8/71.8±1.2 mm Hg, p<0.05), and there was a trend towards a higher BP in athletes training ≥10 h compared with others (121.8±3.8/73.8±2.5 vs 117.6±3.3/66.8±6.9, p=0.058), but overall there was no significant difference in BP between athletes and controls. The prevalence of hypertension varied from 0% to 83%. Some studies showed an association between high BP and LVH. Measurement methods were poorly standardised. BP and prevalence of hypertension in athletes varied considerably partly because of variations in methodology, but type and intensity of training may contribute towards higher BP. High BP may be associated with LVH.


Immune responses at the maternal-fetal interface.
Pregnancy poses an immunological challenge because a genetically distinct (nonself) fetus must be supported within the pregnant female for the required gestational period. Placentation, or the establishment of the fetally derived placenta, is a common strategy used by eutherian mammals to protect the fetus and promote its growth. However, the substantial morphological differences of the placental architecture among species suggest that the process of placentation results from convergent evolution. Although there are considerable similarities in placental function across placental mammals, there are important differences that arise owing to species-specific immunological (and other biological) constraints. This Review focuses on the immunological similarities and differences that occur at the maternal-fetal interface in the context of human and mouse pregnancies. We discuss how the decidua and placenta of these different species form key immunological barriers that sustain maternal tolerance yet generate innate immune responses that prevent microbial infections.


MR imaging: possibility of tissue characterization of brain tumors using T1 and T2 values.
To evaluate the usefulness of T1 and T2 values for tissue characterization of brain tumors, 37 histologically confirmed brain tumors were examined with a 0.5-T superconductive MR system. With spin-echo and inversion-recovery imaging sequences, computed T1 and T2 images were reconstructed, and T1 and T2 values of the tumors were calculated. Relaxation rates (1/T1 and 1/T2), T1/T2 ratios, and malignancy indexes, which were originally designed for gastrointestinal tumors, were also calculated. Values of all these parameters were so wide-ranged that it was impossible to characterize the tumor tissue types.


Supramolecular Semiconductivity through Emerging Ionic Gates in Ion-Nanoparticle Superlattices.
The self-assembly of nanoparticles driven by small molecules or ions may produce colloidal superlattices with features and properties reminiscent of those of metals or semiconductors. However, to what extent the properties of such supramolecular crystals actually resemble those of atomic materials often remains unclear. Here, we present coarse-grained molecular simulations explicitly demonstrating how a behavior evocative of that of semiconductors may emerge in a colloidal superlattice. As a case study, we focus on gold nanoparticles bearing positively charged groups that self-assemble into FCC crystals via mediation by citrate counterions. In silico ohmic experiments show how the dynamically diverse behavior of the ions in different superlattice domains allows the opening of conductive ionic gates above certain levels of applied electric fields. The observed binary conductive/nonconductive behavior is reminiscent of that of conventional semiconductors, while, at a supramolecular level, crossing the "band gap" requires a sufficient electrostatic stimulus to break the intermolecular interactions and make ions diffuse throughout the superlattice's cavities.


Sulfate transport mutants affect hydrogen sulfide and sulfite production during alcoholic fermentation.
Hydrogen sulfide is a common wine fault, with a rotten-egg odour, which is directly related to yeast metabolism in response to nitrogen and sulfur availability. In grape juice, sulfate is the most abundant inorganic sulfur compound, which is taken up by yeast through two high-affinity sulfate transporters, Sul1p and Sul2p, and a low affinity transporter, Soa1p. Sulfate contributes to H2 S production under nitrogen limitation, by being reduced via the Sulfur Assimilation Pathway (SAP). Therefore, yeast strains with limited H2 S are highly desirable. We report on the use of toxic analogues of sulfate following ethyl methane sulfate treatment, to isolate six wine yeast mutants that produce no or reduced H2 S and SO2 during fermentation in synthetic and natural juice. Four amino acid substitutions (A99V, G380R, N588K and E856K) in Sul1p were found in all strains except D25-1 which had heterozygous alleles. Two changes were also identified in Sul2p (L268S and A470T). The Sul1p (G380R) and Sul2p (A470T) mutations were chosen for further investigation as these residues are conserved amongst SLC26 membrane proteins (including sulfate permeases). The mutations were introduced into EC1118 using Crispr cas9 technology and shown to reduce accumulation of H2 S and do not result in increased SO2 production during fermentation of model medium (chemically defined grape juice) or Riesling juice. The Sul1p (G380R) and Sul2p (A470T) mutations are newly reported as causal mutations. Our findings contribute to knowledge of the genetic basis of H2 S production as well as the potential use of these strains for winemaking and in yeast breeding programmes.


Features of small intestinal pathology (epithelial cell kinetics, intraepithelial lymphocytes, disaccharidases) in a primary Giardia muris infection.
In an attempt to correlate host and parasite-related events occurring during the course of a primary Giardia infection in the mouse we have measured epithelial cell kinetics, enzymes, and intraepithelial lymphocytes at different stages of the infection. New methods were developed for the accurate measurement of parasite numbers and distribution within the gut. In jejunum a modest decrease in villus length and intraepithelial lymphocytes at week 1 preceded a pronounced disaccharidase deficiency at week 2, the time of maximum trophozoite numbers, whereas crypt lengthening and increased cell production became maximal at week 3. As trophozoite numbers fell the intraepithelial lymphocyte count and disaccharidase values rose. With the exception of the intraepithelial lymphocyte count, which followed the same pattern as in jejunum but two weeks later, the changes seen in the ileum were the opposite of those in jejunum, suggesting rapid ileal adaptation. The results indicate that the disaccharidase deficiency associated with giardiasis is likely to represent a direct effect of the parasite on the brush border rather than enterocyte immaturity, whereas the intraepithelial lymphocyte response reflects host immunity to the parasite. Profound adaptive changes occur throughout the small intestine in response to a relatively localised insult.


Low health literacy levels in patients with chronic retinal disease.
A patient's health literacy is fundamental for navigating the health system and managing disease. This study aimed to compare the health literacy levels of patients with chronic retinal disease in Denmark. This cross-sectional questionnaire study used the validated HLS-EU-Q16 questionnaire to determine the health literacy of 225 patients with age-related macular degeneration (AMD), diabetic macular edema (DME) or retinal vein occlusion (RVO), receiving intravitreal treatment at the retinal clinic, Zealand University Hospital, Denmark. Patients were consecutively included as participants for the study. All patients had the option of having the survey read aloud to them. Health literacy levels between the patient groups did not differ significantly, however, the proportion of patients with poor health literacy was high-65% of AMD patients, 73% of DME patients, and 63% of patients with RVO. Low health literacy of patients with retinal disease signify a need for more health literacy research in the field of retinal diseases, to secure that patients have the timely and appropriate knowledge and competencies to manage their condition.


Minimally Invasive Embryo Transfer and Embryo Vitrification at the Optimal Embryo Stage in Rabbit Model.
Assisted reproductive techniques (ARTs), such as in vitro embryo culture or embryo cryopreservation, affect natural development patterns with perinatal and postnatal consequences. To ensure the innocuousness of ART applications, studies in animal models are necessary. In addition, as a last step, embryo development studies require evaluation of their capacity to develop full-term healthy offspring. Here, embryo transfer to the uterus is indispensable to perform any ARTs-related experiment. The rabbit has been used as a model organism to study mammalian reproduction for over a century. In addition to its phylogenetic proximity to the human species and its small size and low maintenance cost, it has important reproductive characteristics such as induced ovulation, a chronology of early embryonic development similar to humans and a short gestation that allow us to study the consequences of ART application easily. Moreover, ARTs (such as intracytoplasmic sperm injection, embryo culture, or cryopreservation) are applied with suitable efficiency in this species. Using the laparoscopic embryo transfer technique and the cryopreservation protocol presented in this article, we describe 1) how to transfer embryos through an easy, minimally invasive technique and 2) an effective protocol for long-term storage of rabbit embryos to provide time-flexible logistical capacities and the ability to transport the sample. The outcomes obtained after transferring rabbit embryos at different developmental stages indicate that morula is the ideal stage for rabbit embryo recovery and transfer. Thus, an oviductal embryo transfer is required, justifying the surgical procedure. Furthermore, rabbit morulae are successfully vitrified and laparoscopically transferred, proving the effectiveness of the described techniques.


Self-Fluid Management in Prevention of Kidney Stones: A PRISMA-Compliant Systematic Review and Dose-Response Meta-Analysis of Observational Studies.
Epidemiologic studies have suggested that daily fluid intake that achieves at least 2.5 L of urine output per day is protective against kidney stones. However, the precise quantitative nature of the association between fluid intake and kidney stone risk, as well as the effect of specific types of fluids on such risk, are not entirely clear.We conducted a systematic review and dose-response meta-analysis to quantitatively assess the association between fluid intake and kidney stone risk. Based on a literature search of the PubMed, Embase, and Cochrane Library databases, 15 relevant studies (10 cohort and 5 case-control studies) were selected for inclusion in the meta-analysis with 9601 cases and 351,081 total participants.In the dose-response meta-analysis, we found that each 500 mL increase in water intake was associated with a significantly reduced risk of kidney stone formation (relative risk (RR) = 0.93; 95% CI: 0.87, 0.98; P < 0.01). Protective associations were also found for an increasing intake of tea (RR = 0.96; 95% CI: 0.93, 0.99; P = 0.02) and alcohol (RR = 0.80, 95% CI: 0.75, 0.85; P < 0.01). A borderline reverse association were observed on coffee intake and risk of kidney stone (RR = 0.88; 95% CI: 0.76, 1.00; P = 0.05). The risk of kidney stones was not significantly related to intake of juice (RR = 1.02, 95% CI: 0.95, 1.10; P = 0.64), soda (RR = 1.03; 95% CI: 0.90, 1.17; P = 0.65), or milk (RR = 0.96; 95% CI: 0.88, 1.03; P = 0.21). Subgroup analysis and sensitivity analyses showed inconsistent results on coffee, alcohol, and milk intake.Increased water intake is associated with a reduced risk of kidney stones; increased consumption of tea and alcohol may reduce kidney stone risk. An average daily water intake was recommended for kidney stone prevention.


Determining the effectiveness of mental health services from a consumer perspective: part 1: enhancing recovery.
The routine use of standardized instruments to measure consumer outcomes is now part of mental health policy throughout Australia. However, it has been broadly criticized for (i) not involving consumer input into the design of instruments; and (ii) not reflecting the aspects of care and treatment considered beneficial for recovery by consumers themselves. The importance of the concept of recovery is increasingly considered in the literature. Despite this, there is a paucity of research describing the effectiveness of services in promoting recovery from the perspective of consumers of mental health services. The aim of this study is to explore consumer perspectives in relation to the factors that promote and impede recovery, and the principles that ideally should underpin the evaluation of services. Focus group interviews were conducted with consumers of mental health services (n = 16) from one rural and one metropolitan mental health service in Victoria, Australia. This paper presents Part 1 of the findings, pertaining to aspects of mental health services that enhance recovery. Two main themes arose during the data analysis process: (i) treatment; and (ii) support and social connectedness. Various treatment strategies, including medication and spiritual involvement, were considered helpful. However, support from both staff and peers emerged as a more important and influential factor.


[Purification of flavin-containing liver polyamine oxidase and luminescent properties of the ferment].
A flavin-containing polyamine oxidase was isolated in an electrophoretically homogeneous state from cattle liver cytosol. The molecular mass, subunit composition and flavin content of the enzyme were determined; flavin, was shown to be covalently bound to the protein fragment of the polyamine oxidase molecule. Some optical and luminescent properties of the native and denatured enzyme were investigated. Denaturation and quenching were found to affect the luminescent properties of polyamine oxidase.


dCas9-based epigenome editing suggests acquisition of histone methylation is not sufficient for target gene repression.
Distinct epigenomic profiles of histone marks have been associated with gene expression, but questions regarding the causal relationship remain. Here we investigated the activity of a broad collection of genomically targeted epigenetic regulators that could write epigenetic marks associated with a repressed chromatin state (G9A, SUV39H1, Krüppel-associated box (KRAB), DNMT3A as well as the first targetable versions of Ezh2 and Friend of GATA-1 (FOG1)). dCas9 fusions produced target gene repression over a range of 0- to 10-fold that varied by locus and cell type. dCpf1 fusions were unable to repress gene expression. The most persistent gene repression required the action of several effector domains; however, KRAB-dCas9 did not contribute to persistence in contrast to previous reports. A 'direct tethering' strategy attaching the Ezh2 methyltransferase enzyme to dCas9, as well as a 'recruitment' strategy attaching the N-terminal 45 residues of FOG1 to dCas9 to recruit the endogenous nucleosome remodeling and deacetylase complex, were both successful in targeted deposition of H3K27me3. Surprisingly, however, repression was not correlated with deposition of either H3K9me3 or H3K27me3. Our results suggest that so-called repressive histone modifications are not sufficient for gene repression. The easily programmable dCas9 toolkit allowed precise control of epigenetic information and dissection of the relationship between the epigenome and gene regulation.


Legacy perceptions and interventions for adults and children receiving palliative care: A systematic review.
Legacy has been invoked as a means for strengthening human attachments, continuing bonds, and ensuring that individuals will be remembered; however, little is known about the spectrum of approaches to, outcomes associated with, and best practices for legacy interventions. To systematically review research on legacy perceptions and interventions in pediatric and adult palliative care recipients. A systematic mixed studies review synthesizing quantitative, qualitative, and mixed-methods study findings using PRISMA guidelines. PubMed, PsycINFO, and CINAHL databases were searched on October 1, 2020. GRADE criteria were used to assess quality of quantitative reports, and the Johns Hopkins Evidence Level and Quality Guide was used to rate qualitative, mixed methods, and review articles. Data were synthesized using integrative thematic analysis. The 67 studies reviewed describe a variety of legacy perceptions and interventions with adult and pediatric patients receiving palliative care. Statistically significant improvements in various dimensions of wellbeing are documented, with significant reduction in incidence and symptoms of depression in adults. Studies highlight the utility, feasibility, and perceived benefits of legacy interventions according to adult patients and their caregivers, and parents/caregivers of pediatric patients. Though future research with high-quality, experimental designs is needed, the positive outcomes associated with legacy interventions are documented in adult patient populations; additionally, the application of legacy interventions for children with serious illnesses receiving palliative care is reasonable based on the existing body of evidence. A consistent and operational concept of legacy is still needed for future research and practice.


Validating the Factor Structure of the Psychosocial Assessment Tool Using Internet-Based Data.
The Psychosocial Assessment Tool (PAT) is a well-validated, brief screener of family psychosocial risk. Since 2014 a web-based version of the PAT (WebPAT) has been available for use by clinicians and researchers, but the psychometric properties have not been examined. The objective of this article was to examine the factor structure and internal consistency of the WebPAT, which was administered to caregivers of youth with cancer. The WebPAT was administered to 1,252 caregivers of youth with cancer across 29 institutions. Confirmatory factor analysis (CFA) was used to examine the factor structure of the WebPAT. Internal consistencies of the total and subscale scores were examined via the Kuder-Richardson 20 coefficient. The distribution of total PAT score across the three risk categories of the Pediatric Psychosocial Preventative Health Model (PPPHM) was also examined. The CFA supported the original seven-factor structure of the PAT (Family Structure, Social Support, Child Problems, Sibling Problems, Family Problems, Stress Reactions, and Family Beliefs). Internal consistencies were strong for the total PAT score and four subscales (Social Support, Child Problems, Sibling Problems, and Family Problems). The distribution of total PAT scores across PPPHM risk categories was consistent with prior research. The WebPAT is a psychometrically sound screener of psychosocial risk in families of youth with cancer. Healthcare providers can use the WebPAT to assess families' psychosocial risk and guide the provision of psychosocial care. Future research should evaluate the implementation of the PAT and identify barriers and facilitators to implementation.


How vitamin A metabolizing dendritic cells are generated in the gut mucosa.
CD103(+) dendritic cells (DCs) represent the major migratory DC population in the intestinal lamina propria and are believed to play an essential role in the initiation and regulation of mucosal adaptive immune responses. Small intestine (SI) CD103(+) DCs have an enhanced capacity to generate the vitamin A metabolite, retinoic acid, a property that underlies their ability to induce the gut homing receptors CC chemokine receptor 9 and α4β7 on responding T and B cells, and enhance forkhead box P3(+) T regulatory and IgA plasma cell differentiation in vitro. In this review, we discuss the environmental signals that appear to promote vitamin A metabolising activity in SI CD103(+) DCs in the steady state and thus which may contribute to driving the unique nature of SI immune responses.


Tuberculous appendix: a review of clinical presentations and outcomes.
Tuberculous appendix is surprisingly rare, even in countries where this infection is common. We report our experience with tuberculous appendix over a 15-year period. A search for cases of tuberculous appendix was conducted from January 1995 to December 2009 on the databases of the National Tuberculosis Centre and the Departments of Pathology and Surgery of Raja Isteri Pengiran Anak Saleha Hospital. There were five cases of tuberculous appendix, giving a cumulative incidence of 0.08 percent of all appendectomies (n is 6,593), 0.2 percent of tuberculosis (TB) cases (n is 2,876) and 8.6 percent of abdominal TB (n is 58). Three patients were male and two were female, with a median age of 27 (range 25-48) years. Four patients presented with symptoms of acute appendicitis and one with an appendiceal mass following treatment for acute gastroenteritis. Only one patient had constitutional symptoms. Four patients had appendectomies (one laparoscopic and three open) within the same admission and one had interval appendectomy. Operative findings included perforated appendix (n is 1), appendiceal mass (n is 1) and acute appendicitis (n is 3). In all cases, the diagnoses were made only after review of the histology. None of the patients had pulmonary TB. Delay in initiating anti-TB treatment in one patient resulted in the development of an ileocutaneous fistula following appendectomy. This was resolved by excision, followed by anti-TB treatment. Tuberculous appendix can present as acute appendicitis, and the diagnosis is often made after surgery. Any delay in treatment can lead to significant complications.


Quality of life after traumatic injury: a latent trajectory modeling approach.
It is largely unknown how quality of life (QoL) changes following accidental injuries. Equally, the mechanisms underlying such changes have not yet been identified in detail. This study of injured accident survivors aimed to: (1) detect a model of change which best explains the observed course of QoL, and (2) identify potential predictor variables. 323 injured accident survivors were interviewed within 2 weeks of the trauma, and followed up at 6 and 12 months. Latent trajectory modeling was used to analyze the fit of three potential trajectories regarding the observed course of general QoL as measured by the Questions on Life Satisfaction questionnaire. The trajectory model adopting a negative square-root change fitted the observed data best, meaning that shortly after the accident, general QoL decreased strongly with diminishing negative changes occurring later on. Early and prolonged QoL impairment was largely attributable to the initial level of posttraumatic stress as measured by the Clinician-Administered PTSD Scale. To a lesser extent, depressive symptoms also predicted change in subjective QoL, while injury severity showed no direct effect; rather, its impact on QoL was mediated by initial posttraumatic stress. By contrast, reduced occupational functioning was attributable to injury severity rather than psychopathology. When treating injured accident survivors, clinicians should consider symptoms of posttraumatic stress and comorbid depression in order to prevent or mitigate negative changes in QoL.


Metal removal from acid mine lake using ultrasound-assisted modified fly ash at different frequencies.
Acid mine drainage/lakes (AMD/AMLs) have a low pH with high concentrations of metals and sulfate and have been a major environmental problem in the Can Coal Basin, in northwestern Turkey. In this study, metal removal from Hayirtepe AML by using fly ash (FA) and modified fly ash (MFA) was investigated in batch experiments. The effects of various parameters, such as ultrasonic frequency, dose, contact time, pH, and temperature, were examined to determine the optimum conditions for metal removal from AML. This study also focused on the application of ultrasound-assisted modification by using a 20-kHz ultrasonic probe and a 40-kHz ultrasonic bath to increase the FA surface and improve its adsorption capacity for metal removal. FA modification at 20 kHz showed better results than that at 40 kHz because it produced rapid bubble implosion with acoustic cavitation. The FA and MFAs selectivity for metal removal was 98%-99% for Fe, 96%-99% for Al, 94%-97% for Zn, 90%-95% for Co, 88%-94% for Ni, 77%-92% for Cu, and 74%-92% for Mn according to the determined optimum parameters. Scanning electron microscopy coupled with the energy-dispersive X-ray spectroscopy (SEM/EDX) and X-ray diffractometry of the solid residues (SRs) identified gypsum as a new mineral phase from sulfate removal from the AML. Inductively coupled plasma mass spectrometry and SEM/EDX analysis revealed that the metal content of the SRs increased. The adsorption process fitted the pseudo-second order kinetic model. Thermodynamic parameters showed that the process was exothermic and the randomness of the solid/solution interface increased during adsorption. Reuse experiments indicated that the MFAs were reused more effectively for metal removal from AML compared with the FA. This study showed that the use of MFAs with a high adsorption capacity and surface area is economic and efficient for metal removal from AML.


Mucosal-Associated Invariant T (MAIT) Cells Are Highly Activated and Functionally Impaired in COVID-19 Patients.
Coronavirus disease 2019 (COVID-19), caused by infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), comprises mild courses of disease as well as progression to severe disease, characterised by lung and other organ failure. The immune system is considered to play a crucial role for the pathogenesis of COVID-19, although especially the contribution of innate-like T cells remains poorly understood. Here, we analysed the phenotype and function of mucosal-associated invariant T (MAIT) cells, innate-like T cells with potent antimicrobial effector function, in patients with mild and severe COVID-19 by multicolour flow cytometry. Our data indicate that MAIT cells are highly activated in patients with COVID-19, irrespective of the course of disease, and express high levels of proinflammatory cytokines such as IL-17A and TNFα ex vivo. Of note, expression of the activation marker HLA-DR positively correlated with SAPS II score, a measure of disease severity. Upon MAIT cell-specific in vitro stimulation, MAIT cells however failed to upregulate expression of the cytokines IL-17A and TNFα, as well as cytolytic proteins, that is, granzyme B and perforin. Thus, our data point towards an altered cytokine expression profile alongside an impaired antibacterial and antiviral function of MAIT cells in COVID-19 and thereby contribute to the understanding of COVID-19 immunopathogenesis.


The significance of circadian rhythms and dysrhythmias in critical illness.
Many physiological and cellular processes cycle with time, with the period between one peak and the next being roughly equal to 24 h. These circadian rhythms underlie 'permissive homeostasis', whereby anticipation of periods of increased energy demand or stress may enhance the function of individual cells, organ systems or whole organisms. Many physiological variables related to survival during critical illness have a circadian rhythm, including the sleep/wake cycle, haemodynamic and respiratory indices, immunity and coagulation, but their clinical significance remains underappreciated. Critically ill patients suffer from circadian dysrhythmia, manifesting overtly as sleep disturbance and delirium, but with widespread covert effects on cellular and organ function. Environmental and pharmacological strategies that ameliorate or prevent circadian dysrhythmia have demonstrated clinical benefit. Harnessing these important biological phenomena to match metabolic supply to demand and bolster cell defenses at the apposite time may be a future therapeutic strategy in the intensive care unit.


Directed differential connectivity graph of interictal epileptiform discharges.
In this paper, we study temporal couplings between interictal events of spatially remote regions in order to localize the leading epileptic regions from intracerebral EEG (iEEG). We aim to assess whether quantitative epileptic graph analysis during interictal period may be helpful to predict the seizure onset zone of ictal iEEG. Using wavelet transform, cross-correlation coefficient, and multiple hypothesis test, we propose a differential connectivity graph (DCG) to represent the connections that change significantly between epileptic and nonepileptic states as defined by the interictal events. Postprocessings based on mutual information and multiobjective optimization are proposed to localize the leading epileptic regions through DCG. The suggested approach is applied on iEEG recordings of five patients suffering from focal epilepsy. Quantitative comparisons of the proposed epileptic regions within ictal onset zones detected by visual inspection and using electrically stimulated seizures, reveal good performance of the present method.


Small-molecule targeting AMPA-mediated excitotoxicity has therapeutic effects in mouse models for multiple sclerosis.
While most research and treatments for multiple sclerosis (MS) focus on autoimmune reactions causing demyelination, it is possible that neurodegeneration precedes the autoimmune response. Hence, glutamate receptor antagonists preventing excitotoxicity showed promise in MS animal models, though blocking glutamate signaling prevents critical neuronal functions. This study reports the discovery of a small molecule that prevents AMPA-mediated excitotoxicity by targeting an allosteric binding site. A machine learning approach was used to screen for small molecules targeting the AMPA receptor GluA2 subunit. The lead candidate has potent effects in restoring neurological function and myelination while reducing the immune response in experimental autoimmune encephalitis and cuprizone MS mouse models without affecting basal neurotransmission or learning and memory. These findings facilitate development of a treatment for MS with a different mechanism of action than current immune modulatory drugs and avoids important off-target effects of glutamate receptor antagonists. This class of MS therapeutics could be useful as an alternative or complementary treatment to existing therapies.


Channel modulators affect PGE(2) binding to bovine aortic endothelial cells.
PGE(2), PGF(2alpha) and the thromboxane agonist U-46619 bind to bovine aortic endothelial cells and compete on the same binding site with similar affinity. In addition, binding remains unaffected by prolonged exposure to the ligand. These characteristics differ significantly from those of any known G-coupled prostaglandin receptor. Binding of PGE(2) to the cells is reduced in the presence of the cyclic nucleotides cGMP and cAMP, and is unaffected by protein kinase inhibitors. Removal of permeable cyclic nucleotides from the cell medium results in a fast and complete restoration of PGE(2) binding to the cells, suggesting that both cyclic nucleotides reduce PGE(2) binding by a reversible interaction with the prostaglandin-binding site, without the involvement of second messenger-activated protein kinases. Our data further show that binding of prostaglandins to bovine aortic endothelial cells is sensitive to heavy metals and to activators and blockers of calcium, ATP-sensitive K(+) and chloride channels. Nickel, a specific cyclic nucleotide-gated (CNG) channel activator, decreases PGE(2) binding and so do the CNG channel activators Rp-8-Br-PET-cGMPS and Sp-8-Br-PET-cGMPS. On the other hand, the calcium channel blockers pimozide, diltiazem as well as LY-83,583, a guanylate cyclase inhibitor, which were reported to block CNG channels, enhance PGE(2) binding. The sensitivity of PGE(2) binding to selective CNG channel modifying agents, as well as the rapid and reversible interaction with cyclic nucleotides, may suggest that the common low-affinity prostanoid-binding site on bovine aortic endothelial cells is associated with a molecular entity, which possess several properties of a CNG channel.


Perinatal effects of exposure to PCBs on social preferences in young adult and middle-aged offspring mice.
In social species, social interactions between conspecifics constitute a fundamental component to establish relations, provide best chances to reproduce, and even improve survival rates. In this study, a three-chambered social approach test was used to estimate the level of sociability and level of preference for social novelty in both male and female young adult (postnatal day (PND) 50) and middle-aged (PND 330) offspring mice (n=10 per group) that were perinatally exposed to a mixture of six polychlorinated biphenyls (PCBs), 28, 52, 101, 138, 153, and 180, at environmentally low doses (10 and 1000ng/kg b.w. for dams during gestation and lactation), a profile that closely mimics human exposure to contaminated fish. Our results showed that PCBs bidirectionally modulated social preferences in offspring mice, and the effects were sex and age dependent. However, increased levels of social interactions were rather frequently detected in both assays of the three-chambered test. Reduced social interaction was only induced in 1000ng/kg PCB-exposed middle-aged males, which exhibited similar preferences to social and non-social stimuli when compared to middle-aged controls. Furthermore, results showed that plasma levels of both corticosterone and acetylcholinesterase activity were higher in all PCB-exposed middle-aged males and females than in their control counterparts. In summary, although the effects of PCBs were only of moderate magnitude, our results suggest that a PCB mixture can act as an endocrine disruptor in offspring mice, disturbing the formation of normal social habits.


A central role for decorin during vertebrate convergent extension.
Decorin, an archetypal member of the small leucine-rich proteoglycan gene family, regulates collagen fibrillogenesis and cell growth. To further explore its biological function, we examined the role of Decorin during zebrafish development. Zebrafish Decorin is a chondroitin sulfate proteoglycan that exhibits a high degree of conservation with its mammalian counterpart and displays a unique spatiotemporal expression pattern. Morpholino-mediated knockdown of zebrafish decorin identified a developmental role during medial-lateral convergence and anterior-posterior extension of the body plan, as well as in craniofacial cartilage formation. decorin morphants displayed a pronounced shortening of the head-to-tail axis as well as compression, flattening, and extension of the jaw cartilages. The morphant phenotype was efficiently rescued by zebrafish decorin mRNA. Unexpectedly, microinjection of excess zebrafish decorin mRNA or proteoglycan/protein core into one-cell stage embryos caused cyclopia. The morphant and overexpression phenotype represent a convergent extension defect. Our results indicate a central function for Decorin during early embryogenesis.