[Simultaneous determination of leucovorin and 5-methyl-tetrahydrofolate diastereoisomers in human plasma by high-performance liquid chromatography-tandem mass spectrometry coupled with ultrafiltration centrifugation-based pretreatment and its application to a pharmacokinetic study].
A simple, sensitive, and stable high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was developed and validated for the simultaneous determination of leucovorin and 5-methyltetrahydrofolate diastereomers in human plasma using methotrexate as the internal standard. The analytes and the internal standard were extracted from plasma samples by simple ultrafiltration centrifugation-based extraction. The separation was achieved on a chiral HSA column (150 mm×4 mm, 5 μm) using mobile phases containing 10 mmol pH 8.0 ammonium acetate and acetonitrile in gradient mode. The method showed good linearities in the ranges of 25-5000 μg/L and 12.5-3000 μg/L for leucovorin and 5-methyltetrahydrofolate diastereoisomers, respectively. The method was fully validated with respect to sensitivity, precision, accuracy, matrix effect, extraction recovery, and stability of analytes under various conditions. The method was successfully applied to a pharmacokinetic study of 125 mg/m2 6R,S-leucovorin and 62.5 mg/m2 6S-leucovorin. The results showed that the maximum observed concentrations (Cmax) of 6S-leucovorin and L-5-methyltetrahydrofolate were (3137.917±408.837) and (1679.633±244.132) μg/L, respectively, and the areas under the curve from the time of dosing to the last measurable concentration (AUC0-t) were (7504.883±1185.101) and (14001.214±2868.949) μg/L in the 125 mg/m2 6R,S-leucovorin dose group. The Cmax values of 6S-leucovorin and L-5-methyltetrahydrofolate were (3187.917±387.298) and (1739.204±224.755) μg/L, respectively, and AUC0-t values were (7426.664±854.825) and (14884.331±1843.353) μg/L in the 62.5 mg/m2 6S-leucovorin dose group. There were no significant diffe-rences in the main pharmacokinetic parameters between the two dose groups, and the pharmacokinetic characteristics as well as the rate and extent of absorption were consistent. This method can provide technical support for future bioequivalence studies of sodium leucovorin.


Cytotoxic T cell activation by class I protein on cell-size artificial membranes: antigen density and Lyt-2/3 function.
The role of antigen surface density, and its relationship to the function of the Lyt-2/3 complex, in recognition and triggering of allospecific cytotoxic T lymphocyte (CTL) precursors has been studied by using a novel type of Class I protein-bearing artificial membrane. The cell-size membranes, termed pseudocytes (artificial cells), can be handled like cells but have a well-defined and easily quantitated surface composition. Class I antigen on these membranes stimulated generation of secondary in vitro allogeneic CTL responses as effectively as allogeneic spleen cells, provided that lymphokines were added to the cultures. Antigen density on the pseudocyte surfaces could be varied over a wide range and quantitated by papain cleavage and fluorescence-activated cell sorter analysis. Recognition and triggering of precursor CTL was found to be dramatically dependent on the surface density of antigen and displayed a marked threshold density requirement, below which little or no response occurred. Examination of the effects of anti-Lyt-2 antibody on responses to pseudocytes provided direct evidence for a reciprocal relationship between antigen density and susceptibility to antibody blockade. The results strongly suggest that antigen density is likely to have important biological consequences in control of immune responses. They also show that if Lyt-2/3 functions by interaction with a ligand, then that ligand is the Class I protein.


Transmural mitochondrial differences in myocardium.
Two functional mitochondrial populations with different sedimentation rates (S) were obtained from homogenates of canine myocardium by rate zonal centrifugation using an iso-osmotic Ficoll gradient. To ascertain the origin of these populations, the left ventricular wall of normal myocardium was divided into subepicardial (outer one-third), intermediate (middle on-third), and subendocardial (inner one-third) layers. The slow S mitochondria comprised 75% of the mitochondrial population of the subepicardial layer. In contrast, the fast S mitochondria contributed 65% of the subendocardial population. Intermediate layer mitochondria resembled those of the subepicardium. Mitochondria isolated from the three layers had approximately the same density, as shown by isopycnic zonal centrifugation. These studies indicate that mitochondria from subepicardial and subendocardial layers of normal myocardium differ in size and shape but not in density. Electron micrographs (EM) of the subepicardium showed many mitochondria as long as 4 to 8 sarcomeres. Mitochondria from the outer and inner layers of normal myocardium had the same oxidative phosphorylation parameters. Acute myocardial infarction, lasting 1 or 2 hr, resulted in the selective loss of the fast S mitochondria. Because the fast S mitochondria are prevalent in the subendocardium, these results may explain the greater vulnerability of this layer to anoxia.


Genotype-specific methylation of HPV in cervical intraepithelial neoplasia.
Hypermethylation of human papillomavirus (HPV) and host genes has been reported in cervical cancer. However, the degree of methylation of different HPV types relative to the severity of the cervical lesions remains controversial. Studies of the degree of methylation associated with the host gene and the HPV genome to the severity of cervical lesions are rare. We examined the association of methylation status between host genes and late gene 1 (L1) regions of HPV16, 18, 52, and 58 in cervical brushings. Cervical brushings from 147 HPV-infected patients were obtained. The samples comprised normal (n=28), cervical intraepithelial neoplasia (CIN) 1 (n=45), CIN2 (n=13), and CIN3/carcinoma in situ (n=61). The methylation status of HPV and host genes was measured using bisulfite pyrosequencing and quantitative methylation-specific polymerase chain reaction (PCR). The degree of methylation of L1 in HPV16, 18, and 52 was associated with the severity of the cervical lesion. In HPV52, C-phosphate-G (CpG) sites 6368m, 6405m, and 6443m showed significantly higher methylation in lesions ≥CIN3 (p=0.005, 0.003, and 0.026, respectively). Methylation of most HPV types except HPV52 (r<-0.1) was positively correlated with the degree of methylation of host genes including PAX1 and SOX1 (0.4≤r≤0.7). Combining HPV methylation with PAX1 methylation improved the clustering for ≥CIN2. Our study showed that the degree of L1 methylation of HPV16, 18, and 52 but not 58 is associated with the severity of cervical lesions. The association between HPV methylation and host gene methylation suggests different responses of host cellular epigenetic machinery to different HPV genotypes.


Mechanophenotyping of 3D multicellular clusters using displacement arrays of rendered tractions.
Epithelial tissues mechanically deform the surrounding extracellular matrix during embryonic development, wound repair, and tumor invasion. Ex vivo measurements of such multicellular tractions within three-dimensional (3D) biomaterials could elucidate collective dissemination during disease progression and enable preclinical testing of targeted antimigration therapies. However, past 3D traction measurements have been low throughput due to the challenges of imaging and analyzing information-rich 3D material deformations. Here, we demonstrate a method to profile multicellular clusters in a 96-well-plate format based on spatially heterogeneous contractile, protrusive, and circumferential tractions. As a case study, we profile multicellular clusters across varying states of the epithelial-mesenchymal transition, revealing a successive loss of protrusive and circumferential tractions, as well as the formation of localized contractile tractions with elongated cluster morphologies. These cluster phenotypes were biochemically perturbed by using drugs, biasing toward traction signatures of different epithelial or mesenchymal states. This higher-throughput analysis is promising to systematically interrogate and perturb aberrant mechanobiology, which could be utilized with human-patient samples to guide personalized therapies.


[Synergism of the anti-angina effect of obzidan and finoptin in patients with effort stenocardia].
The effect of propranolol-finoptin combinations was examined in 27 patients with angina of effort. Repeated exercise testing showed this combination to provide an essential antianginal effect that exceeded a mere sum of individual effects of each of the two drugs taken alone in the same doses. The high anti-anginal effect of the combination is suggestive of a synergy in their antianginal action resulting in a further reduction of myocardial oxygen requirement. Side effects necessitating discontinuation of the drugs were only recorded during the early days of the treatment; therefore, patients should be adequately monitored during the first days of the regular use of this combination.


IAP Proteins Antagonist: An Introduction and Chemistry of Smac Mimetics under Clinical Development.
Smac mimetics (also known as IAP antagonist) are a new class of targeted drugs having a goal to suppress the IAPs, reestablishing the apoptotic pathways and inducing cancer cell death. Therefore, development of Smac mimetics was considered an attractive strategy for the development of new anticancer drugs. Lots of reviews have come in yesteryears which mainly discussed the biology of IAPs and their role in cancer development. None of these reviews focused on the chemical synthesis of Smac mimetics. Literature study was done by using standard bibliographic search engines like scifinder, pubmed etc. The characteristic features of screened articles were described in the review. The review gives an introduction of IAP proteins and Smac mimetics. Readers will gain an overview of the development of Smac mimetics with representative examples of both monovalent and bivalent Smac mimetics as anticancer agents and an understanding of their structure-activity relationships. Chemical synthesis of biologically important Smac mimetics was discussed briefly in this review. Small molecules that mimic Smac are continuously progressing towards clinical development. Smac mimetics are generally well tolerated and have demonstrated rapid suppression of their target (the IAPs), activation of apoptosis and anti-tumor activity. Continuous research has been done to generate even more insight into the function of IAP proteins to significantly enhance the therapeutical potential of Smac mimetics.


Population diversification in a yeast metabolic program promotes anticipation of environmental shifts.
Delineating the strategies by which cells contend with combinatorial changing environments is crucial for understanding cellular regulatory organization. When presented with two carbon sources, microorganisms first consume the carbon substrate that supports the highest growth rate (e.g., glucose) and then switch to the secondary carbon source (e.g., galactose), a paradigm known as the Monod model. Sequential sugar utilization has been attributed to transcriptional repression of the secondary metabolic pathway, followed by activation of this pathway upon depletion of the preferred carbon source. In this work, we demonstrate that although Saccharomyces cerevisiae cells consume glucose before galactose, the galactose regulatory pathway is activated in a fraction of the cell population hours before glucose is fully consumed. This early activation reduces the time required for the population to transition between the two metabolic programs and provides a fitness advantage that might be crucial in competitive environments.


Adult colonic intussusception caused by cecum adenoma: successful treatment by emergency laparoscopy: report of a case.
Although intestinal intussusception is relatively common in children, in adults it remains a rare clinical entity. We report an emergency laparoscopy for an adult case of colonic intussusception caused by cecum adenoma. In the present case, owing to a successful reduction via laparoscopy, the extensive intussusception could be treated with a more limited resection (wedge resection) instead of an en-bloc wide resection (right hemicolectomy). Because of the theoretical risk of perforations which could lead to the seeding of tumor cells and microorganisms into the intra-abdominal cavity, most surgeons advocate an en-bloc resection without reduction. The experience gained from the present case suggests that laparoscopy may therefore be a useful diagnostic or therapeutic tool for selected cases of adult intussusception. The choice of using either a laparoscopic approach or an open approach depends on the clinical condition of a patient, the location and extent of intussusception, the possibility of underlying disease, and the availability of surgeons with sufficient laparoscopic expertise. Additional reports may help standardize the management of this uncommon disease.


Staircase electrophoresis profiles of stable low-molecular-weight RNA--a new technique for yeast fingerprinting.
Staircase electrophoresis (SCE) in polyacrylamide gels was used to analyse the stable low-molecular-weight (LMW) RNA profiles of several yeast species and genera. As in prokaryotes, this new electrophoretic technique results in good separation of molecules forming LMW RNA profiles in yeasts. In this study it is reported that, while LMW RNA profiles in prokaryotes include only 5S rRNA, and class 2 and class 1 tRNA, these profiles in eukaryotes also include 5.8S rRNA. Differences in the number and distribution of RNA bands in these profiles allowed identification of differences among the yeast species and genera assayed. LMW RNAs, analysed by SCE, provide a yeast fingerprint that allows them to be clearly differentiated and will in the future enable the rapid assignment of yeast isolates to already described species and the detection of new ones.


Temporal trends of transmitted HIV drug resistance in a multinational seroconversion cohort.
The rate of transmitted drug resistance (TDR) may increase with wider use of antiretroviral therapy and can contribute to therapeutic failure. We analysed time trends in TDR among HIV seroconverters. Using CASCADE data of individuals with well estimated dates of HIV seroconversion, we examined HIV nucleotide sequences collected prior to antiretroviral therapy use from 1996-2012. All samples were taken within 12 months of testing HIV positive. Using logistic regression, we examined the association between TDR and year of seroconversion, adjusting for confounders. Of 4717 individuals seroconverting between 1996 and 2012, median (IQR) age at seroconversion was 33 (27, 39) years. The majority (3839; 92%) were male, mainly exposed through MSM (3767; 80%), and infected with subtype B (3464; 73%). Overall, 515 (11%) individuals had at least one drug resistance-related mutation; 280 individuals with nucleoside reverse transcriptase, 185 with nonnucleoside reverse transcriptase, and 144 with protease inhibitor mutations. Estimated TDR prevalence was 19.4% (8.2, 36.0) in 1996, significantly decreasing to 8.5% (5.9, 11.9) in 2012 [odds ratio (OR; 95% confidence interval (CI)) = 0.92 (0.90, 0.95) per year increase]. Individuals exposed through sex between men and women were significantly less likely to have been infected with a drug-resistant strain [OR (95% CI) = 0.59 (0.41, 0.87) compared with MSM], and there was marginal evidence that sampling during acute infection was associated with higher odds of resistance [OR (95% CI) = 1.20 (0.97, 1.7), P = 0.093] compared with later sampling. TDR has decreased over calendar time although a significant proportion of new infections still carry resistance-related mutations.


The dynamic influence of photoisomerization on optical reorientation in absorbing isotropic liquid crystals.
The induced transient dichroism and the transient optical Kerr effect (OKE) response of azo-doped isotropic liquid crystals were studied in picosecond time scale. Photoisomerization occurs when the azo-dye molecules are resonantly excited. As a result, a significant enhancement of OKE signal was found in the dye doped liquid crystals with respect to the pure liquid crystals. The enhancement was attributed to the extra torque exerted on liquid crystals after anisotropic orientational distribution of trans and cis isomers were generated. A modified mean field theory model was applied to treat the interactions of trans and cis isomers with liquid crystals. A small-pump enhancement factor of 3.8 was obtained, which is comparable with that of the well-known anthraquinone dye doped LC system.


Evidence that type I osteoporosis results from enhanced responsiveness of bone to estrogen deficiency.
Type I osteoporosis occurs within 20 years after menopause and is associated with excessive cancellous bone loss and fractures of the vertebrae and distal radius. We have suggested that it may be caused by estrogen deficiency plus some additional factor predisposing to excessive bone loss. One such factor might be a greater degree of sex steroid deficiency, a possibility that could not be previously excluded because assays of sufficient sensitivity have only recently become available. Thus, we studied 36 women with vertebral fractures due to typical high turnover type I postmenopausal osteoporosis and 36 normal postmenopausal women using new ultrasensitive assays with detection limits of 1 pg/ml for estradiol, 5 pg/ml for estrone and 5 ng/dl for testosterone to test if type I osteoporosis results from enhanced responsiveness of bone to estrogen deficiency. Mean levels of serum sex steroids were identical in both groups, but bone turnover was increased by up to 55% in the women with type I osteoporosis. Moreover, compared with controls, the osteoporotic women had a 51% higher ( P<0.01) serum osteoprotegerin level, which was likely a compensatory response to the increased bone turnover. In the osteoporotic women, 1-year treatment with transdermal estrogen in 14 women reduced total deoxypyridinoline, an index of bone resorption, by 58% as compared with placebo treatment in 17 women ( P<0.001). Thus, as compared to controls, postmenopausal osteoporotic women had comparable sex steroid levels but higher bone turnover levels that were responsive to estrogen therapy. This is consistent with the hypothesis that the greater bone loss in type I osteoporosis is the result of impaired responsiveness of bone to low postmenopausal levels of sex steroids.


Polycystic renal disease: a new experimental model.
A single injection of any one of several long-acting adrenal corticosteroids at birth induces progressive cystic changes in nephrons which develop in the subcapsular zone of the rabbit kidney until 10 days after birth. These cystic lesions enlarge progressively and become visible within a few days after birth. When the animal is 2 weeks old, renal size has become three times as large as that of uninjected littermates. Adrenal corticosteroids prolong the duration of nephrogenic activity in the renal cortex. The cysts are blind and represent dilatation of the developing end of the collecting ducts. When the steroid-induced hypokalemia is prevented with repeated potassium chloride injections, renal cystic disease is almost completely prevented. Certain long-acting steroids induce cystic renal changes without systemic signs of toxicity.


The effect of skin cleansing agents on ethanol results measured with the Du Pont automatic clinical analyzer.
The effect of various prepackaged skin cleansers on the results of serum ethanol analyses performed with the Du Pont automatic clinical analyzer has not been previously reported. When added directly to serum in concentrations of either 0.625 or 1.56% (v/v), neither polyvinyl-pyrrolidone iodine nor benzalkonium chloride affected the ethanol results. The cross-reactivity of isopropanol with the automatic clinical analyzer ethanol procedure was 3.9%. The greatest interference in the measured ethanol concentration was from the addition of green soap tincture, which contained 30% ethanol. The effect of improper phlebotomy technique on ethanol measurements was also investigated by performing venipunctures through a pool of 100% ethanol on the skin. No ethanol was detected in these samples unless an ethanol-soaked sponge was pressed over the venipuncture site while the needle was withdrawn from the skin. When correct phlebotomy technique is used, skin cleansing agents should not affect the results of ethanol measurements determined with the Du Pont automatic clinical analyzer.


Peptide hormone analysis in diagnosis and treatment of Differences of Sex Development: joint position paper of EU COST Action 'DSDnet' and European Reference Network on Rare Endocrine Conditions.
Differences of Sex Development (DSD) comprise a variety of congenital conditions characterized by atypical chromosomal, gonadal, or anatomical sex. Diagnosis and monitoring of treatment of patients suspected of DSD conditions include clinical examination, measurement of peptide and steroid hormones, and genetic analysis. This position paper on peptide hormone analyses in the diagnosis and control of patients with DSD was jointly prepared by specialists in the field of DSD and/or peptide hormone analysis from the European Cooperation in Science and Technology (COST) Action DSDnet (BM1303) and the European Reference Network on rare Endocrine Conditions (Endo-ERN). The goal of this position paper on peptide hormone analysis was to establish laboratory guidelines that may contribute to improve optimal diagnosis and treatment control of DSD. The essential peptide hormones used in the management of patients with DSD conditions are follicle-stimulating hormone, luteinising hormone, anti-Müllerian hormone, and Inhibin B. In this context, the following position statements have been proposed: serum and plasma are the preferred matrices; the peptide hormones can all be measured by immunoassay, while use of LC-MS/MS technology has yet to be implemented in a diagnostic setting; sex- and age-related reference values are mandatory in the evaluation of these hormones; and except for Inhibin B, external quality assurance programs are widely available.


Harnessing PROTAC technology to combat stress hormone receptor activation.
Counteracting the overactivation of glucocorticoid receptors (GR) is an important therapeutic goal in stress-related psychiatry and beyond. The only clinically approved GR antagonist lacks selectivity and induces unwanted side effects. To complement existing tools of small-molecule-based inhibitors, we present a highly potent, catalytically-driven GR degrader, KH-103, based on proteolysis-targeting chimera technology. This selective degrader enables immediate and reversible GR depletion that is independent of genetic manipulation and circumvents transcriptional adaptations to inhibition. KH-103 achieves passive inhibition, preventing agonistic induction of gene expression, and significantly averts the GR's genomic effects compared to two currently available inhibitors. Application in primary-neuron cultures revealed the dependency of a glucocorticoid-induced increase in spontaneous calcium activity on GR. Finally, we present a proof of concept for application in vivo. KH-103 opens opportunities for a more lucid interpretation of GR functions with translational potential.


Autophagy and cell death signaling following dietary sulforaphane act independently of each other and require oxidative stress in pancreatic cancer.
The broccoli isothiocyanate, sulforaphane (SFN), was recently identified as being capable of eliminating highly therapy-resistant pancreatic carcinoma (PC) cells without inducing toxic side effects. While SFN has been shown to stimulate autophagy or 'self-eating', it is unclear whether this catabolic process is a pro- or anti-tumorigenic response. To investigate the role of autophagy in SFN-induced cell death, established PC cell lines were treated with SFN, and the induction of autophagy was evaluated by detecting the abundance of autophagic vesicles by electron microscopy, the increase in converted LC3-II by Western blot analysis and the autophagosome puncta of GFP-LC3 by immunofluorescence. SFN-induced autophagy was suppressed by the autophagy inhibitor chloroquine, while the autophagy inducer rapamycin did not further enhance autophagy in PC cells. Importantly, neither modulator altered SFN cytotoxicity, suggesting that SFN-induced autophagy and cell death act independently of each other. In contrast, the antioxidant N-acetyl-cysteine sustained cell viability and prevented autophagy induction after SFN exposure, indicating that both signaling pathways depend on reactive oxygen species (ROS). Our studies provide a valuable new mechanistic insight into the SFN-induced elimination of PC cells and suggest that an SFN-enriched diet potentially enhances ROS-releasing chemotherapeutic agents.


Diagnosis and management of major trauma.
A regional trauma system involving trained hospital teams as well as physician-manned ambulances and helicopters has been established in Norway. Emphasis is on written guidelines and the continuous evaluation of the results. A grading system of injury severity is used. Despite major advances in the initial treatment of shock and major trauma, posttraumatic pulmonary insufficiency still represents a problem. Our results indicate a beneficial effect of the early prophylactic use of high-dose corticosteroids in major chest trauma. The use of three doses of methylprednisolone was not associated with any increased risk of infection.


Affine-similar shape retrieval: application to multiview 3-D object recognition.
The feasibility of representing a three-dimensional (3-D) object with a small number of standard views is studied. The object boundary of each view is considered as a two dimensional (2-D) shape and is represented by the locations of the maxima of its curvature scale space (CSS) image contours. The idea is to identify an unknown object from an image taken from a random view by using the stored descriptions of the standard views. The CSS image has been selected for MPEG-7 standardization. The maxima of CSS image have already been used to represent 2-D shapes in different applications under similarity transforms. Since the new application involves affine transforms, we first examine the effects of general affine transforms on the representation and show that the locations of the maxima of the CSS image do not move dramatically even under large affine transformations. Our system for shape-based retrieval from large image databases is then applied to multiview 3-D object representation and recognition. Our collection of 3-D objects consists of 18 aircraft of different shapes. Three silhouette contours corresponding to random views are separately used as input for each object. Results indicate that robust and efficient 3-D free-form object recognition through multiview representation can be achieved using the CSS representation.


Hepatic HIF-2 regulates erythropoietic responses to hypoxia in renal anemia.
The kidney is the main physiologic source of erythropoietin (EPO) in the adult and responds to decreases in tissue oxygenation with increased EPO production. Although studies in mice with liver-specific or global gene inactivation have shown that hypoxia-inducible factor 2 (Hif-2) plays a major role in the regulation of Epo during infancy and in the adult, respectively, the contribution of renal HIF-2 signaling to systemic EPO homeostasis and the role of extrarenal HIF-2 in erythropoiesis, in the absence of kidney EPO, have not been examined directly. Here, we used Cre-loxP recombination to ablate Hif-2α in the kidney, whereas Hif-2-mediated hypoxia responses in the liver and other Epo-producing tissues remained intact. We found that the hypoxic induction of renal Epo is completely Hif-2 dependent and that, in the absence of renal Hif-2, hepatic Hif-2 takes over as the main regulator of serum Epo levels. Furthermore, we provide evidence that hepatocyte-derived Hif-2 is involved in the regulation of iron metabolism genes, supporting a role for HIF-2 in the coordination of EPO synthesis with iron homeostasis.


An experimental study on the effect of temperature on piezoelectric sensors for impedance-based structural health monitoring.
The electromechanical impedance (EMI) technique is considered to be one of the most promising methods for developing structural health monitoring (SHM) systems. This technique is simple to implement and uses small and inexpensive piezoelectric sensors. However, practical problems have hindered its application to real-world structures, and temperature effects have been cited in the literature as critical problems. In this paper, we present an experimental study of the effect of temperature on the electrical impedance of the piezoelectric sensors used in the EMI technique. We used 5H PZT (lead zirconate titanate) ceramic sensors, which are commonly used in the EMI technique. The experimental results showed that the temperature effects were strongly frequency-dependent, which may motivate future research in the SHM field.


Detection of enzymatically generated hydrogen peroxide by metal-based fluorescent probe.
We developed a metal-based fluorescent probe for H(2)O(2) called MBFh1, which has an iron complex as a reaction site for H(2)O(2) and a 3,7-dihydroxyphenoxazine derivative as the fluorescent reporter unit. The iron complex reacts quickly with H(2)O(2) to form oxidants, and then the oxidants convert the closely appended nonfluorescent 3,7-dihydroxyphenoxazine moiety to resorufin in an intramolecular fashion. The quick response to H(2)O(2) allows us to plot the enzymatic evolution of H(2)O(2). A combination of N-acetyl-3,7-dihydroxyphenoxazine and horseradish peroxidase has been frequently used to detect enzymatically generated H(2)O(2), but this method has interference with phenol derivatives. The use of MBFh1 overcomes this drawback.


Associations between Adherence to the Taiwan Dietary Reference Intakes of Micronutrients and the Risk of Type 2 Diabetes.
The dietary reference intakes (DRIs) were established as guidance for the intake of micronutrients and other nutrients. However, how DRIs affect disease status has not been thoroughly examined. The aim of this study is to examine the associations between adherence to the DRIs of selected micronutrients and type 2 diabetes. A populational and cross-sectional study was conducted using data from the Nutrition and Health Survey in Taiwan (NAHSIT) 2013-2016. A total of 14 nutrient intakes, including those for vitamin A, C, D, E, B1, B2, B3, B6, B12, iron, magnesium, zinc, calcium, and phosphorus, were evaluated using the 24 h dietary recall method. Type 2 diabetes was defined as a fasting serum HbA1c level of 6.5% or higher and excluded participants who had received treatments for type 2 diabetes or reported a diagnosis of type 2 diabetes by physicians to avoid the possible inverse association. A total of 2685 participants aged 19 and above were included in the final analysis. After adjusting for confounders, we found that adherence to the DRI of vitamin A was associated with a reduced risk of type 2 diabetes among men. The odds ratio (OR) was 0.64 (95% confidence interval (CI) = 0.42-0.99) compared with men who did not adhere to the DRI. As for women, we found that adherence to the DRI of vitamin B1 was associated with a reduced risk of type 2 diabetes. The OR was 0.59 (95% CI = 0.35-0.97) compared with that for women who did not adhere to the DRI. This study showed that adherence to Taiwan DRIs of vitamin A for men and vitamin B1 for women might have beneficial effects on type 2 diabetes.


Chemical excitation of Limulus photoreceptors. II. Vanadate, GTP-gamma-S, and fluoride prolong excitation evoked by dim flashes of light.
We treated Limulus ventral photoreceptors with the phosphatase inhibitors fluoride, vanadate, and GTP-gamma-S [guanosine-5'0-(3-thiotriphosphate)] under various conditions of illumination and external calcium concentrations. In the dark in low-calcium (1 mM) artificial seawater (ASW), fluoride-induced discrete waves cluster together in time. Under these conditions, the intervals between waves were found to be correlated, and there were excess short intervals beyond the number expected from an exponential interval distribution. To assess the effects of the inhibitors on the light response, we stimulated ventral receptors with a series of dim flashes and averaged the current response under voltage clamp. In ASW, vanadate and GTP-gamma-S prolong the decay of the averaged response to dim test flashes, but prolongation does not always accompany the induction of discrete waves in the dark. Prolongation induced by vanadate in normal-calcium (10 mM) ASW was enhanced in low-calcium (1 mM Ca2+) ASW. Many individual response records suggest that prolongation results from extra discrete waves late in the light response, whereas others reveal long-lasting complex waveforms that cannot easily be resolved into discrete waves. The apparent effect of the inhibitors on the light response is to allow a single photoactivated rhodopsin molecule to produce multiple discrete waves and complex long-lasting events. We suggest that both prolongation of the light response and clustering of waves in the dark result from inhibition of a step in the pathway of visual transduction, in which GTP hydrolysis normally helps to turn off the production of both light-evoked and spontaneous waves.


Processed meat and colorectal cancer: a quantitative review of prospective epidemiologic studies.
A tremendous amount of scientific interest has been generated regarding processed meat consumption and cancer risk. Therefore, to estimate the association between processed meat intake and colorectal cancer (CRC), a meta-analysis of prospective studies was conducted. Twenty-eight prospective studies of processed meat and CRC were identified, of which 20 represented independent nonoverlapping study populations. Summary relative risk estimates (SRREs) for high versus low intake and dose-response relationships were calculated. The SRRE for high (vs. low) processed meat intake and CRC was 1.16 [95% confidence interval (CI): 1.10-1.23] for all studies. Summary associations were modified considerably by sex; the SRRE for men was 1.23 (95% CI: 1.07-1.42) and the SRRE for women was 1.05 (95% CI: 0.94-1.16), based on nine and 13 studies, respectively. Sensitivity analyses did not indicate appreciable statistical variation by tumor site, processed meat groups, or study location. The SRRE for each 30-gram increment of processed meat and CRC was 1.10 (95% CI: 1.05-1.15) based on nine studies, and the SRRE for each incremental serving of processed meat per week was 1.03 (95% CI: 1.01-1.05) based on six studies. Overall, summary associations were weak in magnitude (i.e. most less than 1.20), processed meat definitions and analytical comparisons were highly variable across studies, and isolating the independent effects of processed meat intake is difficult, given the likely influence of confounding by other dietary and lifestyle factors. Therefore, the currently available epidemiologic evidence is not sufficient to support a clear and unequivocal independent positive association between processed meat consumption and CRC.


Clinical research on acute hemoptysis treated with kexue ning.
The efficacy of Kexue Ning (stopping hemoptysis) in the treatment of acute hemoptysis, and its comparison with that of the control group (treated with western medicine) are reported in this paper. The results showed that both the time for cessation of hemoptysis and the cure rate were better in the therapeutic group than in the control group. In addition, marked improvement in blood rheology after treatment was also found in the therapeutic group, but not in the control group.


Disparities in obesity and related conditions among Americans with disabilities.
Despite representing nearly 20% of the U.S. population, individuals with disabilities are invisible in obesity surveillance and intervention efforts. The current study (1) compares obesity and extreme obesity prevalence between Americans with and without disabilities and (2) examines the association between BMI category and weight-related chronic disease risk factors in both groups. In 2012, six waves of data from the National Health and Nutrition Examination Survey (NHANES, 1999-2010) were pooled to compare the prevalence of obesity and extreme obesity between adults (aged ≥20 years, N=31,990) with disabilities (n=11,556) versus without disabilities (n= 20,434). Chronic disease risk factors (blood pressure, lipids, C-reactive protein [CRP], glucose) were compared across weight categories, by disability severity, and disability status. Obesity (41.6%) and extreme obesity (9.3%) prevalence among those with disabilities were significantly higher than they were among those without disabilities (29.2% and 3.9%, respectively). Disability severity and disability status negatively affected nearly all chronic disease risk factors. Additionally, there was a disability-by-weight interaction: people with disabilities at all weight categories were significantly more likely to report being told they had hypertension, high cholesterol, or diabetes and to have been prescribed antihypertensive and lipid-lowering medications. The prevalence of obesity (41.6%) and extreme obesity (9.3%) found in individuals with disabilities is high. When compared to obese adults without disabilities, obese adults with disabilities are more likely to have diabetes, high cholesterol, hypertension, and higher CRP. Thus, the study provides convincing evidence of obesity-related health disparities between Americans with and without disabilities.


A Wzz (Cld) protein determines the chain length of K lipopolysaccharide in Escherichia coli O8 and O9 strains.
The modal distribution of O-antigen chain length is determined by the Wzz (Cld/Rol) protein in those cases in which it has been studied. The system of O-antigen synthesis in Escherichia coli serotypes O8 and O9 is different from that reported for most other bacteria, and chain length distribution is thought not to be determined by a Wzz protein. We report the existence in E. coli O8 and O9 strains of wzz genes which are very similar to and have sequences within the range of variation of those which determine the chain length of typical O antigens. We also find that wzz genes previously identified by their effect on O-antigen chain length, when cloned and transferred to O8 and O9 strains, affect the chain length of a capsule-related form of LPS, K(LPS). We conclude that in at least some O8 and O9 strains there is a wzz gene which controls the chain length of K(LPS) but has no effect on the O8 or O9 antigen.


The anxiolytic effects of a Valerian extract is based on valerenic acid.
Valerian is commonly used for the treatment of insomnia and anxiety. Valerian extracts allosterically modulate GABA-A receptors and induced an anxiolytic activity. This activity is closely related to valerenic acid. In the present experiments it was investigated whether acetoxy valerenic acid may interfere with the anxiolytic action of valerenic acid. Situational anxiety was measured using male CD-1 mice in the elevated plus maze test after oral administration of the test substances. In addition the body core temperature was measured. For the 3H-GABA binding assay dissected tissue from frontal cortex of male RjHan:WI rats were used. Statistical evaluation was performed by means of the non-parametric Kruskal-Wallies H-test, followed by the two-tailed Mann-Whitney U-test. Adding of acetoxy valerenic acid abolished the anxiolytic action of valerenic acid. There was no effect on body core temperature. Moreover, the valerian extract did not show any affinity to benzodiazepine binding sites. The determining compound for the observed anxiolytic effect of the valerian extract is its content of valerenic acid.


Nanofiber-structured hydrogel yarns with pH-response capacity and cardiomyocyte-drivability for bio-microactuator application.
Polymeric hydrogels have great potential in soft biological micro-actuator applications. However, inappropriate micro-architecture, non-anisotropy, weak biomechanics, and inferior response behaviors limit their development. In this study, we designed and manufactured novel polyacrylonitrile (PAN)-based hydrogel yarns composed with uniaxially aligned nanofibers. The nanofibrous hydrogel yarns possessed anisotropic architecture and robust mechanical properties with flexibility, and could be assembled into defined scaffold structures by subsequent processes. The as-prepared hydrogel yarns showed excellent pH response behaviors, with around 100% maximum length and 900% maximum diameter changes, and the pH response was completed within several seconds. Moreover, the hydrogel yarns displayed unique cell-responsive abilities to promote the cell adhesion, proliferation, and smooth muscle differentiation of human adipose derived mesenchymal stem cells (HADMSC). Chicken cardiomyocytes were further seeded onto our nanofibrous hydrogel yarns to engineer living cell-based microactuators. Our results demonstrated that the uniaxially aligned nanofibrous networks within the hydrogel yarns were the key characteristics leading to the anisotropic organization of cardiac cells, and improved sarcomere organization, mimicking the cardiomyocyte bundles in the native myocardium. The construct is capable of sustaining spontaneous cardiomyocyte pumping behaviors for 7days. Our PAN-based nanofibrous hydrogel yarns are attractive for creating linear microactuators with pH-response capacity and biological microactuators with cardiomyocyte-drivability. A mechanically robust polyacrylonitrile-based nanofibrous hydrogel yarn is fabricated by using a modified electrospinning setup in combination with chemical modification processes. The as-prepared hydrogel yarn possesses a uniaxially aligned nanofiber microarchitecture and supports a rapid, pH-dependent expansion/contraction response within a few seconds. Embryonic cardiomyocytes-seeded hydrogel yarn improves the sarcomere organization and mimics the cardiomyocyte bundles in the native myocardium, which sustains spontaneous cardiomyocyte pumping behaviors. The nanofibrous hydrogel yarn has several advantages over traditional bulk hydrogel scaffolds in terms of robust biomechanics, anisotropic aligned architecture, and superior pH response behaviors. Our nanofibrous hydrogel yarn holds the potential to be developed into novel linear and biological microactuators for various biomedical applications.


Pharmacological and non-pharmacological interventions to improve cognitive dysfunction and functional ability in clinical depression--a systematic review.
Cognitive dysfunction is of clinical significance and exerts longstanding implication on patients׳ function. Pharmacological and non-pharmacological treatments of cognitive dysfunction are emerging. This review evaluates pharmacological and non-pharmacological treatments of cognitive impairment primarily in the domains of memory, attention, processing speed and executive function in clinical depression. A total of 35 studies were retrieved from Pubmed, PsycInfo and Scopus after applying inclusion and exclusion criteria. Results show that various classes of antidepressants exert improving effects on cognitive function across several cognitive domains. Specifically, studies suggest that SSRIs, the SSRE tianeptine, the SNRI duloxetine, vortioxetine and other antidepressants such as bupropion and moclobemide may exert certain improving effects on cognitive function in depression, such as in learning and memory and executive function. Class-specific cognitive domains or specific dose-response relationships were not identified yet. The few non-pharmacological studies conducted employing cognitive orientated treatments and cognitive remediation therapy show promising results for the improvement of cognitive impairment in depression. However, several methodological constraints of studies limit generalizability of the results and caution the interpretation. Future direction should consider the development of a neuropsychological consensus cognitive battery to support the discovery, clinical assessment, comparison of studies and registration of new agents in clinical depression.


Implementation and sustainability of an enhanced recovery pathway in pediatric bladder reconstruction: Flexibility, commitment, teamwork.
Although enhanced recovery pathways (ERP) provide a safe and effective way to improve the recovery of children undergoing bladder reconstruction, ERPs have not been widely adopted in pediatric urology. We describe a quality improvement initiative and outcomes after implementing a 24-element ERP at a single, freestanding children's hospital. Multiple stakeholder meetings were planned and executed, initially with pediatric practitioners with ERP experience to understand potential implementation barriers then with anesthesiologists, nurses, case managers, and other ancillary staff to draft our institution-specific ERP. A standardized order set was generated to improve ERP adherence. ERP adherence audits and cyclic performance evaluations held every 6-9 months facilitated continuous pathway refinement. Patient outcomes were compared with a pre-ERP historic cohort. Time from initial ERP planning to first implementation was 7 months. ERP was implemented in twenty consecutive patients undergoing bladder reconstruction (median age 11.3 years, range 4.1-21.1) who were compared to twenty consecutive pre-ERP patients (median age 11.4 years, range 7.7-25.1). Median post-operative length of stay (LOS) significantly decreased from 9 days (range 2-31) pre-ERP to 4 days (range 3-29) post-ERP (p < 0.05). A median of 16 (range 12-19) of 24 institutional pathway elements were implemented for each patient. Balancing measures showed no significant increases in highest Clavien complication grade, readmission rate, or unplanned return to the operating room within 30 post-operative days. Implementation of ERP is feasible but requires commitment from multi-disciplinary stakeholders. While we were unable to consistently achieve 80% of the elements, we successfully implemented the pathway and improved our patients' recovery processes (indirectly reflected by a decreased post-operative LOS) with adherence to a median of 67% of elements. Our implementation and effectiveness results are specific to our center and may not be generalizable. However, our experience may offer some insight for others interested in ERP implementation and encourage initiation of their own institutional pathways. Successful ERP implementation at our hospital for children undergoing bladder reconstruction was facilitated by open communication, early stakeholder involvement, and monitoring ERP adherence. ERP implementation significantly decreased LOS without increasing post-operative complications and readmissions (Summary figure).


Three-dimensional mapping of ozone-induced injury in the nasal airways of monkeys using magnetic resonance imaging and morphometric techniques.
Age-related changes in gross and microscopic structure of the nasal cavity may alter local tissue susceptibility as well as the dose of inhaled toxicant delivered to susceptible sites. This article describes a novel method for the use of magnetic resonance imaging, 3-dimensional airway modeling, and morphometric techniques to characterize the distribution and magnitude of ozone-induced nasal injury in infant monkeys. Using this method, we generated age-specific, 3-dimensional, epithelial maps of the nasal airways of infant Rhesus macaques. The principal nasal lesions observed in this primate model of ozone-induced nasal toxicology were neutrophilic rhinitis, along with necrosis and exfoliation of the epithelium lining the anterior maxilloturbinate. These lesions, induced by acute or cyclic (episodic) exposures, were examined by light microscopy, quantified by morphometric techniques, and mapped on 3-dimensional models of the nasal airways. Here, we describe the histopathologic, imaging, and computational biology methods developed to precisely characterize, localize, quantify, and map these nasal lesions. By combining these techniques, the location and severity of the nasal epithelial injury were correlated with epithelial type, nasal airway geometry, and local biochemical and molecular changes on an individual animal basis. These correlations are critical for accurate predictive modeling of exposure-dose-response relationships in the nasal airways, and subsequent extrapolation of nasal findings in animals to humans for determining risk.


Phenomenological Study of Women's Experiences of Neonatal Transport After Childbirth in Korea.
To explore the experiences of women in Korea who were separated from their newborns when they were transported to neonatal intensive care units (NICUs) to receive treatment. Phenomenological. A university hospital with approximately 600 beds in Seoul, Korea. Women who experienced neonatal transport after childbirth from March to May 2021 (N = 9). We collected data through individual in-depth interviews and analyzed them using Colaizzi's procedure. We extracted four overarching themes that represented the experiences of participants: Outsider Left Alone, Enduring in a Different World, The Lost Starting Line, and Running Together. Our findings captured the unique experiences of women whose newborns were transported from the hospitals where they were born to NICUs at other hospitals to receive treatment. It is necessary to develop and apply tailored nursing interventions, such as assessment and support for postpartum blues or depression, to ensure that postnatal care and healthy maternal transition are not hindered.


Receptor subtypes mediating spinal cardiovascular effects of angiotensin II in rat using losartan and PD 123319.
It has previously been shown in this laboratory that intrathecal administration of 10 microg of angiotensin II produces an increase in arterial pressure and heart rate. As two receptor subtypes of angiotensin II, termed AT1 and AT2, have been identified in central nervous tissue this study examines the effects of selective antagonists on the pressor and cardioacceleratory responses to intrathecal administration of 10 microg of angiotensin II to the ninth thoracic spinal cord. The two non-peptide antagonists were losartan (2-n-butyl-4-chloro-5-hydroxymethyl-1-[(2'-(1H)-tetrazol-5-yl)biph enyl-4-yl)methyl]imidazole), which is selective for the angiotensin AT1 receptor, and PD 123319 (1-[[4-(dimethylamino)-3-methylphenyl]methyl]-5-(diphenyacetyl)-4, 5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-6-carboxylic acid, ditrifluoroacetate, dihydrate), which is selective for the angiotensin AT2 receptor. Intravenous administration of losartan blocked both pressor and cardioacceleratory effects of angiotensin II. Intrathecal administration of losartan blocked only the pressor effects, raising the possibility that block of the heart rate response was in the periphery. Intrathecal administration of PD 123319 blocked the pressor effect of angiotensin II but had no effect on the cardioacceleratory response. However, by itself the antagonist produced a transient increase in arterial pressure and a slower increase in heart rate. The data support the involvement of the angiotensin AT1 receptor in mediating the effects of exogenously administered angiotensin II but also indicate a possible role of angiotensin AT2 receptors at the spinal level.


Aneurysmal subarachnoid hemorrhage. Complications and outcome in a hospital population.
We describe total management results in a prospective series of 264 patients with aneurysmal subarachnoid hemorrhage (SAH) admitted to a neurologic unit. Referrals were actively solicited from general physicians as well as neurologists in the area. The diagnosis was based on computed tomography (CT). Secondary deteriorations were analyzed clinically and with serial CT scanning. Outcome was assessed 3 months after SAH. There were no relevant differences between the analyses of patients with and without an aneurysm confirmed by angiography or autopsy. One third of the patients either died within 1 day (12%) or remained in a poor clinical condition that precluded surgery (22%). One third were in good clinical condition, but contraindications to surgery were judged present or serious complications occurred before surgery could be performed. One third of all patients underwent surgery. Overall mortality in our series was 52%; only 26% made a good recovery. The risk of rebleeding was not related to the patients' initial clinical condition, but all other intracranial complications occurred significantly more often in patients graded poor compared with patients in good clinical condition. The most important causes of death and severe disability were hemorrhage (16%), recurrent hemorrhage (18%), and delayed cerebral ischemia (15%). The most important surgical complication was delayed deterioration caused by ischemia (20% of operated patients). We estimated that recognition of 'warning leaks,' surgery in patients over 65, and improvement of our surgical technique could decrease mortality from 52% to approximately 41%.


Agonist-dependent phosphorylation of the mouse delta-opioid receptor: involvement of G protein-coupled receptor kinases but not protein kinase C.
With chronic opiate use, opioid receptor desensitization may be one of the important mechanisms underlying the development of opiate tolerance and addiction. Opioid receptors belong to the G protein-coupled receptor superfamily. In this study, the mouse delta-opioid receptor (delta OR) was used in a model system to investigate the role of opioid receptor phosphorylation in receptor desensitization. When expressed in 293 cells and exposed to agonist, the delta OR underwent receptor-specific desensitization within 10 min. This agonist-induced desensitization corresponded temporally to a 3-fold increase in receptor phosphorylation. Phorbol ester, but not forskolin, also stimulated phosphorylation of the delta OR in 293 cells. Although down-regulation of protein kinase C failed to affect agonist-induced receptor phosphorylation, it abolished phorbol ester-induced receptor phosphorylation. Agonist-induced delta OR phosphorylation must therefore involve kinases other than protein kinase C. Whereas overexpression of a dominant negative mutant (K220R) of beta-adrenergic receptor kinase-1 (beta ARK1) in 293 cells significantly reduced agonist-dependent phosphorylation of the delta OR, overexpression of beta ARK1 or G protein-coupled receptor kinase-5 significantly enhanced this phosphorylation. Concordantly, beta ARK1-K220R overexpression reduced agonist-dependent delta OR desensitization, whereas beta ARK1 overexpression enhanced this densensitization. We conclude that short term desensitization of the delta OR involves phosphorylation of the receptor by one or more G protein-coupled receptor kinases.


Endoscopic microsurgical excision of internal laryngocele.
Internal laryngoceles are relatively rare, and a variety of surgical treatments have been proposed. A case is presented which was treated by endoscopic microsurgical excision.


Neophobia and learning mechanisms: how captive orangutans discover medicinal plants.
Great apes sometimes feed on items of low nutritional value with bioactive secondary compounds. These molecules may be toxic and neophobia is presumed to be an essential factor in avoiding the ingestion of noxious items. The aim of this study is to investigate, in captive orangutans, individual and social learning involved in the discovery and ingestion of new items. We presented novel aromatic plants - 11 fresh plants and 4 infused plants - to 4 captive weaned Bornean orangutans, both under isolated and group conditions, and recorded their behaviour and interactions between group members. All animals tasted by nibbling or ingested most of the plants presented. Regardless of the experimental condition, individual responses did not vary visibly across the sessions, despite numerous close observations, and food transfers between individuals were observed. Our results suggest that a low level of neophobia and a strong propensity to look to conspecifics for information allow Bornean orangutans to expand their diet after weaning. Our results also provide some evidence that olfaction is a key sense in determining food edibility based on previous experience.


Agouti expression in human adipose tissue: functional consequences and increased expression in type 2 diabetes.
It is well recognized that the agouti/melanocortin system is an important regulator of body weight homeostasis. Given that agouti is expressed in human adipose tissue and that the ectopic expression of agouti in adipose tissue results in moderately obese mice, the link between agouti expression in human adipose tissue and obesity/type 2 diabetes was investigated. Although there was no apparent relationship between agouti mRNA levels and BMI, agouti mRNA levels were significantly elevated in subjects with type 2 diabetes. The regulation of agouti in cultured human adipocytes revealed that insulin did not regulate agouti mRNA, whereas dexamethasone treatment potently increased the levels of agouti mRNA. Experiments with cultured human preadipocytes and with cells obtained from transgenic mice that overexpress agouti demonstrated that melanocortin receptor (MCR) signaling in adipose tissue can regulate both preadipocyte proliferation and differentiation. Taken together, these results reveal that agouti can regulate adipogenesis at several levels and suggest that there are functional consequences of elevated agouti levels in human adipose tissue. The influence of MCR signaling on adipogenesis combined with the well-established role of MCR signaling in the hypothalamus suggest that adipogenesis is coordinately regulated with food intake and energy expenditure.


Patterns of family adaptation to childhood asthma.
This study examined patterns in adaptation among parents with a child who had moderate to severe persistent asthma. Specifically, we were interested in examining the differences in adaptation between mothers and fathers in which it was hypothesized that gender effects would be obtained in patterns of coping. Eighty-four parents participated in the study, representing 37 intact families in which both parents were present and 13 single-parent families. Within intact families, mothers exhibited greater efforts than fathers in coping patterns including strategies to acquire social support outside the family, enhance self-worth, and decrease psychological tensions. When compared to mothers in single-parent families, mothers within intact families had a greater tendency to use coping patterns related to family integration and cooperation. Such findings demonstrate a need for additional support for mothers in their role in caring for the chronically ill child. The implications of these findings for clinical practice are discussed.


[Differential diagnosis of lobar (regional, segmental) extent pulmonary tuberculosis].
Two hundred and fifteen patients with pulmonary involvement of lobar (regional, segmental) extent were examined. The examination procedure included clinical and laboratory tests, X-ray tomographic study, and computed tomography, bronchoscopy with biopsy, cyto- and histological studies. The nosological diagnosis was established in 96.5% of cases. Differential diagnosis was found to be difficult in 3.5% of cases.


Eradication of LIG4-deficient glioblastoma cells by the combination of PARP inhibitor and alkylating agent.
Cancer cells often accumulate spontaneous and treatment-induced DNA damage i.e. potentially lethal DNA double strand breaks (DSBs). Targeting DSB repair mechanisms with specific inhibitors could potentially sensitize cancer cells to the toxic effect of DSBs. Current treatment for glioblastoma includes tumor resection followed by radiotherapy and/or temozolomide (TMZ) - an alkylating agent inducing DNA damage. We hypothesize that combination of PARP inhibitor (PARPi) with TMZ in glioblastoma cells displaying downregulation of DSB repair genes could trigger synthetic lethality. In our study, we observed that PARP inhibitor (BMN673) was able to specifically sensitize DNA ligase 4 (LIG4)-deprived glioblastoma cells to TMZ while normal astrocytes were not affected. LIG4 downregulation resulting in low effectiveness of DNA-PK-mediated non-homologous end-joining (D-NHEJ), which in combination with BMN673 and TMZ resulted in accumulation of lethal DSBs and specific eradication of glioblastoma cells. Restoration of the LIG4 expression caused loss of sensitivity to BMN673+TMZ. In conclusion, PARP inhibitor combined with DNA damage inducing agents can be utilized in patients with glioblastoma displaying defects in D-NHEJ.


Complexity: the organizing principle at the interface of biological (dis)order.
The term complexity means several things to biologists.When qualifying morphological phenotype, on the one hand, it is used to signify the sheer complicatedness of living systems, especially as a result of the multicomponent aspect of biological form. On the other hand, it has been used to represent the intricate nature of the connections between constituents that make up form: a more process-based explanation. In the context of evolutionary arguments, complexity has been defined, in a quantifiable fashion, as the amount of information, an informatic template such as a sequence of nucleotides or amino acids stores about its environment. In this perspective, we begin with a brief review of the history of complexity theory. We then introduce a developmental and an evolutionary understanding of what it means for biological systems to be complex.We propose that the complexity of living systems can be understood through two interdependent structural properties: multiscalarity of interconstituent mechanisms and excitability of the biological materials. The answer to whether a system becomes more or less complex over time depends on the potential for its constituents to interact in novel ways and combinations to give rise to new structures and functions, as well as on the evolution of excitable properties that would facilitate the exploration of interconstituent organization in the context of their microenvironments and macroenvironments.


[Effective dose and dose to the crystalline lens during angiographic procedures].
Interventional radiology presents the highest dose levels for the staff and for the patient. For a total of 34 angiographic examinations the doses at the forehead and at the neck of a radiologist have been measured. The effective dose and the dose to the eye lens have been estimated over a period of one year. Doses to the lens of the eye close to the limit of 150 mSv proposed by the ICRP have been observed. There is an imminent need for the radiologist to wear also lead glasses to avoid formation of cataracts at long term.


Derivation and validation of the Denver Human Immunodeficiency Virus (HIV) risk score for targeted HIV screening.
Targeted screening remains an important approach to human immunodeficiency virus (HIV) testing. The authors aimed to derive and validate an instrument to accurately identify patients at risk for HIV infection, using patient data from a metropolitan sexually transmitted disease clinic in Denver, Colorado (1996-2008). With multivariable logistic regression, they developed a risk score from 48 candidate variables using newly identified HIV infection as the outcome. Validation was performed using an independent population from an urban emergency department in Cincinnati, Ohio. The derivation sample included 92,635 patients; 504 (0.54%) were diagnosed with HIV infection. The validation sample included 22,983 patients; 168 (0.73%) were diagnosed with HIV infection. The final score included age, gender, race/ethnicity, sex with a male, vaginal intercourse, receptive anal intercourse, injection drug use, and past HIV testing, and values ranged from -14 to +81. For persons with scores of <20, 20-29, 30-39, 40-49, and ≥50, HIV prevalences were 0.31% (95% confidence interval (CI): 0.20, 0.45) (n = 27/8,782), 0.41% (95% CI: 0.29, 0.57) (n = 36/8,677), 0.99% (95% CI: 0.63, 1.47) (n = 24/2,431), 1.59% (95% CI: 1.02, 2.36) (n = 24/1,505), and 3.59% (95% CI: 2.73, 4.63) (n = 57/1,588), respectively. The risk score accurately categorizes patients into groups with increasing probabilities of HIV infection.


Circulating microRNA‑135a‑3p in serum extracellular vesicles as a potential biological marker of non‑alcoholic fatty liver disease.
Non‑alcoholic fatty liver disease (NAFLD) is a widespread threat to human health. However, the present screening methods for NAFLD are time‑consuming or invasive. The present study aimed to assess the potential of microRNAs (miRNAs/miRs) in serum extracellular vesicles (EVs) as a biomarker of NAFLD. C57BL/6J mice were fed either a 12‑week high‑fat diet (HFD) or standard chow to establish NAFLD and control groups, respectively. Serum samples were obtained from the mouse model of NAFLD, as well as 50 patients with NAFLD and 50 healthy individuals, and EVs were extracted and verified. Using reverse transcription‑quantitative PCR, the mRNA expression level of selected miRNAs in the serum and EVs was analyzed. In order to determine the diagnostic value, receiver operating characteristic (ROC) curves were used. The mice treated with HFD showed notable hepatic steatosis and higher concentrations of serum alanine aminotransferase (ALT). There was also a significant decrease in the expression levels of miR‑135a‑3p, miR‑129b‑5p and miR‑504‑3p, and an increase in miR‑122‑5p expression levels in circulating EVs in mice treated with HFD and patients with NAFLD. There were also similar miR‑135a‑3p and miR‑122‑5p expression patterns in the serum. ROC analysis demonstrated that miR‑135a‑3p in circulating EVs was highly accurate in diagnosing NAFLD, with the area under the curve value being 0.849 (95% CI, 0.777‑0.921; P<0.0001). Bioinformatics analysis indicated that dysregulated miR‑135a‑3p was associated with 'platelet‑derived growth factor receptor signaling pathway' and 'AMP‑activated protein kinase signaling pathway'. In summary, circulating miR‑135a‑3p in EVs may serve as a potential non‑invasive biomarker to diagnose NAFLD. This miRNA was a more sensitive and specific biological marker for NAFLD compared with ALT.


Low physical activity and energy dense Malaysian foods are associated with non-alcoholic fatty liver disease in centrally obese but not in non-centrally obese patients with diabetes mellitus.
To study the dietary intake and level of physical activity (PA) of patients with diabetes mellitus and the association with non-alcoholic fatty liver disease (NAFLD). Consecutive adult patients with type 2 diabetes mellitus seen in our hospital diabetes clinic were enrolled. The Global Physical Activity Questionnaire and a semi-quantitative food-frequency questionnaire were used to assess PA and dietary intake, respectively. Diagnosis of NAFLD was ultrasound-based and following exclusion of significant alcohol intake and other causes of chronic liver disease. Data for 299 patients were analyzed (mean age 63.3±10.5 years old, 41.1% male). Prevalence of NAFLD was 49.2%. Patients with low PA were more likely to have NAFLD (OR=1.75, 95% CI=1.03-2.99, p=0.029). There was no significant difference in energy intake, intake of macronutrients and percentage energy intake from each macronutrient, high sugar food, high cholesterol food and high SFA food between patients with and without NAFLD. Among centrally obese patients, patients with low PA and in the highest quartile of percentage energy intake from fat (OR=4.03, 95% CI=1.12-15.0, p=0.015), high cholesterol food (OR=3.61, 95% CI=1.37-9.72, p=0.004) and high SFA food (OR=2.67, 95% CI=1.08-6.67, p=0.019) were most likely to have NAFLD. Among those who were not centrally obese, PA and percentage energy intake from fat, high cholesterol food and high SFA food was not associated with NAFLD. Low PA and high percentage energy intake from fat, high cholesterol food and high SFA food is associated with NAFLD in centrally obese but not in non-centrally obese patients with diabetes mellitus.


Adenylate cyclase stimulation alters transport in frog retinal pigment epithelium.
The effect of adenylate cyclase inhibitors and activators on enzyme kinetics and cyclic AMP (cAMP) levels was determined in the bullfrog retinal pigment epithelium (RPE). The RPE enzyme has two Km for ATP: 5.2 X 10(-4) and 4.0 X 10(-5) M, with Vmax of 2.5 and 0.25 nmol X mg protein-1 X min-1. Forskolin, the most potent activator, produced a fourfold increase in enzyme activity and, in the presence of isobutylmethylxanthine (IBMX), an inhibitor of phosphodiesterase, caused a 20-fold increase in cAMP levels. Alloxan, a potent inhibitor, blocked the forskolin-induced activation of this enzyme. In the isolated RPE choroid, forskolin (plus IBMX) produced changes in membrane voltage and resistance that were similar in magnitude but slower in time course than those produced by exogenous cAMP. Like exogenous cAMP, forskolin also decreased steady-state fluid and solute transport in isotonic proportions. Therefore, modulation of RPE adenylate cyclase activity plays an important role in the control of RPE transport.


Doxycycline Added to Prednisolone in Outpatient-Treated Acute Exacerbations of COPD: A Cost-Effectiveness Analysis Alongside a Randomised Controlled Trial.
Most patients with mild to severe chronic obstructive pulmonary disease (COPD) experience exacerbations, which are also associated with increased healthcare costs. Despite limited evidence of antibiotics' benefits for exacerbations in outpatients, antibiotics are frequently prescribed. The aim of this study was to investigate whether doxycycline added to prednisolone is cost-effective compared to placebo plus prednisolone for the treatment of COPD acute exacerbations. An economic evaluation from the societal perspective was performed alongside a 2-year randomised trial in 301 COPD patients in the Netherlands. The primary outcome was cost per quality-adjusted life year (QALY). The secondary outcome was cost per exacerbation prevented. Healthcare utilisation and loss of productivity were measured using retrospective questionnaires and clinical report forms. Missing data were imputed using multiple imputations by chained equations. Bootstrapping was employed to estimate statistical uncertainty surrounding cost-effectiveness outcomes. A sensitivity analysis from the healthcare perspective was performed. On average, costs in the doxycycline group were €898 higher than in the placebo group [95% confidence interval (CI) - 2617 to 4409] for the 2 years of follow-up. QALY values were higher in the doxycycline group (0.03; 95% CI - 0.00 to 0.06), but patients in this group suffered 0.01 more exacerbations than patients in the placebo group (95% CI - 0.14 to 0.11). Cost-effectiveness acceptability curves showed that the probability of doxycycline being cost-effective compared to placebo was 61% and 43% at a willingness-to-pay threshold of €34,000 per QALY and per exacerbation avoided, respectively. The sensitivity analysis showed similar results from the healthcare system perspective. In patients with mild to severe COPD treated for exacerbations in an outpatient setting, doxycycline added to prednisolone is not cost-effective compared to prednisolone plus placebo over a 2-year period.


The effect of cycling on muscle activation in the running leg of an Olympic distance triathlon.
The aim of this study was to determine the effect of prior cycling on EMG activity of selected lower leg muscles during running. Ten elite level triathletes underwent two testing sessions at race pace: a 40 km cycle followed by a 2 km run (CR) and a 10 km run followed by a 2 km run (RR). EMG data from selected lower limb muscles were collected at three sections of each run (0 km, 1 km and 2 km) for six strides using a portable data logger. Significant differences (p < 0.05) between condition were found for the level of activation (Lact) for biceps femoris (BF) during stance and vastus lateralis (VL) during flight and stance. Vastus medialis (VM) changed in Lact, during flight, between sections in the 2 km run. Furthermore, significant differences (p < 0.05) between condition were found for BF during stance and for rectus femoris (RF) and VM during flight. There was a significant difference (p < 0.05) in the duration of VL activation (Dact) across sections of the 2 km run. Findings from this investigation highlight changes in muscle function when changing from cycling to running and indicate a need to train specifically for the cycle to run transition. Such training may improve performance and reduce the risk of injury.


Assessment of the sensitivity, specificity, and reproducibility of the indirect immunofluorescent technique for the diagnosis of amebiasis.
The indirect immunofluorescence (IIF) test was used to study antibody titers to Entamoeba histolytica in various manifestations of clinical amebiasis by comparing the results with parasitological and clinical data from confirmed cases. The sensitivity, specificity, and reproducibility of this test for diagnosis were also studied. A high degree of correlation was found between reactivity in the IIF test and confirmed cases of amebic liver abscess. A positivity rate of 96% in sera of confirmed cases of amebic liver abscess in the present study compares favorably with the results of other workers. The IIF test is very useful in the diagnosis of hepatic amebiasis but is less sensitive for amebic dysentery. The test could not be used to differentiate between hepatic amebiasis and amebic dysentery. The IIF test was insensitive for amebic colitis and of no diagnostic value. Specificity and reproducibility of the test were good, as shown by results obtained with sera from patients with parasitic infections other than amebiasis.


Bilioenterostomy enhances biliary carcinogenesis in hamsters.
The aim of this study was to examine whether the type of bilioenterostomy enhances biliary carcinogenesis in the hamster model. Syrian hamsters were divided into the following groups; simple laparotomy (control group), cholecystoduodenostomy with dissection of the extrahepatic bile duct on the distal end of the common duct (CDDB group) and cholecystoileostomy with dissection of the extrahepatic bile duct on the distal end of the common duct (CIDB group). Following these procedures, all hamsters received N-nitrosobis(2-oxopropyl)amine. The diameter of the extrahepatic bile duct and plasma levels of cholecystokinin (CCK) were measured and the number of neoplastic lesions was counted microscopically. Proliferative effect of the procedures on the biliary epithelium was examined by proliferative cell nuclear antigen. In the CDDB group the extrahepatic bile duct was significantly dilated and carcinogenesis of the gall-bladder and extrahepatic bile ducts was enhanced. In the CIDB group the CCK bioactivity was stimulated and intrahepatic biliary duct, but not gall bladder and extrahepatic bile duct, carcinogenesis was promoted more than that observed in the CDDB group. Proliferation of the biliary duct epithelium was enhanced in both the CDDB and CIDB groups. Cholecystoduodenostomy enhanced intra- and extrahepatic bile duct carcinoma, whereas cholecystoileostomy promoted only intrahepatic bile duct carcinoma. Some factors in the intestinal juice seem to play a role in the promotion of biliary tract carcinoma.


Protective effects of ellagic acid against tetrachloride-induced cirrhosis in mice through the inhibition of reactive oxygen species formation and angiogenesis.
Ellagic acid has been proven to have anticancer, antimutation, antimicrobial and antiviral functions. The present study investigated whether treatment with ellagic acid was able to prevent tetrachloride (CCl4)-induced cirrhosis through the inhibition of reactive oxygen species (ROS) formation and angiogenesis. CCl4 diluted in olive oil at a final concentration of 10% was used to induce a cirrhosis model. A total of 40 mice were random allocated into four groups, as follows: Control, cirrhosis model, 7.5 mg/kg ellagic acid and 15 mg/kg ellagic acid groups. In the control group, mice were given normal saline. The results indicated that ellagic acid exerted a protective effect, evidently preventing CCl4-induced cirrhosis. In addition, treatment with ellagic acid significantly inhibited collagen I and inducible nitric oxide synthase protein expression levels in CCl4-induced cirrhosis mice. Oxidative stress and ROS formation were also significantly reduced by ellagic acid treatment. The protein expression levels of vascular endothelial growth factor (VEGF) and VEGF receptor 2 (VEGFR2), and the caspase-3 activity were significantly inhibited by treatment with ellagic acid. In conclusions, these results suggest that ellagic acid exerted protective effects against CCl4-induced cirrhosis through the inhibition of ROS formation and angiogenesis.


A Feasibility Study of the WHO Digital Mental Health Intervention Step-by-Step to Address Depression Among Chinese Young Adults.
Background: Chinese young adults experience barriers to mental health treatment, including the lack of treatment providers and stigma around treatment seeking. Evidence-based digital mental health interventions are promising and scalable alternatives to face-to-face treatment for this population, but lack rigorous evidence to support scale-up in China. Aim: The study was a feasibility study for a large-scale RCT of Step-by-Step, a behavioral activation-based, mental health intervention to address depression and anxiety symptoms in Chinese young adults. It sought to assess feasibility of recruitment and of delivery of Step-by-Step in a University setting, to assess acceptability of the intervention, and to examine potential effectiveness. Method: An uncontrolled, feasibility trial was conducted to assess the feasibility and acceptability of Chinese Step-by-Step for Chinese University students with elevated depressive symptoms (PHQ-9 scores at or above 10) in Macao, China. Data was collected at two different time points (i.e., baseline and 8-weeks after baseline), administered via questionnaires embedded in an interventional mobile application. Participation rate and dropout rate were measured. Depressive and anxiety symptom severity, well-being, and self-defined stress were assessed. Satisfaction with the program was assessed using qualitative interviews. Results: A total of 173 students were screened, 22.0% (n = 38) were eligible, and 63.2% of them (n = 24) started the intervention. The dropout rate by post-test was 45.8%. Results from completers showed that Step-by-Step was potentially effective in reducing depressive and anxiety symptom severity, and self-defined stress. Students were generally satisfied with the program, but also offered suggestions for continued improvement. Qualitative feedback was reported within the RE-AIM framework, covering recruitment, effectiveness, adoption, implementation, and maintenance. Amendments to the program were made according to the feedback (e.g., adding notification for new session, modify the time zone). Conclusion: A minimally guided Step-by-Step protocol and the study procedure were successfully pilot tested for use for Chinese University students. The intervention was acceptable and no adverse events were reported. The results support the potential effectiveness and feasibility of a large-scale evaluation of the program.


Multiscale movement coordination dynamics in collaborative team problem solving.
During collaborative problem solving (CPS), coordination occurs at different spatial and temporal scales. This multiscale coordination should play a functional role in facilitating effective collaboration. To evaluate this, we conducted a study of computer-based CPS with 42 dyadic teams. We used cross-wavelet coherence to examine movement coordination, extracted from videos, at several scales, and tested whether the observed coordination was greater than expected due to chance and due to task demands. We found that coordination at scales less than 2s was greater than chance and at most scales (except 16s, 1m, and 2m), was greater than expected due to task demands. Lastly, we observed that coherence at .25s and 1s scales was predictive of performance. However, when including relative phase, our results suggest that higher in-phase movement coordination at the 1s scale was the strongest predictor of CPS performance. Further, we used growth curve modeling to examine how movement coordination changes across the duration of the task and whether this is moderated by CPS performance. We found that coordination over the duration of the CPS task is quadratic (a U shape) and that better performing teams have higher coordination with a shallower curve. We discuss these findings and their relevance to understanding how low-level movement coordination facilitates CPS.


Comparison of the Body Composition of Caucasian Young Normal Body Mass Women, Measured in the Follicular Phase, Depending on the Carbohydrate Diet Level.
Background and objectives: Some publications indicate the possibility of the influence of meal nutritional value on results of bioelectrical impedance, and of the relation between the long-term carbohydrate intake and body composition. The aim of the presented study was to evaluate the influence of long-term intake of carbohydrates on body composition results assessed using the bioelectrical impedance of Caucasian young women with normal body mass, who were in the follicular phase of their menstrual cycle. Materials and Methods: Body composition was assessed in 100 women (18⁻30 years), according to strict rules, to minimize the influence of disturbing factors and by using two types of bioelectrical impedance device of the same operator to eliminate the influence of measurement (BIA 101/SC and BIA 101/ASE by Akern Srl, Firenze, Italy with the Bodygram 1.31 software and its equations by Akern Srl, Firenze, Italy). The analysis included validation of reproducibility of body composition assessment (fat, fat-free, body cell and muscle mass, water, extracellular water, and intracellular water content), and comparison of body composition for groups characterized by carbohydrate content <50% (n = 55) and >50% of the energy value of the diet (n = 45). Results: Analysis conducted using Bland⁻Altman method, analysis of correlation, analysis of quartile distribution, and weighted κ statistic revealed a positively validated reproducibility, but extracellular water associations were the weakest. Depending on the device, participants characterized by higher carbohydrate intake had significantly higher intracellular water content (p = 0.0448), or close to significantly higher (p = 0.0851) than those characterized by lower carbohydrate intake, whose extracellular water content was close to significantly lower (p = 0.0638) or did not differ. Conclusions: The long-term, moderately reduced, carbohydrate intake may cause the shift of intracellular water to the extracellular space and, as a result, influence the body composition results.


[Basic directions and prospects for international collaboration in gerontology. Role of the International Association of Gerontology].
The overview presents data on the international collaboration in gerontology in the following spheres: 1) organizational basis of collaboration; 2) coordination of research; 3) coordination in the training of specialists; 4) collaboration in the organization of geriatric services; 5) collaboration in social care for the elderly. Role of the International Association of Gerontology and other international organizations in the promotion of gerontology and geriatrics in each of these spheres is shown by the example of Russia.


[Diagnosis of lesions of the central nervous system in patients with systemic lupus erythematosus].
A total of 60 patients with systemic lupus erythematosus (SLE) were under observation; 36 of them had clinical symptoms of the CNS affection and 25 persons included into the control group exhibited no psychic disorders during the clinical examination. Besides, routine clinico-laboratory examinations accepted in rheumatology, the patients were subjected to cranial computer tomography (CT), electroencephalography, examination of cerebral hemodynamics with a radionuclide partechnetate 99mC as well as to psychological testing. Neuropsychic disorders developed during the first four years after the onset of the disease and are grouped in the following way: neurological, border-line, neuropsychic, affective, psychotic, intellectual-mnestic. Moderate affection of the CNS in SLE is characterized by a complex of subjective and objective symptoms: headache, deterioration of memory, insomnia, vertigo, irritability, depressed mood, assymetry of the face innervation, coordinatory disorders. Diffuse widening of the subarchnoidal space, diffuse cerebral changes, interhemispheric assymetry of the venous and arterial phases of cerebral circulation: the most peculiar symptoms of the CNS affection in SLE according to CT and EEG and radionuclide studies of cerebral hemodynamics. Focal changes in the CNS were observed in 50% of the patients with neuropsychic disorders.


Electrostatic influence on rotational mobilities of sol-gel-encapsulated solutes by NMR and EPR spectroscopies.
The rotational mobilities of small solute molecules encapsulated in tetramethyl orthosilicate (TMOS) sol-gels have been investigated by EPR spectroscopy of encapsulated nitroxide probes and by high-resolution NMR spectroscopic measurements of transferred NOE's (trNOE's), of T(1)'s, and of T(1)'s in the rotating frame (T(1)rho). The two spectroscopic methods are sensitive to motions on different time scales and hence, are nicely complementary. Suites of neutral, positively, and negatively charged nitroxide probes (EPR) and of simple diamagnetic small molecules (NMR) were selected to disclose influences of electrostatic interactions with the sol-gel walls and to probe the presence of multiple populations of molecules in distinct regions of the sol-gel pores. For neutral and negatively charged solute probes, both techniques disclose a single population with a significantly increased average rotational correlation time, which we interpret at least in part as resulting from exchange between free-volume and transiently immobilized surface populations. The electrostatic attraction between cationic probes and the negatively charged sol-gel walls causes the positively charged probes to be more effectively immobilized and/or causes a greater percentage of probes to undergo this transient immobilization. The EPR spectra directly disclose a population of cationic probes which are immobilized on the X-band EPR time scale: tau(c) greater than or approximately equal 10(-7) s. However, NMR measurements of trNOE's and of T(1)rho demonstrate that this population does exchange with the free-volume probes on the slower time scale of NMR. This approach is equally applicable to the study of solutes within other types of confined spaces, as well.


Turbans vs. Helmets: A Systematic Narrative Review of the Literature on Head Injuries and Impact Loci of Cranial Trauma in Several Recreational Outdoor Sports.
When in public, faith-based mandates require practising Sikh men to wear a turban which may not be covered by hats or caps. This makes it impossible for practising Sikhs to wear helmets and other protective headwear, mandatory in many countries and facilities for engagement in recreational pursuits (e.g., skiing) and on adventure outdoor recreation camps mandatorily run for school groups. The result is often social exclusion and ostracisation in the case of school children. Despite studies into the efficacy of protective helmets in some recreational outdoor activity settings, virtually nothing is known about the protective potential of turbans. This paper systematically reviews the extant literature on head injuries in several recreational outdoor activities and sports sectors (aerial, water, winter, wheeled and animal-based sports) and finds that the extant literature is of limited value when trying to understand the spatial distribution of trauma on the cranial surface. As the data do not permit to make inferences on the protective potential of turbans, future systematic, evidence-based epidemiological studies derived from hospital admissions and forensic examinations are required. Failure to do so perpetuates social exclusion and discrimination of religious grounds without an evidentiary basis for defensible public health measures.


Development of an amplitude-modulated Nd:YAG pulsed laser with modulation frequency tunability up to 60 GHz by dual seed injection.
A pulsed, modulation frequency tunable, frequency-doubled Nd:YAG laser has been devised for use in target detection through turbid media. A modulated pulse laser radar system offers many advantages in target detection, such as significant signal contrast enhancement, compared with conventional remote-sensing systems. By implementation of the dual-longitudinal-mode seed injection technique, the modulation frequency of the designed Q-switched laser can be tuned from 250 MHz up to 60 GHz in steps of 250 MHz while maintaining a modulation depth of at least 75%. This provides the ability to explore propagation and scattering properties further at previously unattainable high RF modulation frequencies.


Imaging biological structures with fluorescence photoactivation localization microscopy.
Fluorescence photoactivation localization microscopy (FPALM) images biological structures with subdiffraction-limited resolution. With repeated cycles of activation, readout and bleaching, large numbers of photoactivatable probes can be precisely localized to obtain a map (image) of labeled molecules with an effective resolution of tens of nanometers. FPALM has been applied to a variety of biological imaging applications, including membrane, cytoskeletal and cytosolic proteins in fixed and living cells. Molecular motions can be quantified. FPALM can also be applied to nonbiological samples, which can be labeled with photoactivatable probes. With emphasis on cellular imaging, we describe here the adaptation of a conventional widefield fluorescence microscope for FPALM and present step-by-step procedures to successfully obtain and analyze FPALM images. The fundamentals of this protocol may also be applicable to users of similar imaging techniques that apply localization of photoactivatable probes to achieve super-resolution. Once alignment of the setup has been completed, data acquisitions can be obtained in approximately 1-30 min and analyzed in approximately 0.5-4 h.


Phylogeographic structure, demographic history and morph composition in a colour polymorphic lizard.
In polymorphic species, population divergence in morph composition and frequency has the potential to promote speciation. We assessed the relationship between geographic variation in male throat colour polymorphism and phylogeographic structure in the tawny dragon lizard, Ctenophorus decresii. We identified four genetically distinct lineages, corresponding to two polymorphic lineages in the Northern Flinders Ranges and Southern Flinders Ranges/Olary Ranges regions respectively, and a monomorphic lineage in the Mt Lofty Ranges/Kangaroo Island region. The degree of divergence between these three lineages was consistent with isolation to multiple refugia during Pleistocene glacial cycles, whereas a fourth, deeply divergent (at the interspecific level) and monomorphic lineage was restricted to western New South Wales. The same four morphs occurred in both polymorphic lineages, although populations exhibited considerable variation in the frequency of morphs. By contrast, male throat coloration in the monomorphic lineages differed from each other and from the polymorphic lineages. Our results suggest that colour polymorphism has evolved once in the C. decresii species complex, with subsequent loss of polymorphism in the Mt Lofty Ranges/Kangaroo Island lineage. However, an equally parsimonious scenario, that polymorphism arose independently twice within C. decresii, could not be ruled out. We also detected evidence of a narrow contact zone with limited genotypic admixture between the polymorphic Olary Ranges and monomorphic Mt Lofty Ranges regions, yet no individuals of intermediate colour phenotype. Such genetic divergence and evidence for barriers to gene flow between lineages suggest incipient speciation between populations that differ in morph composition.


Disease modifying therapies for relapsing multiple sclerosis.
Multiple sclerosis (MS) is a common, disabling, putatively autoimmune neurological disease with worldwide distribution. It typically begins as a relapsing disorder that later evolves to a secondary progressive phase. Inflammatory and neurodegenerative mechanisms seem to operate in both phases, but their relative contributions and interactions are incompletely understood. Disease modifying therapies (DMTs) approved for relapsing multiple sclerosis interfere with a variety of immunological mechanisms to reduce rates of relapse, accumulation of disease burden measured by magnetic resonance imaging (MRI), and decline in neurological function over the two to three year duration of typical randomized controlled trials. Benefits of longer duration of therapy on disability are less clear, as data beyond three years are largely limited to observational studies. However, current DMTs do not slow accrual of disability once progressive multiple sclerosis is established. This review summarizes the evidence about the use of approved DMTs and examines how to individualize treatment despite the absence of validated biomarkers to guide drug selection. Methods such as stratifying patients on the basis of estimated risk for future disability, weighing patient specific factors and preferences, and using objective outcomes to adjudicate treatment success are discussed. Emerging drug therapies and strategies are also reviewed.


Multicenter pilot study: safety of automated chest percussion in patients at risk for intracranial hypertension.
In the critical care setting, the focus of care during the first few weeks following acute brain injury is prevention of secondary brain injury by optimizing cerebral perfusion. Ensuring adequate oxygenation and perfusion of cerebral tissues requires attention to all of the body systems. Chest percussion therapy (CPT) promotes pulmonary hygiene and optimizes gas exchange by opening the alveoli. However, many patients with brain injury have intracranial pressure (ICP) monitoring, and conventional wisdom supports limiting activities such as CPT that may stimulate the patient and increase ICP. The purpose of this study was to explore the effects of CPT on ICP. Thirty participants were enrolled over a 6-month period. Data were collected at 1-minute intervals for 1 hour. Each patient was randomized to receive automated CPT (using specialty beds) for 10 minutes, starting at 10, 20, 30, or 40 minutes into the hour. There were no differences in mean ICP values before, during, or after CPT. This study provides evidence that it is safe to perform CPT in patients with ICP monitoring in situ.


Association of Hospice Agency Location and Neighborhood Socioeconomic Disadvantage in the U.S.
Despite the growing increase in the utilization of hospice in the U.S, disparities exist in the utilization of hospice. Accumulating evidence has shown that neighborhood characteristics have an impact on availability of hospice agencies. To assess the association between neighborhood social vulnerability and hospice agency availability. Using the Medicare Post-Acute Care and Hospice Provider Utilization and Payment Public Use Files (PAC PUF) for 2019. Hospice agency addresses were geocoded to the census tract level. Multivariable Poisson regression models were used to assess the association between socioeconomic status SVI theme and hospice agency availability adjusting for number of home health agencies, primary care health profession shortage, per cent Black, and Percent Hispanic at the census tract level and rurality. The socioeconomic status SVI subtheme was associated with decreased likelihood of hospice agency availability (adjusted IRR (aIRR), .56; 95% CI, .50- .63; P < .001). Predominantly Black, and predominantly Hispanic neighborhoods had lower rates of hospice agency availability (aIRR, .48; 95% CI, .39-.59; P < .001 and aIRR, .29; 95% CI, .24-.36; P < .001), respectively. Neighborhood socioeconomic disadvantage was associated with lower availability of hospice agencies. Policies aimed at increasing access to hospice should be cognizant of neighborhood socioeconomic disadvantage.


Help-Seeking Buffers the Impact of Grief on Quality of Life: A Study of Loss During the COVID-19 Pandemic.
The COVID-19 pandemic has presented a global challenge for anticipating the support and treatment needs of bereaved individuals. However, no studies have examined how mourners have been coping with grief and which strategies may buffer negative mental health consequences. We examined the various coping strategies being used and which strategies best support quality of life. Participants completed self-report measures of demographic and loss-related characteristics, grief symptoms, quality of life (QOL), and coping strategies used. Despite help-seeking being one of the least endorsed coping strategies used, help-seeking was the only coping strategy that buffered the impact of grief on QOL for individuals with high grief severity. Results support predictions that grief may become a global mental health concern requiring increased accessibility and availability of grief therapies and professional supports for bereaved individuals during and in the aftermath of the pandemic.