Studies on the liver to kidney switch of erythropoietin production.
Although the liver is the major site of erythropoietin (Ep) production in the fetus, this function is assumed by kidneys in the adult. The mechanisms underlying the liver to kidney switch of Ep formation are not understood. We studied the natural progression of this transition in sheep by measuring Ep production in response to anemia in normal and bilaterally nephrectomized fetal and newborn sheep beginning at about 80 d gestation (normal gestation: 140 d). Removal of both kidneys before induction of anemia did not affect Ep formation up to about 120 d of gestation. A significant reduction (29%, P < 0.02) in Ep synthesis was first noted at about 130 d of gestation (initiation of switch). This level of nephrectomy-induced reduction of Ep formation persisted until about 15 d after birth. Thereafter, bilateral nephrectomy caused further significant decreases (P < 0.05) in Ep production, gradually resulting in near total absence of Ep production at about day 40 postpartum (completion of switch). Chronic administration of testosterone (12 mg/wk) or estradiole benzoate (1.5 mg/d, 5 d/wk) to the fetus/newborn beginning at 85-90 d of gestation enhanced or suppressed erythropoiesis, respectively, but failed to affect the time at which the liver to kidney switch was initiated and/or completed. By contrast, a significant delay (P < 0.001) in the onset, but not completion of the switch occurred in animals that were either thyroidectomized or rendered chronically anemic beginning in the second third of the gestation period. Administration of thyroxin (1.2 mg/d, 5 d/wk) to thyroidectomized fetus/newborns not only prevented the delay in the initiation of the switch, but also accelerated the rate at which the switch was completed. These results demonstrate that in sheep (a) the liver to kidney switch of Ep production is initiated in utero during the last third of the gestation period, but is completed after birth, (b) this transition occurs gradually; the assumption of Ep producing capacity by the kidney is not preceded by an abrupt loss of hepatic Ep formation; and (c) the switch is not affected by changes in sex hormone levels during the prenatal-postnatal growth periods, but is profoundly influenced by alterations in thyroid hormone and oxygen supply-demand levels.


Insect cardioactive peptides. I. Distribution and molecular characteristics of two cardioacceleratory peptides in the tobacco hawkmoth, Manduca sexta.
Using an in vitro heart bioassay, the pharmacological and biochemical properties of two cardioactive peptides derived from neural tissues of the moth, Manduca sexta, were analysed. Gel filtration of ventral nerve cords (VNC) from pharate adults identified two cardioacceleratory peptides (CAP1 and CAP2) with apparent molecular weights of 1000 and 500 Da, respectively. Both CAPs were localized to the abdominal perivisceral organs, the major neurohaemal release sites in the insect VNC. Pulse application of CAP1 or CAP2 on the in vitro Manduca heart produced a dose-dependent increase in rate but had no effect on beat amplitude. The threshold dose for the action of each peptide on the isolated heart bioassay was less than 0.05 abdominal nerve cord equivalents. Both CAPs were present in the pharate adult VNC of several other Lepidopteran species. Neither CAP1 nor CAP2 was detected in the prepupal VNC of Manduca sexta.


A simplified procedure to determine the optimal rate of freezing biological systems.
The effect of several cell-level parameters on the predicted optimal cooling rate B(opt) of an arbitrary biological system has been studied using a well-defined water transport model. An extensive investigation of the water transport model revealed three key cell level parameters: reference permeability of the membrane to water L(pg), apparent activation energy E(Lp), and the ratio of the available surface area for water transport to the initial volume of intracellular water (SA/WV). We defined B(opt) as the "highest" cooling rate at which a predefined percent of the initial water volume is trapped inside the cell (values ranging from 5% to 80%) at a predefined end temperature (values ranging from -5 degrees C to -40 degrees C). Irrespective of the choice of the percent of initial water volume trapped and the end temperature, an exact and linear relationship exists between L(pg), SA/WV, and B(opt0. However, a nonlinear and inverse relationship is found between E(Lp) and B(opt). Remarkably, for a variety of biological systems a comparison of the published experimentally determined values of B(opt) agreed quite closely with numerically predicted B(opt) values when the model assumed 5% of initial water is trapped inside the cell at a temperature of -15 degrees C. This close agreement between the experimental and model predicted optimal cooling rates is used to develop a generic optimal cooling rate chart and a generic optimal cooling rate equation that greatly simplifies the prediction of the optimal rate of freezing of biological systems.


Benzodiazepines May be Worse Than Opioids: Negative Medication Effects in Severe Chronic Pain.
Opioid prescription for noncancer pain is increasing in Europe and the United States. Research and guidance have focused on the potential for dependency and medical side effects with high doses. In contrast, benzodiazepines have received little attention in the chronic pain literature, despite evidence for dependency and cognitive impairment in long-term use. We aimed to examine the relationship between these classes of medication use, mood, and functioning. This cross-sectional study included patients (N=229) with disabling chronic pain who were about to start intensive pain rehabilitation. They completed self-report measures of mood, functioning, and responses to pain. We examined each patient's medication use and calculated a single morphine equivalent (ME) dose per person, and a similar diazepam equivalent (DE) dose. We examined the relationship between drug dose, mood, and functioning. Higher DE doses were associated with worse outcomes in most domains. Higher ME doses were more narrowly associated with worse functioning. There was no evidence for any benefit of these drugs; higher doses were not associated with less pain, fear, or disability. Higher ME doses were not more problematic, contrary to our predictions. The combination of opioids and benzodiazepines was associated with particularly poor outcomes for mood. This study is the first to examine both opioid and benzodiazepine use together in chronic pain. We found the anticipated negative effects of opioid medication, and particularly consistent associations between benzodiazepine use and poor well-being. Future guidance on chronic pain prescription should focus on restricting benzodiazepine use.


Dynamic Path Exploration on Mobile Devices.
We present a novel framework for visualizing routes on mobile devices. Our framework is suitable for helping users explore their environment. First, given a starting point and a maximum route length, the system retrieves nearby points of interest (POIs). Second, we automatically compute an attractive walking path through the environment trying to pass by as many highly ranked POIs as possible. Third, we automatically compute a route visualization that shows the current user position, POI locations via pins, and detail lenses for more information about the POIs. The visualization is an animation of an orthographic map view that follows the current user position. We propose an optimization based on a binary integer program (BIP) that models multiple requirements for an effective placement of detail lenses. We show that our path computation method outperforms recently proposed methods and we evaluate the overall impact of our framework in two user studies.


[Enzyme typing of species of the genus Candida isolated from the oral cavity].
The production of phospholipase and proteinase exoenzymes was evaluated in seventy nine samples of Candida isolated from the oral cavity of patients with oral lesions characteristic of candidosis and from individuals presenting a clinically normal mouth, attended at the University of Dentistry of Ribeirão Preto USP. Among the strains of C. albicans isolated from oral lesions, the phospholipase and proteinase were detected in 83.3% and 66.7%, respectively. C. tropicalis and C. parapsilosis produced only proteinase. Regarding the isolated strains from niches without lesions, out of a total of 32 C. albicans, 71.9% presented phospholipase and 68.7% proteinase. C. tropicalis only presented the enzyme proteinase, C. glabrata, C. krusei, C. guilliermondii and Candida spp did not present any of the exoenzymes. Among the samples of C. albicans from both groups, the enzymotype 22 (positive phospholipase and proteinase weakly positive), was prevalent. Different enzymotypes of the same species were detected in samples collected from the same patient.


The mouse elongation factor-2 gene: isolation and characterization of the promoter.
Elongation factor 2 (EF-2) is a protein involved in peptide chain elongation in eukaryotes. We isolated the mouse EF-2 gene and characterized its promoter. We showed that the majority of enhancer elements were located within 500 bp of the flanking sequence and identified a factor binding site sequence (CGTCACGTGACGC) located between nucleotides -58 and -47 containing two CGTCA motifs separated by two nucleotides. The motif represents a half-site for binding of the cAMP response element (CRE) binding protein (CREB). Mutation analysis indicated that the presence of one CGTCA site alone conferred cAMP inducibility, but the presence of one or two CGTCA sites and spacing nucleotides elicited cAMP-independent, constitutive expression. UV cross-linking and DNA affinity chromatography revealed that three 40-, 43-, and 65-kD proteins bound to the CRE-like element. Of these, the 65-kD protein was unique to the CRE-like element. The 40-kD protein was ATF1 and the 43-kD protein with the molecular size of CREB was not CREB, on the basis of reactivity to their respective antibodies. Because ATF1 responds poorly to cAMP induction, it is likely the contributor to the constitutive expression rather than inductive expression of the CRE-like element, and, thus, the EF-2 gene.


Comparison of the sensitivity of the caudal fold skin test and a commercial gamma-interferon assay for diagnosis of bovine tuberculosis.
A study to determine and compare the sensitivity of the caudal fold tuberculin test (CFT) and a commercial gamma-interferon (gamma-IFN) assay for diagnosis of bovine tuberculosis was conducted. A dairy herd with approximately a third of the cattle infected with Mycobacterium bovis was chosen for this study. All cattle from this herd were slaughtered, and tissue specimens for bacteriologic culturing and histologic examination were collected. Results of the CFT and gamma-IFN assay were compared with results of bacteriologic culturing and histologic examination to determine test sensitivity. Results were analyzed, using each of the following 4 standards to classify cattle as infected: positive test result by bacteriologic culturing only; histologic examination only; bacteriologic culturing and histologic examination; and bacteriologic culturing or histologic examination. Sensitivity of the CFT ranged from 80.4 to 84.4%, depending on the standard of comparison. Sensitivity of the gamma-IFN assay ranged from 55.4 to 97.1%, depending on the standard of comparison and on the method of interpretation. The CFT was significantly (P < 0.001) more sensitive than the gamma-IFN assay for diagnosis of bovine tuberculosis when the gamma-IFN assay was conducted and interpreted as instructed by the manufacturer. Maximum overall sensitivity was achieved when results of the CFT and gamma-IFN assay were interpreted in parallel.


A prospective evaluation of a new physical examination in predicting glenoid labral tears.
We studied 62 patients (40 men and 22 women) with an average age of 28 years over a 28-month period who presented with shoulder pain that was refractory to 3 months of conservative management. Patients with a prior glenohumeral dislocation or a rotator cuff tear were excluded. The "crank" test was performed with the arm elevated to 160 degrees in the scapular plane of the body, loaded axially along the humerus, and with maximal internal and external rotation. Although similar tests have been described, the crank test is a new examination previously unreported. Half of the patients (31) had a positive crank test. Arthroscopy performed on all 62 patients revealed glenoid labral tears in 32 patients. Two patients who had positive crank tests did not have labral tears but had partial-thickness, articular-side rotator cuff tears. The sensitivity of the crank test was 91%, the specificity was 93%, the positive predictive value was 94%, and the negative predictive value was 90%. With these data, the crank test fulfills the criteria as a single physical examination test that is highly accurate for the preoperative diagnosis of glenoid labral tears. Accordingly, expensive imaging modalities currently used in this patient population may be employed less in the future.


Low agreement between the modified diet and renal disease formula and the Cockcroft-Gault formula for assessing chronic kidney disease in cognitively impaired elderly outpatients.
Chronic kidney disease is a global public health concern. Glomerular filtration rate (GFR) prediction based on serum creatinine is used to assess renal function in the elderly. The Cockcroft-Gault (CG) formula, based on body surface area (CG/BSA formula), and the Modified Diet and Renal Disease formula (MDRD formula) are commonly used in assessing renal function in clinical practice. The objective of this study was to investigate the agreement between the GFR estimate, CG/BSA formula, and the MDRD formula in elderly outpatients. An outpatient chart review was conducted on consecutive elderly patients aged ≥ 65 years over a 9-month period. Data regarding age, gender, cognitive status, clock drawing, weight, height, and serum creatinine were collected. Pearson's correlation coefficient, Bland-Altman plot, and kappa statistics were used for statistical analysis. Of the 170 patients who participated in the study, 71% were cognitively impaired or had dementia. Using the CG/BSA formula, 79% of the patients had stage 3 renal disease (GFR< 60 mL/min); only 56% were diagnosed as such using the MDRD formula. There was a high correlation between the CG/BSA and MDRD formulas (Pearson correlation coefficient, 0.88; P < 0.0001). However, the kappa statistic was 0.47, indicating low agreement between the 2 formulas. The diagnosis of having stage 3 chronic kidney disease depended on whether the CG/BSA or MDRD formula was used. A 17% discordance rate (9.61 mL/min) was seen between the MDRD and CG/BSA formulas for estimating GFR, and there was low agreement between the 2 formulas. Further studies are needed to assess which predictive formula is appropriate for elderly patients.


Knockdown of glyoxalase 1 mimics diabetic nephropathy in nondiabetic mice.
Differences in susceptibility to diabetic nephropathy (DN) between mouse strains with identical levels of hyperglycemia correlate with renal levels of oxidative stress, shown previously to play a central role in the pathogenesis of DN. Susceptibility to DN appears to be genetically determined, but the critical genes have not yet been identified. Overexpression of the enzyme glyoxalase 1 (Glo1), which prevents posttranslational modification of proteins by the glycolysis-derived α-oxoaldehyde, methylglyoxal (MG), prevents hyperglycemia-induced oxidative stress in cultured cells and model organisms. In this study, we show that in nondiabetic mice, knockdown of Glo1 increases to diabetic levels both MG modification of glomerular proteins and oxidative stress, causing alterations in kidney morphology indistinguishable from those caused by diabetes. We also show that in diabetic mice, Glo1 overexpression completely prevents diabetes-induced increases in MG modification of glomerular proteins, increased oxidative stress, and the development of diabetic kidney pathology, despite unchanged levels of diabetic hyperglycemia. Together, these data indicate that Glo1 activity regulates the sensitivity of the kidney to hyperglycemic-induced renal pathology and that alterations in the rate of MG detoxification are sufficient to determine the glycemic set point at which DN occurs.


HIV infects astrocytes in vivo and egresses from the brain to the periphery.
HIV invades the brain during acute infection. Yet, it is unknown whether long-lived infected brain cells release productive virus that can egress from the brain to re-seed peripheral organs. This understanding has significant implication for the brain as a reservoir for HIV and most importantly HIV interplay between the brain and peripheral organs. Given the sheer number of astrocytes in the human brain and their controversial role in HIV infection, we evaluated their infection in vivo and whether HIV infected astrocytes can support HIV egress to peripheral organs. We developed two novel models of chimeric human astrocyte/human peripheral blood mononuclear cells: NOD/scid-IL-2Rgc null (NSG) mice (huAstro/HuPBMCs) whereby we transplanted HIV (non-pseudotyped or VSVg-pseudotyped) infected or uninfected primary human fetal astrocytes (NHAs) or an astrocytoma cell line (U138MG) into the brain of neonate or adult NSG mice and reconstituted the animals with human peripheral blood mononuclear cells (PBMCs). We also transplanted uninfected astrocytes into the brain of NSG mice and reconstituted with infected PBMCs to mimic a biological infection course. As expected, the xenotransplanted astrocytes did not escape/migrate out of the brain and the blood brain barrier (BBB) was intact in this model. We demonstrate that astrocytes support HIV infection in vivo and egress to peripheral organs, at least in part, through trafficking of infected CD4+ T cells out of the brain. Astrocyte-derived HIV egress persists, albeit at low levels, under combination antiretroviral therapy (cART). Egressed HIV evolved with a pattern and rate typical of acute peripheral infection. Lastly, analysis of human cortical or hippocampal brain regions of donors under cART revealed that astrocytes harbor between 0.4-5.2% integrated HIV gag DNA and 2-7% are HIV gag mRNA positive. These studies establish a paradigm shift in the dynamic interaction between the brain and peripheral organs which can inform eradication of HIV reservoirs.


Significance testing in ridge regression for genetic data.
Technological developments have increased the feasibility of large scale genetic association studies. Densely typed genetic markers are obtained using SNP arrays, next-generation sequencing technologies and imputation. However, SNPs typed using these methods can be highly correlated due to linkage disequilibrium among them, and standard multiple regression techniques fail with these data sets due to their high dimensionality and correlation structure. There has been increasing interest in using penalised regression in the analysis of high dimensional data. Ridge regression is one such penalised regression technique which does not perform variable selection, instead estimating a regression coefficient for each predictor variable. It is therefore desirable to obtain an estimate of the significance of each ridge regression coefficient. We develop and evaluate a test of significance for ridge regression coefficients. Using simulation studies, we demonstrate that the performance of the test is comparable to that of a permutation test, with the advantage of a much-reduced computational cost. We introduce the p-value trace, a plot of the negative logarithm of the p-values of ridge regression coefficients with increasing shrinkage parameter, which enables the visualisation of the change in p-value of the regression coefficients with increasing penalisation. We apply the proposed method to a lung cancer case-control data set from EPIC, the European Prospective Investigation into Cancer and Nutrition. The proposed test is a useful alternative to a permutation test for the estimation of the significance of ridge regression coefficients, at a much-reduced computational cost. The p-value trace is an informative graphical tool for evaluating the results of a test of significance of ridge regression coefficients as the shrinkage parameter increases, and the proposed test makes its production computationally feasible.


Detection of horse allergen around a stable.
Integrating horse stables with built-up areas may lead to conflicts. Dispersion of horse allergen may become a health risk for allergic people. The aim was to measure the dispersion of horse allergen around a stable, considering wind speed and direction and vegetation. The disturbance of staff at a workplace nearby a stable was investigated. Air sampling was performed around a stable (32 horses) at distances of 50-500 m in all directions. Sampling was done with a pump and an IOM sampler. Samples were collected at 50 points during all seasons. Horse allergen levels were determined using ELISA. Disturbance by horses was studied with a questionnaire handed to the employees in an office near the stable. The median horse allergen level at the stable entrance was 316 U/m(3), in the horse fields 40 U/m(3) and in the whole source area 16 U/m(3), which declined to <2 U/m(3) at about 50 m from the source area. Downwind of the prevailing winds low levels of horse allergen (2-4 U/m(3)) could sometimes be detected at up to 500 m. The staff, including those allergic to horses, managed to tolerate horses close to the workplace. At low winds horse allergen spread in ambient air about 50 m from the stable and horse fields. At higher winds low allergen levels were sometimes found in open areas up to 500 m from the source area. These levels were similar to those found in the office after moving away from the stable area. The employees did not report more symptoms of allergy or asthma while working close to the stable compared to after the move.


Training and other predictors of personal protective equipment use in Australian grain farmers using pesticides.
To investigate patterns of use of personal protective equipment (PPE) to reduce pesticide exposure in a sample of Australian farmers and also to assess the influence of possible predictive factors. A cross-sectional survey of 1102 farmers recruited through the Victorian Farmers Federation (VFF) was conducted. A written questionnaire was filled out by participants at VFF meetings attended by a visiting research assistant. Participants answered questions about frequency of pesticide use and PPE items they usually used when doing two different pesticide-related tasks, mixing and application, of each of four classes of pesticides. They also answered questions about personal characteristics, farm characteristics, farming activities, career and health. Nearly all surveyed farmers had ever used pesticides, and over 87% had used Herbicides or Animal Health Products in the previous 12 months. Non-use of PPE was frequently reported, with up to 10-40% of farmers routinely using no PPE at all when using pesticides. Across all pesticide classes, PPE use was higher for pesticide mixing than for application. In multivariate analyses PPE use appeared to be most strongly associated with younger age and farm chemical training. PPE use across all pesticide classes was poor, indicating the possibility of clinically significant pesticide exposure in many farmers. Given that PPE use was found to be associated with farm chemical training, the authors suggest that training is likely to be an important intervention for reducing farmers' pesticide exposure. Poor uptake of farm chemical training by farmers and the aging farming workforce are causes for concern in the light of these findings.


Insights into ALK-driven cancers revealed through development of novel ALK tyrosine kinase inhibitors.
Aberrant forms of the anaplastic lymphoma kinase (ALK) have been implicated in the pathogenesis of multiple human cancers, where ALK represents a rational therapeutic target in these settings. In this study, we report the identification and biological characterization of X-376 and X-396, two potent and highly specific ALK small molecule tyrosine kinase inhibitors (TKIs). In Ambit kinome screens, cell growth inhibition studies, and surrogate kinase assays, X-376 and X-396 were more potent inhibitors of ALK but less potent inhibitors of MET compared to PF-02341066 (PF-1066), an ALK/MET dual TKI currently in clinical trials. Both X-376 and X-396 displayed potent antitumor activity in vivo with favorable pharmacokinetic and toxicity profiles. Similar levels of drug sensitivity were displayed by the three most common ALK fusion proteins in lung cancer (EML4-ALK variants E13;A20, E20;A20, and E6b;A20) as well as a KIF5B-ALK fusion protein. Moreover, X-396 could potently inhibit ALK kinases engineered with two point mutations associated with acquired resistance to PF-1066, L1196M, and C1156Y, when engineered into an E13;A20 fusion variant. Finally, X-396 displayed synergistic growth inhibitory activity when combined with the mTOR inhibitor rapamycin. Our findings offer preclinical proof-of-concept for use of these novel agents to improve therapeutic outcomes of patients with mutant ALK-driven malignancies.


The mouse CD7 gene: identification of a new element common to the human CD7 and mouse Thy-1 promoters.
Human CD7 (hCD7) is a 40 000 Mr member of the immunoglobulin gene superfamily that is expressed early in natural killer (NK) and T-lymphocyte development. CD7 is involved in lymphocyte activation, as ligation of CD7 activates NK and TCRgammadelta T lymphocytes, and ligation of CD7 on TCRalphabeta T lymphocytes induces a non-mitogenic calcium flux. We have previously cloned and characterized the gene for human CD7 (hCD7) and have described its expression in transgenic mice. Recently a mouse cDNA homologous to hCD7 was reported, which we mapped to the corresponding mouse chromosomal location as hCD7. We now report the identification and characterization of a mouse CD7 (mCD7) genomic clone. We demonstrated that the mCD7 gene was similar both in size and structural organization to hCD7. Comparison of the 5' flanking sequences of the mCD7 and hCD7 genes revealed two regions of sequence similarity. Electrophoretic mobility shift assay confirmed both of these regions to be sites of tissue-restricted protein binding in vitro. The more 3' similarity region also shared sequence with a region in the mouse Thy-1 gene 5' flanking region, suggesting that this sequence may be a cis-acting regulatory element common to all three genes. Thus, the promoter regions and exonic organization were similar in the human CD7, mouse CD7, and mouse Thy-1 genes.


Mapping the knowledge structure of research on patient adherence: knowledge domain visualization based co-word analysis and social network analysis.
Patient adherence is an important issue for health service providers and health researchers. However, the knowledge structure of diverse research on treatment adherence is unclear. This study used co-word analysis and social network analysis techniques to analyze research literature on adherence, and to show their knowledge structure and evolution over time. Published scientific papers about treatment adherence were retrieved from Web of Science (2000 to May 2011). A total of 2308 relevant articles were included: 788 articles published in 2000-2005 and 1520 articles published in 2006-2011. The keywords of each article were extracted by using the software Biblexcel, and the synonym and isogenous words were merged manually. The frequency of keywords and their co-occurrence frequency were counted. High frequency keywords were selected to yield the co-words matrix. Finally the decomposition maps were used to comb the complex knowledge structures. Research themes were more general in the first period (2000 to 2005), and more extensive with many more new terms in the second period (2006 to 2011). Research on adherence has covered more and more diseases, populations and methods, but other diseases/conditions are not as hot as HIV/AIDS and have not become specialty themes/sub-directions. Most studies originated from the United States. The dynamic of this field is mainly divergent, with increasing number of new sub-directions of research. Future research is required to investigate specific directions and converge as well to construct a general paradigm in this field.


G-protein-coupled receptor modulation of striatal CaV1.3 L-type Ca2+ channels is dependent on a Shank-binding domain.
Voltage-gated L-type Ca2+ channels are key determinants of synaptic integration and plasticity, dendritic electrogenesis, and activity-dependent gene expression in neurons. Fulfilling these functions requires appropriate channel gating, perisynaptic targeting, and linkage to intracellular signaling cascades controlled by G-protein-coupled receptors (GPCRs). Surprisingly, little is known about how these requirements are met in neurons. The studies described here shed new light on how this is accomplished. We show that D2 dopaminergic and M1 muscarinic receptors selectively modulate a biophysically distinctive subtype of L-type Ca2+ channels (CaV1.3) in striatal medium spiny neurons. The splice variant of these channels expressed in medium spiny neurons contains cytoplasmic Src homology 3 and PDZ (postsynaptic density-95 (PSD-95)/Discs large/zona occludens-1) domains that bind the synaptic scaffolding protein Shank. Medium spiny neurons coexpressed CaV1.3-interacting Shank isoforms that colocalized with PSD-95 and CaV1.3a channels in puncta resembling spines on which glutamatergic corticostriatal synapses are formed. The modulation of CaV1.3 channels by D2 and M1 receptors was disrupted by intracellular dialysis of a peptide designed to compete for the CaV1.3 PDZ domain but not with one targeting a related PDZ domain. The modulation also was disrupted by application of peptides targeting the Shank interaction with Homer. Upstate transitions in medium spiny neurons driven by activation of glutamatergic receptors were suppressed by genetic deletion of CaV1.3 channels or by activation of D2 dopaminergic receptors. Together, these results suggest that Shank promotes the assembly of a signaling complex at corticostriatal synapses that enables key GPCRs to regulate L-type Ca2+ channels and the integration of glutamatergic synaptic events.


Integrated functional networks of process, tissue, and developmental stage specific interactions in Arabidopsis thaliana.
Recent years have seen an explosion in plant genomics, as the difficulties inherent in sequencing and functionally analyzing these biologically and economically significant organisms have been overcome. Arabidopsis thaliana, a versatile model organism, represents an opportunity to evaluate the predictive power of biological network inference for plant functional genomics. Here, we provide a compendium of functional relationship networks for Arabidopsis thaliana leveraging data integration based on over 60 microarray, physical and genetic interaction, and literature curation datasets. These include tissue, biological process, and development stage specific networks, each predicting relationships specific to an individual biological context. These biological networks enable the rapid investigation of uncharacterized genes in specific tissues and developmental stages of interest and summarize a very large collection of A. thaliana data for biological examination. We found validation in the literature for many of our predicted networks, including those involved in disease resistance, root hair patterning, and auxin homeostasis. These context-specific networks demonstrate that highly specific biological hypotheses can be generated for a diversity of individual processes, developmental stages, and plant tissues in A. thaliana. All predicted functional networks are available online at http://function.princeton.edu/arathGraphle.


Immunologic production of antiangiotensin. II. Production and detection of antiangiotensin.
The parenteral administration of benzoylangiotensin II-azo-BGG to rabbits produced an antiserum with antiangiotensin activity. Antiangiotensin inhibited the biological action of p-aminobenzoylangiotensin II, valine(5)-angiotensin II (free acid form), isoleucine(5)-angiotensin II and valine(5)-angiotensin II (amide form), but it was totally inert towards angiotensin 1. Antiangiotensin activity was distinguished from that of serum angiotensinase by the following observations: (a) an angiotensinase-free gamma-globulin fraction contained antiangiotensin, (b) angiotensinase inactivated angiotensin II (both the amide and free acid forms) and angiotensin I in contrast to the remarkable specificity for angiotensin II exhibited by antiangiotensin. Serological demonstration of antiangiotensin included: (a) a comparison of its precipitin reaction with the angiotensin BGG complex with the reaction with BGG alone, (b) the partial inhibition of the precipitin reaction with the angiotensin BGG complex by angiotensin II, (c) a precipitin reaction with a different angiotensin II protein complex (cat serum). Angiotensin II administered parenterally as the free polypeptide was not antigenic.


Exposure to excess glucocorticoids alters dendritic morphology of adult hippocampal pyramidal neurons.
We have used Golgi-impregnated tissue to demonstrate that exposure to excess glucocorticoids alters dendritic morphology in a specific population of neurons in the adult rat hippocampus. Daily injection of 10 mg of corticosterone for 21 days resulted in decreased numbers of apical dendritic branch points and decreased total apical dendritic length measured in a 100-microns-thick section in CA3 pyramidal cells compared to sham-injected and non-injected controls. In contrast, no changes were observed in CA3 pyramidal cell basal dendritic morphology. Furthermore, no changes were observed in the dendritic morphology of CA1 pyramidal cells or granule cells of the dentate gyrus. Cross-sectional cell body area of any of the 3 cell types examined in this study was unaffected by corticosterone treatment. Finally, qualitative analysis of Nissl-stained tissue from the same brains revealed increased numbers of darkly staining, apparently shrunken CA3 pyramidal cells in corticosterone treated compared to control brains. The changes in dendritic morphology we have observed may be indicative of neurons in the early stages of degeneration, as prolonged exposure to high levels of corticosterone has been shown by others to result in a loss of CA3 pyramidal cells. Additionally, these results suggest possible structural alterations which may occur under physiological conditions in which corticosterone levels are chronically elevated such as in aged animals.


Blastic Plasmacytoid Dendritic Cell Neoplasm Is Dependent on BCL2 and Sensitive to Venetoclax.
Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is an aggressive hematologic malignancy with dismal outcomes for which no standard therapy exists. We found that primary BPDCN cells were dependent on the antiapoptotic protein BCL2 and were uniformly sensitive to the BCL2 inhibitor venetoclax, as measured by direct cytotoxicity, apoptosis assays, and dynamic BH3 profiling. Animals bearing BPDCN patient-derived xenografts had disease responses and improved survival after venetoclax treatment in vivo Finally, we report on 2 patients with relapsed/refractory BPDCN who received venetoclax off-label and experienced significant disease responses. We propose that venetoclax or other BCL2 inhibitors undergo expedited clinical evaluation in BPDCN, alone or in combination with other therapies. In addition, these data illustrate an example of precision medicine to predict treatment response using ex vivo functional assessment of primary tumor tissue, without requiring a genetic biomarker. Therapy for BPDCN is inadequate, and survival in patients with the disease is poor. We used primary tumor cell functional profiling to predict BCL2 antagonist sensitivity as a common feature of BPDCN, and demonstrated in vivo clinical activity of venetoclax in patient-derived xenografts and in 2 patients with relapsed chemotherapy-refractory disease. Cancer Discov; 7(2); 156-64. ©2016 AACR.This article is highlighted in the In This Issue feature, p. 115.


Decreased von Willebrand factor protease activity associated with thrombocytopenic disorders.
Recent studies investigating thrombotic thrombocytopenic purpura (TTP) have implicated abnormal plasma von Willebrand factor (vWF)-cleaving metalloprotease activity in this disorder. It has been proposed that a metalloprotease cleaves unusually large (UL) multimers of vWF, which enter the circulation from the endothelium. Abnormal metalloprotease activity could result in ULvWF, which could participate in TTP. However, the diagnostic specificity of abnormalities in the plasma metalloprotease activity has not been established. A prospective study of vWF protease activity was performed using samples from 20 healthy controls, 20 patients with acute TTP, 20 patients with immune idiopathic thrombocytopenic purpura (ITP), 10 patients with disseminated intravascular thrombocytopenia (DIC), 10 patients with systemic lupus erythematosus (SLE,) and 5 thrombocytopenic patients with leukemia. Studies were performed blinded to the diagnosis. Samples from hospitalized patients with normal platelet counts were also tested. The vWF digests and multimer analysis were done using previously described methods. Six laboratory personnel independently scored each of the multimer gels. Reduced protease activity was observed in 9 of 20 patients with TTP. Reduced activity was also observed in 6 of 20 patients with ITP, 6 of 10 patients with DIC, 5 of 10 patients with SLE, 1 of 5 patients with leukemia, 2 of 20 healthy controls, and 3 of 25 hospitalized patients. This study indicates that abnormalities of vWF protease activity are not restricted to patients with the diagnosis of TTP.


Simultaneous single-molecule measurements of phage T7 replisome composition and function reveal the mechanism of polymerase exchange.
A complete understanding of the molecular mechanisms underlying the functioning of large, multiprotein complexes requires experimental tools capable of simultaneously visualizing molecular architecture and enzymatic activity in real time. We developed a novel single-molecule assay that combines the flow-stretching of individual DNA molecules to measure the activity of the DNA-replication machinery with the visualization of fluorescently labeled DNA polymerases at the replication fork. By correlating polymerase stoichiometry with DNA synthesis of T7 bacteriophage replisomes, we are able to quantitatively describe the mechanism of polymerase exchange. We find that even at relatively modest polymerase concentration (∼2 nM), soluble polymerases are recruited to an actively synthesizing replisome, dramatically increasing local polymerase concentration. These excess polymerases remain passively associated with the replisome through electrostatic interactions with the T7 helicase for ∼50 s until a stochastic and transient dissociation of the synthesizing polymerase from the primer-template allows for a polymerase exchange event to occur.


Environmental Conditions Affect Exhalation of H3N2 Seasonal and Variant Influenza Viruses and Respiratory Droplet Transmission in Ferrets.
The seasonality of influenza virus infections in temperate climates and the role of environmental conditions like temperature and humidity in the transmission of influenza virus through the air are not well understood. Using ferrets housed at four different environmental conditions, we evaluated the respiratory droplet transmission of two influenza viruses (a seasonal H3N2 virus and an H3N2 variant virus, the etiologic virus of a swine to human summertime infection) and concurrently characterized the aerosol shedding profiles of infected animals. Comparisons were made among the different temperature and humidity conditions and between the two viruses to determine if the H3N2 variant virus exhibited enhanced capabilities that may have contributed to the infections occurring in the summer. We report here that although increased levels of H3N2 variant virus were found in ferret nasal wash and exhaled aerosol samples compared to the seasonal H3N2 virus, enhanced respiratory droplet transmission was not observed under any of the environmental settings. However, overall environmental conditions were shown to modulate the frequency of influenza virus transmission through the air. Transmission occurred most frequently at 23°C/30%RH, while the levels of infectious virus in aerosols exhaled by infected ferrets agree with these results. Improving our understanding of how environmental conditions affect influenza virus infectivity and transmission may reveal ways to better protect the public against influenza virus infections.


Inhibition of VEGF Signaling Reduces Diabetes-Exacerbated Brain Swelling, but Not Infarct Size, in Large Cerebral Infarction in Mice.
In light of repeated translational failures with preclinical neuroprotection-based strategies, this preclinical study reevaluates brain swelling as an important pathological event in diabetic stroke and investigates underlying mechanism of the comorbidity-enhanced brain edema formation. Type 2 (mild), type 1 (moderate), and mixed type 1/2 (severe) diabetic mice were subjected to transient focal ischemia. Infarct volume, brain swelling, and IgG extravasation were assessed at 3 days post-stroke. Expression of vascular endothelial growth factor (VEGF)-A, endothelial-specific molecule-1 (Esm1), and the VEGF receptor 2 (VEGFR2) was determined in the ischemic brain. Additionally, SU5416, a VEGFR2 inhibitor, was treated in the type 1/2 diabetic mice, and stroke outcomes were determined. All diabetic groups displayed bigger infarct volume and brain swelling compared to nondiabetic mice, and the increased swelling was disproportionately larger relative to infarct enlargement. Diabetic conditions significantly increased VEGF-A, Esm1, and VEGFR2 expressions in the ischemic brain compared to nondiabetic mice. Notably, in diabetic mice, VEGFR2 mRNA levels were positively correlated with brain swelling, but not with infarct volume. Treatment with SU5416 in diabetic mice significantly reduced brain swelling. The study shows that brain swelling is a predominant pathological event in diabetic stroke and that an underlying event for diabetes-enhanced brain swelling includes the activation of VEGF signaling. This study suggests consideration of stroke therapies aiming at primarily reducing brain swelling for subjects with diabetes.


Genome-wide analysis of the diatom cell cycle unveils a novel type of cyclins involved in environmental signaling.
Despite the enormous importance of diatoms in aquatic ecosystems and their broad industrial potential, little is known about their life cycle control. Diatoms typically inhabit rapidly changing and unstable environments, suggesting that cell cycle regulation in diatoms must have evolved to adequately integrate various environmental signals. The recent genome sequencing of Thalassiosira pseudonana and Phaeodactylum tricornutum allows us to explore the molecular conservation of cell cycle regulation in diatoms. By profile-based annotation of cell cycle genes, counterparts of conserved as well as new regulators were identified in T. pseudonana and P. tricornutum. In particular, the cyclin gene family was found to be expanded extensively compared to that of other eukaryotes and a novel type of cyclins was discovered, the diatom-specific cyclins. We established a synchronization method for P. tricornutum that enabled assignment of the different annotated genes to specific cell cycle phase transitions. The diatom-specific cyclins are predominantly expressed at the G1-to-S transition and some respond to phosphate availability, hinting at a role in connecting cell division to environmental stimuli. The discovery of highly conserved and new cell cycle regulators suggests the evolution of unique control mechanisms for diatom cell division, probably contributing to their ability to adapt and survive under highly fluctuating environmental conditions.


Tear production and intraocular pressure in canine eyes with corneal ulceration.
This study aimed to evaluate changes in lacrimation and intraocular pressure (IOP) in dogs with unilateral corneal ulceration using the Schirmer tear test (STT) and rebound (TonoVet®) tonometry. IOP and STT values were recorded in both ulcerated and non-ulcerated (control) eyes of 100 dogs diagnosed with unilateral corneal ulceration. Dogs presented with other ocular conditions as their primary complaint were excluded from this study. The mean ± standard deviation for STT values in the ulcerated and control eyes were 20.2±4.6 mm/min and 16.7±3.5 mm/min respectively. The mean ± standard deviation for IOP in the ulcerated and control eyes were 11.9±3.1 mmHg and 16.7±2.6 mmHg respectively. STT values were significantly higher (p<0.000001) in the ulcerated eye compared to the control eye while IOP was significantly lower (p<0.0001). There is an increase in lacrimation and a decrease in IOP in canine eyes with corneal ulceration. The higher tear production in ulcerated eyes shows the importance of measuring STT in both eyes in cases of corneal ulceration, since this increased lacrimation may mask an underlying keratoconjunctivitis sicca only evident in the contralateral eye. The lower IOP in ulcerated eyes is likely to relate to mild uveitic change in the ulcerated eye with a concomitant increase in uveoscleral aqueous drainage. While these changes in tear production and IOP in ulcerated eyes are widely recognised in both human and veterinary ophthalmology, it appears that this is the first controlled documented report of these changes in a large number of individuals.


Acceleration of cardiovascular-biological age by amphetamine exposure is a power function of chronological age.
Amphetamine abuse is becoming more widespread internationally. The possibility that its many cardiovascular complications are associated with a prematurely aged cardiovascular system, and indeed biological organism systemically, has not been addressed. Radial arterial pulse tonometry was performed using the SphygmoCor system (Sydney). 55 amphetamine exposed patients were compared with 107 tobacco smokers, 483 non-smokers and 68 methadone patients (total=713 patients) from 2006 to 2011. A cardiovascular-biological age (VA) was determined. The age of the patient groups was 30.03±0.51-40.45±1.15 years. This was controlled for with linear regression. The sex ratio was the same in all groups. 94% of amphetamine exposed patients had used amphetamine in the previous week. When the (log) VA was regressed against the chronological age (CA) and a substance-type group in both cross-sectional and longitudinal models, models quadratic in CA were superior to linear models (both p<0.02). When log VA/CA was regressed in a mixed effects model against time, body mass index, CA and drug type, the cubic model was superior to the linear model (p=0.001). Interactions between CA, (CA)2 and (CA)3 on the one hand and exposure type were significant from p=0.0120. The effects of amphetamine exposure persisted after adjustment for all known cardiovascular risk factors (p<0.0001). These results show that subacute exposure to amphetamines is associated with an advancement of cardiovascular-organismal age both over age and over time, and is robust to adjustment. That this is associated with power functions of age implies a feed-forward positively reinforcing exacerbation of the underlying ageing process.


The epidemiology of Staphylococcus aureus skin and soft tissue infection in the southern Barkly region of Australia's Northern Territory in 2017.
The aim of this study was to describe the burden and organism antibiotic resistance patterns of skin and soft tissue infections (SSTI) due to Staphylococcus aureus presenting in a remote Australian Northern Territory community in the Barkly region. We collated reported antibiograms of all skin and superficial soft tissue swab specimens obtained from the town's Indigenous medical clinic from 12 of the 13 months between November 2016 and December 2017. Clinician's notes for the consultation associated with each test request were examined to determine the nature of the clinical problem and to access other relevant data. Amongst 309 tissue swab specimens, S. aureus was cultured in 215 (70%), of which 202 isolations were from Indigenous Australians. Of the 215 S. aureus, 98 [46%, 95% confidence interval (CI) 31-52] were methicillin resistant S. aureus (MRSA) and 117 (54%, 95% CI 48-61) sensitive (MSSA). Significant numbers were also resistant to other frequently used oral antibiotics, with resistance to erythromycin in 52 (24%), clindamycin in 51 (24%), trimethoprim in 22 (10%) and fusidic acid in eight (4%). In the Barkly region of Australia's NT in 2017, community-acquired staphylococcal SSTI needing professional care is equally likely to be caused by MRSA as by MSSA.


Anorexia nervosa manifesting as massive ascites, hypercholesterolemia, and sequential binge eating in an 11-year-old girl: A case report.
Anorexia nervosa (AN) is a serious eating disorder associated with a distorted body image. Hypercholesterolemia has been found in patients with AN but the mechanism of hyperlipidemia in AN remains little known. Ascites in patients with AN has been attributed to hypoalbuminemia and liver diseases, but massive ascites without the aforementioned etiologies has never been reported in AN. An 11-year-old girl was admitted for exclusion of organic underlying diseases due to severe body weight loss (18% within 3 weeks), poor appetite, and hypercholesterolemia (274 mg/dL). She complained of heartburn sensation, nausea, vomiting, constipation, and postprandial dull abdominal pain with fullness. The patient's condition met with all 3 of the Diagnostic and Statistical Manual of Mental Disorders (DSM-5) criteria for diagnosing AN. On admission, her total cholesterol level was 337 mg/dL and hypocomplementemia (C3 55.5 mg/dL) was also found. Abdominal sonography and computed tomography scans showed massive ascites. However, neither proteinuria nor hypoalbuminemia was found. Upper gastroduodenal endoscopy showed chronic superficial gastritis and colonoscopy revealed negative findings. Ascites obtained by paracentesis demonstrated a transudate without bacterial infection, tuberculosis, or pancreatitis. Exploratory laparoscopy showed nonpurulent ascites. However, biopsies from the small intestine, mesentery, and liver showed chronic inflammation and fibrosis. The intensive nutritional therapy by increasing total energy intake stepwise with a combination of high-energy formula and her favorite foods. Her hypercholesterolemia, hypocomplementemia, and massive ascites resolved after her weight was restored. She developed binge eating with continuous weight gain after discharge. Her weight significantly increased to an obese level (body mass index [BMI] 25.9 kg/m) after loss to follow-up for 4 years until she returned to our emergency room due to suicide attempt. Diagnostic crossover between subtypes in anorexia nervosa might be a potential risk factor for illness severity and poor prognosis. AN can manifest as massive ascites with normal albumin concentrations that could possibly be due to chronic inflammation of the intestinal serosa, mesentery, and peritoneal surface of the liver.


A rapid, dispersion-based wavelength-stepped and wavelength-swept laser for optical coherence tomography.
Optical-domain subsampling enables Fourier-domain OCT imaging at high-speeds and extended depth ranges while limiting the required acquisition bandwidth. To perform optical-domain subsampling, a wavelength-stepped rather than a wavelength-swept source is required. This preliminary study introduces a novel design for a rapid wavelength-stepped laser source that uses dispersive fibers in combination with a fast lithium-niobate modulator to achieve wavelength selection. A laser with 200 GHz wavelength-stepping and a sweep rate of 9 MHz over a 94 nm range at a center wavelength of 1550 nm is demonstrated. A reconfiguration of this source design to a continuous wavelength-swept light for conventional Fourier-domain OCT is also demonstrated.


In the eye of the beholder: a survey of models for eyes and gaze.
Despite active research and significant progress in the last 30 years, eye detection and tracking remains challenging due to the individuality of eyes, occlusion, variability in scale, location, and light conditions. Data on eye location and details of eye movements have numerous applications and are essential in face detection, biometric identification, and particular human-computer interaction tasks. This paper reviews current progress and state of the art in video-based eye detection and tracking in order to identify promising techniques as well as issues to be further addressed. We present a detailed review of recent eye models and techniques for eye detection and tracking. We also survey methods for gaze estimation and compare them based on their geometric properties and reported accuracies. This review shows that, despite their apparent simplicity, the development of a general eye detection technique involves addressing many challenges, requires further theoretical developments, and is consequently of interest to many other domains problems in computer vision and beyond.


Osteopathology and selenium deficiency co-occurring in a population of endangered Patagonian huemul (Hippocamelus bisulcus).
About 1,000 endangered Patagonian huemul deer (Hippocamelus bisulcus) remain in Chile and 350-500 in Argentina. Most groups (>100) are not recovering, and prevalence of osteopathology in Argentina was at least 57%. Here I describe relevant cases of osteopathology from a Chilean population which, however, recently also provided data on trace mineral status, supporting the initial hypothesis that nutrition may be a primary etiologic factor. Additionally, recent data on bone chemical composition of Argentine cases and soil analyses are discussed. Fluoride levels in Argentine cases with osteopathology were low and fluorosis was discarded as an etiological factor. Selenium deficiency occurred in 73% of huemul from the Chilean population which exhibited several cases with osteopathology. The pathophysiognomy included extensive erosion; tooth loss; porosification; perforations of palate, maxillar and mandibular bone with frequent exposure of tooth roots; and fractured mandibula. Areas currently used by remaining huemul have mainly acidic volcanic soils, which reduces selenium bioavailability: mean soil selenium levels from areas typically used by extant huemul were very deficient (0.19 ppm), corroborating documented overt selenium deficiency in local livestock and plants. The area of extant huemul is known to result in primary iodine deficiency in livestock which is aggravated by selenium deficiency. Currently the most parsimonious explanation for frequent osteopathology and lack of numerical recovery are the combined effects of selenium and iodine deficiencies based on: osteopathology in a population of selenium deficient huemul; selenium deficient livestock, plants and soils; acidic soils; and regional primary iodine deficiency. The nexus between mineral nutrition and population dynamics of huemul may be due to constraints on their movements to fertile lowlands, including the elimination of historic migratory traditions, and concomitant elimination of source populations.


Observation of the inverse giant piezoresistance effect in silicon nanomembranes probed by ultrafast terahertz spectroscopy.
The anomalous piezoresistance (a-PZR) effects, including giant PZR (GPZR) with large magnitude and inverse PZR of opposite, have exciting technological potentials for their integration into novel nanoelectromechanical systems. However, the nature of a-PZR effect and the associated kinetics have not been clearly determined yet. Even further, there are intense research debates whether the a-PZR effect actually exists or not; although numerous investigations have been conducted, the origin of the effect has not been clearly understood. This paper shows the existence of a-PZR and provides direct experimental evidence through the performance of well-established electrical measurements and terahertz spectroscopy on silicon nanomembranes (Si NMs). The clear inverse PZR behavior was observed in the Si NMs when the thickness was less than 40 nm and the magnitude of the PZR response linearly increased with the decreasing thickness. Observations combined with electrical and optical measurements strongly corroborate that the a-PZR effect originates from the carrier concentration changes via charge carrier trapping into strain-induced defect states.


RF injection locked 18 GHz regeneratively mode-locked semiconductor laser.
In this manuscript, a semiconductor based fiber ring cavity mode-locked laser regeneratively driven at 18 GHz is presented. The optical spectrum of the laser is centered at 1578 nm. The laser is RF injection locked via an external source at 18 GHz. The phase noise of the mode-locked laser is measured and the integrated timing jitter was found to be 10.8 fs (from 100 Hz to 20 MHz) and 13.3 fs (from 100 Hz to Nyquist frequency). The integrated amplitude fluctuation (from 100 Hz to 20 MHz) was less than 0.02%. The laser phase and amplitude noise responses to various injected RF power levels were also investigated. The injection RF power has significant effect on the phase noise and the best jitter value is around 40 dB lower than the cavity regenerated RF power.


Seborrheic keratosis over genitalia masquerading as Buschke Lowenstein tumor.
Seborrheic keratosis (SK) is a common benign condition of the skin. It does not usually appear before middle age. Upper trunk and face are the sites most commonly involved. Lesions are also frequently seen on the extremities. SK of the genitalia is a rare entity. It has been frequently mistaken as genital warts and differentiation is made only on histopathology. We report a case of SK with polypoidal lesions restricted to the skin on and around the genitalia since 10 years. SK should be considered in the differential diagnosis of pedunculated lesions of the penis. The histopathology after shave excision is diagnostic.


Multi-species Ontologies of the Craniofacial Musculoskeletal System.
We created the Ontology of Craniofacial Development and Malformation (OCDM) [1] to provide a unifying framework for organizing and integrating craniofacial data ranging from genes to clinical phenotypes from multi-species. Within this framework we focused on spatio-structural representation of anatomical entities related to craniofacial development and malformation, such as craniosynostosis and midface hypoplasia. Animal models are used to support human studies and so we built multi-species ontologies that would allow for cross-species correlation of anatomical information. For this purpose we first developed and enhanced the craniofacial component of the human musculoskeletal system in the Foundational Model of Anatomy Ontology (FMA)[2], and then imported this component, which we call the Craniofacial Human Ontology (CHO), into the OCDM. The CHO was then used as a template to create the anatomy for the mouse, the Craniofacial Mouse Ontology (CMO) as well as for the zebrafish, the Craniofacial Zebrafish Ontology (CZO).


Evaluation of an immunofluorescence assay for specific detection of immunoglobulin G antibodies directed against Helicobacter pylori, and antigenic cross-reactivity between H. pylori and Campylobacter jejuni.
An immunofluorescence assay (IFA) for the detection of immunoglobulin G antibodies directed against Helicobacter pylori was evaluated by comparing 20 serum specimens from patients with a positive urease test on biopsy material and 20 serum specimens from patients with a negative test and with defined clinical symptoms. The resulting anti-H. pylori titers were classified as follows: negative, less than or equal to 64; borderline, 128; and positive, greater than or equal to 256. By using these criteria, the IFA was subsequently tested, using 100 serum specimens from patients with gastric complaints. Overall, the titers were 71% positive, 10% borderline, and 19% negative. Depending on the patients' biopsy urease test results, the sensitivity and specificity of the assay were calculated to be 96%. Furthermore, these sera were classified into three subgroups on the basis of clinical manifestations: gastritis with 74% positive and 10% borderline titers, duodenal ulcer with 84% positive and 4% borderline titers, and gastric ulcer with 52% positive and 16% borderline titers. A serologic follow-up study was carried out with three patients with gastric ulcers who had been treated with colloidal bismuth subcitrate for 4 weeks and erythromycin for the final 2 weeks. The results indicate that a significant decrease in titer could be expected within 9 to 12 months after successful therapy, as determined by repeated negative CLO tests. Absorption experiments demonstrated that possible cross-reactivity between H. pylori and C. jejuni did not influence serodiagnosis.


Eptifibatide: a review of its use in patients with acute coronary syndromes and/or undergoing percutaneous coronary intervention.
Eptifibatide, a cyclic peptide, is a highly specific, intravenously administered glycoprotein (GP) IIb/IIIa receptor antagonist. By preventing fibrinogen binding to the GP IIb/IIIa receptor, eptifibatide inhibits platelet aggregation and prevents thrombus formation. Clinically, the drug is used as an adjunct to heparin and aspirin. The PURSUIT trial, conducted in >10,000 patients with unstable angina or non-Q-wave myocardial infarction (MI), showed that eptifibatide (180 microg/kg bolus then 2 microg/kg/min infusion for < or =72 hours) reduces the 30-day risk of death or nonfatal MI, with this benefit apparent at 96 hours. The absolute reduction in this end-point of 1.5% persisted at 6 months. The drug is effective in patients undergoing percutaneous coronary intervention (PCI), and, as shown in the North American subgroup, in patients in whom medical management is appropriate. Eptifibatide is also beneficial in patients undergoing PCI, whether or not they have unstable angina or non-Q-wave MI. In a dosage of 135 microg/kg then 0.5 microg/kg/min for 24 hours, eptifibatide reduced the 30-day risk of a combined end-point (death, nonfatal MI and urgent or emergency coronary interventions) by 2.5% (absolute reduction) in patients undergoing PCI in the IMPACT-II trial, when measured by per-protocol (patients treated), but not intent-to-treat, analysis. The drug also decreased the incidence of abrupt vessel closure and ischaemic cardiovascular complications in the first 24 hours (the period of greatest risk). Bleeding episodes are the most common adverse event associated with eptifibatide therapy. Although the incidence of major bleeding is increased with eptifibatide, most bleeding episodes are minor and occur at the vascular access site. The drug is not associated with an excess of intracranial bleeds, stroke or thrombocytopenia, does not appear to increase bleeding risk in patients undergoing coronary artery bypass graft (CABG), and does not cause antibody formation. Limited data suggest that eptifibatide may improve coronary flow when combined with alteplase in patients with acute Q-wave MI, but the possibility of increased bleeding with eptifibatide plus thrombolytics should be borne in mind. Intravenous eptifibatide, when combined with aspirin and heparin, reduces the 30-day risk of ischaemic events in patients with unstable angina and non-Q-wave MI and decreases ischaemic cardiovascular complications at the time of greatest risk in patients undergoing PCI. With its acceptable tolerability profile eptifibatide is a suitable option as a short term adjunct in these clinical settings. Whether eptifibatide in combination with fibrolysis may improve outcome in patients with acute Q-wave MI has yet to be determined.


Increased oxidative stress during hyperglycemic cerebral ischemia.
In this review, we summarize the role of hyperglycemia during cerebral ischemia. Hyperglycemia occurring during experimental and clinical stroke has been associated with increased cerebral damage. Increased oxidative stress resulting from hyperglycemia is believed to contribute to the exacerbated damage. More specifically, superoxide, nitric oxide and peroxynitrite are believed to play an important role in cerebral damage. This also involves increased recruitment of various blood cells to the ischemic zone that contribute to inflammation. We present data from our group and others that demonstrate that free radical production is increased during hyperglycemic stroke in rodents. Recent data suggest that inflammation is an important component of ischemic damage under both normo- and hyperglycemic conditions. We summarize numerous studies that indicate that a variety of antioxidant (inhibition of free radical production, scavenging of free radicals and increasing free radical degradation) and anti-inflammatory strategies decrease cerebral infarction. Finally, we compare the success of some of these strategies in clinical trials compared to the animal models.


Characterization of Parkinson's disease using blood-based biomarkers: A multicohort proteomic analysis.
Parkinson's disease (PD) is a progressive neurodegenerative disease affecting about 5 million people worldwide with no disease-modifying therapies. We sought blood-based biomarkers in order to provide molecular characterization of individuals with PD for diagnostic confirmation and prediction of progression. In 141 plasma samples (96 PD, 45 neurologically normal control [NC] individuals; 45.4% female, mean age 70.0 years) from a longitudinally followed Discovery Cohort based at the University of Pennsylvania (UPenn), we measured levels of 1,129 proteins using an aptamer-based platform. We modeled protein plasma concentration (log10 of relative fluorescence units [RFUs]) as the effect of treatment group (PD versus NC), age at plasma collection, sex, and the levodopa equivalent daily dose (LEDD), deriving first-pass candidate protein biomarkers based on p-value for PD versus NC. These candidate proteins were then ranked by Stability Selection. We confirmed findings from our Discovery Cohort in a Replication Cohort of 317 individuals (215 PD, 102 NC; 47.9% female, mean age 66.7 years) from the multisite, longitudinally followed National Institute of Neurological Disorders and Stroke Parkinson's Disease Biomarker Program (PDBP) Cohort. Analytical approach in the Replication Cohort mirrored the approach in the Discovery Cohort: each protein plasma concentration (log10 of RFU) was modeled as the effect of group (PD versus NC), age at plasma collection, sex, clinical site, and batch. Of the top 10 proteins from the Discovery Cohort ranked by Stability Selection, four associations were replicated in the Replication Cohort. These blood-based biomarkers were bone sialoprotein (BSP, Discovery false discovery rate [FDR]-corrected p = 2.82 × 10-2, Replication FDR-corrected p = 1.03 × 10-4), osteomodulin (OMD, Discovery FDR-corrected p = 2.14 × 10-2, Replication FDR-corrected p = 9.14 × 10-5), aminoacylase-1 (ACY1, Discovery FDR-corrected p = 1.86 × 10-3, Replication FDR-corrected p = 2.18 × 10-2), and growth hormone receptor (GHR, Discovery FDR-corrected p = 3.49 × 10-4, Replication FDR-corrected p = 2.97 × 10-3). Measures of these proteins were not significantly affected by differences in sample handling, and they did not change comparing plasma samples from 10 PD participants sampled both on versus off dopaminergic medication. Plasma measures of OMD, ACY1, and GHR differed in PD versus NC but did not differ between individuals with amyotrophic lateral sclerosis (ALS, n = 59) versus NC. In the Discovery Cohort, individuals with baseline levels of GHR and ACY1 in the lowest tertile were more likely to progress to mild cognitive impairment (MCI) or dementia in Cox proportional hazards analyses adjusting for age, sex, and disease duration (hazard ratio [HR] 2.27 [95% CI 1.04-5.0, p = 0.04] for GHR, and HR 3.0 [95% CI 1.24-7.0, p = 0.014] for ACY1). GHR's association with cognitive decline was confirmed in the Replication Cohort (HR 3.6 [95% CI 1.20-11.1, p = 0.02]). The main limitations of this study were its reliance on the aptamer-based platform for protein measurement and limited follow-up time available for some cohorts. In this study, we found that the blood-based biomarkers BSP, OMD, ACY1, and GHR robustly associated with PD across multiple clinical sites. Our findings suggest that biomarkers based on a peripheral blood sample may be developed for both disease characterization and prediction of future disease progression in PD.


Diagnosis, treatment and outcomes of patients with aortopulmonary window.
Aortopulmonary window (APW) is a communication between the ascending aorta and the pulmonary artery in the presence of two separate semilunar valves and is the rarest of septal defects. To present our experience with the diagnosis and outcome of APW cases. Retrospective cohort study. Between June 2003 and October 2011, thirteen patients were diagnosed with APW. Clinical features of patients, findings of echocardiographic and angiographic examination, results of surgical intervention and follow-up were reviewed retrospectively. Eleven children (10 days to 16 years), underwent surgical correction of APW. In a 12-month-old boy, the defect was repaired by the transcatheter approach. In addition to APW repair, closure of VSD was performed in 2 patients. APW were associated with interruption in two patients; one also had a complex pathology. None of the patients died due to complications of surgical or transcatheter procedures. After a median follow-up period of 40 months, the patients were asymptomatic and none of them required additional medication, except for the patient with complex pathology including an interrupted aortic arch, who underwent balloon angioplasty for recoarctation. In any infant with the findings of congestive heart failure and failure to thrive, APW must be kept in mind as a differential diagnosis. In isolated APW cases before 6 months of age, echocardiography is often sufficient for diagnosis. In complex cases, cardiac catheterisation is performed for the comprehensive evaluation of associated defects. After 6 months, cardiac catheterisation could be utilised to perform vasoreactivity testing and, if possible, to close the defect.


Short communication: Patterns of dairy consumption in free-living children and adolescents.
According to national survey data, dairy food consumption has fallen in recent years and declines further with age, especially from childhood to adolescence. Dietary surveys typically rely on retrospective dietary assessment methods and use broad age groupings (4-10 yr; 11-18 yr), making it challenging to differentiate between middle-childhood and adolescence. Consequently, there is a need to assess dairy food consumption during middle-childhood and adolescence using more robust dietary assessment tools. Therefore, the present study aimed to describe and compare patterns of dairy consumption throughout middle-childhood and adolescence. Dairy food consumption was assessed during school term-time over 4 consecutive days, including 2 weekdays and 2 weekend days, in a sample of free-living children (9-11 yr, n=40) and adolescents (15-18 yr, n=35). For children, free-living dairy intake was evaluated through parental-weighed food records, and for adolescents, a combined weighed self-reported food record and 24-h dietary recall technique was utilized. Food records were explored to determine types, amounts, and frequency of dairy food consumption, and were analyzed for differences between middle-childhood and adolescence using a between group 2×2 (age×sex) ANOVA. Descriptive data suggested that milk was the most popular dairy product consumed by both children and adolescents. Statistical analysis revealed a main effect for sex on total milk consumption (mL) and number of daily milk portions consumed. No interaction or main effect was present for any other variable. The present study indicates that independent of age, boys consumed greater amounts of milk compared with girls. Contrary to existing literature, findings suggest no difference in milk-based dairy consumption between middle-childhood and adolescence.


Multikinase Inhibitors Induce Cutaneous Toxicity through OAT6-Mediated Uptake and MAP3K7-Driven Cell Death.
The use of multikinase inhibitors (MKI) in oncology, such as sorafenib, is associated with a cutaneous adverse event called hand-foot skin reaction (HFSR), in which sites of pressure or friction become inflamed and painful, thus significantly impacting quality of life. The pathogenesis of MKI-induced HFSR is unknown, and the only available treatment options involve dose reduction or discontinuation of therapy, which have negative effects on primary disease management. To investigate the underlying mechanisms by which sorafenib promotes keratinocyte cytotoxicity and subsequent HFSR induction, we performed a transporter-directed RNAi screen in human epidermal keratinocytes and identified SLC22A20 (OAT6) as an uptake carrier of sorafenib. Further investigations into the intracellular mechanism of sorafenib activity through in situ kinome profiling identified the mitogen-activated protein kinase MAP3K7 (TAK1) as a target of sorafenib that induces cell death. Finally, we demonstrate that sorafenib induced keratinocyte injury in vivo and that this effect could be reversed by cotreatment with the OAT6 inhibitor probenecid. Collectively, our findings reveal a novel pathway that regulates the entry of some MKIs into keratinocytes and explains the basis underlying sorafenib-induced skin toxicity, with important implications for the therapeutic management of HFSR.


Experimental and theoretical study on the one- and two-photon absorption properties of novel organic molecules based on phenylacetylene and azoaromatic moieties.
This Article reports a combined experimental and theoretical analysis on the one and two-photon absorption properties of a novel class of organic molecules with a π-conjugated backbone based on phenylacetylene (JCM874, FD43, and FD48) and azoaromatic (YB3p25) moieties. Linear optical properties show that the phenylacetylene-based compounds exhibit strong molar absorptivity in the UV and high fluorescence quantum yield with lifetimes of approximately 2.0 ns, while the azoaromatic-compound has a strong absorption in the visible region with very low fluorescence quantum yield. The two-photon absorption was investigated employing nonlinear optical techniques and quantum chemical calculations based on the response functions formalism within the density functional theory framework. The experimental data revealed well-defined 2PA spectra with reasonable cross-section values in the visible and IR. Along the nonlinear spectra we observed two 2PA allowed bands, as well as the resonance enhancement effect due to the presence of one intermediate one-photon allowed state. Quantum chemical calculations revealed that the 2PA allowed bands correspond to transitions to states that are also one-photon allowed, indicating the relaxation of the electric-dipole selection rules. Moreover, using the theoretical results, we were able to interpret the experimental trends of the 2PA spectra. Finally, using a few-energy-level diagram, within the sum-over-essential states approach, we observed strong qualitative and quantitative correlation between experimental and theoretical results.


Microbial biodiversity of thermophilic communities in hot mineral soils of Tramway Ridge, Mount Erebus, Antarctica.
Tramway Ridge, located near the summit of Mount Erebus in Antarctica, is probably the most remote geothermal soil habitat on Earth. Steam fumaroles maintain moist, hot soil environments creating extreme local physicochemical differentials. In this study a culture-independent approach combining automated rRNA intergenic spacer analysis (ARISA) and a 16S rRNA gene library was used to characterize soil microbial (Bacterial and Archaeal) diversity along intense physicochemical gradients. Statistical analysis of ARISA data showed a clear delineation between bacterial community structure at sites close to fumaroles and all other sites. Temperature and pH were identified as the primary drivers of this demarcation. A clone library constructed from a high-temperature site led to the identification of 18 novel bacterial operational taxonomic units (OTUs). All 16S rRNA gene sequences were deep branching and distantly (85-93%) related to other environmental clones. Five of the signatures branched with an unknown group between candidate division OP10 and Chloroflexi. Within this clade, sequence similarity was low, suggesting it contains several yet-to-be described bacterial groups. Five archaeal OTUs were obtained and exhibited high levels of sequence similarity (95-97%) with Crenarchaeota sourced from deep-subsurface environments on two distant continents. The novel bacterial assemblage coupled with the unique archaeal affinities reinvigorates the hypotheses that Tramway Ridge organisms are relics of archaic microbial lineages specifically adapted to survive in this harsh environment and that this site may provide a portal to the deep-subsurface biosphere.


Commercial Dairy Cow Milk microRNAs Resist Digestion under Simulated Gastrointestinal Tract Conditions.
MicroRNAs are small, gene-regulatory noncoding RNA species present in large amounts in milk, where they seem to be protected against degradative conditions, presumably because of their association with exosomes. We monitored the relative stability of commercial dairy cow milk microRNAs during digestion and examined their associations with extracellular vesicles (EVs). We used a computer-controlled, in vitro, gastrointestinal model TNO intestinal model-1 (TIM-1) and analyzed, by quantitative polymerase chain reaction, the concentration of 2 microRNAs within gastrointestinal tract compartments at different points in time. EVs within TIM-1 digested and nondigested samples were studied by immunoblotting, dynamic light scattering, quantitative polymerase chain reaction, and density measurements. A large quantity of dairy milk Bos taurus microRNA-223 (bta-miR-223) and bta-miR-125b (∼109-1010 copies/300 mL milk) withstood digestion under simulated gastrointestinal tract conditions, with the stomach causing the most important decrease in microRNA amounts. A large quantity of these 2 microRNAs (∼108-109 copies/300 mL milk) was detected in the upper small intestine compartments, which supports their potential bioaccessibility. A protocol optimized for the enrichment of dairy milk exosomes yielded a 100,000 × g pellet fraction that was positive for the exosomal markers tumor susceptibility gene-101 (TSG101), apoptosis-linked gene 2-interacting protein X (ALIX), and heat shock protein 70 (HSP70) and containing bta-miR-223 and bta-miR-125b. This approach, based on successive ultracentrifugation steps, also revealed the existence of ALIX-, HSP70-/low, and TSG101-/low EVs larger than exosomes and 2-6 times more enriched in bta-miR-223 and bta-miR-125b (P < 0.05). Our findings indicate that commercial dairy cow milk contains numerous microRNAs that can resist digestion and are associated mostly with ALIX-, HSP70-/low, and TSG101-/low EVs. Our results support the existence of interspecies transfer of microRNAs mediated by milk consumption and challenge our current view of exosomes as the sole carriers of milk-derived microRNAs.


SBS management in Yb-fiber-amplifiers using multimode seeds and pulse-shaping.
We present a comprehensive analysis of the technique of Longitudinal-Mode-Filling (LMF) to reduce Stimulated Brillouin Scattering (SBS) limitations in Ytterbium Doped Fibre Amplifiers (YDFA), for the generation of nanosecond, temporally shaped pulses. A basic Master-Oscillator-Power-Amplifier (MOPA) system, comprising an output YDFA with 10 µm-core active fibre, is experienced for benchmarking purposes. Input pulse-shaping is operated thanks to direct current modulation in highly multimode laser-diode seeds, either based on the use of Distributed Feed-Back (DFB) or of a Fibre Bragg Grating (FBG). These seeds enable wavelength control. We verify the effectiveness of the combination of LMF, with appropriate mode spacing, in combination with natural chirp effects from the seed to control the SBS threshold in a broad range of output energies, from a few to some tens of µJ. These variations are discussed versus all the parameters of the laser system. In accordance with the proposal of a couple of basic principles and with the addition of gain saturation effects along the active fibre, we develop a full-vectorial numerical model. Fine fits between experimental results and theoretical expectations are demonstrated. The only limitation of the technique arises from broadband beating noise, which is analysed thanks to a simplified, but fully representative description to discuss the signal-to-noise ratio of the amplified pulses. This provides efficient tools for application to the design of robust and cost-effective MOPAs, aiming to the generation of finely shaped and energetic nanosecond pulses without the need for any additional electro-optics.


A controlled study on gastrointestinal nematodes from two Swedish cattle farms showing field evidence of ivermectin resistance.
Anthelmintic resistance (AR) is an increasing problem for the ruminant livestock sector worldwide. However, the extent of the problem is still relatively unknown, especially for parasitic nematodes of cattle. The effect of ivermectin (IVM) (Ivomec inj.®, Merial) was investigated in Swedish isolates of gastrointestinal nematode (GIN) populations showing signs of AR in the field to further characterise the AR status by a range of in vivo and in vitro methods. Three groups, each of 11 calves, were infected with an equal mixture of third stage larvae (L3) of Cooperia oncophora and Ostertagia ostertagi. Group A was inoculated with an IVM-susceptible laboratory isolate and groups B and C with isolates originating from 'resistant' cattle farms. Faecal egg counts (FEC) were monitored from 0 to 45 days post infection (d.p.i.), and L3 were harvested continuously for larval migration inhibition testing (LMIT) and species-specific PCR (ITS2). At 31 d.p.i., one calf from each group was necropsied and adult worms were recovered pre-treatment. At 35 d.p.i., calves from all groups were injected with IVM at the recommended dose (0.2 mg/kg bodyweight). At 45 d.p.i., another two animals from each group were sacrificed and established gastrointestinal worms were collected and counted. A few animals in all three groups were still excreting eggs (50-150 per g faeces) 10 days post IVM injection. However, there was no significant difference in the FEC reductions in groups A (95%; 95% CI 81-99), B (98%; 92-100) and C (99%; 97-100) between 35 and 44 d.p.i. Furthermore, LMIT showed no significant difference between the three groups. Approximately 100 adult O. ostertagi were found in the abomasum of one calf (group B), whereas low to moderate numbers (400-12 200) of C. oncophora remained in the small intestine of the calves in all three groups at 45 d.p.i. PCR on L3 harvested from faecal samples up to 10 days post treatment showed a ratio of 100% C. oncophora in the calves inoculated with isolates A and B, whereas C also had 8% O. ostertagi. Overall, this experiment showed that the animals were successfully treated according to the Faecal egg count reduction test (FECRT) standard (≥ 95% reduction). However, several adult worms of the dose-limiting species C. oncophora demonstrably survived the IVM treatment.


Comparable outcomes for β-lactam/β-lactamase inhibitor combinations and carbapenems in definitive treatment of bloodstream infections caused by cefotaxime-resistant Escherichia coli or Klebsiella pneumoniae.
Extended-spectrum β-lactamase (ESBL) producing Enterobacteriaceae are often susceptible in vitro to β-lactam/β-lactamase inhibitor (BLBLI) combination antibiotics, but their use has been limited by concerns of clinical inefficacy. We aimed to compare outcomes between patients treated with BLBLIs and carbapenems for bloodstream infection (BSI) caused by cefotaxime non-susceptible (likely ESBL- or AmpC β-lactamase-producing) Escherichia coli and Klebsiella pneumoniae. All adult patients with a BSI caused by cefotaxime non-susceptible E. coli or K. pneumoniae were included from May 2012-May 2013. We compared outcomes between patients who had definitive monotherapy with a carbapenem to those who had definitive monotherapy with a BLBLI. There were 92 BSIs that fulfilled the microbiological inclusion criteria. 79 (85.9%) were caused by E. coli and 13 (14.1%) by K. pneumoniae. Four out of 23 (17.4%) patients treated with carbapenem monotherapy and 2 out of 24 (8.3%) patients treated with BLBLI monotherapy died (adjusted HR for survival 0.91, 95% CI 0.13 to 6.28; p = 0.92). The time to resolution of systemic inflammatory response syndrome (SIRS) criteria did not vary between the treatment groups (adjusted HR 0.91, 95% CI 0.32 to 2.59; p = 0.97). The length of hospital admission post-positive blood culture was slightly longer in patients treated with BLBLIs (median duration 15 vs. 11 days), although this was not significant (adjusted HR 0.62; 95% CI 0.27 to 1.42; p = 0.26). There were no significant differences in subsequent isolation of carbapenem resistant organisms (4.3% vs. 4.2%, p = 1.0), C. difficile infection (13.0% vs. 8.3%, p = 0.67) or relapsed BSI (0% vs. 2%, p = 0.23). BLBLIs appear to have a similar efficacy to carbapenems in the treatment of cefotaxime-resistant E. coli and K. pneumoniae bloodstream infections. Directed therapy with a BLBLI, when susceptibility is proven, may represent an appropriate carbapenem-sparing option.


A State of the Art of the Overall Energy Efficiency of Wood Buildings-An Overview and Future Possibilities.
The main goal of this study was to review current studies on the state of the art of wood constructions with a particular focus on energy efficiency, which could serve as a valuable source of information for both industry and scholars. This review begins with an overview of the role of materials in wood buildings to improve energy performance, covering structural and insulation materials that have already been successfully used in the market for general applications over the years. Subsequently, studies of different wood building systems (i.e., wood-frame, post-and-beam, mass timber and hybrid constructions) and energy efficiency are discussed. This is followed by a brief introduction to strategies to increase the energy efficiency of constructions. Finally, remarks and future research opportunities for wood buildings are highlighted. Some general recommendations for developing more energy-efficient wood buildings are identified in the literature and discussed. There is a lack of emerging construction concepts for wood-frame and post-and-beam buildings and a lack of design codes and specifications for mass timber and hybrid buildings. From the perspective of the potential environmental benefits of these systems as a whole, and their effects on energy efficiency and embodied energy in constructions, there are barriers that need to be considered in the future.


The role of sex steroids in white adipose tissue adipocyte function.
With the increasing knowledge that gender influences normal physiology, much biomedical research has begun to focus on the differential effects of sex on tissue function. Sexual dimorphism in mammals is due to the combined effects of both genetic and hormonal factors. Hormonal factors are mutable particularly in females in whom the estrous cycle dominates the hormonal milieu. Given the severity of the obesity epidemic and the fact that there are differences in the obesity rates in men and women, the role of sex in white adipose tissue function is being recognized as increasingly important. Although sex differences in white adipose tissue distribution are well established, the mechanisms affecting differential function of adipocytes within white adipose tissue in males and females remain largely understudied and poorly understood. One of the largest differences in the endocrine environment in males and females is the concentration of circulating androgens and estrogens. This review examines the effects of androgens and estrogens on lipolysis/lipogenesis, adipocyte differentiation, insulin sensitivity and adipokine production in adipocytes from white adipose tissue with a specific emphasis on the sexual dimorphism of adipocyte function in white adipose tissue during both health and disease.


Pulmonary hypertension and other potentially fatal pulmonary complications in systemic juvenile idiopathic arthritis.
Systemic juvenile idiopathic arthritis (JIA) is characterized by fevers, rash, and arthritis, for which interleukin-1 (IL-1) and IL-6 inhibitors appear to be effective treatments. Pulmonary arterial hypertension (PAH), interstitial lung disease (ILD), and alveolar proteinosis (AP) have recently been reported with increased frequency in systemic JIA patients. Our aim was to characterize and compare systemic JIA patients with these complications to a larger cohort of systemic JIA patients. Systemic JIA patients who developed PAH, ILD, and/or AP were identified through an electronic Listserv and their demographic, systemic JIA, and pulmonary disease characteristics as well as their medication exposure information were collected. Patients with these features were compared to a cohort of systemic JIA patients enrolled in the Childhood Arthritis and Rheumatology Research Alliance (CARRA) registry. The patients (n = 25) were significantly (P < 0.05) more likely than the CARRA registry cohort (n = 389) to be female; have more systemic features; and have been exposed to an IL-1 inhibitor, tocilizumab, corticosteroids, intravenous immunoglobulin, cyclosporine, and cyclophosphamide. Twenty patients (80%) were diagnosed with pulmonary disease after 2004. Twenty patients (80%) had macrophage activation syndrome (MAS) during their disease course and 15 patients (60%) had MAS at pulmonary diagnosis. Sixteen patients had PAH, 5 had AP, and 7 had ILD. Seventeen patients (68%) were taking or recently discontinued (<1 month) a biologic agent at pulmonary symptom onset; 12 patients (48%) were taking anti-IL-1 therapy (primarily anakinra). Seventeen patients (68%) died at a mean of 10.2 months from the diagnosis of pulmonary complications. PAH, AP, and ILD are underrecognized complications of systemic JIA that are frequently fatal. These complications may be the result of severe uncontrolled systemic disease activity and may be influenced by medication exposure.


A low dose of an organophosphate insecticide causes dysbiosis and sex-dependent responses in the intestinal microbiota of the Japanese quail (Coturnix japonica).
Organophosphate insecticides have been directly or indirectly implicated in avian populations declining worldwide. Birds in agricultural environments are commonly exposed to these insecticides, mainly through ingestion of invertebrates after insecticide application. Despite insecticide exposure in birds occurring mostly by ingestion, the impact of organophosphates on the avian digestive system has been poorly researched. In this work we used the Japanese quail (Coturnix japonica) as an avian model to study short-term microbial community responses to a single dose of trichlorfon at low concentration in three sample origins of the gastrointestinal tract (GIT): caecum, large intestine and faeces. Using next-generation sequencing of 16S rRNA gene amplicons as bacterial markers, the study showed that ingestion of insecticide caused significant changes in the GIT microbiome. Specifically, microbiota composition and diversity differed between treated and untreated quail. Insecticide-associated responses in the caecum showed differences between sexes which did not occur with the other sample types. In caecal microbiota, only treated females showed significant shifts in a number of genera within the Lachnospiraceae and the Enterobacteriaceae families. The major responses in the large intestine were a significant reduction in the genus Lactobacillus and increases in abundance of a number of Proteobacteria genera. All microbial shifts in faeces occurred in phylotypes that were represented at low relative abundances. In general, changes in microbiota possibly resulted from contrasting responses towards the insecticide, either positive (e.g., biodegrading bacteria) or negative (e.g., insecticide-susceptible bacteria). This study demonstrates the significant impact that organophosphate insecticides have on the avian gut microbiota; showing that a single small dose of trichlorfon caused dysbiosis in the GIT of the Japanese quail. Further research is necessary to understand the implications on birds' health, especially in females.


[Therapy of inherited diseases of platelet function. Interdisciplinary S2K guideline of the Permanent Paediatric Committee of the Society of Thrombosis and Haemostasis Research (GTH e. V.)].
Inherited disorders of platelet function are a heterogeneous group. For optimal prevention and management of bleeding, classification and diagnosis of the underlying defect are highly recommended. An interdisciplinary guideline for a diagnostic approach has been published (AWMF # 086-003 S2K; Hämostaseologie 2014; 34: 201-212). Underlying platelet disorder, platelet count, age and clinical situation modify treatment. Exclusive transfusion of platelet concentrates may be inappropriate as potentially adverse effects can outweigh its benefit. A stepwise and individually adjusted approach for restitution and maintenance of haemostasis is recommended. Administration of antifibrinolytics is generally endorsed, but is of particular use in Quebec disease. Restricted to older children, desmopressin is favourable in storage pool disease and unclassified platelet disorders. Although licensed only for patients with Glanzmann thrombasthenia and alloantibodies, in clinical practice rFVIIa is widely used in inherited platelet disorders with severe bleeding tendency. This guideline aims at presenting the best available advice for the management of patients with inherited platelet function disorders.


Impact of the reperfusion status for predicting the final stroke infarct using deep learning.
Predictive maps of the final infarct may help therapeutic decisions in acute ischemic stroke patients. Our objectives were to assess whether integrating the reperfusion status into deep learning models would improve their performance, and to compare them to current clinical prediction methods. We trained and tested convolutional neural networks (CNNs) to predict the final infarct in acute ischemic stroke patients treated by thrombectomy in our center. When training the CNNs, non-reperfused patients from a non-thrombectomized cohort were added to the training set to increase the size of this group. Baseline diffusion and perfusion-weighted magnetic resonance imaging (MRI) were used as inputs, and the lesion segmented on day-6 MRI served as the ground truth for the final infarct. The cohort was dichotomized into two subsets, reperfused and non-reperfused patients, from which reperfusion status specific CNNs were developed and compared to one another, and to the clinically-used perfusion-diffusion mismatch model. Evaluation metrics included the Dice similarity coefficient (DSC), precision, recall, volumetric similarity, Hausdorff distance and area-under-the-curve (AUC). We analyzed 109 patients, including 35 without reperfusion. The highest DSC were achieved in both reperfused and non-reperfused patients (DSC = 0.44 ± 0.25 and 0.47 ± 0.17, respectively) when using the corresponding reperfusion status-specific CNN. CNN-based models achieved higher DSC and AUC values compared to those of perfusion-diffusion mismatch models (reperfused patients: AUC = 0.87 ± 0.13 vs 0.79 ± 0.17, P < 0.001; non-reperfused patients: AUC = 0.81 ± 0.13 vs 0.73 ± 0.14, P < 0.01, in CNN vs perfusion-diffusion mismatch models, respectively). The performance of deep learning models improved when the reperfusion status was incorporated in their training. CNN-based models outperformed the clinically-used perfusion-diffusion mismatch model. Comparing the predicted infarct in case of successful vs failed reperfusion may help in estimating the treatment effect and guiding therapeutic decisions in selected patients.


Nonlinear effects of micro-cracks on long-wavelength symmetric Lamb waves.
For an elastic medium containing a homogeneous distribution of micro-cracks, an effective one-dimensional stress-strain relation has been determined with finite element simulations. In addition to flat micro-cracks, voids were considered that contain a Hertzian contact, which represents an example for micro-cracks with internal structure. The orientation of both types of micro-cracks was fully aligned or, for flat micro-cracks, totally random. For micro-cracks with Hertzian contacts, the case of random orientation was treated in an approximate way. The two types of defects were found to give rise to different degrees of non-analytic behavior of the effective stress-strain relation, which governs the nonlinear propagation of symmetric (S0) Lamb waves in the long-wavelength limit. The presence of flat micro-cracks causes even harmonics to grow linearly with propagation distance with amplitudes proportional to the amplitude of the fundamental wave, and gives rise to a static strain. The presence of the second type of defects leads to a linear growth of all harmonics with amplitudes proportional to the power 3/2 of the fundamental amplitude, and to a strain-dependent velocity shift. Simple expressions are given for the growth rates of higher harmonics of S0 Lamb waves in terms of the parameters occurring in the effective stress-strain relation. They have partly been determined quantitatively with the help of the FEM results for different micro-crack concentrations.