Assessing the Generalizability of a Deep Learning-based Automated Atrial Fibrillation Algorithm.
Automated detection of atrial fibrillation (AF) from electrocardiogram (ECG) traces remains a challenging task and is crucial for telemonitoring of patients after stroke. This study aimed to quantify the generalizability of a deep learning (DL)-based automated ECG classification algorithm. We first developed a novel hybrid DL (HDL) model using the PhysioNet/CinC Challenge 2017 (CinC2017) dataset (publicly available) that can classify the ECG recordings as one of four classes: normal sinus rhythm (NSR), AF, other rhythms (OR), and too noisy (TN) recordings. The (pre)trained HDL was then used to classify 636 ECG samples collected by our research team using a handheld ECG device, CONTEC PM10 Portable ECG Monitor, from 102 (age: 68 ± 15 years, 74 male) outpatients of the Eastern Heart Clinic and inpatients in the Cardiology ward of Prince of Wales Hospital, Sydney, Australia. The proposed HDL model achieved average test F1-score of 0.892 for NSR, AF, and OR, relative to the reference values, on the CinC2017 dataset. The HDL model also achieved an average F1-score of 0.722 (AF: 0.905, NSR: 0.791, OR: 0.471 and TN: 0.342) on the dataset created by our research team. After retraining the HDL model on this dataset using a 5-fold cross validation method, the average F1-score increased to 0.961. We finally conclude that the generalizability of the HDL-based algorithm developed for AF detection from short-term single-lead ECG traces is acceptable. However, the accuracy of the pre-trained DL model was significantly improved by retraining the model parameters on the new dataset of ECG traces.


Weed control in conventional and transgenic maize with resistance to glyphosate.
The researches were conducted in order to observe the behaviour of conventional and glyphosate resistant transgenic maize to different weed control methods. In this paper, the obtained results are presented. The study was conducted in experimental years 2008-2009 in the frame of Didactical Station USAMVB Timisoara. In order to conduct this study, 4 variants cultivated with conventional maize DKC 5143 and 8 variants cultivated with transgenic maize DKC-MON88017 with resistance against Diabrotica virgifera virgifera and to glyphosate. The efficacy of weed control methods was assessed, as well as the herbicide selectivity to cultivated maize hybrid. The weed coverage degree in control plot (V2) was 304 weeds/sqm in the first year and 465 weeds/sqm in the second year. In the variants cultivated with transgenic maize the control was up to 90% much more than control percent achieved in conventional variants. Although, in order to achieve an efficient control (higher than 95%), even to transgenic maize, two glyphosate sequential treatments has to be done. The yield results were positive correlated to the different control methods. However those were affected by climatic conditions recorded in experimental years.


Reconstitution of active dimeric ribulose bisphosphate carboxylase from an unfoleded state depends on two chaperonin proteins and Mg-ATP.
In vitro reconstitution of active ribulose bisphosphate carboxylase (Rubisco) from unfolded polypeptides is facilitated by the molecular chaperones: chaperonin-60 from Escherichia coli (groEL), yeast mitochondria (hsp60) or chloroplasts (Rubisco sub-unit-binding protein), together with chaperonin-10 from E. coli (groES), and Mg-ATP. Because chaperonins are ubiquitous, a conserved Mg-ATP-dependent mechanism exists that uses the chaperonins to facilitate the folding of some other proteins.


Effects of luminal thymol on epithelial transport in human and rat colon.
Gut lumen is continually exposed to a great variety of agents, including noxious compounds. Chemical receptors that detect the luminal environment are thought to play an important role as sensors and to modulate gastrointestinal functions. Recently, it has been reported that odorant receptors (ORs) are expressed in the small intestinal mucosa and that odorants stimulate serotonin secretion. However, ion transport in the responses to odorants has rarely been discussed, particularly in relation to the large intestine. In the present study, we examined the effects of the OR ligand thymol on ion transport in human and rat colonic epithelia using an Ussing chamber. In the mucosal-submucosal preparations, the mucosal addition of thymol evoked anion secretion concentration dependently. In addition, dextran (4 kDa) permeability was enhanced by the mucosal treatment with thymol. The response to thymol was not affected by tetrodotoxin (TTX) or piroxicam treatments in human or rat colon. Thymol-evoked electrogenic anion secretion was abolished under Ca(2+)-free conditions or mucosal treatment with transient receptor potential (TRP) A1 blocker (HC-030031). Pretreatment of thymol did not affect electrical field stimulation-evoked anion secretion but significantly attenuated short-chain fatty acid-evoked secretion in a concentration-dependent manner. OR1G1 and TRPA1 expression was investigated in isolated colonic mucosa by RT-PCR. The present results provide evidence that the OR ligand thymol modulates epithelial permeability and electrogenic anion secretion in human and rat colon. The anion secretion by luminal thymol is most likely mediated by direct activation of TRPA1 channel. We suggest that the sensing and responding to odorants in the colon also plays a role in maintaining intestinal homeostasis.


Cord blood DNA methylation signatures associated with preeclampsia are enriched for cardiovascular pathways: insights from the VDAART trial.
Preeclampsia has been associated with maternal epigenetic changes, in particular DNA methylation changes in the placenta. It has been suggested that preeclampsia could also cause DNA methylation changes in the neonate. We examined DNA methylation in relation to gene expression in the cord blood of offspring born to mothers with preeclampsia. This study included 128 mother-child pairs who participated in the Vitamin D Antenatal Asthma Reduction Trial (VDAART), where assessment of preeclampsia served as secondary outcome. We performed an epigenome-wide association study of preeclampsia and cord blood DNA methylation (Illumina 450 K chip). We then examined gene expression of the same subjects for validation and replicated the gene signatures in independent DNA methylation datasets. Lastly, we applied functional enrichment and network analyses to identify biological pathways that could potentially be involved in preeclampsia. In the cord blood samples (n = 128), 263 CpGs were differentially methylated (FDR <0.10) in preeclampsia (n = 16), of which 217 were annotated. Top pathways in the functional enrichment analysis included apelin signaling pathway and other endothelial and cardiovascular pathways. Of the 217 genes, 13 showed differential expression (p's < 0.001) in preeclampsia and 11 had been previously related to preeclampsia (p's < 0.0001). These genes were linked to apelin, cGMP and Notch signaling pathways, all having a role in angiogenic process and cardiovascular function. Preeclampsia is related to differential cord blood DNA methylation signatures of cardiovascular pathways, including the apelin signaling pathway. The association of these cord blood DNA methylation signatures with offspring's long-term morbidities due to preeclampsia should be further investigated. VDAART is funded by National Heart, Lung, and Blood Institute grants of R01HL091528 and UH3OD023268. HMK is supported by Jane and Aatos Erkko Foundation, Paulo Foundation, and the Pediatric Research Foundation. HM is supported by K01 award from NHLBI (1K01HL146977-01A1). PK is supported by K99HL159234 from NIH/NHLBI.


Urinary 6 beta-hydroxyprednisolone excretion indicates enhanced prednisolone catabolism.
Patients with enhanced catabolism of prednisolone exhibit a reduction in steroid efficacy. Since prednisolone never reaches steady state at the usual dosage, the clearance rate can only be estimated by determining the area under the plasma concentration-time curve after an i.v. dose of prednisolone. To obviate that time-consuming procedure for screening of patients with enhanced prednisolone metabolism, we developed a quantitative assay for 6 beta-hydroxyprednisolone in urine to determine whether the urinary excretion of 6 beta-hydroxyprednisolone increases concomitantly with the clearance rate of total and of unbound prednisolone in subjects with an enhanced catabolism of prednisolone. The results are given separately for males and females because there is some debate as to whether females compared to males exhibit a higher capacity of the liver to hydroxylate steroids at the sixth position. Before and during the administration of phenytoin the total body clearance of total and of unbound prednisolone and the urinary excretion of prednisolone and of 6 beta-hydroxyprednisolone were determined in six female and eight male volunteers. The fraction of the i.v. dose of prednisolone excreted in urine as 6 beta-hydroxyprednisolone was higher in females than in males (p less than 0.001) and increased during phenytoin dosing from 6.4%-10.4% (range) to 15.6%-20.4% in females and from 2.4%-7.2% to 12.2%-18.3% in males. The ratio of 6 beta-hydroxyprednisolone to prednisolone in urine increased linearly with the nonrenal clearance of both total and unbound prednisolone. This is the first demonstration that the fractional excretion of 6 beta-hydroxyprednisolone increased after the induction of the microsomal liver enzymes. Thus measurements of urinary 6 beta-hydroxyprednisolone excretion may be of potential value to detect patients with enhanced catabolism of prednisolone.


Inhibition of beta-amyloid precursor protein gene in SK-N-SH cells by piperlonguminine/dihydropiperlonguminine components separated from Chinese herbal medicine Futokadsura stem.
Alzheimer disease (AD) is a progressive neurodegenerative disease characterized by progressive cognitive and memory decline. Amyloid precursor protein (APP) is a transmembrane protein, it has been known to play an important role in AD pathogenesis. Previous studies have shown that a Chinese herb Futokadsura stem can selectively inhibit the expression of amyloid precursor protein (APP) gene. We want to find the effective components in Futokadsura stem which have the inhibitory effect. Futokadsura stem was separated and purified with chemical methods, and then different separation components were added on SK-N-SH cells in different concentrations. Using MTT methods, we detected proliferation activity of SK-N-SH cells which were treated with different separation components of Futokadsura stem. Using RT-PCR, Western blot methods, we detected APP gene expression in SK-N-SH cells after they are treated with different Futokadsura stem separation components. We found that piperlonguminine/dihydropiperlonguminine components (1:0.8) separated from Futokadsura stem acetic ether extracts could selectively inhibit the expression of APP gene in SK-N-SH cells in mRNA and protein levels. This inhibition effect is concentration-dependent. Under experimental concentrations, the components did not affect the proliferation activity of SK-N-SH cells. These data suggest that piperlonguminine/dihydropiperlonguminine components are the effective components in Futokadsura stem which can inhibit the expression of APP gene.


Improving biosensor accuracy and speed using dynamic signal change and theory-guided deep learning.
False results and time delay are longstanding challenges in biosensing. While classification models and deep learning may provide new opportunities for improving biosensor performance, such as measurement confidence and speed, it remains a challenge to ensure that predictions are explainable and consistent with domain knowledge. Here, we show that consistency of deep learning classification model predictions with domain knowledge in biosensing can be achieved by cost function supervision and enables rapid and accurate biosensing using the biosensor dynamic response. The impact and utility of the methodology were validated by rapid and accurate quantification of microRNA (let-7a) across the nanomolar (nM) to femtomolar (fM) concentration range using the dynamic response of cantilever biosensors. Data augmentation and cost function supervision based on the consistency of model predictions and experimental observations with the theory of surface-based biosensors improved the F1 score, precision, and recall of a recurrent neural network (RNN) classifier by an average of 13.8%. The theory-guided RNN (TGRNN) classifier enabled quantification of target analyte concentration and false results with an average prediction accuracy, precision, and recall of 98.5% using the initial transient or entire dynamic response, which is indicative of high prediction accuracy and low probability of false-negative and false-positive results. Classification scores were used to establish new relationships among biosensor performance characteristics (e.g., measurement confidence) and design parameters (e.g., inputs and hyperparameters of classification models and data acquisition parameters) that may be used for characterizing biosensor performance.


EEC syndrome sans clefting: variable clinical presentations in a family.
Ectrodactyly, ectodermal dysplasia and cleft palate/lip syndrome (EEC) is a rare autosomal dominant syndrome with varied presentation and is actually a multiple congenital anomaly syndrome leading to intra- and interfamilial differences in severity because of its variable expression and reduced penetrance. The cardinal features include ectrodactyly, sparse, wiry, hypopigmented hair, peg-shaped teeth with defective enamel and cleft palate/lip. A family comprising father, daughter and son presented to us with split hand-split foot deformity (ectrodactyly), epiphora, hair changes and deafness with variable involvement in each family member.


Rhizobia from Lanzarote, the Canary Islands, that nodulate Phaseolus vulgaris have characteristics in common with Sinorhizobium meliloti isolates from mainland Spain.
The stable, low-molecular-weight (LMW) RNA fractions of several rhizobial isolates of Phaseolus vulgaris grown in the soil of Lanzarote, an island of the Canary Islands, were identical to a less-common pattern found within Sinorhizobium meliloti (assigned to group II) obtained from nodules of alfalfa and alfalfa-related legumes grown in northern Spain. The P. vulgaris isolates and the group II LMW RNA S. meliloti isolates also were distinguishable in that both had two conserved inserts of 20 and 46 bp in the 16S-23S internal transcribed spacer region that were not present in other strains of S. meliloti. The isolates from P. vulgaris nodulated bean but not Medicago sativa, while those recovered from Medicago, Melilotus, and Trigonella spp. nodulated both host legumes. The bean isolates also were distinguished from those of Medicago, Melilotus, and Trigonella spp. by nodC sequence analysis. The nodC sequences of the bean isolates were most similar to those reported for S. meliloti bv. mediterranense and Sinorhizobium fredii bv. mediterranense (GenBank accession numbers DQ333891 and AF217267, respectively). None of the evidence placed the bean isolates from Lanzarote in the genus Rhizobium, which perhaps is inconsistent with seed-borne transmission of Rhizobium etli from the Americas to the Canaries as an explanation for the presence of bean-nodulating rhizobia in soils of Lanzarote.


The relation between serum albumin levels and risk of coronary heart disease in the Multiple Risk Factor Intervention Trial.
The relation between serum albumin levels and subsequent incidence of myocardial infarction and coronary heart disease deaths was evaluated using stored serum from the Multiple Risk Factor Intervention Trial (MRFIT). There were 91 coronary heart disease deaths, 113 myocardial infarction patients, and 405 controls matched to cases within 5 years of age, treatment group, and clinic site. There was a highly significant inverse relation between serum albumin level and risk of coronary heart disease. Individuals with a baseline level of serum albumin greater than or equal to 4.7 g/dl had an odds ratio of 0.45 as compared with individuals with a baseline level of serum albumin less than 4.4 g/dl. The relation persisted after adjusting for other cardiovascular risk factors (blood pressure, smoking, and serum cholesterol). The association was stronger for coronary heart disease deaths than for surviving myocardial infarction patients, and for cigarette smokers as compared with cigarette nonsmokers. The deaths studied occurred in the time period at least 6 years after the sera had been obtained and up to 10.5 years of follow-up, and the myocardial infarctions studied occurred within the first 6.5 years of follow-up. There was no consistent relation between time and death due to coronary heart disease or myocardial infarction and albumin levels. Albumin levels are related to the acute phase reaction. Lower albumin levels may be a marker of persistent injury to arteries and progression of atherosclerosis and thrombosis. The consistent relation between albumin and coronary heart disease risk requires further evaluation.


Rapid response to meningococcal disease cluster in Foya district, Lofa County, Liberia January to February 2018.
Early detection of disease outbreaks is paramount to averting associated morbidity and mortality. In January 2018, nine cases including four deaths associated with meningococcal disease were reported in three communities of Foya district, Lofa County, Liberia. Due to the porous borders between Lofa County and communities in neighboring Sierra Leone and Guinea, the possibility of epidemic spread of meningococcal disease could not be underestimated. The county incidence management system (IMS) was activated that coordinated the response activities. Daily meetings were conducted to review response activities progress and challenges. The district rapid response team (DRRT) was the frontline responders. The case based investigation form; case line list and contacts list were used for data collection. A data base was established and analysed daily for action. Tablets Ciprofloxacin were given for chemoprophylaxis. Sixty-seven percent (67%) of the cases were males and also 67% of the affected age range was 3 to 14 years and attending primary school. The attack rate was 7/1,000 population and case fatality rate was 44.4 % with majority of the deaths occurring within 24-48 hours of symptoms onset. Three of the cases tested positive for Neisseria Meningitidis sero-type W while six cases were Epi-linked. None of the cases had recent meningococcal vaccination and no health-worker infections were registered. This cluster of cases of meningococcal disease during the meningitis season in a country that is not traditionally part of the meningitis belt emphasized the need for strengthening surveillance, preparedness and response capacity to meningitis.


Rotational cerebral angiography: procedure and value.
Rotational cerebral angiography provides continuously changing projections and correspondingly excellent visualization of aneurysms or other vascular malformations requiring special or oblique views. Rotation angiography as part of the routine angiography gives a three-dimensional impression with one injection and series of films. Although additional subtraction technique is not possible and the injection time has to be prolonged, the technique is nevertheless advantageous compared with magnification angiography and angiotomography in the preoperative evaluation of cerebral vascular malformations.


Age-dependent and differential effects of Smad7ΔEx1 on neural progenitor cell proliferation and on neurogenesis.
We recently reported that young (3 to 4months old) mice lacking Exon 1 of the Smad7 gene (S7ΔEx1 mice) show enhanced proliferation of neural stem and progenitor cells (NPCs) in the hippocampal dentate gyrus (DG) and in the subventricular zone (SVZ) of the lateral ventricles. It remained unclear, however, whether this phenotype would persist along aging, the latter typically being associated with a profound decrease in neurogenesis. Analysis of NPCs' proliferation based on the cell cycle marker PCNA in 12month-old S7ΔEx1 mice revealed a reversal of the phenotype. Hence, in contrast to their younger counterparts, 12month-old S7ΔEx1 mice had a reduced number of proliferating cells, compared to wildtype (WT) mice. At the same time, the survival of newly generated cells was enhanced in the aged transgenic animals. 12month-old S7ΔEx1 mice further displayed a reduced level of neurogenesis based on the numbers of cells expressing doublecortin (DCX), a marker for newborn neurons. The reduced neurogenesis in aged S7ΔEx1 mice was not due to a stem cell depletion, which might have occurred as a consequence of hyperproliferation in the young mice, since the number of Nestin and Sox2 positive cells was similar in WT and S7ΔEx1 mice. Instead, Nestin positive cells in the DG as well as primary neurosphere cultures derived from 12month-old S7ΔEx1 mice had a reduced capability to proliferate. However, after passaging, when released from their age- and niche-associated proliferative block, neurospheres from aged S7ΔEx1 mice regained the hyperproliferative property. Further, pSmad2 antibody staining intensity was elevated in the DG and SVZ of 12-month old transgenic compared to WT mice, indicating increased intracellular TGF-beta signaling in the aged S7ΔEx1 mice. In summary, this points toward differential effects of S7ΔEx1 on neurogenesis: (i) a hyperproliferation in young animals caused by a cell autonomous mechanism, and (ii) a TGF-beta dependent modulation of neurogenesis in aged S7ΔEx1 animals that abrogates the cell-intrinsic hyperproliferative properties and results in reduced proliferation, increased stem cell quiescence, and enhanced survival of newly generated cells.


Development and measurement properties of the AxEL (attitude toward education and advice for low-back-pain) questionnaire.
Clinician time and resources may be underutilised if the treatment they offer does not match patient expectations and attitudes. We developed a questionnaire (AxEL-Q) to guide clinicians toward elements of first-line care that are pertinent to their patients with low back pain. We used guidance from the COSMIN consortium to develop the questionnaire and evaluated it in a sample of people with low back pain of any duration. Participants were recruited from the community, were over 18 years and fluent in English. Statements that represented first-line care were identified. Semantic scales were used to measure attitude towards these statements. These items were combined to develop the questionnaire draft. Construct validity was evaluated with exploratory factor analysis and hypotheses testing, comparing to the Back Beliefs Questionnaire and modified Pain Self-Efficacy Questionnaire. Reliability was evaluated and floor and ceiling effects calculated. We recruited 345 participants, and had complete data for analysis for 313 participants. The questionnaire draft was reduced to a 3-Factor questionnaire through exploratory factor analysis. Factor 1 comprised 9 items and evaluated Attitude toward staying active, Factor 2 comprised 4 items and evaluated Attitude toward low back pain being rarely caused by a serious health problem, Factor 3 comprised 4 items and evaluated Attitude toward not needing to know the cause of back pain to manage it effectively. There was a strong inverse association between each factor and the Back Beliefs Questionnaire and a moderate positive association with the modified Pain Self-Efficacy Questionnaire. Each independent factor demonstrated acceptable internal consistency; Cronbach α Factor 1 = 0.92, Factor 2 = 0.91, Factor 3 = 0.90 and adequate interclass correlation coefficients; Factor 1 = 0.71, Factor 2 = 0.73, Factor 3 = 0.79. This study demonstrates acceptable construct validity and reliability of the AxEL-Q, providing clinicians with an insight into the likelihood of patients following first-line care at the outset.


A Rat Model with Multivalve Calcification Induced by Subtotal Nephrectomy and High-Phosphorus Diet.
Chronic kidney disease (CKD) with known valve calcification (VC) places individuals at high risk of cardiovascular disease. The study of VC in CKD is challenging due to the lack of a suitable research model. Here, we established a rat model of multivalve calcification induced by subtotal nephrectomy and a high-phosphate (HP) diet and analyzed the valve characteristics. We established a CKD model in Sprague-Dawley rats by performing 5/6 nephrectomy (5/6Nx) followed by feeding with chow containing different phosphate concentrations for 8, 12, or 16 weeks. The rats were divided into 4 groups: sham+normal phosphate (NP, 0.9% P), sham+high phosphate (HP, 2.0% P), 5/6Nx+NP, and 5/6Nx+HP. Serum creatinine (Scr), blood urea nitrogen (BUN), parathyroid hormone (PTH), calcium, phosphorus, and 24-h urine protein levels were investigated. Pathological examinations included histological characterization, safranin staining, Alcian blue staining, and von Kossa staining at different time points. Using nanoanalytical electron microscopy, we examined valves from rats in the 5/6Nx+HP and sham+HP groups and detected spherical particles using energy-dispersive spectroscopy (EDS) to observe microscopic changes in the valves. In addition, the calcified tissues were analyzed for phase and crystallization properties using an X-ray powder diffractometer. The rats in the 5/6Nx+HP and 5/6Nx+NP groups presented with increased levels of Scr, BUN, and 24-h urine protein compared with those of the rats in the sham+HP and sham+NP groups. High levels of PTH were observed, and hematoxylin and eosin staining and immunohistochemistry for proliferating cell nuclear antigen showed parathyroid hyperplasia in rats in the 5/6Nx+HP group but not in the 5/6Nx+NP group. In rats in the 5/6Nx+HP group, extracellular matrix glycosylation was observed in the aortic valve in the 12th week and the mitral valve in the 16th week. In the 16th week, chondrocytes appeared in the aortic valve, as confirmed by immunofluorescence and Western blotting. Calcified particles mainly composed of phosphorus and calcium were observed in both the aortic and mitral valves by transmission electron microscopy and scanning electron microscopy (SEM). The main mineral component of the calcified aortic valve particles was hydroxyapatite [Ca5(PO4)3(OH)], as shown by X-ray diffraction. However, there were no obvious differences in heart function between rats in the 5/6Nx+HP and sham+HP groups. Our findings demonstrate that multivalve calcification is involved in CKD following 16-week HP and that hydroxyapatite [Ca5(PO4)3(OH)] is the main component of the calcified aortic valve particles of rats in the 5/6Nx+HP group.


Unprecedented stabilization of cobalt(II) in a tetrahedral S2O2 environment: the use of a redox-noninnocent ligand.
The reaction of Zn(II) and Co(II) with thiosalicylic acid, o-HSC6H4COOH, and its methyl ester has led to the following complexes: [Zn(SC6H4COO)] (1), (NEt4)Na[Zn(SC6H4COO)2].H2O (2), (NEt4)2Na[Co(SC6H4COO)3].2H2O (3), (NEt4)3Na3[(Co(SC6H4COO)3)2].6MeOH (4), [Zn(SC6H4COOMe)2] (5), and [Co(SC6H4COOMe)n], n = 2 (6), 3 (7). These ligands have not allowed stabilization of Co(II) in a sulfur-oxygen coordination environment. The structures of complexes 2-4 and 7 have been determined crystallographically. Those of 2-4 show significant similarities such as the behavior of the -SC6H4COO- anion as chelating ligand and the involvement of sodium ions as a structural element. Thus, the structure of the [Na(Zn(SC6H4COO)2)(H2O)]- anion in complex 2 can be described as infinite chains of consecutive [Zn(SC6H4COO)2]2- metalloligands linked by [Na(H2O)]+ centers, that of the [Na(Co(SC6H4COO)3(H2O)2)]2(4-) anion in 3 as a centrosymmetric tetranuclear Co2Na2 dimer with a (CoIII(S[symbol: see text]O)3)Na(mu-H2O)2Na(CoIII(S[symbol: see text]O)3) core, and that of the pentanuclear [Na3(Co(SC6H4COO)3)2(MeOH)6]3- anion in 4 as two dinuclear [(CoIII(S[symbol: see text]O)3)Na(MeOH)3] fragments linked to a central sodium ion, which appears to be the first structurally characterized example of a NaS6 site. The use of the o-HSC6H4COOMe ligand allowed the synthesis of [Co(SC6H4COOMe)2] (6) but not its full structural characterization. Instead, [Co(SC6H4COOMe)3] (7) was obtained and structurally characterized. It consists of mononuclear molecules containing an octahedral CoIIIS3O3 core. The selection of 2,2-diphenyl-2-mercaptoacetic acid as ligand with reductive properties has afforded the first mononuclear complex containing a CoIIS2O2 core and thus an unprecedented model for Co(II)-substituted metalloproteins containing tetrahedral MS2O2 active sites. The synthesis and full structural characterization of the isostructural complexes (NEt4)2[Zn(Ph2C(S)COO)2] (8) and (NEt4)2[Co(Ph2C(S)COO)2] (9) show that they consist of discrete [M(Ph2C(S)COO)2]2- anions, with a distorted tetrahedral coordination about the metal. In addition, the stability conferred by the ligand on the CoIIS2O2 core has allowed its characterization in solution by paramagnetic 1D and 2D 1H NMR studies. The longitudinal relaxation times of the hyperfine-shifted resonances and NOESY spectra have led to the assignment of all resonances of the cobalt complex and confirmed that it maintains its tetrahedral geometry in solution. Magnetic measurements (2-300 K) for complex 9 and 9.2H2O are in good agreement with distorted tetrahedral and octahedral environments, respectively.


Strategic reconstruction of macrophage-derived extracellular vesicles as a magnetic resonance imaging contrast agent.
A contrast agent (CA) in magnetic resonance imaging (MRI) is now an essential add-on to obtain high-quality contrast-enhanced anatomical images for disease diagnosis and monitoring the treatment response. However, the rapid elimination of CAs by the immune system and excretion by the renal route has limited its application. As a result, the CA dose for effective contrast is ever-increasing, resulting in toxic side effects such as gadolinium (Gd) related nephrogenic systemic fibrosis (NSF) toxicity. Considering the widespread application of Gd-based CAs, it is now very important to revisit their formulation in order to improve their local concentration and minimize their dose while achieving clinical goals. Therefore, we have adapted a unique strategy to maximize Gd delivery to the target site using macrophage cell-derived extracellular vesicles (EVs) reconstructed with a Gd-conjugated liposomal system herein called gadolinium infused hybrid EVs (Gd-HEVs). We hypothesize that Gd-HEVs, owing to the presence of immune cell-derived EV protein cargo, can effectively disguise themselves as a biological entity, prolong the retention time for contrast enhancement, and show tumor specificity. Incorporation of Gd into nanoformulations can enhance the longitudinal relaxivity r1 by reducing the tumbling rate of paramagnetic metal complexes. Here, Gd-HEVs showed a higher r1 relaxivity of 9.86 mM-1 s-1 compared to 3.98 mM-1 s-1 of Magnevist® at an equivalent Gd concentration, when measured by clinical 3T MRI. This will allow us to reduce the clinically used Gd concentration about three-fold while maintaining contrast in the clinical window thereby supporting our hypothesis. Furthermore, Gd-HEVs showed a preferential cellular interaction and accumulation towards cancer cells compared to non-cancer cells, both in vitro and in vivo. More importantly, Gd-HEVs showed excellent contrast enhancement in the blood vasculature with a higher retention time compared to its counterpart, Magnevist®. Our study successfully showed that the incorporation of Gd in the EV framework can help to enhance the contrast ability, and therefore it can be a platform technology for the development of safer MRI contrast agents.


Chalcogenation of the 1,4-C2P4 ring: oxidation, isomerization, insertion, and ring contraction.
The reaction of H2C(PCl2)2 with 4 equiv of (t)BuMgCl in tetrahydrofuran (THF) produces 1,4-(CH2)2(P(t)Bu)4, 1, in about 65% yield. This six-membered ring reacts directly with elemental sulfur or selenium in toluene at low temperatures to give the mono- and dichalcogenides 1,4-(CH2)2(P(t)BuE)(P(t)Bu)3 (E = S, 2a, E = Se, 2b) and 1,4-(CH2)2-2,5-(P(t)BuE)2(P(t)Bu)2 (E = S, 3a, E = Se, 3b). X-ray structural determinations showed that 3a and 3b are isostructural in the solid state; the six-membered C2P4 ring exhibits a twist-boat geometry with chalcogen substituents in syn positions in each case. Density functional theory (DFT) calculations for the three possible isomers of disubstitution were performed to elucidate the factors that favor the 2,5-isomer. Thermal isomerism was observed in solutions of 3b or 3a in toluene at 60 and 95 °C, respectively, to give the corresponding 2,6-isomers. With an excess of chalcogen in toluene at reflux, the four-membered rings (H2C)(P(t)BuE)2E (E = S, 4a, E = Se, 4b) were obtained and identified by multinuclear NMR spectroscopy and single crystal X-ray crystallography, which showed the (t)Bu groups in a trans orientation with respect to the CP2E ring. With a large excess of chalcogen, the five-membered rings (H2C)(P(t)BuE)2E2 (E = S, 5a, E = Se, 5b) were also observed; the X-ray structure of 5b revealed a half-envelope conformation for the CP2Se2 ring. The direct reaction of 4a with sulfur in boiling toluene does not produce 5a, whereas 5b is formed slowly and in low yields from 4b and selenium under similar conditions. On the basis of DFT calculations of the relative energies of likely intermediates, chalcogen insertion into the P-P bonds of 3a and 3b to give eight-membered C2P4E2 rings, followed by monomerization, is proposed as a feasible pathway for the formation of the four-membered CP2E heterocycles 4a and 4b.


The antidepressant-like effects of sinomenine in mice: a behavioral and neurobiological characterization.
Sinomenine is a bioactive alkaloid extracted from Sinomenium acutum. Here, we investigated the antidepressant effects of sinomenine in mice. The antidepressant actions of sinomenine were first examined in the forced-swim test and the tail-suspension test, and then assessed in the chronic social defeat stress (CSDS) model of depression. Changes in the brain-derived neurotrophic factor (BDNF) signaling pathway after CSDS and sinomenine treatment were also investigated. A tryptophan hydroxylase inhibitor and a BDNF signaling inhibitor were also used to determine the pharmacological mechanisms of sinomenine. It was found that sinomenine induced antidepressant-like effects in the forced-swim test and tail-suspension test without affecting the locomotor activity of mice. Sinomenine also prevented the CSDS-induced depressive-like symptoms. Moreover, sinomenine fully restored the CSDS-induced decrease in the hippocampal BDNF signaling pathway, whereas a BDNF signaling inhibitor, but not a tryptophan hydroxylase inhibitor, blocked the antidepressant effects of sinomenine. In conclusion, sinomenine exerts antidepressant effects in mice by promoting the hippocampal BDNF signaling pathway.


Effects of limit feeding corn or dried distillers grains with solubles at 2 intakes during the growing phase on the performance of feedlot cattle.
Energy density in growing diets may affect carcass quality of cattle; however, few reports have described the impact of energy source. The objectives of this research were to determine effects of source [dried distillers grains with solubles (DDGS) vs. corn] and amount (limit-fed to gain 0.9 vs. 1.4 kg of BW/d) of energy during the growing phase on feedlot performance and marbling. Angus-cross steers (144 head) were blocked by BW (average initial BW = 252 ± 36 kg), allotted within each block to 8 pens (6 steers/pen, 24 pens total), and randomly assigned to 1 of 4 feeding systems in a 2 × 2 factorial arrangement of treatments: 1) 65% DDGS fed to gain 0.9 kg of BW/d, 2) 65% DDGS fed to gain 1.4 kg of BW/d, 3) 65% corn fed to gain 0.9 kg of BW/d, and 4) 65% corn fed to gain 1.4 kg of BW/d. Fecal grab samples were collected on d 52 of the growing phase to determine digestibility of DM, ADF, NDF, ether extract (EE), and CP. After the 98-d growing phase, all steers were fed the same finishing diet. Steers were slaughtered by pen when average BW within the pen was 544, 522, and 499 kg for the large, medium, and small BW blocks, respectively. Average daily gain and DMI differed (P<0.01) by design during the growing phase. Compared with the corn-based diets, digestibilities of DM, NDF, and EE were decreased (P<0.02) when DDGS-based diets were fed during the growing phase, whereas the digestibility of N was increased (P<0.01). The ADG was greatest (P=0.02) during the finishing phase for steers fed to gain 0.9 kg of BW/d initially, but source of energy during the growing phase did not affect (P=0.24) finishing phase ADG. Steers fed to gain 0.9 kg of BW/d during the growing phase also had less backfat (P=0.08), decreased USDA yield grades (P=0.03), and greater LM area (P<0.01) than steers fed to gain 1.4 kg of BW/d. There was an interaction between energy source and amount for marbling scores (P=0.02). Steers fed corn-based diets to gain 0.9 kg of BW/d during the growing phase had the most marbling, whereas those fed to gain 0.9 kg of BW/d on DDGS had the least marbling; the remaining feeding systems were intermediate. Overall ADG and DMI were affected (P < 0.06) by both source and amount of energy fed during the growing phase. Feeding the DDGS-based diet to achieve greater ADG during the growing phase increased marbling, whereas feeding the corn-based diet to increase ADG during the growing phase decreased marbling.


Further studies of the stimulation of DNA synthesis in cultures of spleen cell suspensions by homologous cells in inbred strains of mice and rats.
The early proliferative response previously demonstrated in rabbits has now been shown to follow the mixing of spleen cell suspensions from 2 inbred strains of mice or rats. The size of the response is comparable to that seen in cells from hyperimmune animals exposed to antigen in vitro. Autoradiographs of cells from stimulated cultures showed 1 to 4 per cent of the total population had incorporated thymidine. Modifications in the conditions necessary for the culture of mice and rat spleen cell suspensions and the measurement of thymidine incorporation are described. No responses were observed in isologous mixes. The responses obtained on mixing individual pairs of spleens from different strains showed relatively little variation. Responses were obtained in all of the 21 possible combinations between 7 inbred strains of mice. Responses were obtained when parental cells were mixed with their F1 hybrids. Analysis of these responses showed that, in every case, parental-F1 hybrid responses were less intense than those between the 2 parents. It was shown that there was no inherent defect in the ability of the hybrid cells to respond when mixed with an unrelated strain. The results suggested that the hybrid cells made no response to the parent cells although this was not conclusively established. This has been taken as circumstantial evidence that the response is immunological in nature. The significance of the vigor of the response and the large fraction of the immunologically competent cells that take part is discussed.


Chromatin structure of erythroid-specific genes of immature and mature chicken erythrocytes.
The beta-globin and histone H5 genes are transcriptionally active in immature chicken erythrocytes and potentially active in mature erythrocytes. In both immature and mature erythrocytes, the majority of these erythroid-specific gene sequences are located in two chromatin fractions: the low-salt-insoluble residual nuclear material and the 0.15 M-NaCl-soluble oligo- and poly-nucleosomes. These salt-soluble chromatin fragments are enriched in hyperacetylated species of H4 and H2B, ubiquitinated and polyubiquitinated species of H2A and H2B and are depleted of linker histones H1 and H5. The competent, transcriptionally inactive embryonic epsilon-globin gene, which is part of the DNAase I-sensitive beta-globin domain, is highly enriched in the 0.15 M-NaCl-soluble polynucleosome fraction but not in the insoluble nuclear material. The repressed vitellogenin gene shows no enrichment in either of these fractions. These results suggest that only those genes that are expressed or have the potential for expression are enriched in the low-salt-insoluble nuclear material of immature or mature erythrocytes. The enrichment of active genes in the low-salt-insoluble residual nuclear material of immature erythrocytes is not dependent on on-going transcription, the presence of RNA or changes in the amount of acetylated histone species. Our results are consistent with the hypothesis that active and potentially active genes are insoluble because of the presence of preinitiation transcription complexes.


African American women's views of factors impacting preterm birth.
To explore pregnant African American women's views of factors that may impact preterm birth. Qualitative descriptive exploratory cross-sectional design. A convenience sample of 22 low-risk pregnant African American women participated in focus group interviews. Women were asked questions regarding their belief about why women have preterm birth and factors impacting preterm birth. Data were analyzed using content analysis. Pregnant African American women encounter multiple physical, psychological, and social stressors. The four themes included knowledge of preterm birth, risk factors for preterm birth, protective factors for preterm birth, and preterm birth inevitability. The risk factors for preterm birth were health-related conditions, stressors, and unhealthy behaviors. Stressors included lack of social and financial support, interpersonal conflicts, judging, dangerous neighborhoods, racism, and pregnancy- and mothering-related worries. Protective factors for preterm birth included social support and positive coping/self-care. Clinicians may use the results of this study to better understand women's perceptions of factors that affect preterm birth, to educate women about risk factors for preterm birth, and to develop programs and advocate for policies that have the potential to decrease health disparities in preterm birth.


B-type natriuretic peptide trends after pediatric heart transplantation.
BNP is increasingly utilized in the management of pediatric HT recipients. Performing a retrospective single-center chart review, we sought to describe BNP changes during the first year after HT and identify factors that affect its trend. After exclusion for rejection, 316 BNP levels from 50 patients were evaluated. BNP underwent an exponential decline 120 days after HT followed by a plateau. Log10 BNP decline strongly correlated with time (r = -0.70, p < 0.0001). Initial BNP was less in pretransplant VAD (p = 0.0016) and lower post-HT inotrope use (p = 0.0043). Infant recipients, IT >4 h, and those bridged medically were associated with higher plateau BNP. Multivariable logistic regression demonstrated IT >4 h independently predicted plateau BNP in the upper quartile (OR 7.1, p = 0.02). No significant change in BNP coincided with rejection (N = 6 patients) without severe hemodynamic compromise. BNP correlated modestly with right atrial pressure (r = 0.4652, p < 0.0001) and pulmonary capillary wedge pressure (r = 0.2660, p < 0.001), but poorly with echocardiogram (r = -0.18, p = 0.003). Trending BNP could help provide insight into how the graft recovers after HT and IT >4 h independently predicted higher plateau BNP and may reflect subtle changes in graft performance.


Lifestyle Medicine Around the World: Lifestyle Medicine in Israel.
In 2015, the Centers for Disease Control in the United States declared that 7 of 10 deaths per year are caused by chronic diseases; in Israel, the situation is consistent with this assessment. Healthy Israel 2020, an Israeli Ministry of Health initiative, places physicians at the forefront of health promotion and preventive medicine. In 2012, the Israeli Society of Lifestyle Medicine (ISLM) was established under the auspices of the Israel Association for Family Physicians. This decision was taken because we, the authors, wanted to promote change in the primary care services through the recruitment of key leaders for implementation of lifestyle medicine (LM), including improved attitudes, knowledge, motivation, and skills of primary care physicians and health providers. Today, the ISLM is an active member of the Israeli Medical Association, promoting educational activities, physician's health, and well-being initiatives; developing tools for health-promoting clinics; and more. Our future plans are to incorporate LM as an integral part of daily practice in all sectors of the medical profession in Israel. This is challenging, but we see this as the only way to effectively combat the noncommunicable disease epidemic.


Accelerated solute clearance in Pneumocystis carinii pneumonia.
A radioaerosol clearance test was used to detect lung injury in 12 patients with acquired immune deficiency syndrome (AIDS) and Pneumocystis carinii pneumonia documented by bronchoscopy. The clearance of 99mTc-DTPA was elevated over most regions of interest in all of these patients. In each of 7 subjects who responded to trimethoprim-sulfamethoxazole therapy, clearance returned toward normal. Average clearances were either normal or very slightly above the 95% expected upper limit of normal in each of 6 patients with either AIDS or AIDS-related complex without clinical evidence of Pneumocystis infection. The radioaerosol diffusion test may prove useful for determining whether patients who are predisposed to this infection require bronchoscopy and may provide information concerning their response to therapy.


Linoleic acid and linolenic acid elongation products in muscle tissue of Sncerus caffer and other ruminant species.
The metabolic elongation products of both linoleic acid and linolenic acid were found in muscle tissues of Syncerus caffer and other ruminants. The acids with four double bonds were predominantly in the linoleic acid series, whereas the higher degrees of unsaturation, mainly five double bonds, were in the linolenic acid series. The total linoleic acid and linolenic acid groups were present in the relative proportions of about 4:1, in contrast with the fish oils, where the acids are mainly in the linolenic acid series. The consistent occurrence of members of both groups of acids in the animals studied here suggests to us that both may be important for structural purposes.


REVEILLE2 thermosensitive splicing: a molecular basis for the integration of nocturnal temperature information by the Arabidopsis circadian clock.
Cold stress is one of the major environmental factors that limit growth and yield of plants. However, it is still not fully understood how plants account for daily temperature fluctuations, nor how these temperature changes are integrated with other regulatory systems such as the circadian clock. We demonstrate that REVEILLE2 undergoes alternative splicing after chilling that increases accumulation of a transcript isoform encoding a MYB-like transcription factor. We explore the biological function of REVEILLE2 in Arabidopsis thaliana using a combination of molecular genetics, transcriptomics, and physiology. Disruption of REVEILLE2 alternative splicing alters regulatory gene expression, impairs circadian timing, and improves photosynthetic capacity. Changes in nuclear gene expression are particularly apparent in the initial hours following chilling, with chloroplast gene expression subsequently upregulated. The response of REVEILLE2 to chilling extends our understanding of plants immediate response to cooling. We propose that the circadian component REVEILLE2 restricts plants responses to nocturnal reductions in temperature, thereby enabling appropriate responses to daily environmental changes.


A Single Nucleotide Change in the polC DNA Polymerase III in Clostridium thermocellum Is Sufficient To Create a Hypermutator Phenotype.
Clostridium thermocellum is a thermophilic, anaerobic bacterium that natively ferments cellulose to ethanol and is a candidate for cellulosic biofuel production. Recently, we identified a hypermutator strain of C. thermocellum with a C669Y mutation in the polC gene, which encodes a DNA polymerase III enzyme. Here, we reintroduced this mutation using recently developed CRISPR tools to demonstrate that this mutation is sufficient to recreate the hypermutator phenotype. The resulting strain shows an approximately 30-fold increase in the mutation rate. This mutation is hypothesized to function by interfering with metal ion coordination in the PHP (polymerase and histidinol phosphatase) domain, which is responsible for proofreading. The ability to selectively increase the mutation rate in C. thermocellum is a useful tool for future directed evolution experiments. IMPORTANCE Cellulosic biofuels are a promising approach to decarbonize the heavy-duty-transportation sector. A longstanding barrier to cost-effective cellulosic biofuel production is the recalcitrance of cellulose to solubilization. Native cellulose-consuming organisms, such as Clostridium thermocellum, are promising candidates for cellulosic biofuel production; however, they often need to be genetically modified to improve product formation. One approach is adaptive laboratory evolution. Our findings demonstrate a way to increase the mutation rate in this industrially relevant organism, which can reduce the time needed for adaptive evolution experiments.


Genetic Variations at rs3129891 and rs77005575 are Associated With Reduced Expression of Enteric α-defensins in IBD Patients.
Human enteric antimicrobial peptides composed predominantly of human enteric α-defensins (HD5 and HD6) are important in the mucosal antimicrobial barrier. Previous studies have identified that genetic variations at rs2066844, rs2066845, rs2066847 are associated with diminished enteric α-defensins in ileal Crohn's disease (CD). However, genetic variations associated with enteric antimicrobial peptides in colonic inflammatory bowel disease (IBD) remain unclear. To investigate it, we compared the colonic expression of antimicrobial peptides with respect to genotypes at 22 IBD-associated single-nucleotide polymorphisms (SNPs). In total, 16 controls and 102 colonic IBD patients including 42 ulcerative colitis (UC) and 60 CD were studies. Mutation assay was performed to determine their genotypes at 22 IBD-associated SNPs. Real-time PCR was performed to determine the colonic mRNA expression of HD5, HD6, lysozyme, and secretory phospholipase A2. Mutant genotypes at rs2066844, rs2066845, rs2066847 were not found, and only SNPs rs3129891 and rs77005575 were associated with enteric α-defensin expression in colonic IBD. In both inflamed and noninflamed tissues, colonic expression of HD5 and HD6 was significantly decreased in UC and CD patients carrying rs3129891 homozygous mutant genotype. And their colonic expression was significantly decreased in inflamed but not noninflamed tissues from UC patients carrying rs77005575 homozygous mutant genotype. However, both lysozyme and secretory phospholipase A2 in UC and CD were unaffected by rs3129891 and rs77005575 genotypes. As enteric α-defensins play critical roles in the mucosal antimicrobial barrier, their reduced expression may partly explain the microbial-induced mucosal inflammation in colonic IBD patients, especially in patients carrying rs3129891 and rs77005575 mutant genotypes.


Towards precision apiculture: Traditional and technological insect monitoring methods in strawberry and raspberry crop polytunnels tell different pollination stories.
Over one third of crops are animal pollinated, with insects being the largest group. In some crops, including strawberries, fruit yield, weight, quality, aesthetics and shelf life increase with insect pollination. Many crops are protected from extreme weather in polytunnels, but the impacts of polytunnels on insects are poorly understood. Polytunnels could reduce pollination services, especially if insects have access issues. Here we examine the distribution and activity of honeybees and non-honeybee wild insects on a commercial fruit farm. We evaluated whether insect distributions are impacted by flower type (strawberry; raspberry; weed), or distance from polytunnel edges. We compared passive pan-trapping and active quadrat observations to establish their suitability for monitoring insect distribution and behaviour on a farm. To understand the relative value of honeybees compared to other insects for strawberry pollination, the primary crop at the site, we enhanced our observations with video data analysed using insect tracking software to document the time spent by insects on flowers. The results show honeybees strongly prefer raspberry and weed flowers over strawberry flowers and that location within the polytunnel impacts insect distributions. Consistent with recent studies, we also show that pan-traps are ineffective to sample honeybee numbers. While the pan-traps and quadrat observations tend to suggest that investment in managed honeybees for strawberry pollination might be ineffective due to consistent low numbers within the crop, the camera data provides contrary evidence. Although honeybees were relatively scarce among strawberry crops, camera data shows they spent more time visiting flowers than other insects. Our results demonstrate that a commercial fruit farm is a complex ecosystem influencing pollinator diversity and abundance through a range of factors. We show that monitoring methods may differ in their valuation of relative contributions of insects to crop pollination.


Metabolic Blood Hormones in Young People With Electrocardiographic Signs of Ischemic Myocardial Changes.
Aim To study changes in blood concentrations of metabolic hormones and adipocytokines in people aged 25-44 years with electrocardiographic (ECG) signs of ischemic changes in the myocardium.Material and methods This study was a part of a cross-sectional survey of a random sample of Novosibirsk population aged 25-44 years. The study included 1363 people divided into two groups: group 1, subjects with ECG signs of ischemic changes in the myocardium and group 2, subjects without ECG changes. Blood serum concentrations of adipocytokines and metabolic hormones were measured by multiplex assay on a Luminex MAGPIX flow-through fluorometer.Results The group with ECG signs of myocardial ischemia had higher blood concentrations of adiponectin, resistin, glucagon, and interleukin 6 (IL-6) than in the comparison group. A multivariate logistic regression analysis showed that the glucagon concentration was associated with the presence of ECG signs of myocardial ischemia (OR, 1.019; CI, 1.018-1.034; p=0.017).Conclusion In young people aged 25-44 years, higher blood concentrations of glucagon are associated with the presence of ECG signs of myocardial ischemia.


Fluoro-norchloroepibatidine: preclinical assessment of acute toxicity.
18Fluoro-norchloroepibatidine (exo-2-(6-fluoro-3-pyridyl)-7-azabicyclo-[2.2.1]heptane [NFEP]), a labeled derivative of epibatidine, has shown promise for imaging brain nicotinic acetylcholine receptors with PET. We determined the dose-dependent effects of NFEP in conscious rats. NFEP (1.5 microg/kg; administered intravenously) resulted in 30% mortality. Neither 0.5 microg/kg or 0.25 microg/kg NFEP resulted in any significant changes in cardiorespiratory parameters, but plasma catecholamines increased (2- to 3-fold). Further studies are needed to determine the safety of NFEP that are specifically designed to assess the catecholamine response. Our results suggest that it is not advisable to initiate human PET studies with [18F]-NFEP without further evidence supporting its safety.


A 80-Year-Old Woman with B-Cell Prolymphocytic Leukemia.
Prolymhocytic leukemia (PLL) is a rare subtype of lymphocytic leukemias and its cells are immature lymphocytes. It is divided into 2 subgroups: T-PLL and B-PLL according to the lymphocytic origin of the cells. Discriminating B-PLL from other diseases with clinically-similar features is important because of the different treatment approaches and follow-up programs. Hereby, we report a 80-year-old woman presenting with fatigue, leucocytosis and mild anemia. Her peripheral blood smear evaluation revealed 85% prolymphocytes with moderately condensed nuclear chromatin, prominent nucleoli, and a faintly basophilic cytoplasm. Positron emission tomography-computed tomography showed mediastinal lymph nodes with cervical lymph nodes. There was no pathological FDG involvement in the spleen. Bone marrow aspiration smear exhibit atypical wide lymphocytes with prominent nucleoli and abundant agranular cytoplasm. Flow cytometry analysis revealed positive CD5+, CD19+, CD20+, CD22+, CD11c+, CD25+, CD79a+ and CD79b+. Fluorescence in situ hybridization technique analysis reveals no t(11;14). Bone marrow biopsy revealed interstitially distributed atypical cells with wide nucleus and prominent nucleolus.


Visualizing lignin coalescence and migration through maize cell walls following thermochemical pretreatment.
Plant cell walls are composed primarily of cellulose, hemicelluloses, lignins, and pectins. Of these components, lignins exhibit unique chemistry and physiological functions. Although lignins can be used as a product feedstock or as a fuel, lignins are also generally seen as a barrier to efficient enzymatic breakdown of biomass to sugars. Indeed, many pretreatment strategies focus on removing a significant fraction of lignin from biomass to better enable saccharification. In order to better understand the fate of biomass lignins that remain with the solids following dilute acid pretreatment, we undertook a structural investigation to track lignins on and in biomass cell walls. SEM and TEM imaging revealed a range of droplet morphologies that appear on and within cell walls of pretreated biomass; as well as the specific ultrastructural regions that accumulate the droplets. These droplets were shown to contain lignin by FTIR, NMR, antibody labeling, and cytochemical staining. We provide evidence supporting the idea that thermochemical pretreatments reaching temperatures above the range for lignin phase transition cause lignins to coalesce into larger molten bodies that migrate within and out of the cell wall, and can redeposit on the surface of plant cell walls. This decompartmentalization and relocalization of lignins is likely to be at least as important as lignin removal in the quest to improve the digestibility of biomass for sugars and fuels production.


An assessment of the ability to ingest and excrete microplastics by filter-feeders: A case study with the Mediterranean mussel.
Plastic debris has been recognized as a growing threat to marine biota due to its widespread distribution and possible interactions with marine species. Concerns over the effects of plastic polymers in marine ecosystems is reflected in the high number of toxicological studies, regarding microplastics (<5 mm) and marine fauna. Although several studies reported that organisms ingest and subsequently eliminate microplastics (MP), the potential effects at organ and tissue level remain unclear, especially considering exposure to different microplastic sizes and concentrations. The present study aimed at investigating potential pathophysiological effects of the ingestion of MP by marine filter-feeders. For the purpose, Mediterranean mussel (Mytilus galloprovincialis) was exposed to spherical polystyrene MP (2 and 10 μm Ø) over short- and medium-term exposure periods, under single and combined concentrations that represent high, yet realistic doses (10 and 1000 MP mL-1). Overall, results suggest rapid MP' clearance from water column by filtering, regardless of MP size. Ingestion occurred, identified by MP in the lumen of the gut (mostly in midgut region), followed by excretion through faeces. However, no MP were found in gills or digestive gland diverticula. Biochemical indicators for oxidative stress were generally irresponsive regardless of organ and time of exposure. Small foci of haemocytic infiltration in gastric epithelia were found, albeit not clearly related to MP ingestion. Globally, no evident histopathological damage was recorded in whole-body sections of exposed animals. The present findings highlight the adaptative ability of filter-feeding bivalves to cope with filtration of suspended MP, resulting in rapid elimination and reduced internal damage following ingestion of spherical MP. Nevertheless, the fact that the animals are able to translocate MP to the gut reveals that filter feeding organisms may indeed became a target of concern for fragmented materials with smaller, mixed sizes and sharper edges.


In silico analysis of cytochrome P450 monooxygenases in chronic granulomatous infectious fungus Sporothrix schenckii: Special focus on CYP51.
Sporotrichosis is an emerging chronic, granulomatous, subcutaneous, mycotic infection caused by Sporothrix species. Sporotrichosis is treated with the azole drug itraconazole as ketoconazole is ineffective. It is a well-known fact that azole drugs act by inhibiting cytochrome P450 monooxygenases (P450s), heme-thiolate proteins. To date, nothing is known about P450s in Sporothrix schenckii and the molecular basis of its resistance to ketoconazole. Here we present genome-wide identification, annotation, phylogenetic analysis and comprehensive P450 family-level comparative analysis of S. schenckii P450s with pathogenic fungi P450s, along with a rationale for ketoconazole resistance by S. schenckii based on in silico structural analysis of CYP51. Genome data-mining of S. schenckii revealed 40 P450s in its genome that can be grouped into 32 P450 families and 39 P450 subfamilies. Comprehensive comparative analysis of P450s revealed that S. schenckii shares 11 P450 families with plant pathogenic fungi and has three unique P450 families: CYP5077, CYP5386 and CYP5696 (novel family). Among P450s, CYP51, the main target of azole drugs was also found in S. schenckii. 3D modeling of S. schenckii CYP51 revealed the presence of characteristic P450 motifs with exceptionally large reductase interaction site 2. In silico analysis revealed number of mutations that can be associated with ketoconazole resistance, especially at the channel entrance to the active site. One of possible reason for better stabilization of itraconazole, compared to ketoconazole, is that the more extended molecule of itraconazole may form a hydrogen bond with ASN-230. This in turn may explain its effectiveness against S. schenckii vis-a-vis resistant to ketoconazole. This article is part of a Special Issue entitled: Cytochrome P450 biodiversity and biotechnology, edited by Erika Plettner, Gianfranco Gilardi, Luet Wong, Vlada Urlacher, Jared Goldstone.


Vasodilation mediated by angiotensin II type 2 receptor is impaired in afferent arterioles of young spontaneously hypertensive rats.
Renal vasoconstrictor action of angiotensin II (Ang II) is exaggerated in the spontaneously hypertensive rat (SHR) before development of hypertension. We have recently demonstrated that in the rabbit afferent arteriole (Af-Art) activation of the AT2 receptor causes vasodilation, which modulates the vasoconstrictor action of Ang II mediated by the AT1 receptor. In this study, we tested the hypothesis that vasoconstrictor action of Ang II is exaggerated in SHR Af-Arts due to an impaired function of the AT2 receptor before development of hypertension. Af-Arts were microdissected from the superficial cortex of 4- to 5-week-old SHR or age-matched Wistar Kyoto rats (WKY), and perfused at 60 mm Hg in vitro. Ang II (10(-11) to 10(-8) M) decreased the luminal diameter of Af-Arts of both strains in a dose-dependent manner. However, the constriction was stronger in SHR; at 10(-10) M, the diameter decreased by 34 +/- 4% in SHR (n = 6) compared to 18 +/- 3% in WKY (n = 6; p < 0.01). Pretreatment with PD123319 (PD), an AT2 receptor antagonist, significantly augmented Ang II-induced constriction in WKY but not SHR Af-Arts; at 10(-10) M, the diameter now decreased by 41 +/- 5 and 37 +/- 1% in SHR (n = 6) and WKY (n = 6), respectively. Thus, blockade of the AT2 receptor abolished the difference in Ang II action on Af-Arts between strains. Moreover, with the AT1 receptor blockade Ang II caused dose-dependent dilation of preconstricted Af-Arts only in WKY (27 +/- 5% at 10(-8) M, n = 5), and the dilation was abolished by simultaneous treatment with PD. In contrast, no such dilation was observed in SHR Af-Arts. These results suggest that activation of the AT2 receptor modulates AT1 receptor vasoconstriction in WKY Af-Arts, while impaired modulatory function of AT2 receptor may play a role in the exaggerated vasoconstrictor action of Ang II on the Af-Art of prehypertensive SHR.


Deaths in adults with notified pulmonary tuberculosis 1983-5.
For a study of fatality in pulmonary tuberculosis in England and Wales, a sample (n = 1222) of patients notified in 1983 as having tuberculosis confined to the lungs was selected and their mortality up to the time of finishing chemotherapy calculated. During that time 158 patients died, a case fatality of 12.9%. Stratification by age and radiographic extent of disease confirmed these as important predictors of death. Comparison with a referent population showed that the all cause mortality among people with pulmonary tuberculosis was 10 times greater than that of the age and sex matched general population. This ratio fell with increasing age and with extent of disease. Coefficients derived from a logistic regression analysis allowed probability of death during chemotherapy of patients with pulmonary tuberculosis to be estimated.


Canadian Rheumatology Association Recommendations for the Screening, Monitoring, and Treatment of Juvenile Idiopathic Arthritis-Associated Uveitis.
To develop Canadian recommendations for the screening, monitoring, and treatment of uveitis associated with juvenile idiopathic arthritis (JIA). Recommendations were developed using the Grading of Recommendations Assessment, Development, and Evaluation (GRADE)-ADOLOPMENT approach. A working group of 14 pediatric rheumatologists, 6 ophthalmologists, 2 methodologists, and 3 caregiver/patient representatives reviewed recent American College of Rheumatology (ACR)/Arthritis Foundation (AF) recommendations and worked in pairs to develop evidence-to-decision (EtD) tables. A survey to assess agreement and recommendations requiring group discussion was completed. EtD tables were presented, discussed, and voted upon at a virtual meeting, to produce the final recommendations. A health equity framework was applied to all aspects of the adolopment process including the EtD tables, survey responses, and virtual meeting discussion. The survey identified that 7 of the 19 recommendations required rigorous discussion. Seventy-five percent of working group members attended the virtual meeting to discuss controversial topics as they pertained to the Canadian environment, including timing to first eye exam, frequency of screening, escalation criteria for systemic and biologic therapy, and the role of nonbiologic therapies. Equity issues related to access to care and advanced therapeutics across Canadian provinces and territories were highlighted. Following the virtual meeting, 5 recommendations were adapted, 2 recommendations were removed, and 1 was developed de novo. Recommendations for JIA-associated uveitis were adapted to the Canadian context by a working group of pediatric rheumatologists, ophthalmologists with expertise in the management of uveitis, and parent/patient input, taking into consideration cost, equity, and access.


Clinical and microbiological characterization of carbapenem-resistant Acinetobacter baumannii bloodstream infections.
The incidence of carbapenem-resistant Acinetobacter baumannii infection is increasing, which might be associated with high morbidity and mortality among critically ill patients with limited therapeutic options. This study was conducted to evaluate the clinical and microbiological features of carbapenem-resistant A. baumannii bacteraemia. The medical records of 28 adult patients with this bacteraemia admitted to Korea University Guro Hospital, from January 2005 through December 2010, were reviewed. Using the 28 bloodstream isolates, we intended to detect genes encoding carbapenemases, and investigate the inoculum effect on each of the antimicrobial agents rifampicin, imipenem, colistin and tigecycline. With one blood isolate from a patient with pneumonia, rifampicin-inducible resistance was examined using the experimental mouse pneumonia model. Out of 28 carbapenem-resistant A. baumannii bloodstream infections (BIs), the most common primary focus was the central venous catheter (35.7 %) and then the lung (32.1 %). The 30 day overall mortality was 53.6 %; in most cases (80 %) the patients died within 10 days after the onset of the bacteraemia. By univariate analysis, inappropriate antimicrobial therapy (73.3 vs 30.8 %, P = 0.02), mechanical ventilation (53.3 vs 15.4 %, P = 0.04) and a high Pitt bacteraemia score (4.9±1.9 vs 2.2±1.2, P<0.01) were statistically significant risk factors for mortality, while only a high Pitt bacteraemia score (odds ratio 2.6; 95 % confidence interval 1.1-6.5) was independently associated with 30 day mortality by multivariate analysis. All 28 isolates had the bla(OXA-51)-like gene with upstream ISAbaI, 2 of which additionally had the bla(OXA-58)-like gene and the bla(OXA-23)-like gene. Inoculum effect and rifampicin inducible resistance were not detected. Considering the rapid progression to death in carbapenem-resistant A. baumannii BIs, early empirical antibiotic therapy would be warranted based on the local microbiological data in each hospital.


Effects of oils rich in linoleic and α-linolenic acids on fatty acid profile and gene expression in goat meat.
Alteration of the lipid content and fatty acid (FA) composition of foods can result in a healthier product. The aim of this study was to determine the effect of flaxseed oil or sunflower oil in the goat diet on fatty acid composition of muscle and expression of lipogenic genes in the semitendinosus (ST) muscle. Twenty-one entire male Boer kid goats were fed diets containing different levels of linoleic acid (LA) and α-linolenic acid (LNA) for 100 days. Inclusion of flaxseed oil increased (p < 0.05) the α-linolenic acid (C18:3n-3) concentration in the ST muscle. The diet high in α-linolenic acid (p < 0.05) decreased the arachidonic acid (C20:4n-6) and conjugated linolenic acid (CLA) c-9 t-11 content in the ST muscle. There was a significant (p < 0.05) upregulation of PPARα and PPARγ gene expression and downregulation of stearoyl-CoA desaturase (SCD) gene in the ST muscle for the high α-linolenic acid group compared with the low α-linolenic acid group. The results of the present study show that flaxseed oil as a source of α-linolenic acid can be incorporated into the diets of goats to enrich goat meat with n-3 fatty acids, upregulate the PPARα and PPARγ, and downregulate the SCD gene expression.


Ni3C-assisted growth of carbon nanofibres 300 °C by thermal CVD.
Ni-assisted thermal chemical vapor deposition (TCVD) is one of the most common techniques for the growth of carbon nanofibres/nanotubes (CNFs/CNTs). However, some fundamental issues related to the catalytic growth of CNFs/CNTs, such as the low-limit growth temperature, the limiting steps and the state of Ni, are still controversial. Here, we report the growth of CNFs at 300 °C; that is the lowest temperature for the growth of CNFs by TCVD using Ni as the catalyst so far. The results showed that the Ni existed in rhombohedral Ni3C, not in the normal form of face-centered cubic Ni, and the C atoms for building the CNFs were precipitated from the (001) planes of the faceted Ni3C nanoparticles. The CNFs are believed to be formed by the decomposition-formation cycle of metastable Ni3C that has a low-limit decomposition temperature of about 300 °C. Our results strongly suggest that TCVD is a valuable tool for the synthesis of CNFs/CNTs at temperatures below 400 °C, which is generally considered as the upper-limit temperature for fabricating complementary metal oxide semiconductor devices but is the low-limit temperature for growing CNFs/CNTs by TCVD at present.


The Role of Tinzaparin in Oncology.
Current guidelines recommend low-molecular-weight heparin treatment in patients with cancer with established venous thromboembolism (VTE). The aim of this article was to study the pharmacological properties and effectiveness of tinzaparin in patients with cancer as well as its potential anticancer properties. A search of PubMed and ScienceDirect databases up to March 2016 was carried out to identify published studies that detect the properties and use of tinzaparin in oncology. Protamine sulfate partially (60% to 65%) neutralized tinzaparin's anti-Xa activity. No dose adjustment of tinzaparin is needed even in patients with severe renal impairment and Creatinine Clearance ≥20 mL/min. Tinzaparin demonstrated a statistically significant decline in VTE recurrence at 1 year post the index thromboembolic event. A statistically significant reduction in minor bleeding rates was also described, whereas major bleeding events did not decrease in patients with cancer treated with tinzaparin versus those who received vitamin K antagonists. Tinzaparin treatment in patients suffering from deep vein thrombosis reduced the incidence of postthrombotic syndrome and venous ulcers. Tinzaparin's ability to prevent both metastatic dissemination of cancer cells and tumor angiogenesis has been delineated in preclinical research. Current data show that tinzaparin is safe and efficacious either for short-term or for long-term treatment of VTE in patients with cancer. Clinical trials are needed in order to examine the utility of tinzaparin in primary prevention of VTE and validate its potential anticancer advantages exhibited in preclinical research.


Iditis: protein structure database.
The validation, enrichment and organization of the data stored in PDB files is essential for those data to be used accurately and efficiently for modelling, experimental design and the determination of molecular interactions. The Iditis protein structure database has been designed to allow the widest possible range of queries to be performed across all available protein structures. The Iditis database is the most comprehensive protein structure resource currently available, and contains over 500 fields of information describing all publicly deposited protein structures. A custom-written database engine and graphical user interface provide a natural and simple environment for the construction of searches for complex sequence- and structure-based motifs. Extensions and specialized interfaces allow the data generated by the database to used in conjunction with a wide range of applications.


Personal protective equipment during activity resumption in endoscopy units: waterproof gowns in the daily practice.
Adapting endoscopy units to the resumption of services remains challenging. Published recommendations for the resumption of endoscopy services are logical but, in some cases, may prove difficult to put into practice in the daily workflow of most centers. In relation to the SEED-AEG position statement document, we agree on the use of personal protective equipment during the procedures including footwear, body, hand, face, and hair protection. Since gloves are changed for all interventions, body protection would be the most problematic of all protection equipment as both the forearm and trunk areas can potentially come into contact with the endoscope, which in turn, is inserted into the patient.


Negative cognition, affect, metacognition and dimensions of paranoia in people at ultra-high risk of psychosis: a multi-level modelling analysis.
Paranoia is one of the commonest symptoms of psychosis but has rarely been studied in a population at risk of developing psychosis. Based on existing theoretical models, including the proposed distinction between 'poor me' and 'bad me' paranoia, we aimed to test specific predictions about associations between negative cognition, metacognitive beliefs and negative emotions and paranoid ideation and the belief that persecution is deserved (deservedness). We used data from 117 participants from the Early Detection and Intervention Evaluation for people at risk of psychosis (EDIE-2) trial of cognitive–behaviour therapy, comparing them with samples of psychiatric in-patients and healthy students from a previous study. Multi-level modelling was utilized to examine predictors of both paranoia and deservedness, with post-hoc planned comparisons conducted to test whether person-level predictor variables were associated differentially with paranoia or with deservedness. Our sample of at-risk mental state participants was not as paranoid, but reported higher levels of 'bad-me' deservedness, compared with psychiatric in-patients. We found several predictors of paranoia and deservedness. Negative beliefs about self were related to deservedness but not paranoia, whereas negative beliefs about others were positively related to paranoia but negatively with deservedness. Both depression and negative metacognitive beliefs about paranoid thinking were specifically related to paranoia but not deservedness. This study provides evidence for the role of negative cognition, metacognition and negative affect in the development of paranoid beliefs, which has implications for psychological interventions and our understanding of psychosis.


Achieving Purely Organic Room-Temperature Phosphorescence Mediated by a Host-Guest Charge Transfer State.
Strategies for developing purely organic materials exhibiting both high efficiency and persistent room-temperature phosphorescence (RTP) have remained ambiguous and challenging. Herein, we propose that introducing an intermediate charge transfer (CT) state into the donor-acceptor binary molecular system holds promise for accomplishing this goal. Guest materials showing gradient ionization potentials were selected to fine-tune the intermolecularly formed CT state when doped into the same host material with a large electron affiliation potential. Such a CT intermediate state accelerates the population of the triplet exciton to benefit phosphorescent emission and decreases the phosphorescence lifetime via quenching the long-lived triplet excitons. As a result, a "trade-off" between a long phosphorescence lifetime (595 ms) and a high phosphorescent quantum yield (27.5%) can be obtained by tuning the host-guest energy gap offset. This finding highlights the key role of CT in RTP emission and provides new guidance for developing novel RTP systems.


Energy-linked functional alterations in experimental cardiomyopathies.
Changes in high-energy phosphate content and cardiac contractile function of isolated rat hearts as well as changes in Ca2+ sensitivity and mitochondrial respiration of myocardial skinned fibers were assessed in hereditary cardiomyopathies and in cardiomyopathies induced by chronic treatment with adriamycin or norepinephrine, by autoimmunization, by diabetes, or by creatine deficiency. The sum of ATP and phosphocreatine contents as well as cardiac output at standard load conditions was substantially lower in almost all groups. The common features of cardiac pump failure were mild bradycardia, elevated left ventricular (LV) diastolic pressure, and stiffness that limited cardiac contractile adaptation to volume or resistance loads. The LV diastolic stiffness at maximal functional load was inversely correlated with high-energy phosphate content. Increased myofibrillar sensitivity to Ca2+ and defective function of mitochondrial creatine kinase were found in skinned myocardial fibers. These results suggested that both increased myofibrillar Ca2+ sensitivity and energy deficiency within myofibrils may contribute to increased myocardial stiffness. Increased stiffness limits LV filling but facilitates pressure development, which partly compensates for decreased contractility of cardiomyopathic hearts.


Sex and heredity are determinants of oral oxycodone self-administration in 36 Inbred Rat Strains: correlations with behavioral tests of anxiety and novelty-seeking.
Most individuals affected in the national epidemic of oxycodone abuse began taking oral oxycodone by prescription. We studied vulnerability to oxycodone intake in a rat model of oral drug self-administration (SA), since pharmacokinetics affect abuse potential. Females (33 inbred strains) and males (26) obtained oxycodone at increasing concentrations in operant sessions (FR5; 1-16-h) followed by extinction and reinstatement. Active spout licks were greater in females than males during 4-h and 16-h sessions (p< 0.001 for all). Across all stages of oxycodone SA, intake/session was greater in females (p<0.001). Both sexes escalated intake during 16-h extended access vs 4-h sessions (p<2e-16). Intake and active licks varied greatly by strain. The heritability ( h 2 ) of active licks/4-h at increasing oxycodone dose was larger in males ( h 2 females: 0.30-0.39 vs. males: 0.41-0.53). Under a progressive ratio schedule, breakpoints differed by strain (p<2e-16) and by sex in some strains (p=0.018). For cue-induced reinstatement, active licks were greater in females than males (p<0.001). Behavior in naive rats was assessed using elevated plus maze (EPM), open field (OF) and novel object interaction. (NOI) tests. We correlated these behaviors with 28 parameters of oxycodone SA. EPM-defining traits were most commonly associated with SA in both sexes, whereas more OF and NOI traits were SA-associated in males. Overall, sex and heredity are major determinants of the motivation to take and seek oxycodone, which escalates during extended access. The correlation of EPM, a measure of anxiety, with multiple SA parameters indicates the influence of pleiotropic genes.


Growth changes of the collagen content and architecture in the pectoralis and iliotibialis lateralis muscles of cockerels.
1. Growth changes of the collagen content and architecture in the pectoralis (PT) and iliotibialis lateralis (ITL) muscles were examined using cockerels from 1 to 14 weeks of age. 2. Total collagen content in PT muscle showed little change, but in ITL muscle reached a maximum at 5 weeks and thereafter decreased slightly until 14 weeks. The collagen content was markedly larger in ITL muscle after 5 weeks. Pyridinoline content of collagen increased abruptly from 5 to 14 weeks in both muscles, but no difference between muscle types was detected. 3. The cell size of the endomysial honeycombs increased with the development of myofibres, and the mesh size of the perimysium around the honeycombs enlarged. 4. In both muscles endomysia were an incomplete network of collagen fibrils with many foramina at one week, became a very thin membrane of felt-like fabric in 2 to 5 weeks and thereafter increased in thickness until 11 to 14 weeks. 5. Perimysial width around the secondary fasciculus differed between the muscle types after 5 weeks. In the wider perimysium of ITL muscle, the collagen fibres increased in number and size to make a stack of collagen bands around the fasciculus. In the narrower perimysium of PT muscle, a few platelets of collagen fibres also developed. 6. The perimysial collagen fibre at 1 to 2 weeks had a smooth surface and appeared to be composed of fine collagen fibrils. The fibre at 11 to 14 weeks showed a rugged surface and was composed of coarser collagen bundles that combined with each other into a net-like configuration with very slim meshes. 7. Our results showed that the collagenous components of chicken intramuscular connective tissue changed markedly during the early period of muscle growth in distribution, architecture and quality but with little difference in quantity.


[Balance studies after gastrectomy, with particular reference to the risk factors for coronary heart disease (author's transl)].
Balance studies in 15 patients who had undergone gastrectomy between 1957 and 1966 at the Heidelberg Surgical Hospital confirmed the suggestions in the literature that gastrectomied patients showed a reduced incidence of ischemic myocardiac diseases because of their altered living habits in relation to their age group. Consequently a gastrectomy is associated with a protective effect against the development of coronary heart disease, with underweight and hypotension having a particularly positive effect. This observation is not contradictory to the opinion that increased frequency of severe arteriosclerosis is supposed to be demonstrable in ulcer patients, because total and subtotal gastrectomies were carried out because of carcinoma.


An ideal composite graft donor site for postburn alar rim deficiencies: root of helix.
One of the challenging postburn face deformities is alar rim defects or deformities. Small through-and-through defects of the nasal ala may be managed with a composite graft of auricular tissue, obtained from the earlobe, the helical rim, and the root of the helix. We use usually the root of the helix for postburn alar defect correction as a composite graft donor site. The advantage of these composite grafts is that the procedure is a single-stage operation with excellent contour correction and a well toleration of donor site morbidity by the patient.


Complement: structure, functions, evolution, and viral molecular mimicry.
The complement (C') system has long been recognized as an important mediator of innate immune defense and inflammation. In recent years there is increasing evidence suggesting that complement components may also participate in non-inflammatory and developmental processes. Here we review our current work on the structural-functional aspects of C3-ligand interactions and the rational design of small-sized complement inhibitors. We present a novel, proteomics-based, approach to studying protein-protein interactions within the C' system and discuss our progress in the study of viral immune evasion strategies. Furthermore we discuss the involvement of complement proteins in organ regeneration and hematopoietic development.


Targeting the Tumor Microenvironment in Breast Cancer: Prognostic and Predictive Significance and Therapeutic Opportunities.
Breast cancer is one of the most prevalent tumors among women. Its prognosis and treatment outcomes depend on factors related to tumor cell biology. However, recent studies have revealed the critical role of the tumor microenvironment (TME) in the development, progression, and treatment response of breast cancer. In this review, we explore the different components of the TME and their relevance as prognostic and predictive biomarkers in breast cancer. In addition, techniques for assessing the tumor microenvironment, such as immunohistochemistry or gene expression profiling, and their clinical utility in therapeutic decision-making are examined. Finally, therapeutic strategies targeting the TME are reviewed, highlighting their potential clinical benefits. Overall, this review emphasizes the importance of the TME in breast cancer and its potential as a clinical tool for better patient stratification and the design of personalized therapies.


The effects of β-glucan on iron levels and lipid peroxidation in intra-abdominal sepsis in rats.
Sepsis is defined as a systemic response of organisms to microorganisms and toxins. Sepsis is associated with the enhanced generation of reactive oxygen metabolites, leading to multiple organ dysfunctions. β-glucan is accepted to be one of the most powerful immune response modifiers. The aim of this study was to investigate the putative protective effect of β-glucan on changes of iron and malondialdehyde (MDA) levels in various tissue and blood after experimental sepsis in rats. Sepsis was induced by cecal ligation and perforation (CLP) in 32 male Wistar albino rat. To evaluate this, rats were divided into four groups as sham operated, β-glucan treated sham operated, CLP and β-glucan treated CLP. Sixteen hours after operation, rats were decapitated and MDA and iron levels were measured in the liver, kidney, heart, diaphragm tissues and blood. Also, whole tissue histopathology was evaluated by a light microscope. The results demonstrate that sepsis significantly decreased iron levels of all tissues and blood. The decrease in tissue iron levels and the increase MDA levels demonstrate the role of trace elements and free radicals in sepsis-induced tissue damage. Our results indicate that the given dose of β-glucan was probably insufficient to prevent sepsis-induced organ injury.


Association between gestational cannabis exposure and maternal, perinatal, placental, and childhood outcomes.
Globally, the availability and formulations for the administration of cannabis are changing with decriminalization or legalization of recreational use in some jurisdictions, and the prescription of cannabis also occurring. These changes are likely to affect the prevalence of use, including by women of childbearing age. The effects of in utero and infant alcohol and tobacco exposure are well-documented, but the outcomes of cannabis exposure are less certain. The content of delta-9-tetrahydrocannabinol (THC), the psychoactive component of cannabis has progressively increased over several decades. This review explores the limited knowledge surrounding the epidemiology of gestational and postnatal cannabis exposure and implications for the mother-placenta-fetus/neonate triad. We examine cannabis' effects from antenatal and lactation exposure on (a) pregnancy and perinatal outcomes, (b) placental health, and (c) longer term cardiometabolic and neurodevelopmental risks and outcomes. Though definitive outcomes are lacking, gestational cannabis has been associated with increased risk of other substance use during pregnancy; impaired placental blood flow; increased risk of small for gestational age births; and associated complications. Childhood and adolescent outcomes are sparsely assessed, with suggested outcomes including increased risk of depression and attention-deficit hyperactivity disorder. Cardiometabolic implications of gestational cannabis use may include maternal fatty liver, obesity, insulin resistance, and increased risk of gestational diabetes mellitus (GDM), with potential consequences for the fetus. Clinical implications for pediatric practice were explored in a bid to understand any potential risk or impact on child health and development.


Vasculopathy and disordered angiogenesis in selected rheumatic diseases: rheumatoid arthritis and systemic sclerosis.
Angiogenesis is important in the pathogenesis of systemic inflammatory rheumatic diseases, a family of related disorders that includes rheumatoid arthritis and systemic sclerosis. Rheumatoid arthritis is the rheumatic disease in which the role of angiogenesis has been studied most extensively. However, whereas rheumatoid arthritis is characterized by excessive angiogenesis, the situation is not as clear cut in other rheumatic diseases. For example, systemic sclerosis is characterized by reduced capillary density with insufficient angiogenic responses. Results with angiogenesis inhibitors are controversial, and there is--in parallel--a wide range of upregulated angiogenic factors such as vascular endothelial growth factor. Dysregulation of angiogenesis in systemic sclerosis is accompanied by other pathogenic processes, including fibrosis, autoimmunity and vasculopathy. Animal models with at least partial features of the vasculopathy observed in systemic sclerosis include wound healing models, graft versus host disease models and, in particular, the University of California at Davis line 200 chicken model of systemic sclerosis.


Real-time PCR to identify variola virus or other human pathogenic orthopox viruses.
Variola virus (family Poxviridae, genus Orthopoxvirus) and the closely related cowpox, vaccinia, and monkeypox viruses can infect humans. Efforts are mounting to replenish the smallpox vaccine stocks, optimize diagnostic methods for poxviruses, and develop new antivirals against smallpox, because it is feared that variola virus might be used as a weapon of bioterrorism. We developed an assay for the detection of variola virus DNA. The assay is based on TaqMan chemistry targeting the 14-kD protein gene. For the 1st stage of the assay we used genus consensus primers and a mixture of 2 probes (14-kD POX and 14-kD VAR) spanning the 14-kD protein-encoding gene for detection of all human pathogenic orthopoxviruses. We then tested positive samples with the specific orthopoxvirus-specific probe 14-kD POX to identify monkeypox, cowpox, and vaccinia viruses and with the 14-kD VAR probe to identify variola viruses. The assay was established on 4 different PCR cycler platforms. It was assessed in a study with 85 different orthopoxvirus species and strains that included variola, camelpox, cowpox, monkeypox, and vaccinia viruses at concentrations ranging from 100 ng/L to 1 microg/L. The assay detected as little as 0.05 fg of DNA, corresponding to 25 copies of DNA, and enabled differentiation of variola virus from the other orthopoxviruses. This real-time PCR assay provides a rapid method for the early detection and differentiation of smallpox and other human pathogenic orthopoxvirus infections.


Treatment simplification in HIV-infected adults as a strategy to prevent toxicity, improve adherence, quality of life and decrease healthcare costs.
Since the advent of highly active antiretroviral therapy (HAART), the treatment of human immunodeficiency virus (HIV) infection has become more potent and better tolerated. While the current treatment regimens still have limitations, they are more effective, more convenient, and less toxic than regimens used in the early HAART era, and new agents, formulations and strategies continue to be developed. Simplification of therapy is an option for many patients currently being treated with antiretroviral therapy (ART). The main goals are to reduce pill burden, improve quality of life and enhance medication adherence, while minimizing short- and long-term toxicities, reducing the risk of virologic failure and maximizing cost-effectiveness. ART simplification strategies that are currently used or are under study include the use of once-daily regimens, less toxic drugs, fixed-dose coformulations and induction-maintenance approaches. Improved adherence and persistence have been observed with the adoption of some of these strategies. The role of regimen simplification has implications not only for individual patients, but also for health care policy. With increased interest in ART regimen simplification, it is critical to study not only implications for individual tolerability, toxicity, adherence, persistence and virologic efficacy, but also cost, scalability, and potential for dissemination and implementation, such that limited human and financial resources are optimally allocated for maximal efficiency, coverage and sustainability of global HIV/AIDS treatment.


Mtg16/Eto2 contributes to murine T-cell development.
Mtg16/Eto2 is a transcriptional corepressor that is disrupted by t(16;21) in acute myeloid leukemia. Using mice lacking Mtg16, we found that Mtg16 is a critical regulator of T-cell development. Deletion of Mtg16 led to reduced thymocyte development in vivo, and after competitive bone marrow transplantation, there was a nearly complete failure of Mtg16(-/-) cells to contribute to thymocyte development. This defect was recapitulated in vitro as Mtg16(-/-) Lineage(-)/Sca1(+)/c-Kit(+) (LSK) cells of the bone marrow or DN1 cells of the thymus failed to produce CD4(+)/CD8(+) cells in response to a Notch signal. Complementation of these defects by reexpressing Mtg16 showed that 3 highly conserved domains were somewhat dispensable for T-cell development but required the capacity of Mtg16 to suppress E2A-dependent transcriptional activation and to bind to the Notch intracellular domain. Thus, Mtg16 integrates the activities of signaling pathways and nuclear factors in the establishment of T-cell fate specification.


c-Jun N-terminal kinase mediates lactacystin-induced dopamine neuron degeneration.
Parkinson disease is characterized by the progressive loss of dopaminergic neurons in the substantia nigra pars compacta. It has been proposed that dysfunction of the ubiquitin proteasome system plays an important role in the pathogenesis of Parkinson disease, but the mechanisms underlying ubiquitin proteasome system-related neuron degeneration are unknown. Here, we demonstrate that the proteasome inhibitor lactacystin induces phosphorylation of c-Jun N-terminal kinase (JNK) and c-Jun, the release of cytochrome c, activation of both caspase-9 and caspase-3, and sequential apoptosis of dopaminergic neurons in vitro. Most of these effects can be attenuated by the JNK inhibitor SP600125. Furthermore, infusion of lactacystin in rats in vivo also leads to phosphorylation of JNK before nigral neuron loss; chronic administration of SP600125 also blocks this loss. These results indicate that JNK is involved in proteasome inhibition-induced dopaminergic neuron degeneration through caspase-3-mediated apoptotic pathways, suggesting that this kinase may be a therapeutic target for the prevention of substantia nigra pars compacta degeneration in Parkinson disease patients.


Spinal Fos labeling and penile erection elicited by stimulation of dorsal nerve of the rat penis.
Penile afferents present in the dorsal nerve of the penis (DNP) convey sensory information from the penis to the spinal cord and represent the afferent limb of reflexive erections. Immunocytochemical staining of Fos was used to identify spinal neurons that receive excitatory inputs from the DNP in anesthetized rats. Intracavernous pressure (ICP) was recorded as an index of erection. Dissection as well as stimulation of the DNP elicited a comparable increase in Fos staining. Labeling was present in the dorsal horn, the dorsal gray commissure, and the sacral parasympathetic nucleus, supporting the hypothesis of direct or indirect afferent projection from the penis and penile sheath in these areas. No change in ICP was observed in these rats. Stimulation of the DNP elicited both increased Fos labeling and ICP after spinalization, demonstrating the presence of a supraspinal inhibitory control exerted on the polysynaptic intraspinal circuitry responsible for reflexive penile erection.